Multifunctional immune cell therapies

Abstract

To provide therapeutic methods for hyperproliferative disorders such as cancer.SOLUTION: Provided herein is a multifunctional chimeric antigen receptor (CAR)-based composition and the use thereof for directing immune responses to a target cell. The composition has an application including treatment of a hyperproliferative disorder such as cancer. The provided method generally comprises the use of a CAR cell in combination with an adapter. The adapter confers an ability to modulate, alter and / or redirect the CAR cell-mediated immune response in vitro or in vivo. In some embodiments, the CAR cell comprises genetic modification to reduce or eliminate the expression of a targeted antigenic determinant.SELECTED DRAWING: None

Description

[Background technology]

[0001] Adoptive transfer of genetically modified T cells is a rapidly evolving, innovative treatment for cancer. Chimeric antigen receptor (CAR) genetically modified T cells are renewable agents capable of providing sustained functional immunity. Clinical efficacy has been demonstrated with CD19 CAR T cells in a range of hematological cancers, and promising early clinical data have been reported for other genetically modified CAR T cells in solid tumors. However, significant challenges must be overcome before the full potential of CAR technology can be realized.

[0002] Current clinical trials using CAR T cells have observed a high rate of relapse within a year or less due to the inability of CAR T cells to address antigen escape and the inherent heterogeneity of cancer or tumor phenotype. Furthermore, current CAR technologies cannot adapt to changes in cancer or tumor phenotype. For example, in solid tumors, current CAR technologies demonstrate limited efficacy due to tumor target heterogeneity and the inability to reprogram CAR T cells to recognize an expanded set of antigenic targets expressed over time. In addition to improving the durability and sustainability of clinical responses, other significant obstacles to the use of current CAR cell-based technologies include the time required for CAR T cell generation, suboptimal CAR cell specificity, efficacy, and safety for use in cancers other than leukemia. Therefore, there is a need for CAR cell-based technologies that offer simultaneous and / or sequential targeting multispecificity and the ability to modulate, alter, or redirect in vivo CAR cell-mediated immune responses.

[0003] Bone marrow transplantation (BMT) and hematopoietic stem cell transplantation (HSCT) have the potential to correct any hematologic or immunologic disorder. Czechowicz et al., Blood 128(22):493 (2016). Despite their great potential, the clinical use of BMT and HSCT is significantly limited by significant safety and toxicity risks associated with current high-dose chemotherapy / irradiation conditioning regimens that prepare patients for transplantation and subsequent donor HSC engraftment. The most common toxicities experienced by patients include neutropenia, infections, anemia, mucositis, infertility, and organ damage, particularly in the bone marrow compartment and secondary malignancies. Complete elimination of these toxic conditioning regimens would dramatically improve the safety profile of BMT and HSCT, expanding their potential applications to include many less malignant hematologic disorders, a wide variety of autoimmune disorders, and facilitating solid organ transplantation. Therefore, there is a need for the development of improved transplant conditioning regimens that limit or eliminate the toxicity associated with current high-dose chemotherapy / irradiation while also effectively treating the underlying pathology. [Prior art documents] [Non-patent literature]

[0004] [Non-Patent Document 1] Czechowicz et al.,Blood 128(22):493(2016) Summary of the Invention

[0005] Provided herein are multifunctional chimeric antigen receptor (CAR)-based compositions and their use in inducing immune responses against target cells. The compositions have uses, including the treatment of hyperproliferative disorders such as cancer. The provided methods typically involve the use of CAR cells in combination with an adaptor. The adaptor confers the ability to modulate, alter, and / or redirect CAR cell-mediated immune responses in vitro and in vivo.

[0006] The present disclosure relates to compositions comprising (a) a cell expressing a chimeric antigen receptor (CAR) comprising an antigenic determinant binding domain (ADBD), and (b) a soluble protein ("adapter") comprising (i) an antigenic determinant (AD) and (ii) an ADBD. The present disclosure also provides methods of killing target cells using the compositions provided herein, including therapeutic applications of the compositions provided herein.

[0007] In some embodiments, the present disclosure provides a composition comprising: (a) a cell expressing a CAR comprising (i) an ADBD that binds to a first AD on a target cell, (ii) a transmembrane domain, and (iii) an intracellular domain; and (b) an adaptor comprising an ADBD that binds to (i) the first AD and (ii) a second AD on the target cell.

[0008] In some embodiments, the present disclosure provides a composition comprising: (a) a cell expressing a CAR comprising (i) an ADBD that binds to a first AD on a target cell, (ii) a transmembrane domain, and (iii) an intracellular domain; and (b) an adaptor comprising an ADBD that binds to (i) the first AD and (ii) a second AD on a second target cell.

[0009] In some embodiments, the present disclosure provides a composition comprising: (a) a cell expressing a CAR comprising (i) an ADBD that is an alternative scaffold binding domain (ASBD) that binds to a first AD, (ii) a transmembrane domain, and (iii) an intracellular domain; and (b) an adaptor comprising an ADBD that binds to (i) the first AD and (ii) a second AD on a target cell.

[0010] In some embodiments, the present disclosure provides a composition comprising: (a) a cell expressing a CAR comprising (i) an ADBD that binds to a first AD, (ii) a transmembrane domain, and (iii) an intracellular domain; and (b) an adaptor comprising an ADBD that comprises (i) the first AD and (ii) an ASBD that binds to a second AD on a target cell.

[0011] In some embodiments, the present disclosure provides a composition comprising: (a) a cell expressing a CAR comprising (i) an ADBD that binds to a first AD, (ii) a transmembrane domain, and (iii) an intracellular domain; and (b) an adaptor comprising (i) the first AD and (ii) a D domain that binds to a second AD on a target cell.

[0012] In some embodiments, the present disclosure provides a composition comprising: (a) a cell expressing a CAR comprising (i) a D domain that binds to a first AD, (ii) a transmembrane domain, and (iii) an intracellular domain; and (b) an adaptor comprising an ADBD that binds to (i) the first AD and (ii) a second AD on a target cell.

[0013] In some embodiments, the present disclosure provides a composition comprising: (a) a cell expressing a CAR comprising (i) a first D domain that binds to a first AD, (ii) a transmembrane domain, and (iii) an intracellular domain; and (b) an adaptor comprising (i) the first AD and (ii) a second D domain that binds to a second AD on a target cell.

[0014] In some embodiments, the present disclosure provides a method of killing a target cell comprising contacting a composition comprising the target cell with an adaptor, wherein (a) the composition comprising the target cell further comprises a cell expressing a CAR, the CAR comprising (i) an ADBD that binds to a first AD on said target cell, (ii) a transmembrane domain, and (iii) an intracellular domain; and (b) the adaptor comprises an ADBD that binds to (i) said first AD and (ii) a second AD on said target cell.

[0015] In some embodiments, the present disclosure provides a method of killing one or more target cells comprising contacting a composition comprising a first target cell with an adaptor, wherein (a) the composition comprising the target cell further comprises a cell expressing a CAR, the CAR comprising (i) an ADBD that binds to a first AD on said first target cell, (ii) a transmembrane domain, and (iii) an intracellular domain; and (b) the adaptor comprises an ADBD that binds to (i) said first AD and (ii) a second AD on a second target cell.

[0016] In some embodiments, the present disclosure provides a method of killing a target cell comprising contacting a composition comprising the target cell with an adaptor, wherein (a) the composition comprising the target cell further comprises a cell expressing a CAR, wherein the CAR comprises (i) an ADBD that is an ASBD that binds to a first AD, (ii) a transmembrane domain, and (iii) an intracellular domain; and (b) the adaptor comprises an ADBD that binds to (i) said first AD and (ii) a second AD on said target cell.

[0017] In some embodiments, the present disclosure provides a method of killing a target cell comprising contacting a composition comprising the target cell with an adaptor, wherein (a) the composition comprising the target cell further comprises a cell expressing a CAR, the CAR comprising (i) an ADBD that binds to a first AD, (ii) a transmembrane domain, and (iii) an intracellular domain; and (b) the adaptor comprises an ADBD that is an ASBD that binds to (i) said first AD and (ii) a second AD on said target cell.

[0018] In some embodiments, the present disclosure provides a method of killing a target cell comprising contacting a composition comprising the target cell with a cell expressing a CAR, wherein (a) a first AD is present on the target cell; (b) the composition comprising the first target cell further comprises an adaptor comprising an ADBD that binds to (i) said first AD and (ii) a second AD on the target cell; and (c) the CAR comprises (i) an ADBD that binds to said first AD on the target cell or the adaptor, (ii) a transmembrane domain, and (iii) an intracellular domain.

[0019] In some embodiments, the present disclosure provides a method of killing one or more target cells comprising contacting a composition comprising the target cells with a cell expressing a CAR, wherein (a) a first AD is present on the first target cell; (b) the composition comprising the first target cell further comprises a second target cell and an adaptor comprising (i) the first AD and (ii) a second AD on the second target cell; and (c) the CAR comprises (i) an ADBD that binds to the first AD on the first target cell or the adaptor, (ii) a transmembrane domain, and (iii) an intracellular domain.

[0020] In some embodiments, the present disclosure provides a method of killing a target cell comprising contacting a composition comprising the target cell with a cell expressing a CAR, wherein (a) a first AD is present on the target cell; (b) the composition comprising the target cell further comprises an adaptor comprising (i) an ADBD that binds to said first AD on said target cell, and (ii) a second AD; and (c) the CAR comprises (i) an ADBD that is an ASBD that binds to said second AD, (ii) a transmembrane domain, and (iii) an intracellular domain.

[0021] In some embodiments, the present disclosure provides a method of killing a target cell comprising contacting a composition comprising the target cell with a cell expressing a CAR, wherein (a) a first AD is present on the target cell; (b) the composition comprising the target cell further comprises an adaptor comprising (i) an ADBD that is an ASBD that binds to said first AD on said target cell, and (ii) a second AD; and (c) the CAR comprises (i) an ADBD that binds to said second AD, (ii) a transmembrane domain, and (iii) an intracellular domain.

[0022] In some embodiments, the present disclosure provides a method of transducing an immune response to a target cell comprising contacting a composition comprising the target cell with an adaptor, wherein (a) the composition comprising the target cell further comprises a cell expressing a CAR, the CAR comprising (i) an ADBD that binds to a first AD on said target cell, (ii) a transmembrane domain, and (iii) an intracellular domain; and (b) the adaptor comprises an ADBD that binds to (i) said first AD and (ii) a second AD on said target cell.

[0023] In some embodiments, the present disclosure provides a method of transducing an immune response to one or more target cells comprising contacting a composition comprising a first target cell with an adaptor, wherein (a) the composition comprising the target cell further comprises a cell expressing a CAR, the CAR comprising (i) an ADBD that binds to a first AD on said first target cell, (ii) a transmembrane domain, and (iii) an intracellular domain; and (b) the adaptor comprises an ADBD that binds to (i) said first AD and (ii) a second AD on a second target cell.

[0024] In some embodiments, the present disclosure provides a method of transducing an immune response to a target cell comprising contacting a composition comprising the target cell with an adaptor, wherein (a) the composition comprising the target cell further comprises a cell expressing a CAR, the CAR comprising (i) an ADBD that is an ASBD that binds to a first AD, (ii) a transmembrane domain, and (iii) an intracellular domain; and (b) the adaptor comprises an ADBD that binds to (i) said first AD and (ii) a second AD on said target cell.

[0025] In some embodiments, the present disclosure provides a method of transducing an immune response to a target cell comprising contacting a composition comprising the target cell with an adaptor, wherein (a) the composition comprising the target cell further comprises a cell expressing a CAR, the CAR comprising (i) an ADBD that binds to a first AD, (ii) a transmembrane domain, and (iii) an intracellular domain; and (b) the adaptor comprises an ADBD that is an ASBD that binds to (i) said first AD and (ii) a second AD on said target cell.

[0026] In some embodiments, the present disclosure provides a method of transducing an immune response to a target cell comprising contacting a composition comprising the target cell with a CAR, wherein (a) a first AD is present on the target cell; (b) the composition comprising the first target cell further comprises an adaptor comprising an ADBD that binds to (i) said first AD and (ii) a second AD on the target cell; and (c) the CAR comprises (i) an ADBD that binds to said first AD on the target cell or the adaptor, (ii) a transmembrane domain, and (iii) an intracellular domain.

[0027] In some embodiments, the present disclosure provides a method of transducing an immune response to one or more target cells comprising contacting a composition comprising the target cells with a cell expressing a CAR, wherein (a) a first AD is present on the first target cell; (b) the composition comprising the first target cell further comprises a second target cell and an adaptor comprising (i) the first AD and (ii) a second AD on the second target cell; and (c) the CAR comprises (i) an ADBD that binds to the first AD on the first target cell or the adaptor, (ii) a transmembrane domain, and (iii) an intracellular domain.

[0028] In some embodiments, the present disclosure provides a method of transducing an immune response to a target cell comprising contacting a composition comprising the target cell with a CAR, wherein (a) a first AD is present on the target cell; (b) the composition comprising the target cell further comprises an adaptor comprising (i) an ADBD that binds to said first AD on said target cell, and (ii) a second AD; and (c) the CAR comprises (i) an ADBD that is an ASBD that binds to said second AD, (ii) a transmembrane domain, and (iii) an intracellular domain.

[0029] In some embodiments, the present disclosure provides a method of transducing an immune response to a target cell, comprising contacting a composition comprising the target cell with a CAR, wherein (a) a first antigenic determinant (AD) is present on the target cell; (b) the composition comprising the target cell further comprises (i) an ADBD, an ASBD that binds to said first AD on said target cell, and (ii) an adaptor comprising a second AD; and (c) the CAR comprises (i) an ADBD that binds to said second AD, (ii) a transmembrane domain, and (iii) an intracellular domain.

[0030] In some embodiments, the present disclosure provides a method of killing target cells in a patient comprising administering an adaptor to the patient, wherein (a) the patient has been treated with cells expressing a CAR, the CAR comprising (i) an ADBD that binds to a first AD on said target cell, (ii) a transmembrane domain, and (iii) an intracellular domain; and (b) the adaptor comprises an ADBD that binds to (i) said first AD and (ii) a second AD on said target cell.

[0031] In some embodiments, the present disclosure provides a method of redirecting killing of target cells in a patient comprising administering to the patient an adaptor, wherein (a) the patient has been treated with cells expressing a CAR, the CAR comprising (i) an ADBD that binds to a first AD on a first target cell, (ii) a transmembrane domain, and (iii) an intracellular domain; and (b) the adaptor comprises an ADBD that binds to (i) said first AD and (ii) a second AD on a second target cell.

[0032] In some embodiments, the present disclosure provides a method of killing target cells in a patient comprising administering an adaptor to the patient, wherein (a) the patient has been treated with cells expressing a CAR, the CAR comprising (i) an ADBD that is an ASBD that binds to a first AD, (ii) a transmembrane domain, and (iii) an intracellular domain; and (b) the adaptor comprises an ADBD that binds to (i) said first AD and (ii) a second AD on said target cell.

[0033] In some embodiments, the present disclosure provides a method of killing target cells in a patient comprising administering an adaptor to the patient, wherein (a) the patient has been treated with cells expressing a CAR, the CAR comprising (i) an ADBD that binds to a first AD, (ii) a transmembrane domain, and (iii) an intracellular domain; and (b) the adaptor comprises an ADBD that is an ASBD that binds to (i) said first AD and (ii) a second AD on said target cell.

[0034] In some embodiments, the present disclosure provides a method of killing target cells in a patient comprising administering to the patient cells expressing a CAR, wherein (a) a first AD and a second AD are present on the target cell; (b) the patient has been treated with an adaptor comprising (i) an ADBD that binds to said first AD and (ii) said second AD on said target cell; and (c) the CAR comprises (i) an ADBD that binds to said first AD on the target cell or adaptor, (ii) a transmembrane domain, and (iii) an intracellular domain.

[0035] In some embodiments, the present disclosure provides a method of inducing death of a target cell in a patient comprising administering to the patient cells expressing a CAR, wherein (a) the patient has been treated with an adaptor comprising (i) a first AD and (ii) an ADBD that binds to a second AD on the target cell; and (b) the CAR comprises (i) an ADBD that binds to said first AD on the adaptor, (ii) a transmembrane domain, and (iii) an intracellular domain.

[0036] In some embodiments, the present disclosure provides a method of killing target cells in a patient comprising administering to the patient cells expressing a CAR, wherein (a) a first AD is present on the target cell; (b) the patient has been treated with an adaptor comprising (i) an ADBD that binds to said first AD on said target cell and (ii) a second AD; and (c) the CAR comprises (i) an ADBD that is an ASBD that binds to said second AD, (ii) a transmembrane domain, and (iii) an intracellular domain.

[0037] In some embodiments, the present disclosure provides a method of killing target cells in a patient comprising administering to the patient cells expressing a CAR, wherein (a) a first AD is present on the target cell; (b) the patient has been treated with an adaptor comprising (i) an ADBD that is an ASBD that binds to said first AD on said target cell and (ii) a second AD; and (c) the CAR comprises (i) an ADBD that binds to said second AD, (ii) a transmembrane domain, and (iii) an intracellular domain.

[0038] In some embodiments, the present disclosure provides a method of transducing an immune response to a target cell in a patient comprising administering to the patient an adaptor, wherein (a) the patient has been treated with cells expressing a CAR, the CAR comprising (i) an ADBD that binds to a first AD on said target cell, (ii) a transmembrane domain, and (iii) an intracellular domain; and (b) the adaptor comprises an ADBD that binds to (i) said first AD and (ii) a second AD on said target cell.

[0039] In some embodiments, the present disclosure provides a method of redirecting an immune response to target cells in a patient comprising administering to the patient an adaptor, wherein: (a) the patient is undergoing cell therapy expressing a CAR, the CAR comprising (i) an ADBD that binds to a first AD on a first target cell, (ii) a transmembrane domain, and (iii) an intracellular domain; and (b) the adaptor comprises an ADBD that binds to (i) the first AD and (ii) a second AD on a second target cell.

[0040] In some embodiments, the present disclosure provides a method of transducing an immune response to a target cell in a patient comprising administering to the patient an adaptor, wherein (a) the patient has been treated with cells expressing a CAR, the CAR comprising (i) an ADBD that is an ASBD that binds to a first AD, (ii) a transmembrane domain, and (iii) an intracellular domain; and (b) the adaptor comprises an ADBD that binds to (i) said first AD and (ii) a second AD on said target cell.

[0041] In some embodiments, the present disclosure provides a method of transducing an immune response to a target cell in a patient comprising administering to the patient an adaptor, wherein (a) the patient has been treated with a cell expressing a CAR, the CAR comprising (i) an ADBD that binds to a first AD, (ii) a transmembrane domain, and (iii) an intracellular domain; and (b) the adaptor comprises an ADBD that is an ASBD that binds to (i) said first AD and (ii) a second AD on said target cell.

[0042] In some embodiments, the present disclosure provides a method of transducing an immune response to a target cell in a patient comprising administering to the patient a cell expressing a CAR, wherein (a) a first AD and a second AD are present on the target cell; (b) the patient has been treated with an adaptor comprising (i) an ADBD that binds to said first AD and (ii) said second AD on said target cell; and (c) the CAR comprises (i) an ADBD that binds to said first AD on the target cell or adaptor, (ii) a transmembrane domain, and (iii) an intracellular domain.

[0043] In some embodiments, the present disclosure provides a method of inducing an immune response in a target cell in a patient comprising administering to the patient cells expressing a CAR, wherein (a) the patient has been treated with an adaptor comprising (i) a first AD and (ii) an ADBD that binds to a second AD on the target cell; and (b) the CAR comprises (i) an ADBD that binds to said first AD on the adaptor, (ii) a transmembrane domain, and (iii) an intracellular domain.

[0044] In some embodiments, the present disclosure provides a method of transducing an immune response to a target cell in a patient comprising administering to the patient a cell expressing a CAR, wherein (a) a first AD is present on the target cell; (b) the patient has been treated with an adaptor comprising (i) an ADBD that binds to said first AD on said target cell and (ii) a second AD; and (c) the CAR comprises (i) an ADBD that is an ASBD that binds to said second AD, (ii) a transmembrane domain, and (iii) an intracellular domain.

[0045] In some embodiments, the present disclosure provides a method of transducing an immune response to a target cell in a patient comprising administering to the patient a cell expressing a CAR, wherein (a) a first AD is present on the target cell; (b) the patient has been treated with an adaptor comprising (i) an ADBD that is an ASBD that binds to said first AD on said target cell and (ii) a second AD; and (c) the CAR comprises (i) an ADBD that binds to said second AD, (ii) a transmembrane domain, and (iii) an intracellular domain.

[0046] In some embodiments, the present disclosure provides a method of treating a proliferative disorder, cancer, an autoimmune disease, an infectious disease, or allograft rejection comprising contacting a composition comprising a target cell with a cell expressing a CAR, wherein (a) a first AD is present on the target cell; (b) the composition comprising the target cell further comprises an adaptor comprising (i) an ADBD, an ASBD that binds to said first AD on said target cell, and (ii) a second AD; and (c) the CAR comprises (i) an ADBD that binds to said second AD, (ii) a transmembrane domain, and (iii) an intracellular domain.

[0047] In some embodiments, the present disclosure provides a method of treating a proliferative disorder, cancer, an autoimmune disease, an infectious disease, or allograft rejection comprising administering to a patient an adaptor, wherein (a) the patient is being treated with cells expressing a CAR, the CAR comprising (i) an ADBD that binds to a first AD on said target cell, (ii) a transmembrane domain, and (iii) an intracellular domain; and (b) the adaptor comprises an ADBD that binds to (i) said first AD and (ii) a second AD on said target cell.

[0048] In some embodiments, the present disclosure provides a method of redirecting treatment of a proliferative disorder, cancer, autoimmune disease, infectious disease, or allograft rejection comprising administering to a patient an adaptor, wherein (a) the patient is being treated with cells expressing a CAR, the CAR comprising (i) an ADBD that binds to a first AD on a first target cell, (ii) a transmembrane domain, and (iii) an intracellular domain; and (b) the adaptor comprises an ADBD that binds to (i) said first AD and (ii) a second AD on a second target cell.

[0049] In some embodiments, the present disclosure provides a method of treating a proliferative disorder, cancer, an autoimmune disease, an infectious disease, or allograft rejection comprising administering to a patient an adaptor, wherein (a) the patient is treated with cells expressing a CAR, the CAR comprising (i) an ADBD that is an ASBD that binds to a first AD, (ii) a transmembrane domain, and (iii) an intracellular domain; and (b) the adaptor comprises an ADBD that binds to (i) said first AD and (ii) a second AD on said target cell.

[0050] In some embodiments, the present disclosure provides a method of treating a proliferative disorder, cancer, an autoimmune disease, an infectious disease, or allograft rejection comprising administering to a patient an adaptor, wherein (a) the patient is being treated with cells expressing a chimeric antigen receptor (CAR), the CAR comprising (i) an antigenic determinant binding domain (ADBD) that binds to a first antigenic determinant (AD), (ii) a transmembrane domain, and (iii) an intracellular domain; and (b) the adaptor comprises an ADBD, an ASBD that binds to (i) said first AD and (ii) a second AD on said target cell.

[0051] In some embodiments, the present disclosure provides a method of treating a proliferative disorder, cancer, an autoimmune disease, an infectious disease, or allograft rejection comprising administering to a patient cells expressing a CAR, wherein (a) a first AD and a second AD are present on a target cell; (b) the patient has been treated with an adaptor comprising (i) an ADBD that binds to said first AD and (ii) said second AD on said target cell; and (c) the CAR comprises (i) an ADBD that binds to said first AD on the target cell or the adaptor, (ii) a transmembrane domain, and (iii) an intracellular domain.

[0052] In some embodiments, the present disclosure provides a method of inducing treatment of a proliferative disorder, cancer, autoimmune disease, infectious disease, or allograft rejection comprising administering cells expressing a CAR to a patient, wherein (a) the patient has been treated with an adaptor comprising (i) a first AD and (ii) an ADBD that binds to a second AD on a target cell; and (b) the CAR comprises (i) an ADBD that binds to said first AD on the adaptor, (ii) a transmembrane domain, and (iii) an intracellular domain.

[0053] In some embodiments, the present disclosure provides a method of treating a proliferative disorder, cancer, an autoimmune disease, an infectious disease, or allograft rejection comprising administering to a patient cells expressing a CAR, wherein (a) a first AD is present on a target cell; (b) the patient has been treated with an adaptor comprising (i) an ADBD that binds to said first AD on said target cell and (ii) a second AD; and (c) the CAR comprises (i) an ADBD that is an ASBD that binds to said second AD, (ii) a transmembrane domain, and (iii) an intracellular domain.

[0054] In some embodiments, the present disclosure provides a method of treating a proliferative disorder, cancer, an autoimmune disease, an infectious disease, or allograft rejection comprising administering to a patient cells expressing a CAR, wherein (a) a first AD is present on a target cell; (b) the patient has been treated with an adaptor comprising (i) an ADBD, which is an ASBD that binds to said first AD on said target cell, and (ii) a second AD; and (c) the CAR comprises (i) an ADBD that binds to said second AD, (ii) a transmembrane domain, and (iii) an intracellular domain.

[0055] In some embodiments, the present disclosure provides a method of treating a hematological cancer comprising contacting a composition comprising a target cell with a cell expressing a CAR, wherein (a) a first AD is present on the target cell; (b) the composition comprising the target cell further comprises an adaptor comprising (i) an ADBD, which is an ASBD that binds to said first AD on said target cell, and (ii) a second AD; and (c) the CAR comprises (i) an ADBD that binds to said second AD, (ii) a transmembrane domain, and (iii) an intracellular domain.

[0056] In some embodiments, the present disclosure provides a method of treating a hematological cancer comprising administering an adaptor to a patient, wherein (a) the patient is being treated with cells expressing a CAR, the CAR comprising (i) an ADBD that binds to a first antigenic determinant (AD) on said target cell, (ii) a transmembrane domain, and (iii) an intracellular domain; and (b) the adaptor comprises an ADBD that binds to (i) said first AD and (ii) a second AD on said target cell.

[0057] In some embodiments, the present disclosure provides a method of redirecting treatment of a hematological cancer comprising administering to a patient an adaptor, wherein (a) the patient is being treated with cells expressing a CAR, the CAR comprising (i) an ADBD that binds to a first AD on a first target cell, (ii) a transmembrane domain, and (iii) an intracellular domain; and (b) the adaptor comprises an ADBD that binds to (i) said first AD and (ii) a second AD on a second target cell.

[0058] In some embodiments, the present disclosure provides a method of treating a hematological cancer comprising administering an adaptor to a patient, wherein (a) the patient is being treated with cells expressing a CAR, the CAR comprising (i) an ADBD that is an ASBD that binds to a first AD, (ii) a transmembrane domain, and (iii) an intracellular domain; and (b) the adaptor comprises an ADBD that binds to (i) said first AD and (ii) a second AD on said target cell.

[0059] In some embodiments, the present disclosure provides a method of treating a hematological cancer comprising administering an adaptor to a patient, wherein (a) the patient is being treated with cells expressing a CAR, the CAR comprising (i) an ADBD that binds to a first AD, (ii) a transmembrane domain, and (iii) an intracellular domain; and (b) the adaptor comprises an ADBD that is an ASBD that binds to (i) said first AD and (ii) a second AD on said target cell.

[0060] In some embodiments, the present disclosure provides a method of treating a hematological cancer comprising administering to a patient cells expressing a CAR, wherein (a) a first AD and a second AD are present on a target cell; (b) the patient has been treated with an adaptor comprising (i) an ADBD that binds to said first AD and (ii) said second AD on said target cell; and (c) the CAR comprises (i) an ADBD that binds to said first AD on the target cell or the adaptor, (ii) a transmembrane domain, and (iii) an intracellular domain.

[0061] In some embodiments, the present disclosure provides a method of inducing treatment of a hematological cancer comprising administering to a patient cells expressing a CAR, wherein (a) the patient has been treated with an adaptor comprising (i) a first AD and (ii) an ADBD that binds to a second AD on a target cell; and (b) the CAR comprises (i) an ADBD that binds to said first AD on the adaptor, (ii) a transmembrane domain, and (iii) an intracellular domain.

[0062] In some embodiments, the present disclosure provides a method of treating a hematological cancer comprising administering to a patient cells expressing a CAR, wherein (a) a first AD is present on a target cell; (b) the patient has been treated with an adaptor comprising (i) an ADBD that binds to said first AD on said target cell and (ii) a second AD; and (c) the CAR comprises (i) an ADBD that is an ASBD that binds to said second AD, (ii) a transmembrane domain, and (iii) an intracellular domain.

[0063] In some embodiments, the present disclosure provides a method of treating a hematological cancer comprising administering to a patient cells expressing a CAR, wherein (a) a first AD is present on a target cell; (b) the patient has been treated with an adaptor comprising (i) an ADBD, which is an ASBD that binds to said first AD on said target cell, and (ii) a second AD; and (c) the CAR comprises (i) an ADBD that binds to said second AD, (ii) a transmembrane domain, and (iii) an intracellular domain.

[0064] In some embodiments, the present disclosure provides genetically modified human immune effector cells comprising: (a) a chimeric antigen receptor (CAR) comprising (1) an antigenic determinant binding domain (ADBD) that specifically binds to a human CD45 antigenic determinant (AD), (2) a transmembrane domain, and (3) an intracellular domain; and (b) a genetic modification that abolishes expression of human CD45 AD on the genetically modified cell, wherein the genetically modified cell is capable of inducing an immune response against CD45 AD-expressing cells in an in vitro assay, and wherein the genetically modified cell does not express CD45 AD.

[0065] In some embodiments, the present disclosure provides a method of killing human CD45-expressing target cells, comprising contacting the target cells with genetically modified human immune effector cells, wherein the genetically modified human immune effector cells comprise: (a) a chimeric antigen receptor (CAR) comprising (1) an antigenic determinant binding domain (ADBD) that specifically binds a human CD45 antigenic determinant (AD), (2) a transmembrane domain, and (3) an intracellular domain; and (b) a genetic modification that abolishes expression of the human CD45 AD on the genetically modified cells.

[0066] In some embodiments, the present disclosure provides a method of transducing an immune response to a human CD45-expressing target cell, comprising contacting the target cell with a genetically modified human immune effector cell, wherein the genetically modified human immune effector cell comprises: (a) a chimeric antigen receptor (CAR) comprising (1) an antigenic determinant binding domain (ADBD) that specifically binds a human CD45 antigenic determinant (AD), (2) a transmembrane domain, and (3) an intracellular domain; and (b) a genetic modification that abolishes expression of the human CD45 AD on the genetically modified cell.

[0067] In some embodiments, the present disclosure provides a method of treating a proliferative disorder, cancer, an autoimmune disease, an infectious disease, or allograft rejection in a subject comprising administering to a subject in need thereof a therapeutically effective amount of genetically modified human immune effector cells, wherein the genetically modified human immune effector cells comprise: (a) a chimeric antigen receptor (CAR) comprising (1) an antigenic determinant binding domain (ADBD) that specifically binds a human CD45 antigenic determinant (AD), (2) a transmembrane domain, and (3) an intracellular domain; and (b) a genetic modification that abolishes expression of human CD45 AD on the genetically modified cells.

[0068] In some embodiments, the present disclosure provides genetically modified human immune effector cells comprising: (a) a chimeric antigen receptor (CAR) comprising: (1) a first antigenic determinant binding domain (ADBD) that specifically binds a first antigenic determinant (AD), (2) a transmembrane domain, and (3) an intracellular domain; (b) a genetic modification that abolishes expression of at least one human CD45 AD on the genetically modified cell, wherein the first AD is not the at least one human CD45 AD; and wherein the genetically modified cell, used in combination with an adaptor in an in vitro assay, is capable of inducing an immune response against CD45-expressing cells, wherein the adaptor comprises a second ADBD that specifically binds the first AD and the at least one human CD45 AD, and wherein the genetically modified cell does not express the at least one human CD45 AD.

[0069] In some embodiments, the present disclosure provides a method of killing a target cell, comprising contacting the target cell with a genetically modified human immune effector cell and an adaptor, wherein the target cell expresses CD45, and the genetically modified human immune effector cell comprises (a) a chimeric antigen receptor (CAR) comprising (1) a first antigenic determinant binding domain (ADBD) that specifically binds a first antigenic determinant (AD), (2) a transmembrane domain, and (3) an intracellular domain; and (b) a genetic modification that abolishes expression of at least one human CD45 AD on the genetically modified cell, and the adaptor comprises the first AD and a second ADBD that specifically binds the human CD45 AD.

[0070] In some embodiments, the present disclosure provides a method of transducing an immune response to a target cell, comprising contacting the target cell with a genetically modified human immune effector cell and an adaptor, wherein the target cell expresses CD45, and the genetically modified human immune effector cell comprises: (a) a chimeric antigen receptor (CAR) comprising (1) a first antigenic determinant binding domain (ADBD) that specifically binds a first antigenic determinant (AD), (2) a transmembrane domain, and (3) an intracellular domain; and (b) a genetic modification that abolishes expression of at least one human CD45 AD on the genetically modified cell, and the adaptor comprises the first AD and a second ADBD that specifically binds the human CD45 AD.

[0071] In some embodiments, the present disclosure provides methods of inducing an immune response against CD45-expressing target cells in a subject, comprising: (a) (i) administering to a subject in need thereof a therapeutically effective amount of genetically modified human immune effector cells comprising: (1) a first antigenic determinant binding domain (ADBD) that specifically binds a first antigenic determinant (AD), (2) a transmembrane domain, and (3) an intracellular domain; and (ii) a genetic modification that abolishes expression of at least one human CD45 AD on the genetically modified cells; and (b) administering to the subject a therapeutically effective amount of an adaptor comprising the first AD and a second ADBD that specifically binds the human CD45 AD.

[0072] In some embodiments, the present disclosure provides methods of inducing an immune response against CD45-expressing target cells in a subject comprising administering to a subject in need thereof a therapeutically effective amount of genetically modified human immune effector cells comprising: (i) a chimeric antigen receptor (CAR) comprising: (1) a first antigenic determinant binding domain (ADBD) that specifically binds a first antigenic determinant (AD), (2) a transmembrane domain, and (3) an intracellular domain; and (ii) a genetic modification that abolishes expression of at least one human CD45 AD on the genetically modified cells, wherein the subject has been administered an adaptor comprising the first AD and a second ADBD that specifically binds the human CD45 AD.

[0073] In some embodiments, the present disclosure provides a method of inducing an immune response against CD45-expressing target cells in a subject comprising administering to a subject in need thereof a therapeutically effective amount of an adapter comprising a first AD and a second ADBD that specifically binds to a human CD45 AD, wherein the subject has been administered (i) a chimeric antigen receptor (CAR) comprising (1) a first antigenic determinant binding domain (ADBD) that specifically binds to the first antigenic determinant (AD), (2) a transmembrane domain, and (3) an intracellular domain; and (ii) genetically modified human immune effector cells comprising a genetic modification that abolishes expression of at least one human CD45 AD on the genetically modified cells.

[0074] In some embodiments, the present disclosure provides methods of treating a proliferative disorder, cancer, an autoimmune disease, an infectious disease, or allograft rejection in a subject, comprising: (a) (i) administering to a subject in need thereof a therapeutically effective amount of genetically modified human immune effector cells comprising: (1) a first antigenic determinant binding domain (ADBD) that specifically binds a first antigenic determinant (AD), (2) a transmembrane domain, and (3) an intracellular domain; and (ii) a genetic modification that abolishes expression of at least one human CD45 AD on the genetically modified cells; and (b) administering to the subject a therapeutically effective amount of an adaptor comprising the first AD and a second ADBD that specifically binds the human CD45 AD.

[0075] In some embodiments, the present disclosure provides a method of treating a proliferative disorder, cancer, autoimmune disease, infectious disease, or allograft rejection in a subject comprising administering to a subject in need thereof a therapeutically effective amount of genetically modified human immune effector cells comprising: (i) a chimeric antigen receptor (CAR) comprising: (1) a first antigenic determinant binding domain (ADBD) that specifically binds a first antigenic determinant (AD), (2) a transmembrane domain, and (3) an intracellular domain; and (ii) a genetic modification that abolishes expression of at least one human CD45 AD on the genetically modified cells, wherein the subject has been administered an adaptor comprising the first AD and a second ADBD that specifically binds the human CD45 AD.

[0076] In some embodiments, the present disclosure provides a method of treating a proliferative disorder, cancer, autoimmune disease, infectious disease, or allograft rejection in a subject comprising administering to a subject in need thereof a therapeutically effective amount of an adapter comprising a first AD and a second ADBD that specifically binds to a human CD45 AD, wherein the subject has been administered (i) a chimeric antigen receptor (CAR) comprising (1) a first antigenic determinant binding domain (ADBD) that specifically binds to the first antigenic determinant (AD), (2) a transmembrane domain, and (3) an intracellular domain; and (ii) genetically modified human immune effector cells comprising a genetic modification that abolishes expression of at least one human CD45 AD on the genetically modified cells.

[0077] In some embodiments, the present disclosure provides genetically modified human immune effector cells comprising: (a) a chimeric antigen receptor (CAR) comprising two or more antigenic determinants (ADBDs) comprising (i) a first antigenic determinant binding domain (ADBD) that binds to a first AD, (ii) a transmembrane domain, and (iii) an intracellular domain; and (b) a genetic modification that abolishes expression of the first AD on the genetically modified cell, wherein the genetically modified cell does not express the first AD.

[0078] In some embodiments, the present disclosure provides a method of killing a target cell, comprising: (a) contacting the target cell with a genetically modified human immune effector cell comprising: (i) two or more chimeric antigen receptor (CAR) comprising a first antigenic determinant (ADBD), the two or more ADBDs comprising a first antigenic determinant binding domain (ADBD) that binds to the first AD, (ii) a transmembrane domain, and (iii) an intracellular domain; and (b) a genetic modification that abolishes expression of the first AD on the genetically modified cell.

[0079] In some embodiments, the present disclosure provides methods of inducing an immune response against target cells in a subject comprising administering to a subject in need thereof a therapeutically effective amount of genetically modified human immune effector cells comprising: (a) a chimeric antigen receptor (CAR) comprising two or more antigenic determinant binding domains (ADBDs) comprising: (i) a first antigenic determinant binding domain (ADBD) that binds a first antigenic determinant (ADBD), (ii) a transmembrane domain, and (iii) an intracellular domain; and (b) a genetic modification that abolishes expression of the first AD on the genetically modified cells.

[0080] In some embodiments, the present disclosure provides methods of treating a proliferative disorder, cancer, an autoimmune disease, an infectious disease, or allograft rejection in a subject comprising administering to a subject in need thereof a therapeutically effective amount of (a) a chimeric antigen receptor (CAR) comprising: (i) two or more antigenic determinant binding domains (ADBDs) comprising a first antigenic determinant (ADBD) that binds to the first AD, (ii) a transmembrane domain, and (iii) an intracellular domain; and (b) a genetic modification that abolishes expression of the first AD on the genetically modified cell.

[0081] In some embodiments, the present disclosure provides methods of inducing an immune response against target cells in a subject, comprising: (a) (i) administering to a subject in need thereof a therapeutically effective amount of a chimeric antigen receptor (CAR) comprising: (1) two or more antigenic determinant (ADBD) comprising a first antigenic determinant binding domain (ADBD) that binds to a first AD, (2) a transmembrane domain, and (3) an intracellular domain; and (ii) a genetic modification that abolishes expression of the first AD on the genetically modified cell; and (b) administering to the subject a therapeutically effective amount of an adaptor comprising the AD recognized by the CAR and a second ADBD capable of binding to a second AD on the target cell.

[0082] In some embodiments, the present disclosure provides genetically modified human immune effector cells comprising: (a) a chimeric antigen receptor (CAR) comprising: (i) a first antigenic determinant binding domain (ADBD) that specifically binds to a first antigenic determinant (AD), (ii) a transmembrane domain, and (iii) an intracellular domain; and (b) a genetic modification that abolishes expression of a second AD on the genetically modified cell, wherein the genetically modified cell used in combination with an adaptor is capable of inducing an immune response against cells expressing the second AD in an in vitro assay, wherein the adaptor comprises the second ADBD that specifically binds the first AD and the second AD, and wherein the genetically modified cell does not express the second AD.

[0083] In some embodiments, the present disclosure provides methods of inducing an immune response against target cells in a subject, comprising: (a) (i) administering to a subject in need thereof a therapeutically effective amount of genetically modified human immune effector cells comprising: (1) a first antigenic determinant binding domain (ADBD) that specifically binds to a first antigenic determinant (AD), (2) a transmembrane domain, and (3) an intracellular domain; and (ii) a genetic modification that abolishes expression of a second AD on the genetically modified cell; and (b) administering to the subject a therapeutically effective amount of an adaptor comprising the first AD recognized by the CAR and a second ADBD capable of binding to the first AD recognized by the CAR and the second AD expressed on the target cell.

[0084] In some embodiments, the present disclosure provides genetically modified human immune effector cells comprising: (a) two or more chimeric antigen receptors (CARs) comprising (i) a first antigenic determinant (ADBD) that specifically binds a first AD, (ii) a transmembrane domain, and (iii) an intracellular domain; and (b) a genetic modification that abolishes expression of the second AD on the genetically modified cells, wherein the genetically modified cells used in combination with an adaptor are capable of inducing an immune response against cells expressing the second AD in an in vitro assay, wherein the adaptor comprises the second ADBD that specifically binds the first AD and the second AD, and the genetically modified cells do not express the second AD.

[0085] In some embodiments, the present disclosure provides methods of inducing an immune response against target cells in a subject, comprising: (a) administering to a subject in need thereof a therapeutically effective amount of a chimeric antigen receptor (CAR) comprising: (1) two or more antigenic determinant (ADBD) comprising a first antigenic determinant binding domain (ADBD) that specifically binds to a first AD, (2) a transmembrane domain, and (3) an intracellular domain; and (ii) a genetic modification that abolishes expression of the second AD on the genetically modified cell; and (b) administering to the subject a therapeutically effective amount of an adaptor comprising the first AD recognized by the CAR and a second ADBD capable of binding to the second AD expressed on the target cell.

[0086] In some embodiments, the present disclosure provides: [1] A composition comprising: (a) a cell expressing a chimeric antigen receptor (CAR) comprising: (i) an antigenic determinant binding domain (ADBD) that binds to a first antigenic determinant (AD) on a target cell; (ii) a transmembrane domain; and (iii) an intracellular domain; and (b) an adaptor comprising an ADBD that binds to (i) the first AD and (ii) a second AD on the target cell. [2] A composition comprising: (a) a cell expressing a chimeric antigen receptor (CAR) comprising: (i) an antigenic determinant binding domain (ADBD) that binds to a first antigenic determinant (AD) on a target cell; (ii) a transmembrane domain; and (iii) an intracellular domain; and (b) an adaptor comprising an ADBD that binds to (i) the first AD and (ii) a second AD on a second target cell. [3] A composition comprising: (a) a cell expressing a chimeric antigen receptor (CAR) comprising: (i) an antigenic determinant binding domain (ADBD), which is an alternative scaffold binding domain (ASBD) that binds to a first antigenic determinant (AD); (ii) a transmembrane domain; and (iii) an intracellular domain; and (b) an adaptor comprising an ADBD that binds to (i) the first AD and (ii) a second AD on a target cell. [4] A composition comprising: (a) a cell expressing a chimeric antigen receptor (CAR) comprising: (i) an antigenic determinant binding domain (ADBD) that binds to a first antigenic determinant (AD), (ii) a transmembrane domain, and (iii) an intracellular domain; and (b) an adaptor comprising an ADBD comprising an ASBD that binds to (i) the first AD and (ii) a second AD on a target cell. [5] A composition comprising: (a) a cell expressing a chimeric antigen receptor (CAR) comprising: (i) an antigenic determinant binding domain (ADBD) that binds to a first antigenic determinant (AD), (ii) a transmembrane domain, and (iii) an intracellular domain; and (b) an adaptor comprising a D domain that binds to (i) the first AD and (ii) a second AD on a target cell. [6] A composition comprising: (a) a cell expressing a chimeric antigen receptor (CAR) comprising (i) a D domain that binds to a first antigenic determinant (AD), (ii) a transmembrane domain, and (iii) an intracellular domain; and (b) an adaptor comprising an ADBD that binds to (i) the first AD and (ii) a second AD on a target cell. [7] A composition comprising: (a) a cell expressing a chimeric antigen receptor (CAR) comprising (i) a first D domain that binds to a first antigenic determinant (AD), (ii) a transmembrane domain, and (iii) an intracellular domain; and (b) an adaptor comprising a second D domain that binds to (i) the first AD and (ii) a second AD on a target cell. [8] The composition according to any one of [1] to [7], wherein the CAR comprises a single-chain variable fragment (scFv) ADBD. [9] The composition according to any one of [1] to [7], wherein the CAR comprises an ADBD that is an alternative scaffold binding domain (ASBD).

[10] . The composition of [9], wherein the CAR comprises a D domain, and optionally, the CAR comprises a sequence selected from the group: SEQ ID NOs: 17-26, and 27 or SEQ ID NOs: 44-1078, and 1079.

[11] . The composition according to any one of [1] to

[10] , wherein the CAR comprises two ADBDs.

[12] . The composition of

[11] , wherein the CAR comprises an ASBD and an scFv.

[13] . The composition of

[11] , wherein the CAR comprises a D domain and an scFv.

[14] . The composition according to

[11] , wherein the CAR comprises two ASBDs.

[15] . The composition described in

[11] , wherein the CAR comprises a 2D domain.

[16] The composition according to any one of [1] to

[15] , wherein the intracellular domain of CAR is a signal transduction domain.

[17] . The composition of

[16] , wherein the intracellular domain of the CAR comprises a primary signaling domain.

[18] . The composition described in

[16] , wherein the intracellular domain of the CAR comprises a CD3ζ primary signaling domain.

[19] . The composition of

[17] or

[18] , wherein the intracellular domain of the CAR further comprises a costimulatory signaling domain.

[20] . The composition of

[19] , wherein the intracellular domain of the CAR comprises a costimulatory signaling domain selected from CD28, 41BB, CD27, and CD134.

[21] . The composition described in

[20] , wherein the intracellular domain of the CAR comprises a 41BB costimulatory signaling domain.

[22] The composition described in any of [1] to

[21] , wherein the CAR binds to an antigen selected from CD19, CD22, CD123, BCMA, CS1, HER2, TACI, BAFFR, and PDL1.

[23] . The composition of

[22] , wherein the CAR binds to BCMA, and optionally the CAR comprises a sequence selected from the group: SEQ ID NOs: 44-338, and 339.

[24] . The composition of

[22] , wherein the CAR binds to CD123, and optionally the CAR comprises a sequence selected from the group: SEQ ID NOs: 340-77 and 773.

[25] . The composition of

[22] , wherein the CAR binds to CD19, and optionally the CAR comprises a sequence selected from the group: SEQ ID NOs: 1030-1058 and 1059.

[26] . The composition of

[22] , wherein the CAR binds to CD22, and optionally the CAR comprises a sequence selected from the group: SEQ ID NOs: 1060-1068, and 1069.

[27] . The composition of

[22] , wherein the CAR binds to CS1, and optionally the CAR comprises a sequence selected from the group: SEQ ID NOs: 780-794 and 795.

[28] . The composition of

[22] , wherein the CAR binds to HER2, and optionally the CAR comprises a sequence selected from the group: SEQ ID NOs: 800-839 and 840.

[29] . The composition of

[22] , wherein the CAR binds to PDL1, and optionally the CAR comprises a sequence selected from the group: SEQ ID NOs: 1010-1016, 1074-1078, and 1079.

[30] . The composition of any of [1] to

[21] , wherein the CAR binds to AFP p26, and optionally the CAR comprises a sequence selected from the group: SEQ ID NOs: 841 to 983, and 984.

[31] The composition described in any of [1] to

[30] , wherein the CAR comprises two ADBDs that bind to different targets.

[32] The composition of

[31] , wherein the CAR binds to CD19 and CD123.

[33] The composition of

[31] , wherein CAR binds to BCMA and CS1.

[34] The composition described in

[31] , wherein the CAR binds to CD22 and CD123.

[35] The composition described in

[31] , wherein CAR binds to PDL1 and CD123.

[36] The composition of

[32] , wherein the CAR comprises a first ASBD that binds to CD19 and a second ASBD that binds to CD123.

[37] The composition of

[36] , wherein the CAR comprises a first D domain that binds to CD19 and a second D domain that binds to CD123.

[38] The composition described in

[32] , wherein the CAR comprises a D domain that binds to CD19 and an scFv that binds to CD123.

[39] The composition of

[33] , wherein the CAR comprises a first ASBD that binds to BCMA and a second ASBD that binds to CS1.

[40] The composition of

[39] , wherein the CAR comprises a first D domain that binds to BCMA and a second D domain that binds to CS1.

[41] The composition of

[33] , wherein the CAR comprises a D domain that binds to CS1 and an scFv that binds to BCMA.

[42] The composition of

[39] , wherein the CAR comprises a D domain that binds to BCMA and an scFv that binds to CS1.

[43] The composition of

[34] , wherein the CAR comprises a first ASBD that binds to CD22 and a second ASBD that binds to CD123.

[44] The composition of

[43] , wherein the CAR comprises a first D domain that binds to CD22 and a second D domain that binds to CD123.

[45] The composition described in

[34] , wherein the CAR comprises a D domain that binds to CD22 and an scFv that binds to CD123.

[46] The composition of

[35] , wherein the CAR comprises a first ASBD that binds to PDL1 and a second ASBD that binds to CD123.

[47] The composition of

[46] , wherein the CAR comprises a first D domain that binds to PDL1 and a second D domain that binds to CD123.

[48] The composition described in

[35] , wherein the CAR comprises a D domain that binds to PDL1 and an scFv that binds to CD123.

[49] The composition described in

[32] , wherein the CAR comprises an ASBD that binds to CD19 and an scFv that binds to CD123.

[50] . The composition of any of [1] to

[49] , wherein the adapter comprises an AD of a tumor antigen, and optionally the tumor antigen is selected from the group: BCMA, CD123, CD19, CD22, CS1, HER2, TACI, BAFFR, and PDL1.

[51] . The composition according to any one of [1] to

[50] , wherein the adaptor comprises an AD that is an epitope of AFP p26 or AFP, and optionally comprises amino acid residues of SEQ ID NO: 16 or 1117 to 1123.

[52] The composition according to any one of [1] to

[51] , comprising an ADBD in which the adaptor is an scFv.

[53] The composition according to any one of [1] to

[51] , wherein the adaptor comprises an ADBD that is an ASBD.

[54] . The composition of

[53] , wherein the adapter comprises a D domain, and optionally, the adapter comprises a sequence selected from the group: SEQ ID NOs: 17-26, and 27, or SEQ ID NOs: 44-1078, and 1079.

[55] . The composition according to any one of [1] to

[54] , wherein the adaptor comprises two ADBDs.

[56] The composition described in

[55] , wherein the two ADBDs are (a) the same, (b) bind to the same antigenic determinant, (c) bind to different ADs of the same antigen, or (d) bind to different antigens on the same cell, or (e) bind to different antigens on different cells.

[57] The composition of

[55] or

[56] , wherein the adaptor comprises two ASBDs.

[58] . The composition of any one of

[55] to

[57] , wherein the adapter comprises two D domains, and optionally the adapter comprises an amino acid sequence selected from the group: SEQ ID NOs: 44 to 1079.

[59] The composition of any one of

[55] or

[56] , wherein the adaptor comprises an ADBD that is an scFv and an ADBD that is an ASBD.

[60] The composition of

[59] , wherein the adaptor comprises an ADBD that is an scFv and an ADBD that is a D domain.

[61] The composition described in any of [1] to

[60] , wherein the adaptor comprises an ADBD that binds to a member selected from BCMA, CD123, CD19, CD22, CS1, HER2, TACI, BAFFR, and PDL1.

[62] The composition of

[61] , wherein the adaptor comprises an ADBD that binds to BCMA, and optionally the ADBD comprises a sequence selected from the group: SEQ ID NOs: 44-338, and 339.

[63] The composition of

[61] , wherein the adaptor comprises an ADBD that binds to CS1, and optionally the ADBD comprises a sequence selected from the group: SEQ ID NOs: 780-794, and 795.

[64] The composition of

[61] , wherein the adaptor comprises an ADBD that binds to CD123, and optionally the ADBD comprises a sequence selected from the group: SEQ ID NOs: 340-772, and 773.

[65] The composition of

[61] , wherein the adaptor comprises an ADBD that binds to CD19, and optionally the ADBD comprises a sequence selected from the group: SEQ ID NOs: 1030-1058, and 1059.

[66] The composition of

[61] , wherein the adaptor comprises an ADBD that binds to CD22, and optionally the ADBD comprises a sequence selected from the group: SEQ ID NOs: 1060-1068, and 1069.

[67] The composition of

[61] , wherein the adaptor comprises an ADBD that binds to HER2, and optionally the ADBD comprises a sequence selected from the group: SEQ ID NOs: 800-839, and 840.

[68] The composition of

[61] , wherein the adaptor comprises an ADBD that binds to TACI or BAFFR.

[69] The composition of

[61] , wherein the adaptor comprises an ADBD that binds to PDL1, and optionally the ADBD comprises a sequence selected from the group: SEQ ID NOs: 1010-1016, 1074-1078, and 1079.

[70] The composition according to any one of [1] to

[69] , wherein the adaptor is bispecific.

[71] The composition of

[70] , wherein the adaptors comprise an ADBD that binds to CD19 and an ADBD that binds to CD123.

[72] The composition of

[70] , wherein the adaptor comprises an ADBD that binds to BCMA and an ADBD that binds to CS1.

[73] The composition of

[70] , wherein the adaptors comprise an ADBD that binds to CD22 and an ADBD that binds to CD123.

[74] The composition of

[70] , wherein the adaptors comprise an ADBD that binds to PDL1 and an ADBD that binds to CD123.

[75] The composition according to any one of [1] to

[74] , wherein the target cells are tumor cells.

[76] . The composition of

[75] , wherein the tumor cells are selected from acute leukemia, chronic leukemia, polycythemia vera, lymphoma, Hodgkin's disease, non-Hodgkin's lymphoma, multiple myeloma, Waldenstrom's macroglobulinemia, heavy chain disease, myelodysplastic syndrome, hairy cell leukemia, myelodysplasia, breast cancer, and ovarian cancer.

[77] . The composition according to

[75] , wherein the tumor cells are multiple myeloma cells.

[78] The composition described in

[75] , wherein at least one target cell is a tumor cell.

[79] . The composition described in

[75] , wherein the first and second target cells are tumor cells.

[80] . The composition of

[79] , wherein the first and second target cells are of the same type.

[81] The composition of

[79] , wherein the first and second target cells are of different types.

[82] The composition of

[80] or

[81] , wherein the tumor cells are selected from acute leukemia, chronic leukemia, polycythemia vera, lymphoma, Hodgkin's disease, non-Hodgkin's lymphoma, multiple myeloma, Waldenstrom's macroglobulinemia, heavy chain disease, myelodysplastic syndrome, hairy cell leukemia, myelodysplasia, breast cancer, and ovarian cancer.

[83] The composition according to

[82] , wherein the tumor cells are multiple myeloma cells.

[84] The composition according to any one of [1] to

[83] , wherein the CAR-expressing cells are immune effector cells.

[85] The composition described in

[84] , wherein the immune effector cells are T cells.

[86] The composition according to

[84] , wherein the immune effector cells are NK cells.

[87] The composition according to any one of [1] to

[86] , wherein the CAR-expressing cells kill target cells.

[88] The composition according to any one of [1] to

[87] , wherein binding of the adapter to the antigenic determinant blocks the activity of the antigen, including AD.

[89] . A method for killing a target cell, comprising contacting the target cell with any one of the compositions [1] to

[88] .

[90] . A method for killing target cells, comprising contacting a composition comprising the target cells with an adaptor, (a) the composition comprising the target cell further comprises a cell expressing a CAR comprising (i) an antigenic determinant binding domain (ADBD) that binds to a first AD on the target cell, (ii) a transmembrane domain, and (iii) an intracellular domain; and (b) the adaptor comprises an ADBD that binds to (i) the first AD and (ii) a second AD on the target cell.

[91] . A method of killing one or more target cells, comprising contacting a composition comprising a first target cell with an adaptor, (a) the composition comprising the target cell further comprises a cell expressing a chimeric antigen receptor (CAR) comprising (i) an antigenic determinant binding domain (ADBD) that binds to a first AD on the first target cell, (ii) a transmembrane domain, and (iii) an intracellular domain; and (b) the adaptor comprises an ADBD that binds to (i) the first AD and (ii) a second AD on a second target cell.

[92] . A method for killing target cells, comprising contacting a composition comprising the target cells with an adaptor, (a) the composition comprising the target cells further comprises cells expressing a chimeric antigen receptor (CAR) comprising: (i) an antigenic determinant binding domain (ADBD) that is an ASBD that binds to a first AD, (ii) a transmembrane domain, and (iii) an intracellular domain; and (b) the adaptor comprises an ADBD that binds to (i) the first AD and (ii) a second AD on the target cell.

[93] A method for killing target cells, comprising contacting a composition comprising the target cells with an adaptor, (a) the composition comprising the target cells further comprises cells expressing a chimeric antigen receptor (CAR) comprising (i) an antigenic determinant binding domain (ADBD) that binds to the first AD, (ii) a transmembrane domain, and (iii) an intracellular domain; and (b) the adaptor comprises an ADBD that is an ASBD that binds to (i) the first AD and (ii) a second AD on the target cell.

[94] A method of killing a target cell, comprising contacting a composition comprising the target cell with a cell expressing a chimeric antigen receptor (CAR), (a) a first antigenic determinant (AD) is present on the target cell; (b) the composition comprising the first target cell further comprises an adaptor comprising an antigenic determinant binding domain (ADBD) that binds to (i) the first AD and (ii) a second AD on the target cell; and (c) The method, wherein the CAR comprises (i) an ADBD that binds to the first AD on a target cell or an adapter, (ii) a transmembrane domain, and (iii) an intracellular domain.

[95] . A method of killing one or more target cells, comprising contacting a composition comprising the target cells with a cell expressing a chimeric antigen receptor (CAR), (a) a first AD is present on a first target cell; (b) the composition comprising the first target cell further comprises a second target cell and an adaptor comprising an antigenic determinant binding domain (ADBD) that binds to (i) the first AD and (ii) a second AD on the second target cell; and (c) The method, wherein the CAR comprises (i) an ADBD that binds to said first AD on a first target cell or adapter, (ii) a transmembrane domain, and (iii) an intracellular domain.

[96] A method of killing a target cell, comprising contacting a composition comprising the target cell with a cell expressing a chimeric antigen receptor (CAR), (a) a first antigenic determinant (AD) is present on the target cell; (b) the composition comprising the target cell further comprises an adaptor comprising (i) an antigenic determinant binding domain (ADBD) that binds to the first AD on the target cell and (ii) a second AD; and (c) the CAR comprises (i) an ADBD that is an ASBD that binds to said second AD, (ii) a transmembrane domain, and (iii) an intracellular domain.

[97] A method of killing a target cell, comprising contacting a composition comprising the target cell with a cell expressing a chimeric antigen receptor (CAR), (a) a first antigenic determinant (AD) is present on the target cell; (b) the composition comprising the target cell further comprises an adaptor comprising (i) an antigenic determinant binding domain (ADBD), which is an ASBD that binds to the first AD on the target cell, and (ii) a second AD; and (c) the method, wherein the CAR comprises (i) an ADBD that binds to the second AD, (ii) a transmembrane domain, and (iii) an intracellular domain.

[98] . A method for transmitting an immune response to a target cell, comprising contacting a composition comprising the target cell with an adaptor, (a) the composition comprising the target cell further comprises a cell expressing a CAR comprising (i) an antigenic determinant binding domain (ADBD) that binds to a first AD on the target cell, (ii) a transmembrane domain, and (iii) an intracellular domain; and (b) the adaptor comprises an ADBD that binds to (i) the first AD and (ii) a second AD on the target cell.

[99] . A method of transducing an immune response to one or more target cells, comprising contacting a composition comprising a first target cell with an adaptor, (a) the composition comprising the target cell further comprises a cell expressing a chimeric antigen receptor (CAR) comprising (i) an antigenic determinant binding domain (ADBD) that binds to a first AD on the first target cell, (ii) a transmembrane domain, and (iii) an intracellular domain; and (b) the adaptor comprises an ADBD that binds to (i) the first AD and (ii) a second AD on a second target cell.

[0100] 1. A method of transducing an immune response to a target cell, comprising contacting a composition comprising the target cell with an adaptor, (a) the composition comprising the target cells further comprises cells expressing a chimeric antigen receptor (CAR) comprising: (i) an antigenic determinant binding domain (ADBD) that is an ASBD that binds to a first AD, (ii) a transmembrane domain, and (iii) an intracellular domain; and (b) the adaptor comprises an ADBD that binds to (i) the first AD and (ii) a second AD on the target cell.

[0101] 1. A method of transducing an immune response to a target cell, comprising contacting a composition comprising the target cell with an adaptor, (a) the composition comprising the target cells further comprises cells expressing a chimeric antigen receptor (CAR) comprising (i) an antigenic determinant binding domain (ADBD) that binds to the first AD, (ii) a transmembrane domain, and (iii) an intracellular domain; and (b) the adaptor comprises an ADBD that is an ASBD that binds to (i) the first AD and (ii) a second AD on the target cell.

[0102] 1. A method of transducing an immune response in a target cell, comprising contacting a composition comprising the target cell with a cell expressing a chimeric antigen receptor (CAR), (a) a first antigenic determinant (AD) is present on the target cell; (b) the composition comprising the first target cell further comprises an adaptor comprising an antigenic determinant binding domain (ADBD) that binds to (i) the first AD and (ii) a second AD on the target cell; and (c) The method, wherein the CAR comprises (i) an ADBD that binds to the first AD on a target cell or an adapter, (ii) a transmembrane domain, and (iii) an intracellular domain.

[0103] 1. A method of delivering an immune response to one or more target cells, comprising contacting a composition comprising the target cells with cells expressing a chimeric antigen receptor (CAR), (a) a first AD is present on a first target cell; (b) the composition comprising the first target cell further comprises a second target cell and an adaptor comprising an antigenic determinant binding domain (ADBD) that binds to (i) the first AD and (ii) a second AD on the second target cell; and (c) The method, wherein the CAR comprises (i) an ADBD that binds to said first AD on a first target cell or adapter, (ii) a transmembrane domain, and (iii) an intracellular domain.

[0104] 1. A method of transducing an immune response in a target cell, comprising contacting a composition comprising the target cell with a cell expressing a chimeric antigen receptor (CAR), (a) a first antigenic determinant (AD) is present on the target cell; (b) the composition comprising the target cell further comprises an adaptor comprising (i) an antigenic determinant binding domain (ADBD) that binds to the first AD on the target cell and (ii) a second AD; and (c) the CAR comprises (i) an ADBD that is an ASBD that binds to said second AD, (ii) a transmembrane domain, and (iii) an intracellular domain.

[0105] 1. A method of transducing an immune response in a target cell, comprising contacting a composition comprising the target cell with a cell expressing a chimeric antigen receptor (CAR), (a) a first antigenic determinant (AD) is present on the target cell; (b) the composition comprising the target cell further comprises an adaptor comprising (i) an antigenic determinant binding domain (ADBD), which is an ASBD that binds to the first AD on the target cell, and (ii) a second AD; and (c) the method, wherein the CAR comprises (i) an ADBD that binds to the second AD, (ii) a transmembrane domain, and (iii) an intracellular domain.

[0106] 1. A method of killing target cells in a patient comprising administering to the patient an adapter, (a) the patient is treated with cells expressing a chimeric antigen receptor (CAR) comprising (i) an antigenic determinant binding domain (ADBD) that binds to a first antigenic determinant (AD) on said target cell, (ii) a transmembrane domain, and (iii) an intracellular domain; (b) the adaptor comprises an ADBD that binds to (i) the first AD and (ii) a second AD on the target cell.

[0107] 1. A method of redirecting target cell killing in a patient comprising administering to the patient an adapter, (a) a patient is treated with cells expressing a chimeric antigen receptor (CAR) comprising (i) an antigenic determinant binding domain (ADBD) that binds to a first antigenic determinant (AD) on a first target cell, (ii) a transmembrane domain, and (iii) an intracellular domain; (b) the adaptor comprises an ADBD that binds to (i) the first AD and (ii) a second AD on a second target cell.

[0108] 1. A method of killing target cells in a patient comprising administering to the patient an adapter, (a) a patient is treated with cells expressing a chimeric antigen receptor (CAR) that includes (i) an antigenic determinant binding domain (ADBD), which is an ASBD that binds to a first antigenic determinant (AD), (ii) a transmembrane domain, and (iii) an intracellular domain; (b) the adaptor comprises an ADBD that binds to (i) the first AD and (ii) a second AD on the target cell.

[0109] 1. A method of killing target cells in a patient comprising administering to the patient an adapter, (a) a patient is treated with cells expressing a chimeric antigen receptor (CAR) comprising (i) an antigenic determinant binding domain (ADBD) that binds to a first antigenic determinant (AD), (ii) a transmembrane domain, and (iii) an intracellular domain; (b) the adaptor comprises an ADBD that is an ASBD that binds to (i) the first AD and (ii) a second AD on the target cell.

[0110] 1. A method of killing target cells in a patient comprising administering to the patient cells expressing a chimeric antigen receptor (CAR), (a) a first antigenic determinant (AD) and a second AD are present on a target cell; (b) the patient is treated with an adaptor comprising an antigenic determinant binding domain (ADBD) that binds to (i) the first AD and (ii) the second AD on the target cell; (c) The method, wherein the CAR comprises (i) an ADBD that binds to the first AD on a target cell or an adapter, (ii) a transmembrane domain, and (iii) an intracellular domain.

[0111] 1. A method of inducing target cell death in a patient comprising administering to the patient cells expressing a chimeric antigen receptor (CAR), (a) a patient is treated with an adaptor comprising an antigenic determinant binding domain (ADBD) that binds to (i) a first antigenic determinant (AD) and (ii) a second AD on a target cell; (b) the CAR comprises (i) an ADBD that binds to the first AD on the adaptor, (ii) a transmembrane domain, and (iii) an intracellular domain.

[0112] 1. A method of killing target cells in a patient comprising administering to the patient cells expressing a chimeric antigen receptor (CAR), (a) a first antigenic determinant (AD) is present on the target cell; (b) the patient is treated with an adaptor comprising (i) an antigenic determinant binding domain (ADBD) that binds to the first AD on the target cell and (ii) a second AD; (c) the CAR comprises (i) an ADBD that is an ASBD that binds to said second AD, (ii) a transmembrane domain, and (iii) an intracellular domain.

[0113] 1. A method of killing target cells in a patient comprising administering to the patient cells expressing a chimeric antigen receptor (CAR), (a) a first antigenic determinant (AD) is present on the target cell; (b) the patient is treated with an adaptor comprising (i) an antigenic determinant binding domain (ADBD), which is an ASBD that binds to the first AD on the target cell, and (ii) a second AD; (c) the method, wherein the CAR comprises (i) an ADBD that binds to the second AD, (ii) a transmembrane domain, and (iii) an intracellular domain.

[0114] 1. A method of delivering an immune response to a target cell in a patient, comprising administering to the patient an adaptor, (a) the patient is treated with cells expressing a chimeric antigen receptor (CAR) comprising (i) an antigenic determinant binding domain (ADBD) that binds to a first antigenic determinant (AD) on said target cell, (ii) a transmembrane domain, and (iii) an intracellular domain; (b) the adaptor comprises an ADBD that binds to (i) the first AD and (ii) a second AD on the target cell.

[0115] 1. A method of redirecting an immune response to a target cell in a patient, comprising administering to the patient an adaptor, (a) a patient is treated with cells expressing a chimeric antigen receptor (CAR) comprising (i) an antigenic determinant binding domain (ADBD) that binds to a first antigenic determinant (AD) on a first target cell, (ii) a transmembrane domain, and (iii) an intracellular domain; (b) the adaptor comprises an ADBD that binds to (i) the first AD and (ii) a second AD on a second target cell.

[0116] 1. A method of delivering an immune response to a target cell in a patient, comprising administering to the patient an adaptor, (a) a patient is treated with cells expressing a chimeric antigen receptor (CAR) that includes (i) an antigenic determinant binding domain (ADBD), which is an ASBD that binds to a first antigenic determinant (AD), (ii) a transmembrane domain, and (iii) an intracellular domain; (b) the adaptor comprises an ADBD that binds to (i) the first AD and (ii) a second AD on the target cell.

[0117] 1. A method of delivering an immune response to a target cell in a patient, comprising administering to the patient an adaptor, (a) a patient is treated with cells expressing a chimeric antigen receptor (CAR) comprising (i) an antigenic determinant binding domain (ADBD) that binds to a first antigenic determinant (AD), (ii) a transmembrane domain, and (iii) an intracellular domain; (b) the adaptor comprises an ADBD that is an ASBD that binds to (i) the first AD and (ii) a second AD on the target cell.

[0118] 1. A method of delivering an immune response to target cells in a patient comprising administering to the patient cells expressing a chimeric antigen receptor (CAR), (a) a first antigenic determinant (AD) and a second AD are present on a target cell; (b) the patient is treated with an adaptor comprising an antigenic determinant binding domain (ADBD) that binds to (i) the first AD and (ii) the second AD on the target cell; (c) The method, wherein the CAR comprises (i) an ADBD that binds to the first AD on a target cell or an adapter, (ii) a transmembrane domain, and (iii) an intracellular domain.

[0119] 1. A method of inducing an immune response against target cells in a patient, comprising administering to the patient cells expressing a chimeric antigen receptor (CAR), (a) a patient is treated with an adaptor comprising an antigenic determinant binding domain (ADBD) that binds to (i) a first antigenic determinant (AD) and (ii) a second AD on a target cell; (b) the CAR comprises (i) an ADBD that binds to the first AD on the adaptor, (ii) a transmembrane domain, and (iii) an intracellular domain.

[0120] 1. A method of delivering an immune response to target cells in a patient comprising administering to the patient cells expressing a chimeric antigen receptor (CAR), (a) a first antigenic determinant (AD) is present on the target cell; (b) the patient is treated with an adaptor comprising (i) an antigenic determinant binding domain (ADBD) that binds to the first AD on the target cell and (ii) a second AD; (c) the CAR comprises (i) an ADBD that is an ASBD that binds to said second AD, (ii) a transmembrane domain, and (iii) an intracellular domain.

[0121] 1. A method of delivering an immune response to target cells in a patient comprising administering to the patient cells expressing a chimeric antigen receptor (CAR), (a) a first antigenic determinant (AD) is present on the target cell; (b) the patient is treated with an adaptor comprising (i) an antigenic determinant binding domain (ADBD), which is an ASBD that binds to the first AD on the target cell, and (ii) a second AD; (c) the method, wherein the CAR comprises (i) an ADBD that binds to the second AD, (ii) a transmembrane domain, and (iii) an intracellular domain.

[0122] 1. A method of treating a proliferative disorder, cancer, autoimmune disease, infectious disease, or allograft rejection comprising contacting a composition comprising target cells with cells expressing a chimeric antigen receptor (CAR), (a) a first antigenic determinant (AD) is present on the target cell; (b) the composition comprising the target cell further comprises an adaptor comprising (i) an antigenic determinant binding domain (ADBD), which is an ASBD that binds to the first AD on the target cell, and (ii) a second AD; and (c) the method, wherein the CAR comprises (i) an ADBD that binds to the second AD, (ii) a transmembrane domain, and (iii) an intracellular domain.

[0123] A method of treating a proliferative disorder, cancer, autoimmune disease, infectious disease, or allograft rejection comprising administering to a patient an adapter, (a) the patient is treated with cells expressing a chimeric antigen receptor (CAR) comprising (i) an antigenic determinant binding domain (ADBD) that binds to a first antigenic determinant (AD) on said target cell, (ii) a transmembrane domain, and (iii) an intracellular domain; (b) the adaptor comprises an ADBD that binds to (i) the first AD and (ii) a second AD on the target cell.

[0124] 1. A method of redirecting treatment of a proliferative disorder, cancer, autoimmune disease, infectious disease, or allograft rejection comprising administering to a patient an adapter, (a) a patient is treated with cells expressing a chimeric antigen receptor (CAR) comprising (i) an antigenic determinant binding domain (ADBD) that binds to a first antigenic determinant (AD) on a first target cell, (ii) a transmembrane domain, and (iii) an intracellular domain; (b) the adaptor comprises an ADBD that binds to (i) the first AD and (ii) a second AD on a second target cell.

[0125] A method of treating a proliferative disorder, cancer, autoimmune disease, infectious disease, or allograft rejection comprising administering to a patient an adapter, (a) a patient is treated with cells expressing a chimeric antigen receptor (CAR) that includes (i) an antigenic determinant binding domain (ADBD), which is an ASBD that binds to a first antigenic determinant (AD), (ii) a transmembrane domain, and (iii) an intracellular domain; (b) the adaptor comprises an ADBD that binds to (i) the first AD and (ii) a second AD on the target cell.

[0126] A method of treating a proliferative disorder, cancer, autoimmune disease, infectious disease, or allograft rejection comprising administering to a patient an adapter, (a) a patient is treated with cells expressing a chimeric antigen receptor (CAR) comprising (i) an antigenic determinant binding domain (ADBD) that binds to a first antigenic determinant (AD), (ii) a transmembrane domain, and (iii) an intracellular domain; (b) the adaptor comprises an ADBD that is an ASBD that binds to (i) the first AD and (ii) a second AD on the target cell.

[0127] 1. A method of treating a proliferative disorder, cancer, autoimmune disease, infectious disease, or allograft rejection comprising administering to a patient cells expressing a chimeric antigen receptor (CAR), (a) a first antigenic determinant (AD) and a second AD are present on a target cell; (b) the patient is treated with an adaptor comprising an antigenic determinant binding domain (ADBD) that binds to (i) the first AD and (ii) the second AD on the target cell; (c) The method, wherein the CAR comprises (i) an ADBD that binds to the first AD on a target cell or an adapter, (ii) a transmembrane domain, and (iii) an intracellular domain.

[0128] 1. A method of inducing treatment of a proliferative disorder, cancer, autoimmune disease, infectious disease, or allograft rejection comprising administering to a patient cells expressing a chimeric antigen receptor (CAR), (a) a patient is treated with an adaptor comprising an antigenic determinant binding domain (ADBD) that binds to (i) a first antigenic determinant (AD) and (ii) a second AD on a target cell; (b) the CAR comprises (i) an ADBD that binds to the first AD on the adaptor, (ii) a transmembrane domain, and (iii) an intracellular domain.

[0129] 1. A method of treating a proliferative disorder, cancer, autoimmune disease, infectious disease, or allograft rejection comprising administering to a patient cells expressing a chimeric antigen receptor (CAR), (a) a first antigenic determinant (AD) is present on the target cell; (b) the patient is treated with an adaptor comprising (i) an antigenic determinant binding domain (ADBD) that binds to the first AD on the target cell and (ii) a second AD; (c) the CAR comprises (i) an ADBD that is an ASBD that binds to said second AD, (ii) a transmembrane domain, and (iii) an intracellular domain.

[0130] 1. A method of treating a proliferative disorder, cancer, autoimmune disease, infectious disease, or allograft rejection comprising administering to a patient cells expressing a chimeric antigen receptor (CAR), (a) a first antigenic determinant (AD) is present on the target cell; (b) the patient is treated with an adaptor comprising (i) an antigenic determinant binding domain (ADBD), which is an ASBD that binds to the first AD on the target cell, and (ii) a second AD; (c) the method, wherein the CAR comprises (i) an ADBD that binds to the second AD, (ii) a transmembrane domain, and (iii) an intracellular domain.

[0131] .A method according to any one of

[0122] to

[0130] , wherein the proliferative disorder or cancer is selected from acute leukemia, chronic leukemia, polycythemia vera, lymphoma, Hodgkin's disease, non-Hodgkin's lymphoma, multiple myeloma, Waldenstrom's macroglobulinemia, heavy chain disease, myelodysplastic syndrome, hairy cell leukemia, myelodysplasia, breast cancer, and ovarian cancer.

[0132] .A method according to any one of

[0122] to

[0130] , wherein the proliferative disorder or cancer is multiple myeloma.

[0133] 1. A method of treating a hematological cancer comprising contacting a composition comprising target cells with cells expressing a chimeric antigen receptor (CAR), (a) a first antigenic determinant (AD) is present on the target cell; (b) the composition comprising the target cell further comprises an adaptor comprising (i) an antigenic determinant binding domain (ADBD), which is an ASBD that binds to the first AD on the target cell, and (ii) a second AD; and (c) the method, wherein the CAR comprises (i) an ADBD that binds to the second AD, (ii) a transmembrane domain, and (iii) an intracellular domain.

[0134] A method of treating hematological cancer comprising administering to a patient an adapter, (a) the patient is treated with cells expressing a chimeric antigen receptor (CAR) comprising (i) an antigenic determinant binding domain (ADBD) that binds to a first antigenic determinant (AD) on said target cell, (ii) a transmembrane domain, and (iii) an intracellular domain; (b) the adaptor comprises an ADBD that binds to (i) the first AD and (ii) a second AD on the target cell.

[0135] 1. A method of redirecting treatment for hematological cancer comprising administering to a patient an adapter, (a) a patient is treated with cells expressing a chimeric antigen receptor (CAR) comprising (i) an antigenic determinant binding domain (ADBD) that binds to a first antigenic determinant (AD) on a first target cell, (ii) a transmembrane domain, and (iii) an intracellular domain; (b) the adaptor comprises an ADBD that binds to (i) the first AD and (ii) a second AD on a second target cell.

[0136] A method of treating hematological cancer comprising administering to a patient an adapter, (a) a patient is treated with cells expressing a chimeric antigen receptor (CAR) that includes (i) an antigenic determinant binding domain (ADBD), which is an ASBD that binds to a first antigenic determinant (AD), (ii) a transmembrane domain, and (iii) an intracellular domain; (b) the adaptor comprises an ADBD that binds to (i) the first AD and (ii) a second AD on the target cell.

[0137] A method of treating hematological cancer comprising administering to a patient an adapter, (a) a patient is treated with cells expressing a chimeric antigen receptor (CAR) comprising (i) an antigenic determinant binding domain (ADBD) that binds to a first antigenic determinant (AD), (ii) a transmembrane domain, and (iii) an intracellular domain; (b) the adaptor comprises an ADBD that is an ASBD that binds to (i) the first AD and (ii) a second AD on the target cell.

[0138] 1. A method of treating hematological cancer comprising administering to a patient cells expressing a chimeric antigen receptor (CAR), (a) a first antigenic determinant (AD) and a second AD are present on a target cell; (b) the patient is treated with an adaptor comprising an antigenic determinant binding domain (ADBD) that binds to (i) the first AD and (ii) the second AD on the target cell; (c) The method, wherein the CAR comprises (i) an ADBD that binds to the first AD on a target cell or an adapter, (ii) a transmembrane domain, and (iii) an intracellular domain.

[0139] 1. A method of inducing treatment for hematological cancer comprising administering to a patient cells expressing a chimeric antigen receptor (CAR), (a) a patient is treated with an adaptor comprising an antigenic determinant binding domain (ADBD) that binds to (i) a first antigenic determinant (AD) and (ii) a second AD on a target cell; (b) the CAR comprises (i) an ADBD that binds to the first AD on the adaptor, (ii) a transmembrane domain, and (iii) an intracellular domain.

[0140] 1. A method of treating hematological cancer comprising administering to a patient cells expressing a chimeric antigen receptor (CAR), (a) a first antigenic determinant (AD) is present on the target cell; (b) the patient is treated with an adaptor comprising (i) an antigenic determinant binding domain (ADBD) that binds to the first AD on the target cell and (ii) a second AD; (c) the CAR comprises (i) an ADBD that is an ASBD that binds to said second AD, (ii) a transmembrane domain, and (iii) an intracellular domain.

[0141] 1. A method of treating hematological cancer comprising administering to a patient cells expressing a chimeric antigen receptor (CAR), (a) a first antigenic determinant (AD) is present on the target cell; (b) the patient is treated with an adaptor comprising (i) an antigenic determinant binding domain (ADBD), which is an ASBD that binds to the first AD on the target cell, and (ii) a second AD; (c) the method, wherein the CAR comprises (i) an ADBD that binds to the second AD, (ii) a transmembrane domain, and (iii) an intracellular domain.

[0142] .A method according to any one of

[0133] to

[0141] , wherein the blood cancer is selected from acute leukemia, chronic leukemia, polycythemia vera, lymphoma, Hodgkin's disease, non-Hodgkin's lymphoma, multiple myeloma, Waldenstrom's macroglobulinemia, heavy chain disease, myelodysplastic syndrome, hairy cell leukemia, and myelodysplasia.

[0143] .A method according to any one of

[0133] to

[0141] , wherein the blood cancer is multiple myeloma.

[0144] A method according to any one of

[90] to

[0131] , wherein the CAR comprises a single-chain variable fragment (scFv) ADBD.

[0145] A method according to any one of

[90] to

[0143] , wherein the CAR comprises an ADBD that is an alternative scaffold binding domain (ASBD).

[0146] The method described in

[0145] , wherein the CAR comprises a D domain, and optionally the CAR comprises an array selected from the group: SEQ ID NOs: 17 to 26, and 27 or SEQ ID NOs: 44 to 1078, and 1079.

[0147] .A method according to any one of

[90] to

[0146] , wherein the CAR comprises two ADBDs.

[0148] The method described in

[0147] , wherein the CAR comprises an ASBD and an scFv.

[0149] The method described in

[0147] , wherein the CAR comprises a D domain and an scFv.

[0150] The method described in

[0147] , wherein the CAR contains two ASBDs.

[0151] The method described in

[0147] , wherein the CAR contains two D domains.

[0152] .A method according to any one of

[90] to

[0151] , wherein the intracellular domain of CAR is a signal transduction domain.

[0153] The method described in

[0152] , wherein the intracellular domain of CAR comprises a primary signaling domain.

[0154] The method described in

[0152] , wherein the intracellular domain of the CAR comprises the CD3ζ primary signaling domain.

[0155] The method described in

[0153] or

[0154] , wherein the intracellular domain of the CAR further comprises a costimulatory signaling domain.

[0156] The method described in

[0155] , wherein the intracellular domain of the CAR comprises a costimulatory signaling domain selected from CD28, 41BB, CD27, and CD134.

[0157] .The method described in

[0156] , wherein the intracellular domain of CAR comprises a 41BB costimulatory signaling domain.

[0158] A method according to any one of

[90] to

[0157] , wherein the CAR binds to an antigen selected from CD19, CD22, CD123, BCMA, CS1, HER2, TACI, BAFFR, and PDL1.

[0159] The method described in

[0158] , wherein the CAR binds to BCMA, and optionally the CAR comprises an array selected from the group: SEQ ID NOs: 44 to 338, and 339.

[0160] The method described in

[0158] , wherein the CAR binds to CS1, and optionally the CAR comprises an array selected from the group: SEQ ID NOs: 780 to 794, and 795.

[0161] The method described in

[0158] , wherein the CAR binds to CD123, and optionally the CAR comprises an array selected from the group: SEQ ID NOs: 340 to 772, and 773.

[0162] The method described in

[0158] , wherein the CAR binds to CD19, and optionally the CAR comprises an array selected from the group: SEQ ID NOs: 1030 to 1058, and 1059.

[0163] The method described in

[0158] , wherein the CAR binds to CD22, and optionally the CAR comprises an array selected from the group: SEQ ID NOs: 1060 to 1068, and 1069.

[0164] The method described in

[0158] , wherein the CAR binds to HER2, and optionally the CAR comprises an array selected from the group: SEQ ID NOs: 800 to 839, and 840.

[0165] The method described in

[0158] , wherein the CAR binds to PDL1, and optionally the CAR comprises an array selected from the group: SEQ ID NOs: 1010-1016, 1074-1078, and 1079.

[0166] A method according to any one of

[90] to

[0157] , wherein the CAR binds to AFP p26, and optionally the CAR comprises a sequence selected from the group: SEQ ID NOs: 841 to 983, and 984.

[0167] A method according to any one of

[90] to

[0166] , wherein the CAR comprises two ADBDs that bind to separate targets.

[0168] .The method described in

[0167] , wherein CAR binds to CD19 and CD123.

[0169] .The method described in

[0167] , wherein CAR binds to CD22 and CD123.

[0170] .The method described in

[0167] , wherein CAR binds to PDL1 and CD123.

[0171] .The method described in

[0167] , wherein CAR binds to CS1 and BCMA.

[0172] The method described in

[0168] , wherein the CAR comprises a first ASBD that binds to CD19 and a second ASBD that binds to C123.

[0173] The method described in

[0172] , wherein the CAR comprises a first D domain that binds to CD19 and a second D domain that binds to CD123.

[0174] The method described in

[0168] , wherein the CAR comprises a D domain that binds to CD19 and an scFv that binds to CD123.

[0175] The method described in

[0169] , wherein the CAR comprises a first ASBD that binds to CD22 and a second ASBD that binds to CD123.

[0176] The method described in

[0175] , wherein the CAR comprises a first D domain that binds to CD22 and a second D domain that binds to CD123.

[0177] The method described in

[0169] , wherein the CAR comprises a D domain that binds to CD22 and an scFv that binds to CD123.

[0178] The method described in

[0170] , wherein the CAR comprises a first ASBD that binds to PDL1 and a second ASBD that binds to CD123.

[0179] The method described in

[0178] , wherein the CAR comprises a first D domain that binds to PDL1 and a second D domain that binds to CD123.

[0180] The method described in

[0170] , wherein the CAR comprises a D domain that binds to PDL1 and an scFv that binds to CD123.

[0181] The method described in

[0168] , wherein the CAR comprises an ASBD that binds to CD19 and an scFv that binds to CD123.

[0182] The method described in

[0171] , wherein the CAR comprises a first ASBD that binds to BCMA and a second ASBD that binds to CS1.

[0183] The method described in

[0182] , wherein the CAR comprises a first D domain that binds to BCMA and a second D domain that binds to CS1.

[0184] The method described in

[0171] , wherein the CAR comprises a D domain that binds to BCMA and an scFv that binds to CS1.

[0185] The method described in

[0171] , wherein the CAR comprises a D domain that binds to CS1 and an scFv that binds to BCMA.

[0186] .A method according to any one of

[90] to

[0185] , wherein the adapter comprises an AD of a tumor antigen, and optionally the tumor antigen is selected from the group: BCMA, CD123, CD19, CD22, CS1, HER2, TACI, BAFFR, and PDL1.

[0187] The method described in

[90] to

[0186] , wherein the adapter comprises an AD that is an epitope of AFP p26 or AFP, and optionally comprises amino acid residues of SEQ ID NO: 16 or 1117 to 1123.

[0188] .A method according to any one of

[90] to

[0187] , comprising an ADBD in which the adaptor is an scFv.

[0189] .A method according to any one of

[90] to

[0187] , wherein the adaptor comprises an ADBD that is an ASBD.

[0190] The method described in

[0189] , wherein the adapter comprises a D domain, and optionally the adapter comprises an array selected from the group: SEQ ID NOs: 17 to 26, and 27, or SEQ ID NOs: 44 to 1078, and 1079.

[0191] .A method according to any one of

[90] to

[0190] , wherein the adapter comprises two ADBDs.

[0192] The method described in

[0191] , wherein the two ADBDs are (a) the same, (b) bind to the same antigenic determinant, (c) bind to different ADs of the same antigen, or (d) bind to different antigens on the same cell, or (e) bind to different antigens on different cells.

[0193] .A method described in

[0191] or

[0192] , wherein the adapter comprises two ASBDs.

[0194] .A method described in any one of

[0191] ,

[0192] , or

[0193] , wherein the adapter comprises two D domains, and optionally the adapter comprises an amino acid sequence selected from the group: SEQ ID NOs: 44 to 1079.

[0195] .A method described in

[0191] or

[0192] , wherein the adapter comprises an ADBD that is an scFv and an ADBD that is an ASBD.

[0196] The method described in

[0195] , wherein the adapter comprises an ADBD that is an scFv and an ADBD that is a D domain.

[0197] .A method according to any one of

[90] to

[0196] , wherein the adaptor comprises an ADBD that binds to a member selected from BCMA, CS1, HER2, CD123, CD19, CD22, TACI, BAFFR, and PDL1.

[0198] The method described in

[0197] , wherein the adapter comprises an ADBD that binds to BCMA, and optionally the ADBD comprises an array selected from the group consisting of SEQ ID NOs: 44 to 338, and 339.

[0199] The method described in

[0197] , wherein the adapter comprises an ADBD that binds to CS1, and optionally the ADBD comprises an array selected from the group: SEQ ID NOs: 780 to 794, and 795.

[0200] The method described in

[0197] , wherein the adapter comprises an ADBD that binds to CD123, and optionally the ADBD comprises an array selected from the group consisting of SEQ ID NOs: 340 to 772, and 773.

[0201] The method described in

[0197] , wherein the adapter comprises an ADBD that binds to CD19, and optionally the ADBD comprises an array selected from the group: SEQ ID NOs: 1030 to 1058, and 1059.

[0202] The method described in

[0197] , wherein the adapter comprises an ADBD that binds to CD22, and optionally the ADBD comprises an array selected from the group: SEQ ID NOs: 1060 to 1068, and 1069.

[0203] The method described in

[0197] , wherein the adapter comprises an ADBD that binds to HER2, and optionally, the ADBD comprises an array selected from the group: SEQ ID NOs: 800 to 839, and 840.

[0204] The method described in

[0197] , wherein the adapter comprises an ADBD that binds to TACI or BAFFR.

[0205] The method described in

[0197] , wherein the adapter comprises an ADBD that binds to PDL1, and optionally the ADBD comprises an array selected from the group: SEQ ID NOs: 1010-1016, 1074-1078, and 1079.

[0206] .A method according to any one of

[90] to

[0205] , wherein the adapter is bispecific.

[0207] The method described in

[0206] , wherein the adaptor comprises an ADBD that binds to BCMA and an ADBD that binds to CS1.

[0208] The method described in

[0206] , wherein the adaptor comprises an ADBD that binds to CD19 and an ADBD that binds to CD123.

[0209] The method described in

[0206] , wherein the adaptor comprises an ADBD that binds to CD22 and an ADBD that binds to CD123.

[0210] The method described in

[0206] , wherein the adaptor comprises an ADBD that binds to PDL1 and an ADBD that binds to CD123.

[0211] .A method according to any one of

[90] to

[0210] , wherein the target cells are tumor cells.

[0212] The method of

[0211] , wherein the tumor cells are selected from acute leukemia, chronic leukemia, polycythemia vera, lymphoma, Hodgkin's disease, non-Hodgkin's lymphoma, multiple myeloma, Waldenstrom's macroglobulinemia, heavy chain disease, myelodysplastic syndrome, hairy cell leukemia, myelodysplasia, breast cancer, and ovarian cancer.

[0213] .The method described in

[0211] , wherein the tumor cells are multiple myeloma.

[0214] .The method described in

[0211] , wherein at least one target cell is a tumor cell.

[0215] .The method described in

[0211] , wherein the first and second target cells are tumor cells.

[0216] .The method described in

[0215] , wherein the first and second tumor cells are of the same type.

[0217] .The method described in

[0215] , wherein the first and second tumor cells are of different types.

[0218] The method of

[0214] or

[0215] , wherein the tumor cells are selected from acute leukemia, chronic leukemia, polycythemia vera, lymphoma, Hodgkin's disease, non-Hodgkin's lymphoma, multiple myeloma, Waldenstrom's macroglobulinemia, heavy chain disease, myelodysplastic syndrome, hairy cell leukemia, myelodysplasia, breast cancer, and ovarian cancer.

[0219] .A method according to

[0214] or

[0215] , wherein the tumor cells are multiple myeloma.

[0220] .A method according to any one of

[90] to

[0219] , wherein the CAR-expressing cells are immune effector cells.

[0221] .The method described in

[0220] , wherein the immune effector cells are T cells.

[0222] .The method described in

[0220] , wherein the immune effector cells are NK cells.

[0223] .A method according to any one of

[90] to

[0222] , in which CAR-expressing cells kill target cells.

[0224] Genetically modified human immune effector cells, (a) a chimeric antigen receptor (CAR) comprising (1) an antigenic determinant binding domain (ADBD) that specifically binds to a human CD45 antigenic determinant (AD), (2) a transmembrane domain, and (3) an intracellular domain; and (b) Genetically modified human immune effector cells comprising a genetic modification that abolishes expression of human CD45 AD on the genetically modified cells, wherein the genetically modified cells are capable of inducing an immune response against CD45 AD-expressing cells in an in vitro assay, and wherein the genetically modified cells do not express CD45 AD.

[0225] A method for killing target cells, comprising contacting the genetically modified cells described in

[0224] with the target cells, wherein the target cells express human CD45.

[0226] A method for transmitting an immune response to a target cell, comprising contacting the genetically modified cell described in

[0224] with the target cell, wherein the target cell expresses human CD45.

[0227] A method for inducing an immune response against target cells in a subject, comprising administering a therapeutically effective amount of the genetically modified cells described in

[0224] to a subject in need thereof.

[0228] .A method for treating a proliferative disorder, cancer, autoimmune disease, infectious disease, or allograft rejection in a subject, comprising administering to a subject in need thereof a therapeutically effective amount of the genetically modified cells described in

[0224] .

[0229] A method for depleting lymphocytes, comprising administering an effective amount of the genetically modified cells described in

[0224] to a subject in need thereof.

[0230] A method for depleting memory T cells, comprising administering an effective amount of the genetically modified cells described in

[0224] to a subject in need thereof.

[0231] .A method for treating an autoimmune disease or disorder, comprising administering a therapeutically effective amount of the genetically modified cells described in

[0224] to a subject in need thereof.

[0232] A method for pretreating a subject for transplantation, comprising administering to a subject in need thereof an effective amount of the genetically modified cells described in

[0224] .

[0233] .A method for treating blood cancer, comprising administering a therapeutically effective amount of the genetically modified cells described in

[0224] to a subject in need thereof.

[0234] Genetically modified human immune effector cells, (a) a chimeric antigen receptor (CAR) comprising: (1) a first antigenic determinant binding domain (ADBD) that specifically binds to a first antigenic determinant (AD), (2) a transmembrane domain, and (3) an intracellular domain; and (b) comprising a genetic modification that abolishes expression of at least one human CD45 AD on the genetically modified cells; and a genetically modified human immune effector cell, wherein the first AD is not at least one human CD45 AD, and the genetically modified cell used in combination with the adapter in an in vitro assay is capable of inducing an immune response against CD45 AD-expressing cells, the adapter comprising a second ADBD that specifically binds to the first AD and at least one human CD45 AD, and the genetically modified cell does not express at least one human CD45 AD.

[0235] A method for killing target cells, comprising contacting a genetically modified cell described in

[0234] with an adapter and a target cell, wherein the target cell expresses CD45 and the adapter comprises a first AD and a second ADBD that specifically binds to human CD45 AD.

[0236] A method for transmitting an immune response to a target cell, comprising contacting a genetically modified cell described in

[0234] with an adapter and a target cell, wherein the target cell expresses CD45 and the adapter comprises a first AD and a second ADBD that specifically binds to human CD45 AD.

[0237] A method for treating hematological cancer, comprising contacting a genetically modified cell described in

[0234] with an adapter and cancer cells, wherein the cancer cells express CD45 and the adapter comprises a first AD and a second ADBD that specifically binds to human CD45 AD.

[0238] 1. A method of inducing an immune response against a target cell in a subject, comprising: (a) administering a therapeutically effective amount of a genetically modified cell according to any one of claims 1 to 3 to a subject in need thereof; and (b) administering to the subject a therapeutically effective amount of an adaptor comprising a first AD and a second ADBD that specifically binds to a human CD45 AD.

[0239] A method for inducing an immune response against target cells in a subject, comprising administering a therapeutically effective amount of the genetically modified cells described in

[0234] to a subject in need thereof, wherein the subject has been administered an adaptor comprising a first AD and a second ADBD that specifically binds to human CD45 AD.

[0240] A method for inducing an immune response against target cells in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of an adapter comprising a first AD and a second ADBD that specifically binds to human CD45 AD, wherein the subject has been administered a genetically modified cell described in

[0234] .

[0241] 1. A method of treating a proliferative disorder, cancer, autoimmune disease, infectious disease, or allograft rejection in a subject, comprising: (a) administering a therapeutically effective amount of a genetically modified cell according to any one of claims 1 to 3 to a subject in need thereof; and (b) administering to the subject a therapeutically effective amount of an adaptor comprising a first AD and a second ADBD that specifically binds to a human CD45 AD.

[0242] .A method for treating a proliferative disorder, cancer, autoimmune disease, infectious disease, or allograft rejection in a subject, comprising administering to a subject in need thereof a therapeutically effective amount of the genetically modified cells described in

[0234] , wherein the subject has been administered an adapter comprising a first AD and a second ADBD that specifically binds to human CD45 AD.

[0243] A method for treating a proliferative disorder, cancer, autoimmune disease, infectious disease, or allograft rejection in a subject, comprising administering to a subject in need thereof a therapeutically effective amount of an adapter comprising a first AD and a second ADBD that specifically binds to human CD45 AD, wherein the subject has been administered a genetically modified cell described in

[0234] .

[0244] A method for lymphocyte depletion comprising: (a) administering a therapeutically effective amount of a genetically modified cell according to any one of claims 1 to 3 to a subject in need thereof; and (b) administering to the subject a therapeutically effective amount of an adaptor comprising a first AD and a second ADBD that specifically binds to a human CD45 AD.

[0245] A method for lymphocyte depletion comprising administering a therapeutically effective amount of the genetically modified cells described in

[0234] to a subject in need thereof, wherein the subject has been administered an adaptor comprising a first AD and a second ADBD that specifically binds to human CD45 AD.

[0246] A method for lymphocyte depletion comprising administering to a subject in need thereof a therapeutically effective amount of an adapter comprising a first AD and a second ADBD that specifically binds to human CD45 AD, wherein the subject has been administered a genetically modified cell described in

[0234] .

[0247] A method for depleting memory T cells, comprising: (a) administering a therapeutically effective amount of a genetically modified cell according to any one of claims 1 to 3 to a subject in need thereof; and (b) administering to the subject a therapeutically effective amount of an adaptor comprising a first AD and a second ADBD that specifically binds to a human CD45 AD.

[0248] A method for depleting memory T cells, comprising administering to a subject in need thereof a therapeutically effective amount of an adapter comprising a first AD and a second ADBD that specifically binds to human CD45 AD, wherein the subject has been administered a genetically modified cell described in

[0234] .

[0249] A method for depleting memory T cells, comprising administering to a subject in need thereof a therapeutically effective amount of an adapter comprising a first AD and a second ADBD that specifically binds to human CD45 AD, wherein the subject has been administered a genetically modified cell described in

[0234] .

[0250] 1. A method of treating an autoimmune disease or disorder, comprising: (a) administering a therapeutically effective amount of a genetically modified cell according to any one of claims 1 to 3 to a subject in need thereof; and (b) administering to the subject a therapeutically effective amount of an adaptor comprising a first AD and a second ADBD that specifically binds to a human CD45 AD.

[0251] A method for treating an autoimmune disease or disorder, comprising administering to a subject in need thereof a therapeutically effective amount of an adapter comprising a first AD and a second ADBD that specifically binds to human CD45 AD, wherein the subject has been administered a genetically modified cell described in

[0234] .

[0252] A method for treating an autoimmune disease or disorder, comprising administering to a subject in need thereof a therapeutically effective amount of an adapter comprising a first AD and a second ADBD that specifically binds to human CD45 AD, wherein the subject has been administered a genetically modified cell described in

[0234] .

[0253] 1. A method of preparing a subject for transplantation, comprising: (a) administering a therapeutically effective amount of a genetically modified cell according to any one of claims 1 to 3 to a subject in need thereof; and (b) administering to the subject a therapeutically effective amount of an adaptor comprising a first AD and a second ADBD that specifically binds to a human CD45 AD.

[0254] A method for pretreating a subject for transplantation, comprising administering to a subject in need thereof a therapeutically effective amount of the genetically modified cells described in

[0234] , wherein the subject has been administered an adaptor comprising a first AD and a second ADBD that specifically binds to human CD45 AD.

[0255] A method for pretreating a subject for transplantation, comprising administering to a subject in need thereof a therapeutically effective amount of an adaptor comprising a first AD and a second ADBD that specifically binds to human CD45 AD, wherein the subject has been administered a genetically modified cell described in

[0234] .

[0256] 1. A method for treating hematological cancer, comprising: (a) administering a therapeutically effective amount of a genetically modified cell according to any one of claims 1 to 3 to a subject in need thereof; and (b) administering to the subject a therapeutically effective amount of an adaptor comprising a first AD and a second ADBD that specifically binds to a human CD45 AD.

[0257] .A method for treating hematological cancer comprising administering a therapeutically effective amount of the genetically modified cells described in

[0234] to a subject in need thereof, wherein the subject has been administered an adapter comprising a first AD and a second ADBD that specifically binds to human CD45 AD.

[0258] A method for treating hematological cancer, comprising administering to a subject in need thereof a therapeutically effective amount of an adapter comprising a first AD and a second ADBD that specifically binds to human CD45 AD, wherein the subject has been administered a genetically modified cell described in

[0234] .

[0259] Genetically modified human immune effector cells, (a) a chimeric antigen receptor (CAR) comprising: (i) two or more first antigen determinant binding domains (ADBDs) each comprising a first antigen determinant (ADBD) that binds to an AD; (ii) a transmembrane domain; and (iii) an intracellular domain; and (b) comprising a genetic modification that abolishes expression of the first AD on the genetically modified cell; Genetically modified human immune effector cells, wherein the genetically modified cells do not express the first AD.

[0260] A method for killing a target cell, comprising contacting the genetically modified cell described in

[0259] with the target cell.

[0261] .A method for transmitting an immune response to a target cell, comprising contacting the target cell with a genetically modified cell described in

[0259] .

[0262] A method for inducing an immune response against target cells in a subject, comprising administering a therapeutically effective amount of the genetically modified cells described in

[0259] to a subject in need thereof.

[0263] .A method for treating a proliferative disorder, cancer, autoimmune disease, infectious disease, or allograft rejection in a subject, comprising administering to a subject in need thereof a therapeutically effective amount of the genetically modified cells described in

[0259] .

[0264] A method for depleting lymphocytes, comprising administering an effective amount of the genetically modified cells described in

[0259] to a subject in need thereof.

[0265] A method for pretreating a subject for transplantation, comprising administering to a subject in need thereof an effective amount of the genetically modified cells described in

[0259] .

[0266] .A method for treating blood cancer, comprising administering a therapeutically effective amount of the genetically modified cells described in

[0259] to a subject in need thereof.

[0267] A method for killing a target cell, comprising contacting a genetically modified cell described in

[0259] with an adapter and the target cell, wherein the adapter comprises a CAR and an AD recognized by a second ADBD.

[0268] A method for transmitting an immune response to a target cell, comprising contacting a genetically modified cell described in

[0259] with an adapter and a target cell, wherein the adapter comprises a CAR and an AD recognized by a second ADBD capable of binding to a second AD on the target cell.

[0269] A method for treating hematological cancer comprising contacting a genetically modified cell described in

[0259] with an adapter and cancer cells, wherein the adapter comprises a CAR and an AD recognized by a second ADBD capable of binding to a second AD on the target cell.

[0270] 1. A method of inducing an immune response against a target cell in a subject, comprising: (a) administering a therapeutically effective amount of the genetically modified cells described in

[0259] to a subject in need thereof; and (b) administering to a subject a therapeutically effective amount of an adapter comprising a CAR and an AD recognized by a second ADBD capable of binding to a second AD on a target cell.

[0271] A method for inducing an immune response against target cells in a subject, comprising administering a therapeutically effective amount of the genetically modified cells described in

[0259] to a subject in need thereof, wherein the subject has been administered an adapter comprising a CAR and an AD recognized by a second ADBD capable of binding to a second AD on the target cell.

[0272] A method for inducing an immune response against target cells in a subject, comprising administering to a subject in need thereof a therapeutically effective amount of an adapter comprising a CAR and an AD recognized by a second ADBD capable of binding to a second AD on the target cell, wherein the subject has been administered a genetically modified cell described in

[0259] .

[0273] 1. A method of treating a proliferative disorder, cancer, autoimmune disease, infectious disease, or allograft rejection in a subject, comprising: (a) administering a therapeutically effective amount of the genetically modified cells described in

[0259] to a subject in need thereof; and (b) administering to a subject a therapeutically effective amount of an adapter comprising a CAR and an AD recognized by a second ADBD capable of binding to a second AD on a target cell.

[0274] A method for treating a proliferative disorder, cancer, autoimmune disease, infectious disease, or allograft rejection in a subject, comprising administering to a subject in need thereof a therapeutically effective amount of a genetically modified cell described in

[0259] , wherein the subject has been administered an adapter comprising a CAR and an AD recognized by a second ADBD capable of binding to a second AD on a target cell.

[0275] A method for treating a proliferative disorder, cancer, autoimmune disease, infectious disease, or allograft rejection in a subject, comprising administering to a subject in need thereof a therapeutically effective amount of an adapter comprising a CAR and an AD recognized by a second ADBD capable of binding to a second AD on a target cell, wherein the subject has been administered a genetically modified cell described in

[0259] .

[0276] A method for lymphocyte depletion comprising: (a) administering a therapeutically effective amount of the genetically modified cells described in

[0259] to a subject in need thereof; and (b) administering to a subject a therapeutically effective amount of an adapter comprising a CAR and an AD recognized by a second ADBD capable of binding to a second AD on a target cell.

[0277] A method for depleting lymphocytes, comprising administering a therapeutically effective amount of the genetically modified cells described in

[0259] to a subject in need thereof, wherein the subject has been administered an adapter comprising a CAR and an AD recognized by a second ADBD capable of binding to a second AD on a target cell.

[0278] A method for depleting lymphocytes, comprising administering to a subject in need thereof a therapeutically effective amount of an adapter comprising a CAR and an AD recognized by a second ADBD capable of binding to a second AD on a target cell, wherein the subject has been administered a genetically modified cell described in

[0259] .

[0279] 1. A method of preparing a subject for transplantation, comprising: (a) administering a therapeutically effective amount of the genetically modified cells described in

[0259] to a subject in need thereof; and (b) administering to a subject a therapeutically effective amount of an adapter comprising a CAR and an AD recognized by a second ADBD capable of binding to a second AD on a target cell.

[0280] A method for pretreating a subject for transplantation, comprising administering to a subject in need thereof a therapeutically effective amount of the genetically modified cells described in

[0259] , wherein the subject has been administered an adapter comprising a CAR and an AD recognized by a second ADBD capable of binding to a second AD on the target cell.

[0281] A method for pretreating a subject for transplantation, comprising administering to a subject in need thereof a therapeutically effective amount of an adapter comprising a CAR and an AD recognized by a second ADBD capable of binding to a second AD on a target cell, wherein the subject has been administered a genetically modified cell described in

[0259] .

[0282] 1. A method for treating hematological cancer, comprising: (a) administering a therapeutically effective amount of the genetically modified cells described in

[0259] to a subject in need thereof; and (b) administering to a subject a therapeutically effective amount of an adapter comprising a CAR and an AD recognized by a second ADBD capable of binding to a second AD on a target cell.

[0283] A method for treating hematological cancer comprising administering a therapeutically effective amount of the genetically modified cells described in

[0259] to a subject in need thereof, wherein the subject has been administered an adapter comprising a CAR and an AD recognized by a second ADBD capable of binding to a second AD on a target cell.

[0284] A method for treating hematological cancer, comprising administering to a subject in need thereof a therapeutically effective amount of an adapter comprising a CAR and an AD recognized by a second ADBD capable of binding to a second AD on a target cell, wherein the subject has been administered a genetically modified cell described in

[0259] .

[0285] Genetically modified human immune effector cells, (a) a chimeric antigen receptor (CAR) comprising: (i) a first antigenic determinant binding domain (ADBD) that specifically binds to a first antigenic determinant (AD), (ii) a transmembrane domain, and (iii) an intracellular domain; and (b) comprising a genetic modification that abolishes expression of a second AD on the genetically modified cells; The genetically modified human immune effector cells, wherein the genetically modified cells used in combination with the adaptor are capable of inducing an immune response against cells expressing a second AD in an in vitro assay, the adaptor comprising a second ADBD that specifically binds to the first AD and the second AD, and the genetically modified cells do not express the second AD.

[0286] Genetically modified human immune effector cells, (a) a chimeric antigen receptor (CAR) comprising: (i) two or more first antigen determinant binding domains (ADBDs) each comprising a first antigen determinant (ADBD) that specifically binds to the first antigen determinant (AD), (ii) a transmembrane domain, and (iii) an intracellular domain; and (b) comprising a genetic modification that abolishes expression of a second AD on the genetically modified cells; The genetically modified human immune effector cells, wherein the genetically modified cells used in combination with the adaptor are capable of inducing an immune response against cells expressing a second AD in an in vitro assay, the adaptor comprising a second ADBD that specifically binds to the first AD and the second AD, and the genetically modified cells do not express the second AD.

[0287] A method for inducing an immune response against target cells in a subject, comprising administering a therapeutically effective amount of genetically modified cells described in

[0285] or

[0286] to a subject in need thereof.

[0288] A method for treating a proliferative disorder, cancer, autoimmune disease, infectious disease, or allograft rejection in a subject, comprising administering to the subject in need thereof a therapeutically effective amount of a genetically modified cell described in

[0285] or

[0286] .

[0289] A method for depleting lymphocytes, comprising administering an effective amount of the genetically modified cells described in

[0285] or

[0286] to a subject in need thereof.

[0290] A method for pretreating a subject for transplantation, comprising administering to a subject in need thereof an effective amount of genetically modified cells described in

[0285] or

[0286] .

[0291] A method for treating blood cancer, comprising administering a therapeutically effective amount of the genetically modified cells described in

[0285] or

[0286] to a subject in need thereof.

[0292] A method for killing a target cell, comprising contacting a genetically modified cell described in

[0285] or

[0286] with an adapter and a target cell, wherein the adapter comprises a first AD and a second ADBD that specifically binds to the second AD, and the second AD is expressed on the target cell.

[0293] A method for transmitting an immune response to a target cell, comprising contacting a genetically modified cell described in

[0285] or

[0286] with an adapter and a target cell, wherein the adapter comprises a first AD and a second ADBD that specifically binds to the second AD, and the second AD is expressed on the target cell.

[0294] A method for treating hematological cancer, comprising contacting a genetically modified cell described in

[0285] or

[0286] with an adapter and cancer cells, wherein the adapter comprises a first AD and a second ADBD that specifically binds to the second AD, and the second AD is expressed on the cancer cells.

[0295] 1. A method of inducing an immune response against a target cell in a subject, comprising: (a) administering a therapeutically effective amount of a genetically modified cell according to

[0285] or

[0286] to a subject in need thereof; and (b) administering to a subject a therapeutically effective amount of an adapter comprising a first AD and a second ADBD that specifically binds to the second AD, wherein the second AD is expressed on a target cell.

[0296] A method for inducing an immune response against target cells in a subject, comprising administering a therapeutically effective amount of a genetically modified cell described in

[0285] or

[0286] to a subject in need thereof, wherein the subject has been administered an adapter comprising a first AD and a second ADBD that specifically binds to the second AD, and the second AD is expressed on the target cells.

[0297] A method for inducing an immune response against target cells in a subject, comprising administering to a subject in need thereof a therapeutically effective amount of an adapter comprising a first AD and a second ADBD that specifically binds to the second AD, wherein the subject has been administered a genetically modified cell described in

[0285] or

[0286] , and the second AD is expressed on the target cells.

[0298] 1. A method of treating a proliferative disorder, cancer, autoimmune disease, infectious disease, or allograft rejection in a subject, comprising: (a) administering a therapeutically effective amount of a genetically modified cell according to

[0285] or

[0286] to a subject in need thereof; and (b) administering to a subject a therapeutically effective amount of an adapter comprising a first AD and a second ADBD that specifically binds to the second AD, wherein the second AD is expressed on target cells associated with a proliferative disorder, cancer, an autoimmune disease, an infectious disease, or allograft rejection.

[0299] A method for treating a proliferative disorder, cancer, autoimmune disease, infectious disease, or allograft rejection in a subject, comprising administering a therapeutically effective amount of a genetically modified cell described in

[0285] or

[0286] to a subject in need thereof, wherein the subject has been administered an adapter comprising a first AD and a second ADBD that specifically binds to the second AD, and the second AD is expressed on target cells associated with the proliferative disorder, cancer, autoimmune disease, infectious disease, or allograft rejection.

[0300] A method for treating a proliferative disorder, cancer, autoimmune disease, infectious disease, or allograft rejection in a subject, comprising administering to a subject in need thereof a therapeutically effective amount of an adapter comprising a first AD and a second ADBD that specifically binds to the second AD, wherein the subject has been administered a genetically modified cell described in

[0285] or

[0286] , and the second AD is expressed on target cells associated with the proliferative disorder, cancer, autoimmune disease, infectious disease, or allograft rejection.

[0301] A method for lymphocyte depletion comprising: (a) administering a therapeutically effective amount of a genetically modified cell according to

[0285] or

[0286] to a subject in need thereof; and (b) administering to a subject a therapeutically effective amount of an adapter comprising a first AD and a second ADBD that specifically binds to the second AD, wherein the second AD is expressed on a lymphocyte target cell.

[0302] A method for depleting lymphocytes, comprising administering a therapeutically effective amount of a genetically modified cell described in

[0285] or

[0286] to a subject in need thereof, wherein the subject has been administered an adapter comprising a first AD and a second ADBD that specifically binds to the second AD, and the second AD is expressed on lymphocyte target cells.

[0303] A method for depleting lymphocytes, comprising administering to a subject in need thereof a therapeutically effective amount of an adapter comprising a first AD and a second ADBD that specifically binds to the second AD, wherein the subject has been administered a genetically modified cell described in

[0285] or

[0286] , and the second AD is expressed on lymphocyte target cells.

[0304] 1. A method of preparing a subject for transplantation, comprising: (a) administering a therapeutically effective amount of a genetically modified cell according to

[0285] or

[0286] to a subject in need thereof; and (b) administering to the subject a therapeutically effective amount of an adapter comprising a first AD and a second ADBD that specifically binds to the second AD, wherein the second AD is expressed on target cells associated with the transplant.

[0305] A method for pretreating a subject for transplantation, comprising administering to a subject in need thereof a therapeutically effective amount of genetically modified cells described in

[0285] or

[0286] , wherein the subject has been administered an adapter comprising a first AD and a second ADBD that specifically binds to the second AD, and the second AD is expressed on target cells associated with the transplant.

[0306] A method for pretreating a subject for transplantation, comprising administering to a subject in need thereof a therapeutically effective amount of an adaptor comprising a first AD and a second ADBD that specifically binds to the second AD, wherein the subject has been administered a genetically modified cell described in

[0285] or

[0286] , and the second AD is expressed on target cells associated with the transplant.

[0307] 1. A method for treating hematological cancer, comprising: (a) administering a therapeutically effective amount of a genetically modified cell according to

[0285] or

[0286] to a subject in need thereof; and (b) administering to a subject a therapeutically effective amount of an adapter comprising a first AD and a second ADBD that specifically binds to the second AD, wherein the second AD is expressed on a target cell associated with a hematological cancer.

[0308] A method for treating hematological cancer, comprising administering a therapeutically effective amount of a genetically modified cell described in

[0285] or

[0286] to a subject in need thereof, wherein the subject has been administered an adapter comprising a first AD and a second ADBD that specifically binds to the second AD, and the second AD is expressed on target cells associated with the hematological cancer.

[0309] A method for treating hematological cancer, comprising administering to a subject in need thereof a therapeutically effective amount of an adapter comprising a first AD and a second ADBD that specifically binds to the second AD, wherein the subject has been administered a genetically modified cell described in

[0285] or

[0286] , and the second AD is expressed on target cells associated with the hematological cancer.

[0310] An isolated adaptor polypeptide comprising (1) an antigenic determinant (AD) and (b) one or more antigenic determinant binding domains (ADBDs), An isolated adaptor polypeptide, wherein at least one ADBD specifically binds to human CD45 AD, and wherein the adaptor is capable of inducing an immune response by the genetically modified cells against target cells in an in vitro assay by contacting the adaptor with CD45 AD-expressing target cells in the presence of the genetically modified cells described in

[0234] . [Brief explanation of the drawings]

[0087] [Figure 1]We compared the cytolytic activity of CD123-specific (cg06) and BCMA-specific (bc-40) CARs to a CAR with no known target specificity (α3D) for a range of tumors using effector cell:target cell ratios ranging from 1:4 to 1:64. Briefly, 20,000 CAR T cells expressing bc40, cg06, or α3D were incubated with increasing numbers of CD123+ / BCMA- tumor targets (MOLM13) (Figure 1A); CD123- / BCMA+ tumor targets (H929) (Figure 1B); CD123- tumor targets (RAJI) (Figure 1C); or CD123+ / BCMA- tumor targets (MOLM13) (Figure 1D). After 16 h, cells were washed and luciferase activity was assessed. [Figure 2] Adapter binding correlates the binding specificity of the adapter and CAR. Jurkat NFAT luciferase reporter cells were transduced with a negative control CAR (α3D), an AFP (p26)-binding CAR (af03), or a BCMA-binding CAR (bc40). In Figure 2A, CAR-transduced Jurkat cells were incubated with 0.5 μg of adapter protein (20 min at 4°C), washed, and then stained with anti-His PE (clone J095G46, 20 min at 4°C). Figure 2B shows CAR expression by FLAG staining (clone L5) compared to mock-transduced Jurkat cells. [Figure 3-1] We demonstrate that adaptor binding of matched CAR:adapter and target:adapter specificities promotes target cell lysis. In Figure 3A, 40,000 CD123+BCMA-MOLM13-GFP / luciferase cells were incubated with various adaptors for 16 hours in the presence or absence of 20,000 T cells transduced with a BCMA-binding CAR (bc40) (E:T ratio = 1:2, donor D14-053017, day 7). After 16 hours, cells were washed and luciferase activity was assessed. % lysis was assessed relative to MOLM13-GFP / luciferase wells cultured in the absence of T cells or adaptor protein. Control CD123-specific CAR T cells (cg06) cultured at the same ratio were used as a positive control for lysis. [Figure 3-2] We demonstrate that adaptor ligation of matched CAR:adapter and target:adapter specificities promotes target cell lysis. In Figure 3B, 40,000 CD123+BCMA-MOLM13-GFP / luciferase cells were incubated with the CD123(cg06)-AFP(p26) adaptor in the presence or absence of 20,000 T cells transduced with AFP-conjugated CARs (Af03 or Af05) (E:T ratio = 1:2, donor D16-061317, day 7) for 16 hours. After 16 hours, cells were washed and luciferase activity was assessed. % lysis was assessed relative to MOLM13-GFP / luciferase wells cultured in the absence of T cells or adaptor protein. Control CD123-specific CAR T cells (cg06) cultured at the same ratio served as a positive control for lysis. In Figure 3C, 40,000 BCMA+NCI H929-GFP / luciferase cells were incubated with the bc40-AFP(p26) adaptor for 16 hours in the presence or absence of 10,000 T cells (E:T ratio = 1:4, donor D15-062017, day 8) mock-transduced or transduced with an AFP-conjugated CAR (Af03 or Af05). After 16 hours, cells were washed and luciferase activity was assessed. % lysis was assessed relative to NCI-H929-GFP / luciferase wells cultured in the absence of T cells or adaptor protein. [Figure 4-1] We demonstrate that adaptor ligation of matched specificities, CAR:adapter and target:adapter, promotes cytokine production by CAR T cells. In Figures 4A and 4B, donor D14-053017 T cells transduced with a BCMA-binding CAR (bc40) were cultured overnight with various adaptors in the presence or absence of CD123+BCMA-MOLM13 cells (25,000 T cells and target cells). Culture supernatants were collected and evaluated for IL-2 (Figure 4A) and IFN-γ (Figure 4B) production. [Figure 4-2]We demonstrate that adaptor ligation of matched specificities, CAR:adapter and target:adapter, promotes cytokine production by CAR T cells. In Figures 4C and 4D, donor D15-062017 T cells transduced with AFP-conjugated CARs (Af03 and Af05) were cultured overnight with the cg06-AFP(p26) adaptor in the presence or absence of CD123+BCMA-MOLM13 cells (25,000 T cells and target cells). Culture supernatants were collected and evaluated for IL-2 (Figure 4C) and IFN-γ (Figure 4D) production. [Figure 5] Adapter ligation of matched specificities, CAR:adapter and target:adapter, promotes CAR T cell proliferation. Donor D16-062717 cells transduced with an AFP-conjugated CAR (Af03) were CFSE-labeled (0.5 μM for 10 min) and then cultured for 72 h with or without mitomycin C-treated CD123-BCMA+ NCI-H929 cells (25,000 cells) in the presence or absence of CD123-specific or BCMA-specific adapters. At 72 h, cells were stained for CD3, and absolute CD3+ cell counts were subsequently analyzed by flow cytometry. [Figure 6] We demonstrate that matched CAR:adapter and target:adapter adapter ligation promotes signaling by CAR-expressing Jurkat NFAT-luciferase reporter cells. In Figure 6A, 50,000 reporter cells previously transduced with a BCMA-binding CAR (bc40) were cultured for 5 hours in the presence or absence of 50,000 CD123+BCMA-MOLM14 cells in the presence of various adapter proteins, and then luciferase activity was assessed. In Figure 6B, 50,000 reporter cells previously transduced with an AFP(p26)-binding CAR (af03) were cultured for 5 hours in the presence or absence of 50,000 BCMA+NCI-H929 cells in the presence of a nonspecific α3D-adapter or a BCMA-specific Bc40-adapter protein, and then luciferase activity was assessed. [Figure 7]This shows that a CD123-specific adaptor bearing a BCMA antigenic determinant can function together with a BCMA-specific D-domain CAR (bc40) or a BCMA-specific scFv CAR (c11D5-3). In Figure 7A, 40,000 CD123+ BCMA-MOLM13-GFP / luciferase cells were incubated with the Cg06-BCMA adaptor for 16 hours in the presence or absence of 20,000 T cells transduced with a nonspecific CAR (α3D), a BCMA-binding D-domain CAR (bc40), or a BCMA-binding scFv CAR (c11D5-3) (E:T ratio = 1:2, donor D14-062717, day 9). After 16 hours, cells were washed and luciferase activity was assessed. % lysis was assessed relative to the MOLM13-GFP / luciferase wells cultured in the absence of T cells or adaptor protein. The solid line shows the calculated three-parameter nonlinear curve, and the dotted line is for c11D5-3 and is presented for illustrative purposes only. In Figure 7B, CD123+BCMA-MOLM13-GFP / luciferase cells were cultured with transduced T cells in the absence of adapter protein in the same experiment as in Figure 7A. [Figure 8] This shows that AFP-specific CARs can simultaneously bind to both CD123 and BCMA through incubation with multiple adaptor proteins. 10 Jurkat NFAT luciferase cells transduced with AFP(p26)-binding CAR (af03) were incubated with 0.5 μg of BCMA-specific adaptor protein at various ratios (4°C for 20 min). After washing, they were incubated with CD123-Fc and biotinylated BCMA (0.5 μg each) for 20 min at 4°C. After washing, binding was detected with anti-Fc A488 and streptavidin-PE. Figure 8A shows flow cytometry analysis of CD123 and BCMA binding to their respective target proteins, and Figure 8B shows a comparison of the mean fluorescence intensity (MFI) of the A488 MFI (CD123 binding, left axis) and PE MFI (BCMA binding, right axis) flow cytometry data presented in Figure 8A. [Figure 9] Compared to single-binding domain adaptor proteins, dual-binding domain adaptor proteins promote enhanced signaling in CAR-expressing Jurkat NFAT luciferase reporter cells. In Figure 9A, 50,000 reporter cells pre-transduced with AFP (p26 domain)-conjugated CAR (af03) were cultured for 5 hours in the presence of 50,000 CD123+ or CD123-deficient MOLM13 cells in the presence of either the CD123-specific Cg06-adapter (Cg06-p26) or the Cg06-dual adaptor protein (Cg06-p26-Cg06), and then luciferase activity was assessed. CD123-deficient cells were generated using CRISPR / Cas9 genetic engineering technology. In Figure 9B, 50,000 reporter cells pre-transduced with AFP (p26 domain)-conjugated CAR (af03) were cultured for 5 hours in the presence or absence of 50,000 BCMA+ U266 cells in the presence of the BCMA-specific Bc40-adapter (Bc40-p26) or Bc40-dual adaptor protein (Bc40-p26-Bc40), and luciferase activity was then assessed. [Figure 10] Figure 1 shows that binding of truncated and full-length p26 to human FcRn is pH dependent. [Figure 11-1] We show that adaptors containing CS1 (SLAMF7, CRACC, CD319)-specific ADBDs regulate intracellular signaling and killing of CS1-positive tumors. In Figure 11A, the cc02 and cc08 ADBDs exhibit the most potent NFAT signaling when cultured in the presence of af59-CAR-expressing JNL10 cells and the CS1-positive tumor cell line, MM.1S (Figure 11A). Figure 11B shows that the bispecific bc98-p26-cc02 adaptor, which can bind both CS1 and BCMA, was more potent in its ability to signal than the monospecific BCMA-binding bc98-p26-α3DQ19E adaptor and the monospecific CS1-binding α3DQ19E-p26-cc02 adaptor. [Figure 11-2]We demonstrate that CS1 (SLAMF7, CRACC, CD319)-specific ADBD-containing adapters regulate intracellular signaling and killing of CS1-positive tumors. Figures 11C and 11D show that the bispecific bc98-p26-cc02 is an effective adapter in terms of killing HT929 (high expression of both BCMA and CS1, Figure 11C) and MM.1S (high expression of BCMA, low expression of CS1; Figure 11D). [Figure 12A] Adapters containing HER2-binding ADBDs induce signaling in Af59-CAR-expressing JNL10-cells cultured with HER2-positive SKBR3 tumors. Figure 12A shows that eb08 HER2-binding ADBD was the most potent stimulator in this assay. [Figure 12B] Adapters containing HER2-binding ADBD induce signaling in Af59-CAR-expressing JNL10 cells cultured with HER2-positive SKBR3 tumors. Figure 12B shows that NFAT signaling in JNL10 cells mediated by adapters containing eb08 is greater than that mediated by adapters containing zHER:4, comparable to that mediated by adapters containing 9.29, and less than that of G3 and zHER2:342. [Figure 12C] Adapters containing HER2-binding ADBD induce signaling in Af59-CAR-expressing JNL10-cells cultured with HER2-positive SKBR3 tumors. Figure 12C shows that adapters containing HER2-binding eb08 or eb04 modulate tumor lysis in a dose-dependent manner. DETAILED DESCRIPTION OF THE INVENTION

[0088] I. Definition Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs.

[0089] Whenever an embodiment is described herein with the term "comprising," other similar embodiments are also provided that are described with the terms "consisting of" and / or "consisting essentially of." However, when used as transitional phrases in the claims, each should be construed separately and in the appropriate legal and factual context (e.g., "comprising" is considered the more open-ended phrase, "consisting of" is considered more exclusive, and "consisting essentially of" is considered intermediate).

[0090] As used herein, the singular forms "a," "an," and "the" include plural references unless specifically stated otherwise.

[0091] The term "and / or" as used in phrases such as "A and / or B" is intended herein to include both A and B; A or B; A alone; and B alone. Similarly, the term "and / or" as used in phrases such as "A, B and / or C" encompasses each of the following embodiments: A, B, and C; A, B, or C; A or C; A or B; B or C; A and C; A and B; B and C; A alone; B alone; and C alone.

[0092] As used herein, the term "about," when referring to a measurable value, such as an amount, a time period, and other measurable values known in the art, is intended to encompass a ±20% variation from the specified value, or in some embodiments, a ±10% variation, or in some embodiments, a ±5% variation, or in some embodiments, a ±1% variation, or in some embodiments, a ±0.1% variation, as such variations are appropriate for practicing the disclosed methods.

[0093] The terms "protein" and "polypeptide" are used interchangeably herein to refer to a biological polymer comprising units derived from amino acids joined via peptide bonds; a protein may be composed of two or more polypeptide chains.

[0094] "Cell surface receptor" refers to molecules and complexes of molecules that can receive signals and transmit such signals across the plasma membrane of a cell. An example of a cell surface receptor provided herein is an activated integrin receptor, for example, an activated αβ integrin receptor on a metastatic cell. As used herein, "cell surface receptor" also encompasses molecules expressed on the cell surface, including CARs that can bind to a target antigenic determinant. The term "receptor" refers to a cell-associated protein that binds to or interacts with a molecule (e.g., a ligand) and mediates the action of the ligand on the cell. In some embodiments, the molecule that interacts with the receptor is a bioactive molecule. Membrane-associated cell surface receptors are characterized by a multidomain structure, which includes a ligand-binding domain, a transmembrane domain, and an intracellular effector domain that is typically involved in signal transduction.

[0095] As used herein, the term "chimeric antigen receptor" or "CAR" or "CARs" refers to a genetically engineered chimeric polypeptide that transfers antigen or target specificity to a cell, such as an immune cell (e.g., a T cell, such as a naive T cell, a central memory T cell, an effector memory T cell, an NK cell, an NKT, or a plurality or combination thereof). CARs may also be referred to herein as artificial T cell receptors, chimeric T cell receptors, or chimeric immune receptors. CARs share structural or functional properties with cellular immune function receptors or adapter molecules. Upon binding to a cognate antigen, CARs can activate or inactivate resident cytotoxic cells, modulate cellular anti-tumor activity, or otherwise modulate cellular immune responses. In some embodiments, CARs comprise one or more elements (e.g., domains) from a T cell receptor (TCR, e.g., the zeta chain associated with the T cell receptor complex) or from a natural killer cell receptor (NKR). In some embodiments, a CAR comprises (1) an antigenic determinant binding domain (ADBD) that specifically binds to an antigenic determinant (AD), (2) a transmembrane domain, and (3) an intracellular domain. In some embodiments, a CAR comprises two or more antigenic determinant binding domains. In some embodiments, a CAR comprises two or more antigenic determinant binding domains that bind to different antigenic determinants of the same antigen, different antigenic determinants on different antigens, or antigenic determinants expressed by different target cells.

[0096] As used herein, the term "immune cell" refers to a cell of the mammalian immune system, including, but not limited to, antigen-presenting cells, B cells, basophils, cytotoxic T cells, dendritic cells, eosinophils, granulocytes, helper T cells, leukocytes, lymphocytes, macrophages, mast cells, memory cells, monocytes, natural killer cells, neutrophils, phagocytes, plasma cells, and T cells.

[0097] The terms "T cells" and "T lymphocytes" are interchangeable and are used synonymously herein. Examples include, but are not limited to, naive T cells, central memory T cells, effector memory T cells, or combinations thereof.

[0098] "Autologous" as the term is used herein means any material derived from the same individual to whom it is later readministered.

[0099] "Allogeneic," as the term is used herein, refers to any material derived from a different animal of the same species as the individual to which it is introduced. Two or more individuals are said to be allogeneic to one another if their genes are not identical at one or more loci. In some embodiments, allogeneic material derived from individuals of the same species may be sufficiently genetically different to interact antigenically.

[0100] The term "effector cell" refers to a leukocyte that expresses one or more FcRs and performs effector function. Preferably, the cell expresses at least FcRIII and performs ADCC effector function. Examples of human leukocytes that mediate ADCC include peripheral blood mononuclear cells (PBMCs), natural killer (NK) cells, monocytes, cytotoxic T cells, and neutrophils; in certain embodiments, PBMCs and NK cells are preferred. Effector cells can be isolated from their natural sources, such as blood or PBMCs, as described herein or known in the art. In certain embodiments, the effector cells are human effector cells.

[0101] The term "effector function" refers to a specialized function of a differentiated cell. Effector functions of T cells can be, for example, cytolytic activity or helper activity, including secretion of cytokines.

[0102] As used herein, the term "immune response" refers to immunity, including but not limited to innate immunity, humoral immunity, cellular immunity, immunity, inflammatory response, acquired immunity, autoimmunity, and / or hyperactive immunity. Indicators of immune response may include cytokine secretion by immune cells, proliferation of immune cell populations, antibody production, degranulation of cytotoxic cells, and target cell killing. Such indicators can be measured in a routine manner using readily available assays, such as ELISA or ELISPOT, known in the art.

[0103] As used herein, the term "adapter" refers to a multidomain soluble protein that includes an antigenic determinant (AD) and an antigenic determinant binding domain (ADBD) that binds to a second AD. In addition to the AD and ADBD, the adapter may include additional ADs, additional ADBDs, and / or other additional domains.

[0104] The terms "antibody" and "immunoglobulin," as used interchangeably herein, include whole antibodies. Whole antibodies contain at least two heavy (H) chains and two light (L) chains interconnected by disulfide bonds. Each heavy chain consists of a heavy chain variable region (abbreviated as VH) and a heavy chain constant region. The heavy chain constant region consists of three domains, CH1, CH2, and CH3. Each light chain consists of a light chain variable region (abbreviated as VL) and a light chain constant region. The light chain constant region consists of one domain, Cl. The VH and VL regions can be further subdivided into regions of hypervariability called complementarity-determining regions (CDRs) interspersed with more conserved regions called framework regions (FWs). Each VH and VL consists of three CDRs and four FWs, arranged from amino terminus to carboxy terminus in the following order: FW1, CDR1, FW2, CDR2, FW3, CDR3, FW4. The variable regions of the heavy and light chains contain a binding domain that interacts with an antigen, and the constant regions of the antibody can mediate the binding of the immunoglobulin to host tissues or factors, including various cells of the immune system (e.g., effector cells) and the first component (C1q) of the classical complement system.

[0105] As used herein, the term "antibody fragment" or the like includes any functional domain of an antibody, such as an antigen-binding fragment or single chain thereof, an effector domain, a salvage receptor-binding epitope, or portions thereof. Antibody fragments described herein can exist in various forms. Examples of antibody fragments include, but are not limited to, Fab, Fab', F(ab')2, and Fv fragments, disulfide-linked Fv (sdFv), Fd fragments consisting of VH and CH1 domains, scFv, minibodies, BiTEs, Tandbabs, diabodies ((VL-VH)2 or (VH-VL)2), single-domain antibodies (e.g., sdAbs (VL or VH) such as nanobodies, and camelid VHH domains), and multispecific antibodies formed from antibody fragments. In some embodiments, an "antibody fragment" corresponds to the antigen-binding or epitope-binding site of an antibody. In other embodiments, the "antibody fragment" corresponds to other functional regions of the antibody, such as an effector domain or portion thereof, or a salvage receptor binding epitope or portion thereof.

[0106] As used herein, the term "single-chain variable fragment," or "scFv" antibody, refers to a form of antibody (e.g., antibody fragment) that contains only the variable regions of the heavy and light chains joined by a linker peptide. scFvs can contain VL-linker-VH or VH-linker-VL. scFv antibodies are typically 220-250 amino acids in length and contain a linker that is 10-25 amino acids in length.

[0107] As used herein, the term "Fc region" or simply "Fc" is understood to mean the carboxyl-terminal portion of an immunoglobulin chain constant region, preferably an immunoglobulin heavy chain constant region, or a portion thereof. For example, an immunoglobulin Fc region can include (1) a CH1 domain, a CH2 domain, and a CH3 domain, (2) a CH1 domain and a CH2 domain, (3) a CH1 domain and a CH3 domain, (4) a CH2 domain and a CH3 domain, or (5) a combination of two or more domains and an immunoglobulin hinge region. In a preferred embodiment, the immunoglobulin Fc region includes at least an immunoglobulin hinge region, a CH2 domain, and a CH3 domain, and preferably lacks a CH1 domain. In one embodiment, the immunoglobulin class from which the heavy chain constant region is derived is IgG (Igγ) (γ subclass 1, 2, 3, or 4). Other classes of immunoglobulins, such as IgA (Igα), IgD (Igδ), IgE (Igε), and IgM (Igμ), may also be used. The selection of appropriate immunoglobulin heavy chain constant regions is discussed in detail in U.S. Patent Nos. 5,541,087 and 5,726,044, each of which is incorporated herein by reference in its entirety. The selection of specific immunoglobulin heavy chain constant region sequences from specific immunoglobulin classes and subclasses to achieve particular results is considered to be within the level of skill in the art. The portion of the DNA construct encoding the immunoglobulin Fc region preferably includes at least a portion of the hinge domain and preferably at least a portion of the CH3 domain of Fc gamma or a homologous domain of either IgA, IgD, IgE, or IgM. Furthermore, substitutions or deletions of amino acids within the immunoglobulin heavy chain constant region may be useful in practicing the methods and compositions disclosed herein. One example would be to introduce amino acid substitutions in the upstream CH2 region to generate Fc variants with reduced affinity for Fc receptors (Cole, J. Immunol. 159:3613 (1997)).

[0108] "Antibody-dependent cellular cytotoxicity" or "ADCC" refers to a cell-mediated reaction in which nonspecific cytotoxic cells expressing Fc receptors (FcRs) (e.g., natural killer (NK) cells, neutrophils, and macrophages) recognize bound antibodies on target cells and subsequently lyse (or cause other cytotoxic effects on) the target cells. To assess ADCC activity of a molecule of interest, any in vitro ADCC assay known in the art can be used, such as those described in U.S. Pat. Nos. 5,500,362 or 5,821,337. Useful effector cells for such assays include, but are not limited to, peripheral blood mononuclear cells (PBMCs) and natural killer (NK) cells. Alternatively, or additionally, ADCC activity of a molecule of interest can be assessed in vivo using an animal model such as that disclosed in Clynes et al., PNAS 95:652-656 (1998).

[0109] The term "antigenic determinant binding domain" or "ADBD" is used herein to refer to a polypeptide sequence (e.g., an adapter or CAR) sufficient to confer recognition and specific binding to a target antigenic determinant (AD). In some embodiments, the ADBD is an antigen-binding antibody fragment, scFv, or an antigen-binding peptide not based on an antibody or antibody fragment sequence (e.g., a D domain or affibody). In some embodiments, the ADBD comprises a non-antibody-based binding scaffold (e.g., a D domain, affibody, fibronectin domain, nanobody, lipocalin domain, ankyrin domain, maxibody, protein A domain, or affilin domain). In some embodiments, the ADBD is a D domain. In some embodiments, the ADBD is an antibody-based binding sequence. In some embodiments, the ADBD is an scFv or domain antibody (dAb). In some embodiments, the ADBD is capable of binding to a target antigen on the surface of a cell. In some embodiments, the ADBD is capable of binding to a target antigen on the surface of an immune effector cell. In some embodiments, the ADBD is capable of binding to a growth factor receptor or hormone receptor.

[0110] In certain embodiments, the ADBD is a non-antibody scaffold-based polypeptide sequence sufficient to confer recognition and specific binding to a target antigenic determinant. In some embodiments, the non-antibody-based ADBD is a polypeptide capable of binding to a target antigen on the surface of a cell. In some embodiments, the non-antibody-based ADBD is capable of binding to a growth factor receptor or hormone receptor. In some embodiments, the ADBD is a D-domain-based polypeptide. In certain embodiments, the ADBD is a D-domain-based polypeptide sufficient to confer recognition and specific binding to a target antigenic determinant. In some embodiments, the ADBD is a D-domain-based polypeptide capable of binding to a target antigen on the surface of a cell. In some embodiments, the ADBD is a D-domain-based polypeptide capable of binding to a growth factor receptor or hormone receptor. In some embodiments, the ADBD is a D-domain-based polypeptide capable of binding to a target antigen on a serum protein.

[0111] The terms "specifically bind" or "having selective affinity for" mean that a binding substance, such as an adapter or CAR, reacts or binds to an epitope, protein, or target molecule more frequently, more rapidly, for a longer period of time, with greater affinity for the target epitope, or some combination thereof, than to other substances, including unrelated proteins. Due to sequence identity between homologous proteins in different species, specific binding, in some embodiments, includes binding substances that recognize proteins or targets in more than one species. Similarly, due to homology within sequence regions of particular polypeptides of different proteins, specific binding can include binding substances that recognize more than one protein or target. In certain embodiments, a binding substance that specifically binds to a first target may or may not specifically bind to a second target. Thus, "specific binding" does not necessarily require (although it can include) exclusive binding, e.g., binding to a single target. Thus, a binding substance may, in certain embodiments, specifically bind to more than one target. In certain embodiments, multiple targets may be bound by the same antigen-binding site on a binding substance.

[0112] The terms "linker," "spacer," and "hinge" are used interchangeably herein to refer to a peptide or other chemical linkage between two or more otherwise independent functional domains of an adapter or CAR. For example, a linker can be placed between the antigenic determinant domain and the antigenic determinant binding domain of an adapter. Similarly, a linker can be placed between two antigenic determinant binding domains of a CAR, or between the antigen binding domain and the transmembrane domain. Suitable linkers for connecting two or more domains of an adapter are described herein and / or will be apparent to those skilled in the art for other reasons.

[0113] As used herein, the term "operably linked" refers to the association of two molecules such that each retains at least some level of functional activity that each molecule had alone. In some embodiments, where one molecule does not have a functional activity, it is operably linked to another molecule if the other molecule has at least some level of that functional activity. Operably linked can also refer to the linkage of two non-functional molecules. Two molecules can be "operably linked" whether they are directly or indirectly (e.g., via a linker).

[0114] "Target" means any molecule or combination of molecules that can be bound by an adaptor or CAR, or an element of an adaptor or CAR, e.g., an antigenic determinant binding domain.

[0115] As used herein, the term "target cell" refers to a cell that is involved in a disease and can be targeted by a CAR, adapter, and / or CAR / adapter composition provided herein. Target cells include any cell in a subject (e.g., a human or animal) that can be targeted by a CAR, adapter, and / or CAR / adapter composition. A target cell can be a cell that expresses or overexpresses a target specifically bound by a CAR, adapter, and / or CAR / adapter composition.

[0116] Terms such as "binding affinity to a target," "binding to a target," and similar terms known in the art refer to a property of a polypeptide that can be directly measured by determining the affinity constant, for example, the amount of adaptor that binds and dissociates at a given antigen concentration. Other methods, including but not limited to, competitive analysis, equilibrium analysis, and microcalorimetry, and real-time interaction analysis based on surface plasmon resonance interactions (e.g., using a Biacore® device), can be used to characterize intermolecular interactions. These methods are well known to those skilled in the art and are described, for example, in Neri et al., Tibtech 14:465-470 (1996), and Jansson et al., J. Biol. Chem. 272:8189-8197 (1997).

[0117] The terms "antigenic determinant" and "epitope" are used interchangeably herein and refer to a portion of any molecule (e.g., a target or adapter) that can be recognized and specifically bound by a specific binding agent (e.g., an adapter or CAR). When the recognized molecule is a polypeptide, an epitope is formed from contiguous and non-contiguous amino acids and / or chemically active surface groups of other molecules (such as carbohydrates) juxtaposed by tertiary folding of the protein. Epitopes formed from contiguous amino acids are typically retained upon protein denaturation, whereas epitopes formed by tertiary folding are typically lost upon protein denaturation. Epitopes typically comprise at least three amino acids, more commonly at least five or 8-10 amino acids, in a unique spatial conformation.

[0118] As used herein, the term "derived from" refers to a relationship between a first and a second molecule. It generally refers to a structural similarity between the first and second molecules and does not imply or imply a limitation on the process or source of the first molecule from which the second molecule is derived. For example, in the case of an intracellular signaling domain derived from the CD3ζ molecule, the intracellular signaling domain retains the appropriate CD3ζ structure such that it has the required function, i.e., the ability to generate a signal under appropriate conditions. It does not imply or imply a limitation on the particular process by which the intracellular signaling domain is generated; for example, it does not mean starting with the CD3ζ sequence and deleting or mutating undesired sequences to arrive at the intracellular signaling domain.

[0119] The term "native" when used in reference to biological materials such as nucleic acid molecules, polypeptides, antigenic determinants, and host cells means that which is found in nature and has not been modified by humans. Conversely, the terms "non-natural" or "synthetic" when used in reference to biological materials means that which is not found in nature and has been modified by humans.

[0120] As used herein, a "modification" relative to a reference sequence includes substitutions, deletions, insertions, and / or additions to the sequence compared to the corresponding amino acid positions in the reference sequence.

[0121] A "substitution" relative to a reference sequence refers to the substitution of a particular amino acid residue with a different amino acid residue at the corresponding amino acid position in the reference sequence.

[0122] A "conservative" amino acid substitution is one in which one amino acid residue is replaced with another amino acid residue having a similar side chain. Families of amino acid residues having similar side chains have been defined in the art, including basic side chains (e.g., lysine (K), arginine (R), histidine (H)), acidic side chains (e.g., aspartic acid (D), glutamic acid (E)), uncharged polar side chains (e.g., glycine (G), asparagine (N), glutamine (Q), serine (S), threonine (T), tyrosine (Y), cysteine (C), and cysteine (D). Conservative substitutions include: alanine (A), valine (V), leucine (L), isoleucine (I), proline (P), phenylalanine (F), methionine (M), tryptophan (W)), non-polar side chains (e.g., alanine (A), valine (V), leucine (L), isoleucine (I), proline (P), phenylalanine (F), methionine (M), tryptophan (W)), beta-branched side chains (e.g., threonine (T), valine (V), isoleucine (I)), and aromatic side chains (e.g., tyrosine (Y), phenylalanine (F), tryptophan (W), histidine (H)). For example, substitution of tyrosine for phenylalanine is a conservative substitution. In one embodiment, conservative substitutions in the sequence of an adaptor or CAR retain specific binding of the adaptor or CAR containing the substitution to the binding target. Methods for identifying nucleotide and amino acid conservative and non-conservative substitutions that confer, alter, or maintain selective binding affinity are known in the art (see, e.g., Brummell, Biochem. 32:1180-1187 (1993); Kobayashi, Protein Eng. 12(10):879-884 (1999); and Burks, PNAS 94:412-417 (1997)).

[0123] A "non-conservative" amino acid substitution is one in which one amino acid residue is replaced with another amino acid residue having a heterologous side chain. In one embodiment, a non-conservative substitution in the sequence of an adapter or CAR retains the specific binding of the adapter or CAR containing the substitution to the binding target.

[0124] The terms "unnatural amino acid," "amino acid analog," and "non-standard amino acid residue" are used interchangeably herein. Unnatural amino acids that can be substituted in the adaptors provided herein are known in the art. In one embodiment, the unnatural amino acid is 4-hydroxyproline, which can be substituted for proline, 5-hydroxylysine, which can be substituted for lysine, 3-methylhistidine, which can be substituted for histidine, homoserine, which can be substituted for serine, and ornithine, which can be substituted for lysine. Additional examples of unnatural amino acids that can be substituted in adaptors include, but are not limited to, molecules such as D-isomers of the common amino acids, 2,4-diaminobutyric acid, α-aminoisobutyric acid, A-aminobutyric acid, Abu, 2-aminobutyric acid, γ-Abu, ε-Ahx, 6-aminohexanoic acid, Aib, 2-aminoisobutyric acid, 3-aminopropionic acid, ornithine, norleucine, norvaline, hydroxyproline, sarcosine, citrulline, homocitrulline, cysteic acid, t-butylglycine, t-butylalanine, phenylglycine, cyclohexylalanine, β-alanine, lanthionine, dehydroalanine, γ-aminobutyric acid, selenocysteine, and pyrrolidine fluoroamino acids, designer amino acids such as β-methyl amino acids, Cα-methyl amino acids, and Nα-methyl amino acids, or combinations of unnatural amino acids. Further additional unnatural amino acids can include 4-aminobutyric acid, 4-amino-3-hydroxy-5-phenylpentanoic acid, 4-amino-3-hydroxy-6-methylheptanoic acid, 2-thienylalanine, and / or D-isomers of amino acids. As discussed herein, in some embodiments, the unnatural amino acid or amino acid analog can include one or more amino acid deletions from the sequence.

[0125] The terms "polynucleotide" and "nucleic acid," used interchangeably herein, refer to polymeric forms of nucleotides of any length, ribbon nucleotides, or deoxynucleotides. These terms include, but are not limited to, DNA, RNA, cDNA (complementary DNA), mRNA (messenger RNA), rRNA (ribosomal RNA), shRNA (small hairpin RNA), snRNA (small nuclear RNA), snoRNA (small nucleolar RNA), miRNA (microRNA), genomic DNA, synthetic DNA, synthetic RNA, and / or tRNA. In some embodiments, the isolated polynucleotide is a modified mRNA containing unnatural nucleosides or nucleotides. In some embodiments, the modified mRNA contains 2-thiouridine, pseudouridine, or 1-methylpseudouridine.

[0126] As used herein, the term "naked DNA" refers to DNA (e.g., histone-free DNA) encoding a protein such as an adapter or CAR cloned into a suitable expression vector (e.g., a plasmid) in the appropriate orientation for expression. Viral vectors that can be used include, but are not limited to, SIN lentiviral vectors, retroviral vectors, foamy virus vectors, adenoviral vectors, adeno-associated virus (AAV) vectors, hybrid vectors, and / or plasmid transposons (e.g., the Sleeping Beauty transposon system), or integrase-based vector systems. Other vectors that can be used in connection with the production and use of adapters and CARs are described herein or known in the art.

[0127] As used herein, the terms "vector," "cloning vector," and "expression vector" refer to a vehicle by which a nucleic acid sequence (e.g., an adapter or CAR coding sequence) can be maintained or propagated in or introduced into a host cell (e.g., a cloning vector) so as to transform the host and promote expression (e.g., transcription and translation) of the introduced sequence. Vectors include plasmids, phages, viruses, etc.

[0128] A "host cell" includes an individual cell or cell culture that can be or has been a recipient of a nucleic acid encoding an adapter or CAR. Host cells include, but are not limited to, viral particles, phagemids, bacteria, yeast, plant, animal, and mammalian cells. A host cell includes the progeny of a single host cell, which progeny need not be exactly identical (in morphology or total DNA complement) to the original parent cell due to natural, accidental, or deliberate mutations and / or alterations. A host cell includes cells transfected or infected with a nucleic acid encoding an adapter or CAR in vivo, in vitro, or ex vivo. In some examples, the host cell is capable of expressing the adapter. In some examples, the host cell is capable of expressing and secreting the adapter. In some examples, the host cell is capable of expressing the CAR. In some examples, the host cell is capable of expressing and displaying the CAR on its surface. "Expression" includes transcription and / or translation.

[0129] As used herein, the terms "pharmaceutically acceptable" or "physiologically acceptable" and grammatical variations thereof, when referring to compositions, carriers, diluents, and reagents, are used interchangeably to indicate that the substance can be administered to a human without producing therapeutically prohibited undesirable physiological effects, such as nausea, dizziness, acute gastric peristalsis, and other therapeutically prohibited undesirable physiological effects known in the art.

[0130] "Parenteral" administration of the immunogenic compositions includes, for example, subcutaneous (sc), intravenous (iv), intramuscular (im), or intrasternal injection or infusion techniques.

[0131] The term "stimulate" or "stimulation" refers to a primary response induced by binding of a stimulatory molecule with its cognate ligand, thereby inducing a signal transduction event, such as, but not limited to, signal transduction through an appropriate receptor, such as, for example, a T receptor or an NK receptor.

[0132] "Modulate" or "modulation" refers to the regulation or control of magnitude, frequency, extent, or activity. In another related aspect, such modulation can be positive (e.g., an increase in frequency, extent, or activity) or negative (e.g., a decrease in frequency, extent, or activity). In some embodiments, the positive or negative modulation is relative to the function of the cell, tissue, or organ prior to administration of the therapeutic agent. In further embodiments, the positive or negative modulation is relative to the function of a normal, healthy cell, tissue, or organ.

[0133] An "effective amount" of a CAR cell, adaptor, and / or CAR cell / adaptor composition provided herein is an amount sufficient to achieve a specifically stated purpose, such as causing an observable change in one or more biological activities associated with the target to which the CAR cell and / or adaptor binds. In certain embodiments, the change increases the level of target activity. In other embodiments, the change decreases the level of target activity. An "effective amount" can be determined empirically and routinely with respect to the stated purpose. The term "therapeutically effective amount" refers to an amount of CAR cells and / or adaptors or other therapeutic agent effective to "treat" (e.g., reduce symptoms of a disease or disorder) a disease or disorder in a subject (mammal). A "prophylactically effective amount" refers to an amount effective, at dosages and for periods of time necessary, to achieve the desired prophylactic result.

[0134] The terms "patient," "subject," "animal," and "mammal" are used interchangeably and refer to mammals, such as human patients and non-human primates, as well as laboratory animals, such as rabbits, rats, mice, and other animals. Animals include all vertebrates, e.g., mammals and non-mammals, such as chickens, amphibians, and reptiles. As used herein, "mammal" refers to any member of the class Mammalia, including, but not limited to, humans and non-human primates, e.g., chimpanzees and other apes and monkey species; livestock animals, such as cows, sheep, pigs, goats, and horses; domestic mammals, such as dogs and cats; laboratory animals, such as rodents, such as mice, rats, guinea pigs, and other members of the class Mammalia known in the art. In certain embodiments, the patient is a human. The term does not denote a particular age or sex. Thus, adult and neonatal / pup subjects, as well as fetuses / pups, regardless of male or female, are intended to be included within the scope of this term.

[0135] As used herein, the terms "treat," "treatment," and "treating" refer to both therapeutic and prophylactic or preventative treatment, where the objective is to prevent or delay (reduce or slow) symptoms, complications, or biochemical manifestations of a disease, condition, or disorder, or to alleviate or arrest or inhibit the further development of a disease, condition, or disorder. "Treatment" can target a pathology; prophylactic (preventing or delaying the onset of the disease or preventing the onset of clinical or asymptomatic symptoms thereof) or therapeutic suppression or alleviation of symptoms after the onset of a disease, condition, or disorder; prevent a pathology; pursue or obtain a beneficial outcome; or reduce the likelihood of the development of a particular condition, even if the treatment is ultimately unsuccessful. Subjects in need of treatment include those already with the condition, as well as those prone to the condition or those in whom the condition is to be prevented. Treatment can involve the use of CAR cells, adaptors, and / or CAR cell / adaptor compositions, alone or in combination with additional therapeutic agents. In some embodiments, the terms "treat," "treatment," and "treating" as used herein refer to both therapeutic and prophylactic or preventative treatment, where the objective is to prevent or slow (reduce or delay) a symptom, complication, or biochemical manifestation of a proliferative disorder, or to ameliorate one or more symptoms (preferably one or more identifiable symptoms) of a proliferative disorder. In certain embodiments, the terms "treat," "treatment," and "treating" refer to the amelioration of at least one measurable physical parameter of a proliferative disorder, such as tumor growth, which may not necessarily be discernible by the patient. In other embodiments, the terms "treat," "treatment," and "treating" refer to the inhibition of progression of a proliferative disorder, either physically, e.g., by stabilization of a discernible symptom, or physiologically, e.g., by stabilization of a physical parameter, or both.In another embodiment, the terms "treat", "treatment" and "treating" refer to the reduction or stabilization of tumor, tumor cell growth or survival, or cancer cell numbers.

[0136] "Cancer," "tumor," or "malignant tumor" are used interchangeably and refer to any of a number of diseases characterized by uncontrolled, abnormal proliferation of cells, spread of infected cells locally or to other parts of the body via the bloodstream and lymphatic system (metastasis), and numerous distinctive structural and / or molecular features. As used herein, "tumor" refers to all neoplastic cell growth and proliferation, and all pre-cancerous and cancerous cells and tissues, whether malignant or benign. A "cancerous tumor," or "malignant cell," is understood to be a cell that has specific structural characteristics, lacks differentiation, and is capable of invasion and metastasis. Cancers that can be treated using the CAR cells, adaptors, and / or CAR cell / adaptor compositions provided herein include, but are not limited to, breast cancer, lung cancer, brain cancer, cervical cancer, skin cancer, bone cancer, liver cancer, pancreatic cancer, colorectal cancer, renal cancer, head and neck cancer, ovarian cancer, hematopoietic cancers (e.g., leukemia), and prostate cancer, as well as lymphoma. Other types of cancers and tumors that can be treated using CAR cells, adaptors, and / or CAR cell / adaptor compositions are described herein or are known in the art. A particular "type" is understood to refer to a cancer, tumor, or tumor cell characterized by a particular disease. For example, in some embodiments, the first and second cancers of the same type are mixed-cell type Hodgkin lymphoma and lymphocyte-rich Hodgkin lymphoma. In other embodiments, the first and second cancers of the same type are precursor B-cell acute lymphoblastic leukemia (ALL) and mature B-cell ALL. Examples of different types of first and second cancers include, for example, Hodgkin lymphoma and ALL.

[0137] The term "tumor antigen" refers to an antigen common to a particular hyperproliferative disorder, such as cancer. The terms "tumor antigen" and "cancer antigen" are used interchangeably herein. In certain embodiments, the antigen is derived from a cancer, including, but not limited to, primary or metastatic melanoma, thymoma, lymphoma, sarcoma, lung cancer (e.g., NSCLC or SCLC), liver cancer, non-Hodgkin's lymphoma, Hodgkin's lymphoma, leukemia, multiple myeloma, glioblastoma, neuroblastoma, uterine cancer, cervical cancer, renal cancer, thyroid cancer, bladder cancer, kidney cancer, mesothelioma, and adenocarcinomas such as breast cancer, prostate cancer, ovarian cancer, pancreatic cancer, colon cancer, and other cancers known in the art. In some embodiments, the cancer is one or more of B-cell acute lymphoblastic leukemia ("BALL"), T-cell acute lymphoblastic leukemia ("TALL"), acute lymphoblastic leukemia (ALL), acute myeloid leukemia (AML); one or more chronic leukemias, such as, but not limited to, chronic myeloid leukemia (CML), chronic lymphocytic leukemia (CLL); B-cell prolymphocytic leukemia, blastic plasmacytoid dendritic cell neoplasm, Burkitt's lymphoma, diffuse large intestinal lymphoma, and leukemia. and additional blood cancers or hematological conditions including: chronic large B-cell lymphoma, follicular lymphoma, hairy cell leukemia, small cell or large cell follicular lymphoma, malignant lymphoproliferative conditions, MALT lymphoma, mantle cell lymphoma, marginal zone lymphoma, multiple myeloma, myelodysplasia and myelodysplastic syndromes, non-Hodgkin's lymphoma, plasmablastic lymphoma, blastic plasmacytoid dendritic cell neoplasm, and Waldenstrom's hypergammaglobulinemia.

[0138] Tumor and cancer antigens may be further defined as "tumor-specific antigens (TSAs)," "cancer-specific antigens (CSAs)," "tumor-associated antigens (TAAs)," or "cancer-associated antigens (CAAs)." TSAs are antigens unique to tumor cells and not found on other cells in the body. TAAs are antigens found on both tumors and some normal cells. TAAs may be expressed on normal cells under conditions that do not induce a state of immune tolerance to the antigen. Expression of TAAs on tumors can occur under conditions that allow the immune system to respond to the antigen. TAAs are expressed on normal cells during fetal development, when the immune system is immature and unable to respond, or may normally be present at very low levels on normal cells but are expressed at much higher levels on tumor cells. Due to the dynamic nature of tumors, in some cases, tumor cells may express antigens unique to certain stages, and in other cases, express antigens that are also expressed on non-tumor cells. Therefore, the inclusion of a particular marker as a TAA does not exclude it from being considered a TSA. In some embodiments, the TAA and / or TSA comprising an antigenic determinant specifically bound by the CAR cells, adaptors, and / or CAR cell / adaptor compositions provided herein is selected from the following: BCMA, CD19, CD20, CD22, CD30, CD33 / 1L3Ra, CD70, CD123, CD171 (L1-CAM), CS1, EGFRvIII, GD2, Lewis γ、ROR 1, IL13Ra2, cMet, PSMA, free radical α(FR-α), CEA, ErbB2( HER-2 / neu); EGFR(HER), PSCA, PSA, MUC1, MUC16, CD44v6, CD 44v6 / 7, CD44v7 / 8, CD55, IL11Ra, EphA2, EGP40, TAG72, CAIX, HMW-MAA(CSPG4), MAGEA4, NKG2D, β-HCG, Glycolipid F77, HLA-A2(NY-ESO-1), HMW-MAA, GD3, TCR, MAGE A3, MARTI, WT1, Mulberry, gp100(Pmel 17) TRP1, TRP2, HLA-A1, MAGE1, MAGE3, BAGE, GAGE1, GAGE 2, pi5, p53, Ras, BCR-ABL, E2A-PRL, H4-RET, IGH-IGK, MYL-RAR;VE GFR2, FAP, FAR, EBVA, HPV vaccine E6, HPV vaccine E7, TSP-180, MAGE4, MAGE5, M AGE6, RAGE, pl85erbB2, pl80erbB3, nm-23Hl, CA19-9, CA72-4, CAM 17.1 NuMa, K-ras, β-linkage, CDK4, Mum-1, p15, p 16, 43-9F, α-protein, BCA225, BTAA, CA125, CA 15-3, CA 27.29(BCAA), CA195, CA242, CA50, CAM43, CD68, CO-029, FGF5, G250, HTgp-175 344. MA50, MG7-Ag, NB / 70K, NY-CO-1, RCAS1, SDCCAG16, M2BP, TAAL6, TLP, およ, 3TPS.

[0139] As used herein, the term "CS1" refers to an NK cell receptor that regulates immune function and is expressed on B cells, T cells, dendritic cells, NK-T cells, and monocytes alike. CS1 is overexpressed in multiple myeloma and has been successfully targeted for immunotherapy of multiple myeloma (Malaer & Mathew, Am J Cancer Res. 7(8):1637-1641(2017)). CS1 is also known as SLAM7, protein 19A, CRACC, and CD319. The term "CS1" includes variants, isoforms, homologs, orthologs, and paralogs. CS1 is a transmembrane protein with various alternatively spliced isoforms. In some embodiments, the amino acid sequence of human CS1 comprising a 22 amino acid residue N-terminal signal sequence (MAGSPTCLTLIYILWQLTGSAA, SEQ ID NO: 1119) and an extracellular domain comprising 226 N-terminal residues (SEQ ID NO: 1120) has GenBank Accession Number NP_067004 (SEQ ID NO: 1121). In some embodiments, the amino acid sequence of human CS1 has GenBank Accession Number NP_001269517, NP_001269518, NP_001269519, NP_001269520, NP_001269521, NP_001269522, NP_001269523, NP_001269524, or NP_001269525.

[0140] As used herein, the term "autoimmune disease" is defined as a disorder resulting from an autoimmune response. Autoimmune diseases are the result of an inappropriate or excessive response to self-antigens. Examples of autoimmune diseases include, but are not limited to, Addison's disease, alopecia areata, ankylosing spondylitis, autoimmune hepatitis, autoimmune parotitis, Crohn's disease, diabetes mellitus (type 1), dystrophic epidermolysis bullosa, epididymitis, glomerulonephritis, Graves' disease, Guillain-Barré syndrome, Hashimoto's disease, hemolytic anemia, systemic lupus erythematosus, multiple sclerosis, myasthenia gravis, pemphigus vulgaris, psoriasis, rheumatic fever, rheumatoid arthritis, sarcoidosis, scleroderma, Sjögren's syndrome, spondyloarthropathy, thyroiditis, vasculitis, vitiligo, myxedema, pernicious anemia, and ulcerative colitis.

[0141] As used herein, the term "transduction" refers to the introduction of foreign nucleic acid into a cell using a viral vector. As used herein, the term "gene transfer" refers to the introduction of foreign nucleic acid into a cell using recombinant DNA technology. The term "transformation" refers to the introduction of a "foreign" (e.g., exogenous, extracellular, or otherwise non-endogenous) nucleic acid (DNA or RNA) sequence into a host cell, where the host cell expresses the introduced nucleic acid to produce a substance, such as a protein or enzyme, encoded by the desired introduced coding sequence. The introduced nucleic acid sequence may also be referred to as a "cloned" or "foreign" gene or sequence and may include regulatory or control sequences, such as start, stop, promoter, signal, secretion, or other sequences used by the cell's genetic machinery. The nucleic acid sequence may also include nonfunctional sequences with no known function. A host cell that receives and expresses introduced nucleic acid (e.g., DNA or RNA) has been "transformed" and is a "transformant" or "clone." The DNA or RNA introduced to a host cell can come from any source, including cells of the same genus or species as the host cell, or cells of a different genus or species, or can be non-naturally occurring.

[0142] The term "D domain" refers to a target-binding polypeptide that shares the specific sequence and structural features of the following reference scaffold sequence: MGSWAEFKQRLAAIK TRLQALGGSEAELAAFEKEIAAFESELQAYKGKGNPEVEALRK EAAAIRDELQAYRHN (SEQ ID NO: 1) (see International Publication Nos. WO 2016 / 164305 and WO 2016 / 164308, which are incorporated herein by reference in their entireties). The reference scaffold is a non-natural, target-free antiparallel three-helical bundle reference polypeptide variant originally engineered for protein folding training (see Walsh et al., PNAS 96:5486-5491 (1999)), which is incorporated herein by reference in its entirety). It has been discovered that polypeptides containing modifications of the target-free reference scaffold having the amino acid sequence of SEQ ID NO: 1 can specifically bind to the target. Thus, D domains or molecules containing D domains can specifically (or intentionally) bind to target molecules. Without being bound by theory, it is believed that the structural constraints placed on surface-exposed residues (which may be modified) in the design of D domains confer the ability of the surface-exposed residues to specifically bind to the target.

[0143] As used herein, "co-express" refers to the expression of two or more protein-coding sequences by the same cell or population of cells. The coding sequences may be, for example, nucleic acids that each encode a single protein or a chimeric protein as a single polypeptide chain.

[0144] As used herein, "antigen loss escape variant" refers to a cell that exhibits reduced or lost expression of the target antigen, the antigenic determinant of which is targeted by the adapter or CAR.

[0145] II. Antigenic determinants (AD) An antigenic determinant (AD) is an epitope that can be recognized and specifically bound by an antigenic determinant binding domain (ADBD) (e.g., an antigen-binding fragment (e.g., a D domain) of an antibody or an alternative scaffold binding domain (ASBD). The ADs in the adapters and on target cells provided herein can be bound by CARs, as discussed below.

[0146] In some embodiments, the AD (e.g., in the adaptor and / or on the target cell) is an AD present in a naturally occurring protein or other molecule. In some embodiments, the AD is an AD that is endogenous to humans.

[0147] In some embodiments, the AD in the adaptor is an AD present on a target cell.

[0148] In some embodiments, the AD (e.g., in the adaptor and / or on the target cell) is an AD present in a transmembrane protein, e.g., an AD present in the extracellular portion of a transmembrane protein. In some embodiments, the AD is a tumor antigen. In some embodiments, the AD is a tumor-associated antigen. In some embodiments, the AD is a tumor-specific antigen.

[0149] In some embodiments, the AD (e.g., in the adapter and / or on the target cell) is a cancer antigen. In some embodiments, the AD is a cancer-associated antigen. In some embodiments, the AD is a cancer-specific antigen.

[0150] In some embodiments, the AD (e.g., in the adaptor and / or on the target cell) is an epitope of BCMA. In further embodiments, the AD comprises 5-25, 5-50, 5-75, 5-100, 5-125, or 5-150 amino acid residues, more than 150 amino acid residues, or all amino acid residues of SEQ ID NO:5.

[0151] In some embodiments, the AD is an epitope of CD 19. In further embodiments, the AD comprises 5 to 25, 5 to 50, 5 to 75, 5 to 100, 5 to 125, or 5 to 150 amino acid residues, more than 150 amino acid residues, or all amino acid residues of SEQ ID NO:3.

[0152] In some embodiments, the AD is an epitope of CD20. In further embodiments, the AD comprises 5-25, 5-50, 5-75, 5-100, 5-125, or 5-150 amino acid residues, more than 150 amino acid residues, or all amino acid residues of SEQ ID NOs: 6-9, or 10.

[0153] In some embodiments, the AD is an epitope of CD22. In further embodiments, the AD comprises 5-25, 5-50, 5-75, 5-100, 5-125, or 5-150 amino acid residues, more than 150 amino acid residues, or all amino acid residues of SEQ ID NO:41.

[0154] In some embodiments, the AD is an epitope of CD 123. In further embodiments, the AD comprises 5 to 25, 5 to 50, 5 to 75, 5 to 100, 5 to 125, or 5 to 150 amino acid residues, more than 150 amino acid residues, or all amino acid residues of SEQ ID NO:11.

[0155] In some embodiments, the AD is an epitope of CD37. In further embodiments, the AD comprises 5 to 25, 5 to 50, 5 to 75, 5 to 100, 5 to 125, or 5 to 150 amino acid residues, more than 150 amino acid residues, or all amino acid residues of SEQ ID NO: 12 or 13.

[0156] In some embodiments, the AD is an epitope of CS1. In further embodiments, the AD comprises 5-25, 5-50, 5-75, 5-100, 5-125, or 5-150 amino acid residues, more than 150 amino acid residues, or all amino acid residues of SEQ ID NO: 1139. In further embodiments, the AD is an epitope of CS1 bound by elotuzumab.

[0157] In some embodiments, the AD is an epitope of HER2. In further embodiments, the AD comprises 5 to 25, 5 to 50, 5 to 75, 5 to 100, 5 to 125, or 5 to 150 amino acid residues, more than 150 amino acid residues, or all amino acid residues of SEQ ID NO:42.

[0158] In some embodiments, the AD is an epitope of AFP. In further embodiments, the AD comprises 5 to 25, 5 to 50, 5 to 75, 5 to 100, 5 to 125, or 5 to 150 amino acid residues, more than 150 amino acid residues, or all amino acid residues of SEQ ID NO: 15.

[0159] In some embodiments, the AD is an epitope of AFP p26. In further embodiments, the AD comprises 5 to 25, 5 to 50, 5 to 75, 5 to 100, 5 to 125, or 5 to 150 amino acid residues, more than 150 amino acid residues, or all amino acid residues of SEQ ID NO: 16. In further embodiments, the AD comprises 5 to 25, 5 to 50, 5 to 75, 5 to 100, 5 to 125, or 5 to 150 amino acid residues, more than 150 amino acid residues, or all amino acid residues of SEQ ID NO: 1117. In further embodiments, the AD comprises amino acid residues of SEQ ID NO: 16, 1117, 1118, 1119, 1120, 1121, 1122, or 1123.

[0160] In some embodiments, the AD (eg, in the adaptor and / or on the target cell) is expressed on the surface of an immune effector cell.

[0161] In some embodiments, the AD is an epitope of the extracellular domain (ECD) of human CD45. In further embodiments, the AD comprises 5-25, 5-50, 5-75, 5-100, 5-125, or 5-150 amino acid residues, more than 150 amino acid residues, or all of residues 29-766 of SEQ ID NO: 1106.

[0162] In some embodiments, the AD is an epitope of human CD45 bound by the UCHL-1, A6, or ODP4 antibody. In some embodiments, the AD is an epitope of human CD45 bound by the 4KB5, MB1, KiB3, 2H4, or MT2 antibody.

[0163] In some embodiments, the AD is an epitope of CD26. In further embodiments, the AD comprises 5-25, 5-50, 5-75, 5-100, 5-125, or 5-150 amino acid residues, more than 150 amino acid residues, or all of residues 29-766 of SEQ ID NO: 1113.

[0164] In some embodiments, the AD is an epitope of CD30. In further embodiments, the AD comprises 5-25, 5-50, 5-75, 5-100, 5-125, or 5-150 amino acid residues, more than 150 amino acid residues, or all of residues 19-379 of SEQ ID NO: 1114.

[0165] In some embodiments, the AD is an epitope of CD33. In further embodiments, the AD comprises 5-25, 5-50, 5-75, 5-100, 5-125, or 5-150 amino acid residues, more than 150 amino acid residues, or all of residues 18-259 of SEQ ID NO: 1115.

[0166] In some embodiments, the AD is an epitope of CD38. In further embodiments, the AD comprises 5-25, 5-50, 5-75, 5-100, 5-125, or 5-150 amino acid residues, more than 150 amino acid residues, or all of residues 43-300 of SEQ ID NO: 1116.

[0167] In some embodiments, the AD is an epitope of a human intracellular protein. In further embodiments, the AD is an epitope of the intracellular portion of a membrane-bound receptor protein selected from the group: cytokine receptor, chemokine receptor, T cell receptor, B cell receptor, NK cell receptor, myeloid cell receptor, endothelial cell receptor, and epithelial cell receptor. In some embodiments, the AD is an epitope of the intracellular portion of CD3, CD137, CD279, CD223, CD152, CD28, and VEGFR-2. In some embodiments, the AD is an epitope of a human nuclear protein.

[0168] In some embodiments, the AD (e.g., in the adapter and / or on the target cell) is an epitope of a tumor antigen associated with a malignancy. In some embodiments, the AD is an epitope of a tissue-specific antigen from melanoma. In some embodiments, the AD is an epitope of a tissue-specific melanoma antigen selected from MART-1, tyrosinase, and GP100. In some embodiments, the AD is an epitope of a tissue-specific antigen from prostate cancer. In some embodiments, the tissue-specific prostate cancer antigen is selected from prostatic acid phosphatase (PAP) and prostate-specific antigen (PSA). In some embodiments, the AD is an epitope of a transformation-associated molecule. In further embodiments, the AD is an epitope of ErbB2 (HER2). In some embodiments, the AD is an epitope of a carcinoembryonic antigen. In some embodiments, the AD is an epitope of a carcinoembryonic antigen (CEA). In some embodiments, the AD is an epitope of a B-cell lymphoma-specific idiotypic immunoglobulin. In some embodiments, the AD is an epitope of a B-cell differentiation antigen. In some embodiments, the AD is an epitope of a B cell differentiation antigen selected from CD19, CD20, and CD37. In some embodiments, the AD is an epitope of an antigen on a myeloid cell. In some embodiments, the AD is an epitope of a myeloid cell antigen selected from TSLPR and IL-7R. In some embodiments, the AD is an epitope of a cancer-testis (CT) antigen. In some embodiments, the AD is an epitope of a cancer-testis (CT) antigen selected from NY-ESO-1 and LAGE-1a. In some embodiments, the AD is an epitope of an antigen selected from CS1, CD38, CD138, MUC1, HM1.24, CYP1B1, SP17, PRAME, Wilms' tumor 1 (WT1), and heat shock protein gp96 on multiple myeloma cells.

[0169] In some embodiments, the AD (e.g., in the adapter and / or on the target cell) is an epitope of a TSA or TAA. In some embodiments, the AD is an epitope of a tumor differentiation antigen. In some embodiments, the AD is an epitope of a tumor differentiation antigen selected from MART1 / MelanA, gp100 (Pmel 17), tyrosinase, TRP1, and TRP2. In some embodiments, the AD is an epitope of a tumor-specific multilineage antigen. In some embodiments, the AD is an epitope of a tumor-specific multilineage antigen selected from MAGE1, MAGE3, BAGE, GAGE1, GAGE2, and p15. In some embodiments, the AD is an epitope of an overexpressed embryonic antigen. In some embodiments, the AD is an epitope of CEA. In some embodiments, the AD is an epitope of an overexpressed oncogene or mutated tumor suppressor gene product. In some embodiments, the AD is an epitope of an overexpressed oncogene or mutated tumor suppressor gene product selected from p53, Ras, and HER2 / neu. In some embodiments, the AD is an epitope of a unique tumor antigen resulting from a chromosomal translocation. In some embodiments, the AD is an epitope of a unique tumor antigen resulting from a chromosomal translocation selected from BCR-ABL, E2A-PRL, H4-RET, IGH-IGK, and MYL-RAR. In some embodiments, the AD is an epitope of a viral antigen. In some embodiments, the AD is an epitope of the Epstein-Barr virus antigen EBVA. In some embodiments, the AD is an epitope of the human papillomavirus (HPV) antigen E6 or E7. In some embodiments, the AD is an epitope of a large protein-based antigen.

[0170] In some embodiments, the AD (e.g., in the adapter and / or on the target cell) is an epitope of a hematological tumor antigen. In some embodiments, the AD is an epitope of BCMA, CD19, CD20, CD22, CD30, CD138, CD33, CD38, CD123, CS1, ROR1, Lewis Y, an epitope of an antigen selected from Ig kappa light chain, TCR, BCMA, TACI, BAFFR (CD268), and NKG2DL ligand.

[0171] In some embodiments, the AD (e.g., in the adapter and / or on the target cell) is an epitope of a solid tumor antigen, hi some embodiments, the AD is an epitope of an antigen selected from disialoganglioside (GD2), o-acetyl GD2, EGFRvIII, HER2 (ErbB2), VEGFR2, FAP, mesothelin, IL13Ra2 (glioma), cMET, PSMA, folate receptor alpha, L1CAM, carcinoembryonic antigen (CEA), and EGFR.

[0172] In some embodiments, the AD (e.g., in the adaptor and / or on the target cell) is an epitope of an antigen selected from the group consisting of CD137, PDL1, CTLA4, CD47, KIR, TNFRSF10B (DR5), TIM3, PD1, cMet, glycolipid F77, EGFRvIII, HLAA2 (NY-ESO-1), LAG3, CD134 (OX40), HVEM, BTLA, TNFRSF25 (DR3), CD133, MAGE A3, PSCA, MUC1, CD44v6, CD44v6 / 7, CD44v7 / 8, IL11Ra, ephA2, CAIX, MNCAIX, CSPG4, MUC16, EPCAM (EGP2), TAG72, EGP40, ErbB receptor family, ErbB2 (HER2), ErbB3 / 4, RAGE1, GD3, FAR, Lewis Y, NCAM, HLAA1 / MAGE1, MAGEA1, MAGEA3, MAGE-A4, B7H3, WT1, MelanA (MART1), HPV E6, HPV E7, thyroglobulin, tyrosinase, PSA, CLL1GD3, Tn Ag, FLT3, KIT, PRSS21, CD24, PDGFR-β, SSEA4, prostase, PAP, ELF2M, ephB2, IGF1, IGFII, IGF1 receptor, LMP2, gp100, bcr-ab1, fucosyl-GM1, sLe, GM3, TGS5, folate receptor β, TEM1 (CD248), TEM7R, CLDN6, TSHR, GPRC5D, CXORF61, CD97, CD7a, HLE, CD179a, ALK, sialic acid, PLAC1, GloboH, NY-BR-1, UPK2, HAVCR1, ADRB3, PANX3, GPR20, LY6K, OR51E2, TARP, LAGE1a, legumain, E7, ETV6-AML, sperm protein 17, XAGE1, Tie 2, MAD-CT1, MAD-CT2, Fos-related antigen 1, p53, p53 mutant, prostein, survivin, telomerase, PCTA1 (galectin 8), Ras mutant, hTERT, sarcoma translocation breakpoint, ML-IAP, ERG (TMPRSS2 ETS fusion gene), NA17, PAX3, androgen receptor, cyclin B1, MYCN, RhoC, TRP2, CYP1B1, BORIS, SART3, PAX5, OY-TES1, LCK, AKAP4, SSX2, reverse transcriptase, RU1, RU2, intestinal carboxylesterase, neutrophil elastase, mut hsp70-2, CD79a, CD79b, CD72, LAIR1, FCAR, LILRA2, CD300LF, CLEC12A, BST2, EMR2, LY75, GPC3, FCRLS, I GLL1, TSP-180, MAGE4, MAGE5, MAGE6, VEGFR1, IGF1R, hepatocyte growth factor receptor, p185ErbB2, p180ErbB-3, nm-23H1, CA 19-9, CA 72-4, CAM 17.1, NuMa, K-ras, β-catenin, CDK4, Mum1, p15, p16, 43-9F, 5T4, 791Tgp72, β-human chorionic gonadotropin, BCA225, BTAA, CA125, CA15-3, CA 27.29 (BCAA), CA195, CA242, CA-50, CAM43, CD68, CO-029, FGF5, G250, HTgp-175, M344, MA50, MG7-Ag, MOV18, NB / 70K, NY-CO1, RCAS1, SDCCAG16, M2BP, TAAL6, TLP, and TPS, glioma-associated antigen, alpha-fetoprotein (AFP), p26 fragment of AFP or its mutants, lectin-reactive AFP, and TLR4.

[0173] In some embodiments, the AD (e.g., in the adaptor and / or on the target cell) is an epitope of a TSA or TAA. In some embodiments, the AD is an epitope of an antigen selected from PTGER4, ITGA4, CD37, CD52, CD62L (L-selectin), CXCR4, CD69, EVI2B (CD361), SLC39A8, MICB, LRRC70, CLELC2B, HMHA1, LST1, and CMTM6 (CKLFSF6). In some embodiments, the AD is an epitope of BCMA. In some embodiments, the AD is an epitope of CS1.

[0174] In some embodiments, the AD (e.g., in the adapter and / or on the target cell) is an epitope of an antigen selected from the following: PDGFRA, VEGFR1, VEGFR3, neuropilin 1 (NRP1), neuropilin 2 (NRP2), betacellulin, PLGF, RET (rearranged during gene transfer), TIE1, TIE2 (TEK), CA125, CD3, CD4, CD7, CD10, CD13, CD25.CD32, CD32b, CD44 (e.g., CD44v6), CD47, CD49e (integrin α5), CD54 (ICAM), CD55, CD64, CD74, CD80, CD90, CD200, CD147, CD166, CD200, ESA, SHH, DHH, IHH, patched 1 (PTCH1), smoothened 1 (SMO), WNT1, WNT2B, WNT3A, WNT4, WNT4A, WNT5A, WNT5B, WNT7B, WNT8A, WNT10A, WNT10B, WNT16B, LKP5, LRP5 , LRP6, FZD1, FZD2, FZD4, FZD5, FZD6, FZD7, FZD8, Notch, Notch1, Notch3, Notch4, DLL4, Jagged, Jagged1, Jagged2, Jagged3, TNFRSF1A (TNFR1, p55, p60), TNFRSF1B (TNFR2), TNFRSF6 (Fas, CD95), TNFRSF6B (DcR3), TNFRSF7 (CD27), TNFSF9 (41BB ligand), TNFRSF8 (CD30), TNFRSF10A (TRAILRl, DR4), TNF RSF11A(RANK), TNFRSF12(TWEAKR), TNFRSF19L(KELT), TNFRSF19(TROY), TNFRSF21(DR6), ILIRI, 1L1R2, IL2R, IL5R, IL6R, 1L8R, IL10R, IL1 2R, IL13R, IL15R, IL18R, IL19R, IL21R, IL23R, XAG1, XAG3, REGIV, FGFR1, FGFR2, FGFR3, ALK, ALK1, ALK7, ALCAM, Axl, TGFb, TGFb2, TGFb3, TG FBR1, IGFIIR, BMPRI, N-cadherin, E-cadherin, VE-cadherin, ganglioside GM2, ganglioside GD3, PSGR, DCC, CDCP1, CXCR2, CXCR7, CCR3, CCR4, CCR5, CCR7, CCR10, claudin 1, claudin 2, claudin 3, claudin 4, TMEFF2, neuregulin, MCSF, CSF, CSFR(fms), GCSF, GCSFR, BCAM, BRCA1, BRCA2, HLA-DR, ABCC3, ABCB5, HM 1.24, LFA1, LYNX, S100A8, S100A9, SCF, von Willebrand factor, Lewis Y6 receptor, CAG250 (CA9), CRYPTO, VLA5, HLADR, MUCl8, mucin CanAg, EGFL7, integrin avb3, integrin α5β, activin Blα, leukotriene B4 receptor (LTB4R), neurotensin NT receptor (NTR), 5T4 carcinoembryonic antigen, tenascin-C, MMP, MMP2, MMP7, MMP9, MMP12, MMP14, MMP26, cathepsin G, SULF1, SULF2, MET, CA9, TM4SF1, syndecan (SDCl), ephrin B4, TEM1, TGFβ1, and TGFBRII.

[0175] In some embodiments, the AD (e.g., in the adapter and / or on the target cell) is an epitope of an antigen associated with an autoimmune disorder, associated with an inflammatory or other disorder of the immune system, or associated with modulation of the immune response.

[0176] In some embodiments, the AD (e.g., in the adaptor and / or on the target cell) is an epitope of an immunosuppressive target. In some embodiments, the AD is an epitope of an immunosuppressive target selected from IL1Ra, IL6R, CD26L, CD28, CD80, FcGamma RIIB. In some embodiments, the AD in the adaptor is an epitope of an immunosuppressive target selected from CD25, CD28, CTLA4, PD1, B7H1 (PDL1), B7H4 TGFbeta, TNFRSF4 (OX40), TNFRSF5 (CD40), TNFRSF9 (41BB, CD137), TNFRSF14 (HVEM), TNFRSF25 (DR3), and TNFRSF18 (GITR).

[0177] In some embodiments, the AD (e.g., in the adapter and / or on the target cell) is a target epitope selected from IL1Rb, C3AR, C5AR, CXCR1, CXCR2, CCR1, CCR3, CCR7, CCR8, CCR9, CCR10, ChemR23, MPL, GP130, TLR2, TLR3, TLR4, TLR5, TLR7, TLR8, TLR9, TREM1, TREM2, CD49a (integrin alpha 1), integrin a5b3, alpha 4 integrin subunit, A4B7 integrin, cathepsin G, or TNF RSF3(LTBR), TNFRSF6(Fas, CD95), TNFRSF6B(DcR3), TNFRSF8(CD30), TNFRSF11A(RANK), TNFRSF16(NGFR), TNFR SF19L(RELT), TNFRSF19(TROY), TNFRSF21(DR6), CD14, CD23, CD36, CD36L, CD39, CD91, CD153, CD164, CD200, CD20 0R, B71(CD80), B72(CD86), B7h, B7DC(PDL2), ICOS, ICOSL, MHC, CD, B7H2, B7H3, B7x, SLAM, KIM1, SLAMF2, SLAMF3 , SLAMF4, SLAMF5, SLAMF6, SLAMF7, TNFRSF1A(TNFR1, p55, p60), TNFRSF1B(TNFR2), TNFRSF7(CD27), TNFRSF12(TW EAKR), TNFRSF5(CD40), IL1R, IL2R, IL4Ra, IL5R, IL6RIL15R, IL17R, IL17Rb, IL17RC, IL22RA, IL23R, TSLPR, B7RP 1, cKit, GMCSF, GMCSFR, CD2, CD4, CD11a, CD18, CD30, CD40, CD86, CXCR3, CCR2, CCR4, CCR5, CCR8, RhD, IgE, and Rh.

[0178] In some embodiments, the AD (eg, in the adaptor and / or on the target cell) is an epitope of an antigen associated with a neurological disorder.

[0179] In some embodiments, the AD (e.g., in the adaptor and / or on the target cell) is an epitope of an antigen selected from amyloid beta (Abeta), beta amyloid, PLP, ROBO4, ROBO, LINGO, gpIIB, gpIIIa, integrin a2bB3, AOC3, TNFRSF19L (RELT), TNFRSF19 (TROY), and sclerostin.

[0180] The above targets and those described elsewhere herein are intended to be exemplary, not limiting.

[0181] In some embodiments, the AD (e.g., in the adapter and / or on the target cell) is bound by a chimeric antigen receptor (CAR). In some embodiments, the AD is bound by a cell expressing a chimeric antigen receptor. In some embodiments, the AD is bound by an scFv. In some embodiments, the AD is bound by an alternative scaffold binding domain (ASBD). In some embodiments, the AD is bound by a D domain. In some embodiments, the AD is bound by an antibody or antigen-binding fragment thereof.

[0182] III. Antigenic determinant binding domain (ADBD) A protein domain that binds to an antigenic determinant (AD) (e.g., as described in Sections II and XI) is referred to herein as an "antigenic determinant binding domain" or "ADBD." In some embodiments, the ADBD is sufficient to confer recognition and specific binding to a target. The ADBDs described herein can be present in an adapter (e.g., as described in Sections V and XI) and / or in a chimeric antigen receptor (CAR) (e.g., as described in Sections VI and XI).

[0183] The target specifically bound by an ADBD (e.g., of an adaptor and / or CAR) can be any molecule to which it is desirable for the adaptor and / or CAR to bind, for example, to any of the ADs described herein (e.g., as described in Sections II and XI). In some embodiments, the target specifically bound by an ADBD can be a target for purification, manufacture, formulation, treatment, diagnostic or prognostic relevance or prognosis determination. In some embodiments, the target of an ADBD can be natural or synthetic. In some embodiments, the target of an ADBD can be an extracellular component, an intracellular component, a soluble factor (e.g., an enzyme, hormone, cytokine, growth factor, toxin, venom, contaminant, etc.), or a transmembrane protein (e.g., a cell surface receptor).

[0184] In some embodiments, the ADBD (e.g., of the adaptor and / or CAR) specifically binds to a target on the surface of a target cell. In some embodiments, the ADBD specifically binds to a cell surface receptor. In some embodiments, the ADBD specifically binds to a target that is a member of a family selected from phosphatase receptors, growth factor receptors, tyrosine kinase receptors, TNF family receptors, G protein-coupled receptors, and chemokine receptors. In some embodiments, the ADBD binds to multiple members of the same family (e.g., to the TNF receptors TRAILR1 and TRAILR2). In some embodiments, the ADBD binds to members from different families. Thus, for example, in some embodiments, the ADBD can bind to a growth factor receptor and a TNF receptor, or a G protein-coupled receptor and a chemokine receptor.

[0185] In some embodiments, the ADBD (e.g., of the adaptor and / or CAR) binds to a tumor antigen. In some embodiments, the ADBD binds to a tumor-associated antigen. In some embodiments, the ADBD binds to a tumor-specific antigen.

[0186] In some embodiments, the ADBD (e.g., of the adaptor and / or CAR) binds to a cancer antigen. In some embodiments, the ADBD binds to a cancer-associated antigen. In some embodiments, the ADBD binds to a cancer-specific antigen.

[0187] In some embodiments, the ADBD (e.g., of the adaptor and / or CAR) binds to an antigen expressed on the surface of an immune effector cell.

[0188] In some embodiments, the target bound by the ADBD (e.g., of the adapter and / or CAR) is a human protein. In one embodiment, the ADBD binds to a human protein target and its monkey (e.g., cynomolgus), mouse, rabbit, hamster, and / or rabbit orthologs.

[0189] In another embodiment, the ADBD (e.g., of the adapter and / or CAR) binds to a peptide tag present on the target. Such peptide tags provide a useful means for detecting, monitoring, and / or attaching one or more additional moieties to the adapter. In one embodiment, the ADBD binds to a peptide tag selected from a hexahistidyl (His6) tag, a nucleotide tag, and a FLAG tag. Other peptide tags are described herein or known in the art.

[0190] The affinity requirements for a given ADBD binding event depend on a variety of factors, including, but not limited to, the composition and complexity of the binding matrix, the valency and density of both the ADBD and the target molecule, and the functional application of the ADBD. In one embodiment, the ADBD binds to the target at a binding affinity of 5x10 -3 M, 10 -3 Medium, 5x10 -4 M, 10 -4 Medium, 5x10 -5 M, or 10 -5 In a further embodiment, the ADBD binds to the target with a dissociation constant (KD) of 5x10 -6 M, 10 -6Medium, 5x10 -7 M, 10 -7 Medium, 5x10 -8 M, or 10 -8 In a further embodiment, the ADBD binds to the target with a KD of 5x10 -9 M, 10 -9 Medium, 5x10 -10 M, 10 -10 Medium, 5x10 -11 M, 10 -11 Medium, 5x10 -12 M, 10 -12 Medium, 5x10 -13 M, 10 -13 Medium, 5x10 -14 M, 10 -14 Medium, 5x10 -15 M, or 10 -15 In some embodiments, the ADBDs produced by the methods disclosed herein bind with a KD of 10 M or less. -4 M and 10 -5 Between M and 10 -5 M and 10 -6 Between M and 10 -6 M and 10 -7 Between M and 10 -7 M and 10 -8 Between M and 10 -8 M and 10 -9 Between M and 10 -9 M and 10 -10 Between M and 10 -10 M and 10 -11 Between M and 10 -11 M and 10 -12 M has a dissociation constant selected from the group

[0191] In one embodiment, the ADBD binds to the target in an active form. In one embodiment, the ADBD reversibly binds to the target in an active form and releases the binding target in an active form. In one embodiment, the ADBD binds to the target in a native form. In a particular embodiment, the ADBD binds to the target in a 10 -10 seconds -1 , 5x10 -9 seconds -1 , 10 -9 seconds -1 , 5x10 -8 seconds -1 , 10-8 second -1 、 5 x 10 -7 seconds -1 、 10 -7 seconds -1 、 5 x 10 -6 seconds -1 、 10 -6 seconds -1 、 5 x 10 -5 seconds -1 、 10 -5 seconds -1 、 5 x 10 -4 seconds -1 、 10 -4 seconds -1 、 5 x 10 -3 seconds -1 、 10 -3 seconds -1 、 5 x 10 -2 seconds -1 、 10 -2 seconds -1 、 5 x 10 -1 seconds -1 、 or 10 -1 seconds -1 bind to the target specifically with a dissociation rate constant or Koff of 10 seconds or more.

[0192] Binding experiments to determine KD and the dissociation rate constant can be carried out under many conditions including, but not limited to, [pH 6.0, 0.01% Tween 20], [pH 6.0, 0.1% gelatin], [pH 5.0, 0.01% Tween 20], [pH 9.0, 0.1% Tween 20], [pH 6.0, 15% ethylene glycol, ......

[0193] In one embodiment, the ADBD is 0.1 to 10 -7 seconds -1 , 10 -2 ~10 -7 seconds -1 , or 0.5x10 -2 ~10 -7 seconds -1 In certain embodiments, the ADBD specifically binds to the target with a Koff in the range of 5x10 -2 seconds -1 , 10 -2 seconds -1 , 5x10 -3 seconds -1 , or 10 -3 seconds -1 In a further embodiment, the ADBD binds specifically with a dissociation rate constant (Koff) of less than 5x10 -4 seconds -1 , 10 -4 seconds -1 , 5x10 -5 seconds -1 , or 10 -5 seconds -1 , 5x10 -6 seconds -1 , 10 -6 seconds -1 , 5x10 -7 seconds -1 , or 10 -7 seconds -1 It binds specifically with a dissociation rate constant (Koff) less than

[0194] In one embodiment, the ADBD is 10 3 ~10 7 M -1 seconds -1 , 10 3 ~10 6 M -1 seconds -1 , or 10 3 ~10 5 M -1 seconds -1 In another specific embodiment, the ADBD specifically binds to the target with a KOn in the range of 10 3 M -1 seconds -1 , 5x10 3 M -1 seconds -1 , 104 M -1 seconds -1 , or 5x10 4 M -1 seconds -1 In a further embodiment, the ADBD specifically binds with a rate (KOn) greater than 10 5 M -1 seconds -1 , 5x10 5 M -1 seconds -1 , 10 6 M -1 seconds -1 , or 5x10 6 M -1 seconds -1 , or 10 7 M -1 seconds -1 Combine with the target with a superior KOn.

[0195] In some embodiments, the ADBD (e.g., of the adaptor and / or CAR) is an antibody or antigen-binding fragment thereof. In some embodiments, the ADBD is an scFv. In some embodiments, the ADBD is an alternative scaffold binding domain. In some embodiments, the ADBD is a D domain.

[0196] IIIa. Antibody-derived antigenic determinant binding domains (ADBDs) In some embodiments, one or more ADBDs (e.g., of the adapter and / or CAR) can be derived from an antibody molecule, such as a monoclonal antibody, a polyclonal antibody, a recombinant antibody, a human antibody, a humanized antibody, or a single-domain antibody, such as a heavy chain variable domain (VH), a light chain variable domain (VL), and / or a variable domain (VHH), for example, from a human or camelid origin. In some embodiments, the ADBD is derived from the same species from which the adapter or CAR will ultimately be used, for example, in human applications. It is advantageous for the adapter and / or CAR to comprise a human or humanized ADBD. Compositions and techniques for routinely producing such ADBDs are known in the art.

[0197] In some embodiments, the ADBD (e.g., of the adapter and / or CAR) comprises an antibody fragment sufficient to confer recognition and specific binding to a target antigen. Examples of antibody fragments include, but are not limited to, Fab, Fab', F(ab')2, and Fv fragments, scFv antibody fragments, linear antibodies, single domain antibodies such as sdAb (VL or VH), camelid VHH domains, and multispecific antibodies formed from antibody fragments.

[0198] In some embodiments, the ADBD (e.g., of the adapter and / or CAR) is an "scFv," which can comprise a fusion protein comprising the VL and VH chains of an antibody, where the VH and VL are linked, for example, via a flexible polypeptide linker, e.g., a linker described herein. scFvs can be produced according to methods routine in the art (see, e.g., Bird et al., Science 242:423-426 (1988) and Huston et al., Proc. Natl. Acad. Sci. USA 85:5879-5883 (1988)).

[0199] In some embodiments, the ADBD (e.g., of the adapter and / or CAR) is a single-domain antigen-binding (SDAB) molecule. SDAB molecules include molecules containing a complementarity-determining region that is part of a single-domain polypeptide. Examples include, but are not limited to, heavy chain variable domains, binding molecules that naturally lack light chains, single domains derived from traditional four-chain antibodies, genetically engineered domains other than those derived from antibodies, and single-domain scaffolds. SDAB molecules can be derived from any species, including, but not limited to, mouse, human, camel, llama, fish, shark, goat, rabbit, and cow. The term also includes naturally occurring single-domain antibody molecules from species other than camelids and sharks.

[0200] In some embodiments, the ADBD (e.g., of the adapter and / or CAR) comprises a human antibody or fragment thereof. In some embodiments, the ADBD (e.g., of the adapter and / or CAR) comprises a humanized antibody or fragment thereof.

[0201] Antibody humanization is well known in the art and is essentially the same as that described by Winter and co-workers (Jones et al., Nature 321:522-525 (1986); Riechmann et al., Nature 332:323-327 (1988); Verhoeyen et al., Science 239:1534-1536 (1988)), by substituting the corresponding sequences of a human antibody with rodent CDRs or CDR sequences, i.e., CDR grafting (European Patent No. 239,400; International Publication No. WO 91 / 09967; and U.S. Pat. Nos. 4,816,567; 6,331,415; 5,225,539; 5,530,101; 5,585,089; and 6,548,640; the contents of which are incorporated herein by reference in their entireties). Antibody humanization can also be achieved by veneering or resurfacing (EP 592,106; EP 519,596; Padlan, 1991, Molecular Immunology, 28(4 / 5):489-498; Studnicka et al., Protein Engineering 7(6):805-814 (1994); and Roguska et al., PNAS 91:969-973 (1994)) or chain shuffling (U.S. Pat. No. 5,565,332), the contents of which are incorporated herein by reference in their entireties.

[0202] IIIb. Alternative Scaffold Binding Domains In some embodiments, the ADBD (e.g., of an adaptor and / or CAR) is an alternative scaffold binding domain. As used herein, an "alternative scaffold binding domain" or "ASBD" is an antigenic determinant binding domain derived from or corresponding to a non-antibody-based binding scaffold.

[0203] In some embodiments, the present disclosure provides a CAR comprising an ADBD that is an ASBD. In some embodiments, the present disclosure provides a cell comprising a CAR comprising an ADBD that is an ASBD. In further embodiments, an immune effector cell comprising a CAR comprising an ASBD is provided. In some embodiments, the present disclosure provides an adaptor comprising an ADBD that is an ASBD.

[0204] In a further embodiment, the present disclosure provides a composition comprising an adaptor and a CAR, each comprising an ASBD.

[0205] In some embodiments, binding of the ASBD (e.g., of the adaptor and / or CAR) to the target AD is mediated by a secondary structure of the binding scaffold, such as an alpha helix or a beta sheet. In some embodiments, the ASBD is a three helix bundle-based binding domain. In some embodiments, the ASBD is a D domain-based binding domain. In some embodiments, the ASBD is a Z domain (affibody)-based binding domain.

[0206] In some embodiments, the ASBD (e.g., of the adapter and / or CAR) is a D-domain (novel binding domain)-based AD binding domain. D-domain scaffold-based binding domains typically consist of 70-75 amino acid residues, in which substitutions at up to 20 positions corresponding to constrained, surface-exposed residues in a non-natural antiparallel, three-helical bundle reference scaffold (SEQ ID NO: 1) confer target recognition and binding specificity for a target of interest (AD). D-domain scaffold-based binding domains are further disclosed in WO2016164308, the contents of which are incorporated herein by reference in their entirety. In one embodiment, the D-domain comprises an amino acid sequence that differs (e.g., by amino acid modification) by up to 20 substitutions from that of the reference scaffold having the sequence of SEQ ID NO: 1. In some embodiments, the D-domain comprises a sequence selected from the group: SEQ ID NOs: 17, 18, and 19. In some embodiments, the D-domain comprises a sequence selected from the group: SEQ ID NOs: 20-26, and 27. In some embodiments, the D domain comprises a sequence selected from the group: SEQ ID NOs: 44-1078, and 1079.

[0207] In some embodiments, the ASBD (e.g., of the adaptor and / or CAR) is a Z domain scaffold (affibody)-based AD binding domain. Z domain scaffold-based binding domains typically consist of 58 amino acid residues, in which substitutions at up to 13 positions, located in the first and second of three alpha helices, confer target (AD) recognition and binding specificity for the target (AD) of interest. In further embodiments, the Z domain ASBD comprises a sequence selected from SEQ ID NOs: 28 and 29. Z domain (affibody) scaffold-based binding domains are further described in U.S. Patent No. 5,831,012, the entire contents of which are incorporated herein by reference.

[0208] Additional examples of ASBDs that exhibit secondary structure-mediated target binding include DARPins, affilins, and armadillo repeat-based binding scaffolds.

[0209] In some embodiments, the ASBD (e.g., of the adaptor and / or CAR) is a DARPin-based AD-binding domain. DARPin-based binding domains typically contain 2-3 repeats of the sequence of SEQ ID NO: 30 between the N- and C-terminal capping repeats (e.g., the sequences MRGSHHHHHHGSDLGKKLLEAARAGQDDEVRILMANGA DVNAX, respectively). 33 (SEQ ID NO: 31) and the sequence QDKFGKTAFDISIDNGNEDLAEILQ (SEQ ID NO: 32), the first Gln being at consensus repeat position X of the preceding repeat 33 Each internal repeat contains 27 framework residues and up to six substituted non-framework residues, forming a β-turn followed by two antiparallel helices and a loop connecting the β-turn of the next repeat. The collective substitutions and structure of DARPins confer target (AD) recognition and binding specificity. [Table 1]

[0210] In some embodiments, the binding specificity of an ASBD (e.g., of an adaptor and / or CAR) for a target AD is mediated by amino acids in an exposed loop on the ASBD. Examples of scaffolds with these binding properties include adnectins, lipocalins, avimers, knottins, finomers, atrimers, Kunitz domain-based binders, and CTLA4-based binding scaffolds.

[0211] In some embodiments, the ASBD is an Adnectin-based AD binding domain. The Adnectin-based binding domain is derived from the 10th domain of fibronectin type III (10Fn3). This ADBD is a 94 amino acid binding domain that typically adopts a beta-sandwich fold, comprising seven chains connected by six loops. Substitutions in three surface-exposed loops on either side of the Adnectin domain generate target (AD)-specific binding moieties.

[0212] In some embodiments, the ASBD (e.g., of the adaptor and / or CAR) is a lipocalin-, affilin-, or anticalin-based AD-binding domain. The anticalin scaffold exhibits a conserved β-barrel structure composed of eight antiparallel β-strands and typically consists of 160-180 amino acids. The ligand-binding pocket of anticalin-based scaffolds is composed of four loops, each containing up to 24 substitutions, which collectively confer target (AD) recognition and binding specificity.

[0213] In some embodiments, the ASBD (e.g., of the adaptor and / or CAR) is an avimer scaffold-based AD binding domain. The avimer scaffold-based binding domain is derived from the A domain of a cell surface receptor and is typically 35 amino acids in length. The structure of the avimer-based binding domain is maintained by 12 conserved amino acids. Substitution of all of the remaining 23 residues in the binding domain confers target (AD) recognition and binding specificity. In some embodiments, the avimer scaffold-based binding domain has the sequence EFX3CX5NGX8CIPX 12 X 13 WX 15 CDGX 19 DDCGDX 25 SDE, and X is any amino acid (SEQ ID NO: 33). Avimer scaffold-based binding domains are further described in U.S. Patent Application Publication Nos. 20040175756, 20050053973, 20050048512, and 20060008844, each of which is incorporated herein by reference in its entirety.

[0214] In some embodiments, the ASBD (e.g., of the adapter and / or CAR) is a phenomer scaffold-based AD-binding domain. A phenomer-binding domain is typically 60-75 amino acids long and is composed of a pair of antiparallel beta sheets connected by two flexible loops. Substitutions / insertions in the loops confer AD target recognition and binding specificity. In some embodiments, the phenomer-based AD-binding domain has the sequence GVTLFV ALYDYX. 12 X13 X 14 X 15 X 16 X 17 X 18 X 19 X 20 X 21 X 22 LSFHKGEKFQILSTHEYEX 41 X 42 X 43 X 44 X 45 X 46 X 47 X 48 WEARSLTTGETGX 61 X is any amino acid; 13 ~X 21 and X 42 ~X 46 may optionally be absent (SEQ ID NO: 34). In some embodiments, the phenomer-based AD binding domain has the sequence GVTLFVALYDYX 12 X 13 X 14 X 15 X 16 X 17 X 18 X 19 X 20 X 21 X 22 X is any amino acid; 16 ~X 21 is optionally absent (SEQ ID NO: 35).

[0215] In some embodiments, the ASBD (e.g., of the adaptor and / or CAR) is a knottin scaffold-based AD-binding domain, which corresponds to a 30 amino acid protein fold composed of three antiparallel β-strands of variable length and linked by multiple disulfide-bonded loops.

[0216] In some embodiments, the ASBD (e.g., of the adaptor and / or CAR) is a Kunitz domain-based AD-binding domain. Kunitz domain-based binding domains are derived from the active motif of Kunitz-type protease inhibitors and are typically approximately 60 amino acids in length. The hydrophobic core of this ADBD is composed of a twisted, two-stranded antiparallel β-sheet and two α-helices stabilized by three disulfide bond pairs. Substitutions and insertions in three loops confer AD target recognition and binding specificity. In some embodiments, the Kunitz domain-based AD-binding domain has the sequence MHSFCAFKADX 11 GX 13 CX 15 X 16 X 17 X 18 X 19 RFFFNIFTRQCEEFX 34 YGGCX 39 X 40 NQNRFESLEECKKMCTRDGA (SEQ ID NO: 36), which is at least 85% identical to this sequence at positions other than X; 11 is one of: A, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y; X 13 is one of: A, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y; X 15 is one of: A, D, E, F, G, H, I, K, L, M, N, Q, R, S, T, V, W, Y; X 16 is one of: A, G, E, D, H, T; X 17 is one of: A, D, E, F, G, H, I, K, L, M, N, Q, R, S, T, V, W, Y; X 18 is one of: A, D, E, F, G, H, I, K, L, M, N, Q, R, S, T, V, W, Y; X 19 is one of: A, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y; X 34 is one of: A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y; X 39is one of: A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y; and X 40 is one of: G, and A. Kunitz scaffold-based binding domains are further described in WO 2004063337, which is incorporated herein by reference in its entirety.

[0217] In some embodiments, the ASBD (e.g., of the adaptor and / or CAR) is a WW domain-based AD-binding domain. WW domain-based binding scaffolds are typically 30-35 amino acids in length. In some embodiments, the WW domain-based AD-binding domain has the sequence KLPPGWX7KX9WSX 12 X 13 X 14 GRVX 18 YX 20 NX 22 ITX 25 AX 27 QWERP (SEQ ID NO: 37), 12 , X 13 , X 14 , X 18 , X 20 , X 22 , X 25 , and X 27 is any amino acid, and X 14 is optional and may not be present.

[0218] In some embodiments, the WW domain-based AD binding scaffold has the sequence KLPPGWX7KX9WSX 12 X 13 GRVX 17 YX 19 NX 21 ITX 24 AX 26 QWERP (SEQ ID NO: 38), 12 , X 13 , X 17 , X 19 , X 21 , X 24 , and X 26 is any amino acid, and X 14 is optional and may not be present. [Table 2]

[0219] IV. Linker Linker refers to a peptide or other chemical linking material that connects domains that would otherwise be two or more independent functional domains of an adaptor or CAR.

[0220] Suitable linkers that operably link two or more functional domains of an adaptor in a single amino acid sequence include, but are not limited to, polypeptide linkers such as a glycine linker, a serine linker, a mixed glycine / serine linker, a glycine and serine rich linker, or a linker composed of a mostly polar polypeptide fragment.

[0221] In one embodiment, the linker is composed mostly of amino acids selected from glycine, alanine, proline, asparagine, glutamine, and lysine. In one embodiment, one or more linkers of an adaptor or CAR are composed mostly of amino acids selected from glycine, alanine, proline, asparagine, aspartic acid, threonine, glutamine, and lysine. In one embodiment, one or more linkers of an adaptor or CAR are composed mostly of one or more amino acids selected from glycine, alanine, proline, asparagine, aspartic acid, threonine, glutamine, and lysine. In another embodiment, one or more linkers of an adaptor or CAR are composed mostly of amino acids that are sterically unhindered. In another embodiment, the linker is composed mostly of glycine, serine, and / or alanine amino acids. In some embodiments, one or more linkers of an adaptor or CAR linker comprise polyglycine (e.g., (Gly)5 (SEQ ID NO: 1099)), and (Gly)8 (SEQ ID NO: 1100), poly(Gly-Ala), and polyalanine. In some embodiments, a peptide linker comprises the sequence Gly-Gly-Gly-Gly-Thr-Gly-Gly-Gly-Gly-Ser (SEQ ID NO: 39). In some embodiments, one or more linkers of an adaptor or CAR comprise the sequence Gly-Gly-Gly-Gly-Asp-Gly-Gly-Gly-Gly-Ser (SEQ ID NO: 40).

[0222] In one embodiment, the adapter or CAR comprises an ADBD directly attached (i.e., without a linker) to another component of the adapter or CAR, respectively. In one embodiment, the adapter or CAR comprises at least two, at least three, at least four, or at least five ADBDs directly attached to another component of the adapter or CAR, respectively.

[0223] In another embodiment, the ADBD can be operably linked to another component of the adaptor or CAR via a linker. The adaptor or CAR can comprise a single linker, multiple linkers, or no linker. In one embodiment, the adaptor or CAR comprises an ADBD operably linked to another component of the adaptor or CAR via a linker peptide, respectively. In one embodiment, the adaptor or CAR comprises at least two, at least three, at least four, or at least five ADBDs operably linked to another domain of the adaptor or CAR via the same or different linkers, respectively.

[0224] Linkers can be of any size or composition, so long as they operably link the functional domains of an adaptor or CAR such that the functional domains function (e.g., function in the ability of the binding domains to bind to the target). In some embodiments, the linker is about 1 to about 100 amino acids, about 1 to about 50 amino acids, about 1 to about 20 amino acids, about 1 to about 15 amino acids, about 1 to about 10 amino acids, about 1 to about 5 amino acids, about 2 to about 20 amino acids, about 2 to about 15 amino acids, about 2 to about 10 amino acids, or about 2 to about 5 amino acids. It should be clear that the length, degree of flexibility, and / or other characteristics of the linker can have some effect on the properties of the final polypeptide of the invention, including, but not limited to, affinity, specificity, or avidity for the target or for one or more other target proteins of interest. When more than one linker is used in an adaptor or CAR, these linkers can be the same or different. In the context and disclosure provided herein, one of skill in the art would be able to routinely determine the optimal linker composition and length for operably linking the functional domains of an adapter or CAR.

[0225] The linker may also be a non-peptide linker such as an alkyl linker or a PEG linker. For example, an alkyl linker such as -NH-(CH2)sC(0)- (s = 2 to 20) can be used. These alkyl linkers may be further substituted with a non-sterically hindering group such as lower alkyl (e.g., C1-C6), lower acyl, halogen (e.g., Cl, Br), CN, NH2, phenyl, etc. A representative non-peptide linker is a PEG linker. In certain embodiments, the PEG linker has a molecular weight of about 100 to 5,000 kDa, or about 100 to 500 kDa.

[0226] Suitable linkers for attaching adaptor or CAR functional domains by chemical cross-linking include, but are not limited to, homobifunctional chemical cross-linking compounds such as glutaraldehyde, imidoesters such as dimethyl adipimidate (DMA), dimethyl suberimidate (DMS), and dimethyl pimelimide (DMP), or N-hydroxysuccinimide (NHS) esters such as dithiobis(succinimidyl propionate) (DSP) and dithiobis(sulfosuccinimidyl propionate) (DTSSP). Examples of suitable linkers for attaching adaptor or CAR functional domains include, but are not limited to, cross-linkers with one amine-reactive end and a sulfhydryl-reactive moiety at the other end, or cross-linkers with an NHS ester and an SH-reactive group (e.g., maleimide or pyridyl) at one end.

[0227] In further embodiments, one or more linkers of the adaptor or CAR are cleavable. Examples of cleavable linkers include, but are not limited to, various types of peptide sequences recognized by proteases (in vitro or in vivo), such as Tev, thrombin, Factor Xa, plasmin (a blood protease), metalloproteases, cathepsins (e.g., GFLG, etc.), and proteases found in other body compartments.

[0228] In some embodiments, the linker is a "cleavable linker" that facilitates release of the adaptor's functional domain or cytotoxic agent in or on the cell surface. For example, an acid-labile linker (e.g., hydrazone), protease-sensitive (e.g., peptidase-sensitive) linker, photolabile linker, dimethyl linker, or disulfide-containing linker (see, e.g., Chari, Can. Res. 52:127-131 (1992); U.S. Pat. No. 5,208,020; and U.S. Patent Application Publication No. 20090110753; the contents of which are incorporated herein by reference in their entireties) can be used, and it is desirable for the covalent bond between the adaptor and the cytotoxic agent to be cleaved intracellularly upon internalization of the composition into the cell. The terms "cleaved intracellularly" and "intracellular cleavage" refer to a metabolic process or intracellular reaction on an adapter-drug conjugate, whereby the covalent bond, i.e., the bond via the linker, between the adapter and the cytotoxic agent is broken, resulting in the free adapter and / or cytotoxic agent being released intracellularly.

[0229] In further embodiments, one or more linkers in the CAR are cleavable. Examples of cleavable linkers include, but are not limited to, various types of peptide sequences recognized by proteases (in vitro or in vivo), such as Tev, thrombin, Factor Xa, plasmin (blood proteases), metalloproteases, cathepsins (e.g., GFLG, etc.), and proteases found in other body compartments.

[0230] In some embodiments, short oligo- or polypeptide linkers of about 1 to 100 amino acids in length are used to link either domain of the CAR together. The linker can be composed of flexible residues such as glycine and serine (or any other amino acid) to allow adjacent protein domains to move freely relative to each other. The amino acid sequence composition of the linker can be selected to minimize the potential immunogenicity of the CAR. Longer linkers can be used if it is desirable to ensure that two adjacent domains do not sterically interfere with each other.

[0231] In some embodiments, a linker between the transmembrane domain and the cytoplasmic signaling domain of the CAR is preferably 2-10 amino acids in length. In further embodiments, the linker is 10-15 amino acids in length, or 15-20 amino acids in length, or 20-30 amino acids in length, or 30-60 amino acids in length, or 60-100 amino acids in length (or any range between the recited lengths). In further embodiments, the linker is a glycine-serine doublet sequence. In some embodiments, the ESD corresponds to the human cell surface glycoprotein CD8 alpha chain ESD region (e.g., amino acid residues 138-182 CD8 alpha chain: SwissProt Accession Number P01732). In some embodiments, the ESD corresponds to a CD8 ESD region that has been further modified by amino acid substitution to improve expression function or immunogenicity. In further embodiments, the ESD corresponds to the CD28 ESD or a sequence containing modifications of the CD28 ESD to confer improved expression function or immunogenicity.

[0232] Linker optimization can be assessed using techniques described herein and / or other techniques known in the art. In some embodiments, the linker does not interfere with the ability of the adaptor or CAR to bind to its target antigenic determinant and / or the ability of another adaptor or CAR functional domain to function properly (e.g., the ability of an adaptor effector function domain to elicit effector function, or the ability of an adaptor FcRn binding domain to bind to FcRn).

[0233] V. Adaptor - Soluble Proteins Provided herein are multidomain soluble adaptor proteins. The adaptor comprises an antigenic determinant (AD) (e.g., as described in Sections II and XI) and an antigenic determinant binding domain (ADBD) (e.g., as described in Sections III and XI). The adaptor can further comprise additional ADs, additional ADBDs, and / or other additional domains.

[0234] In the adapters provided herein, the AD can be N-terminal to the ADBD. Alternatively, the ADBD can be N-terminal to the AD. In some embodiments, the AD and ADBD are fused directly. In some embodiments, the AD and ADBD are fused via a linker (a protein linker or a chemical linker) or via another protein domain (e.g., a functional domain).

[0235] In some embodiments, the adaptor comprises a linker positioned between the ADBD and another functional domain of the adaptor. In some embodiments, the linker is positioned between two ADBDs of the adaptor. In some embodiments, the linker is positioned between the AD and ADBD of the adaptor. Suitable linkers for connecting two or more functional domains of the adaptor will be apparent to those of skill in the art and may generally be any linker used in the art to connect peptides, proteins, or other organic molecules. Exemplary linkers are provided in Section IV. In certain embodiments, the linker is suitable for constructing proteins or polypeptides intended for pharmaceutical use.

[0236] In addition to an AD (or ADs) and an ADBD (or ADBDs), the adapters provided herein can further comprise an additional domain or additional domains, for example, a domain that confers extended half-life.

[0237] In some embodiments, the adaptor, or the ADBD in the adaptor, is deimmunized.

[0238] The adapters provided herein have uses including, but not limited to, diagnostic, analytical, and therapeutic applications. In certain embodiments, the adapters are used in combination with a chimeric antigen receptor (CAR) (e.g., as described in Sections VI and XI) expressed on the surface of a cell (e.g., as described in Sections VII and XI), for example, to kill a target cell.

[0239] Va. Antigenic determinant (AD) The adapters provided herein comprise at least one antigenic determinant (AD). In some embodiments, the adapter comprises a single AD. In some embodiments, the adapter comprises two or more ADs. When the adapter comprises two or more ADs, the ADs can be the same or different.

[0240] In the adapters provided herein, the AD can be any AD or combination of multiple ADs (e.g., as described in Section II and Section XI).

[0241] In some embodiments, the adaptor comprises the extracellular domain of BCMA (e.g., a polypeptide comprising the sequence of SEQ ID NO: 5). In some embodiments, the adaptor comprises 5 to 25, 5 to 50, 5 to 75, 5 to 100, 5 to 125, or 5 to 150 amino acid residues, more than 150 amino acid residues, or all amino acid residues of SEQ ID NO: 5.

[0242] In some embodiments, the adaptor comprises the extracellular domain of CD123 (e.g., a polypeptide comprising the sequence of SEQ ID NO: 11). In some embodiments, the adaptor comprises 5 to 25, 5 to 50, 5 to 75, 5 to 100, 5 to 125, or 5 to 150 amino acid residues, more than 150 amino acid residues, or all amino acid residues of SEQ ID NO: 11.

[0243] In some embodiments, the adaptor comprises the extracellular domain of CD19 (e.g., a polypeptide comprising the sequence of SEQ ID NO: 2 or 3). In some embodiments, the adaptor comprises 5 to 25, 5 to 50, 5 to 75, 5 to 100, 5 to 125, or 5 to 150 amino acid residues, more than 150 amino acid residues, or all amino acid residues of SEQ ID NO: 2 or 3.

[0244] In some embodiments, the adaptor comprises the extracellular domain of CD20. In some embodiments, the adaptor comprises 5 to 25, 5 to 50, 5 to 75, 5 to 100, 5 to 125, or 5 to 150 amino acid residues, more than 150 amino acid residues, or all amino acid residues of SEQ ID NOs: 6-9, or 10.

[0245] In some embodiments, the adaptor comprises the extracellular domain of CD22 (e.g., a polypeptide comprising the sequence of SEQ ID NO: 41). In some embodiments, the adaptor comprises 5 to 25, 5 to 50, 5 to 75, 5 to 100, 5 to 125, or 5 to 150 amino acid residues, more than 150 amino acid residues, or all amino acid residues of SEQ ID NO: 41.

[0246] In some embodiments, the adaptor comprises the extracellular domain of CD37 (e.g., a polypeptide comprising the sequence of SEQ ID NO: 12 or 13). In some embodiments, the adaptor comprises 5 to 25, 5 to 50, 5 to 75, 5 to 100, 5 to 125, or 5 to 150 amino acid residues, more than 150 amino acid residues, or all amino acid residues of SEQ ID NO: 12 or 13.

[0247] In some embodiments, the adaptor comprises the extracellular domain of CS1 (e.g., a polypeptide comprising the sequence of SEQ ID NO: 1138). In some embodiments, the adaptor comprises between 5 and 25, between 5 and 50, between 75, between 5 and 100, between 5 and 125, or between 5 and 150 amino acid residues, more than 150 amino acid residues, or all amino acid residues of SEQ ID NO: 1138.

[0248] In some embodiments, the adaptor comprises the extracellular domain of HER2 (e.g., a polypeptide comprising the sequence of SEQ ID NO: 42). In some embodiments, the adaptor comprises 5 to 25, 5 to 50, 5 to 75, 5 to 100, 5 to 125, or 5 to 150 amino acid residues, more than 150 amino acid residues, or all amino acid residues of SEQ ID NO: 42.

[0249] In some embodiments, the adaptor comprises the extracellular domain of CD45 (e.g., a polypeptide comprising the sequence of residues 29-766 of SEQ ID NO: 1106). In some embodiments, the adaptor comprises 5-25, 5-50, 5-75, 5-100, 5-125, or 5-150 amino acid residues, more than 150 amino acid residues, or all of residues 29-766 of SEQ ID NO: 1106.

[0250] In some embodiments, the adaptor comprises the extracellular domain of CD26, CD30, CD33, or CD38. In some embodiments, the adaptor comprises 5 to 25, 5 to 50, 5 to 75, 5 to 100, 5 to 125, or 5 to 150 amino acid residues, more than 150 amino acid residues, or all amino acid residues of the extracellular domain of CD26, CD30, CD33, or CD38.

[0251] In some embodiments, the AD is an epitope of AFP. In further embodiments, the AD comprises 5 to 25, 5 to 50, 5 to 75, 5 to 100, 5 to 125, or 5 to 150 amino acid residues, more than 150 amino acid residues, or all amino acid residues of SEQ ID NO: 15.

[0252] In some embodiments, the AD is an epitope of AFP p26. In further embodiments, the AD comprises 5 to 25, 5 to 50, 5 to 75, 5 to 100, 5 to 125, or 5 to 150 amino acid residues, more than 150 amino acid residues, or all amino acid residues of SEQ ID NO: 16. In further embodiments, the AD comprises 5 to 25, 5 to 50, 5 to 75, 5 to 100, 5 to 125, or 5 to 150 amino acid residues, more than 150 amino acid residues, or all amino acid residues of SEQ ID NO: 1117. In further embodiments, the AD comprises amino acid residues of SEQ ID NO: 16, 1117, 1118, 1119, 1120, 1121, 1122, or 1123.

[0253] In some embodiments, the adapter comprises a p26 protein (e.g., a polypeptide comprising the sequence of SEQ ID NO: 16, 1117, 1118, 1119, 1120, 1121, 1122, or 1123). Fusion proteins comprising such p26 sequences have surprisingly been discovered herein to have long serum half-lives. In some embodiments, the adapter has an in vivo plasma half-life of at least 1 hour, at least 2 hours, at least 4 hours, at least 8 hours, at least 16 hours, at least 32 hours, at least 64 hours, or more than 2 hours. In some embodiments, the adapter has an in vivo plasma half-life in mice of at least 1 hour, at least 2 hours, at least 4 hours, at least 8 hours, at least 16 hours, at least 32 hours, at least 64 hours, or greater than 65 hours, 1-10 hours, 2-10 hours, 4-10 hours, 6-10 hours, or 6-9 hours. In some embodiments, the adapter has an in vivo plasma half-life in humans of at least 1 hour, at least 2 hours, at least 4 hours, at least 8 hours, at least 16 hours, at least 32 hours, at least 64 hours, or more than 65 hours, 1-10 hours, 2-10 hours, 4-10 hours, 6-10 hours, or 6-9 hours.

[0254] In some embodiments, the disclosure provides methods for improving the in vivo half-life (e.g., in mice or humans) of an adapter comprising a p26 protein (e.g., having the sequence of SEQ ID NO: 16, 1117, 1118, 1119, 1120, 1121, 1122, or 1123). In some embodiments, the adapter comprises one or more target-binding DDpps. In some embodiments, the half-life of the adapter is extended or shortened by substituting or deleting one or more amino acid residues normally found in human p26 proteins, or by inserting one or more amino acid residues not normally found in human p26 proteins. In another embodiment, the p26 sequence of the adapter is modified by making 1, 2, 3, 5, 5, 10, or 1-20, 1-10, 3-10, or 3-5 amino acid substitutions (conservative and / or non-conservative substitutions), deletions, and / or insertions to extend or shorten the in vivo half-life of the adapter. In a specific embodiment, the amino acid residue corresponding to glutamine (Gln, Q) at position 217 of p26 (SEQ ID NO: 16) is substituted with another amino acid residue. In a further embodiment, the substitution is Gln217Pro. In another embodiment, the p26 sequence of the adapter is modified by deleting 1 to 150, 1 to 100, 1 to 50, 1 to 25, or 1 to 10 amino acid residues to increase or decrease the in vivo half-life of the adapter. In a further embodiment, the p26 sequence of the adapter is modified by 1, 2, 3, 5, 5, 10, or 1 to 20, 1 to 10, 3 to 10, or 3 to 5 amino acid substitutions (conservative and / or non-conservative substitutions), deletions, and / or insertions to increase or decrease the interaction of the adapter with FcRn.

[0255] In some embodiments, the AD (e.g., in the adaptor and / or on the target cell) is an AD present in a naturally occurring protein or other molecule. In some embodiments, the AD is an AD that is endogenous to humans.

[0256] In some embodiments, the AD is an epitope of a human intracellular protein. In further embodiments, the AD is an epitope of a human intracellular protein selected from elastin, Tyk2, Jak1, Jak2, Jak3, LCK, ZAP-70, and GRB2. In further embodiments, the AD comprises 5-25, 5-50, 5-75, 5-100, 5-125, or 5-150 amino acid residues, more than 150 amino acid residues, or all amino acid residues of the intracellular protein.

[0257] In some embodiments, the target specifically bound by the ADBD of an adapter is itself the AD of another adapter having a different sequence.

[0258] Vb. Antigenic determinant binding domain (ADBD) The adapters provided herein comprise at least one antigenic determinant binding domain (ADBD). In some embodiments, the adapter comprises one ADBD. In some embodiments, the adapter comprises at least two, three, four, or five, or more than five ADBDs. In some embodiments, the adapter comprises one to three, one to four, one to five, or more than five different ADBDs. In some embodiments, the adapter comprises at least two, three, four, or five, or more than five different ADBDs. Thus, the adapter can comprise a monomeric ADBD (i.e., comprising one antigenic determinant binding domain) or a multimeric ADBD (i.e., two or more antigenic determinant binding domains operably linked in tandem, optionally with a linker). In some embodiments, the use of a multimeric adapter results in enhanced (e.g., synergistic) target binding. In further embodiments, the use of a multimeric adapter allows for targeting of two or more targets using a single adapter construct (e.g., bispecific, trispecific, etc.).

[0259] Multimeric adapters may be homomultimeric (i.e., containing two or more identical ADBDs, optionally joined by a linker) adapters (e.g., homodimers, homotrimers, homotetramers, etc.) or heteromultimeric (i.e., containing two or more antigenic determinant binding domains, where at least two different antigenic determinant binding domains are present). The number of ADBDs contained in any particular adapter may vary depending on the embodiment and is determined, at least in part, by the expression system in which the adapter is produced. However, in some embodiments, the fusion protein may contain multimers of about 5 to about 10 ADBDs, about 10 to about 15 ADBDs, about 15 to about 20 ADBDs, about 20 to about 25 ADBDs, or about 25 to about 30 ADBDs (including intermediate numbers and endpoints of the recited numbers). Furthermore, the multidomains of an adapter may contain the same or different ADBDs. In some embodiments, 2, 3, 4, 5, or more than 5 domains are present in tandem.

[0260] In one embodiment, the adapter comprises two or more operably linked ADBDs. In one embodiment, the adapter comprises two ADBDs that bind to the same or different ADs on the target antigen. The combination of two or more identical ADBDs that bind to the same target antigen results in a multivalent molecule that provides distinct advantages (e.g., increased avidity, target clustering, and receptor activation) over compositions comprising only one ADBD for a target antigen. In another embodiment, the adapter comprises two ADBDs that bind to different antigens. In some embodiments, the adapter comprises two ADBDs that bind to different antigens on the same cell. In some embodiments, the adapter comprises two ADBDs that bind to different antigens on different cells. The combination of two or more ADBDs results in a multivalent and multispecific adapter that has the potential to bind to two or more target antigens, separately or simultaneously. In some embodiments, the multivalent adapter can simultaneously bind to the same target antigen. In some embodiments, the multivalent adapter can simultaneously bind to different target antigens. In some embodiments, the adapter comprises two or more operably linked ADBDs separated by antigenic determinants. In some embodiments, the antigenic determinant is at least 50, 75, 100, 150, 200, 250, 300, 350, 400, 450, or 500 amino acids in length. In some embodiments, the antigenic determinant is 5 to 500, 5 to 400, 10 to 300, 5 to 200, 50 to 100, 5 to 50, 10 to 500, 10 to 400, 10 to 300, 10 to 200, 10 to 100, 10 to 50, 50 to 500, 50 to 400, 50 to 300, 50 to 200, 50 to 100, 50 to 75, 100 to 500, 100 to 400, 100 to 300, 100 to 200, or 100 to 150 amino acids in length. In a further embodiment, the adapter comprises two or more operably linked ADBDs separated by a BCMA antigenic determinant. In a further embodiment, the adapter comprises two or more operably linked ADBDs separated by an AFP p26 antigenic determinant. In a further embodiment, the adapter comprises two or more operably linked ADBDs separated by a CD45 antigenic determinant.In a further embodiment, the adapter comprises two or more operably linked ADBDs separated by a CD26, CD30, CD33, or CD38 antigenic determinant.

[0261] The ADBD in the adapters provided herein can bind to any AD (e.g., as described in Sections II and XI). In some embodiments, the ADBD binds to BCMA (e.g., a polypeptide comprising the sequence of SEQ ID NO: 5). In some embodiments, the ADBD binds to CD123 (e.g., a polypeptide comprising the sequence of SEQ ID NO: 11). In some embodiments, the ADBD binds to CD22 (e.g., a polypeptide comprising the sequence of SEQ ID NO: 41). In some embodiments, the ADBD binds to CD19 (e.g., a polypeptide comprising the sequence of SEQ ID NO: 3). In some embodiments, the ADBD binds to CD20 (e.g., a polypeptide comprising the sequence of SEQ ID NO: 6-9 or 10). In some embodiments, the ADBD binds to CD37 (e.g., a polypeptide comprising the sequence of SEQ ID NO: 12 or 13). In some embodiments, the ADBD binds to CS1 (e.g., a polypeptide comprising the sequence of SEQ ID NO: 1138). In some embodiments, the ADBD binds to HER2 (e.g., a polypeptide comprising the sequence of SEQ ID NO: 42). In some embodiments, the ADBD binds to CD45. In some embodiments, the ADBD in the adapters provided herein binds to the AD of human CD26, CD30, CD33, or CD38. Adapters can be "monospecific" or "multispecific." A "multispecific" (e.g., bispecific, trispecific, or even more multispecific) adapter recognizes and binds to two or more different epitopes present on one or more different molecules.

[0262] In some embodiments, the adaptor comprises a domain (e.g., extracellular domain) of BCMA (e.g., a polypeptide comprising the sequence of SEQ ID NO: 4). In some embodiments, the adaptor comprises a domain (e.g., extracellular domain) of CD123 (e.g., a polypeptide comprising the sequence of SEQ ID NO: 11). In some embodiments, the adaptor comprises a domain (e.g., extracellular domain) of CD22 (e.g., a polypeptide comprising the sequence of SEQ ID NO: 24). In some embodiments, the adaptor comprises a domain (e.g., extracellular domain) of CD19 (e.g., a polypeptide comprising the sequence of SEQ ID NO: 3). In some embodiments, the adaptor comprises a domain (e.g., extracellular domain) of CS1 (e.g., a polypeptide comprising the sequence of SEQ ID NO: 1138). In some embodiments, the adaptor comprises a domain (e.g., extracellular domain) of HER2 (e.g., a polypeptide comprising the sequence of SEQ ID NO: 42). In some embodiments, the adaptor comprises a domain (e.g., extracellular domain) of CD45. In some embodiments, the adaptor comprises a domain (e.g., extracellular domain) of CD26, CD30, CD33, or CD38. In some embodiments, the adaptor comprises a fragment of a domain. In further embodiments, the adaptor comprises a fragment of a domain having an amino acid sequence selected from the group: SEQ ID NO: 4 or 5, SEQ ID NO: 11, SEQ ID NO: 24, and SEQ ID NO: 3. In some embodiments, the adaptor comprises a fragment of a domain that is at least 50, 75, 100, 150, 200, 250, 300, 350, 400, 450, or 500 amino acids in length. In some embodiments, the antigenic determinant is 5 to 500, 5 to 400, 10 to 300, 5 to 200, 50 to 100, 5 to 50, 10 to 500, 10 to 400, 10 to 300, 10 to 200, 10 to 100, 10 to 50, 50 to 500, 50 to 400, 50 to 300, 50 to 200, 50 to 100, 50 to 75, 100 to 500, 100 to 400, 100 to 300, 100 to 200, or 100 to 150 amino acids in length.

[0263] In some embodiments, the adapter comprises at least two ADBDs that bind to and bridge one or more target antigens bound by the ADBDs and / or complexes containing the target antigens. In some embodiments, the bridged antigens are on the same cell. In some embodiments, the bridged antigens are on different cells. In some embodiments, the adapter comprises two or more operably linked ADBDs separated by an antigenic determinant (e.g., a domain described above). In some embodiments, the antigenic determinant is at least 50, 75, 100, 150, 200, 250, 300, 350, 400, 450, or 500 amino acids in length. In some embodiments, the antigenic determinant is 5-500, 5-400, 10-300, 5-200, 50-100, 5-50, 10-500, 10-400, 10-300, 10-200, 10-100, 10-50, 50-500, 50-400, 50-300, 50-200, 50-100, 50-75, 100-500, 100-400, 100-300, 100-200, or 100-150 amino acids in length. In further embodiments, the adapter comprises two or more operably linked ADBDs separated by a BCMA antigenic determinant. In further embodiments, the adapter comprises two or more operably linked ADBDs separated by a CD45 antigenic determinant. In a further embodiment, the adapter comprises two or more operably linked ADBDs separated by a CD26, CD30, CD33, or CD38 antigenic determinant, hi a further embodiment, the adapter comprises two or more operably linked ADBDs separated by an AFP p26 antigenic determinant.

[0264] In some embodiments, the adapter comprises at least two identical ADBDs (i.e., multivalent). In some embodiments, the multivalent adapter is capable of simultaneously binding to two or more of the same target antigens. In some embodiments, the multivalent adapter is multivalent and capable of simultaneously binding to the same target antigen. In some embodiments, the multivalent adapter comprises two or more operably linked ADBDs separated by an antigenic determinant. In some embodiments, the antigenic determinant is at least 50, 75, 100, 150, 200, 250, 300, 350, 400, 450, or 500 amino acids in length. In some embodiments, the antigenic determinant is 5-500, 5-400, 10-300, 5-200, 50-100, 5-50, 10-500, 10-400, 10-300, 10-200, 10-100, 10-50, 50-500, 50-400, 50-300, 50-200, 50-100, 50-75, 100-500, 100-400, 100-300, 100-200, or 100-150 amino acids in length. In further embodiments, the multivalent adapter comprises two or more operably linked ADBDs separated by a BCMA antigenic determinant. In further embodiments, the multivalent adapter comprises two or more operably linked ADBDs separated by a CD45 antigenic determinant. In a further embodiment, the multivalent adaptor comprises two or more operably linked ADBDs separated by an AFP p26 antigenic determinant.

[0265] In some embodiments, the adapter comprises at least two ADBDs that bind to different antigens (i.e., are multispecific). In some embodiments, the multispecific adapter can simultaneously bind to different target antigens. In some embodiments, the adapter is also multivalent and can simultaneously bind to the same target antigen. In some embodiments, the multispecific adapter comprises two or more operably linked ADBDs separated by an antigenic determinant. In some embodiments, the antigenic determinant is at least 50, 75, 100, 150, 200, 250, 300, 350, 400, 450, or 500 amino acids in length. In some embodiments, the antigenic determinant is 5-500, 5-400, 10-300, 5-200, 50-100, 5-50, 10-500, 10-400, 10-300, 10-200, 10-100, 10-50, 50-500, 50-400, 50-300, 50-200, 50-100, 50-75, 100-500, 100-400, 100-300, 100-200, or 100-150 amino acids in length. In further embodiments, the adapter comprises two or more operably linked ADBDs separated by a BCMA antigenic determinant. In further embodiments, the adapter comprises two or more operably linked ADBDs separated by a CD45 antigenic determinant. In a further embodiment, the adapter comprises two or more operably linked ADBDs separated by an AFP p26 antigenic determinant.

[0266] In one embodiment, the multispecific adapter comprises at least two ADBDs that bind to at least two different epitopes on a single target (i.e., multi-epitopic to the same target antigen). In further embodiments, the multispecific adapter comprises at least one ADBD that specifically binds to one epitope on a target and at least one other ADBD that specifically binds to a different epitope on the same target antigen. In one embodiment, the multispecific adapter comprises at least one ADBD that specifically binds to an epitope on a first target and at least one ADBD that specifically binds to an epitope on a second antigen. In some embodiments, the adapter comprises at least one ADBD that specifically binds to an epitope on a first target antigen on a cell and at least one ADBD that specifically binds to an epitope on a second antigen on the same cell. In some embodiments, the adapter comprises at least one ADBD that specifically binds to an epitope on a first target antigen on a cell and at least one ADBD that specifically binds to an epitope on a second antigen on a different cell.

[0267] In further embodiments, the adapter comprises two or more ADBDs operably linked to other heterologous proteins (or subdomains thereof), thereby conferring the multivalency, multispecificity, and / or functional properties (e.g., pharmacokinetics, such as extended half-life) of the fusion partner to the adapter fusion protein. Examples of adapter fusion partners include, but are not limited to, antibodies, antibody subdomains (e.g., scFv or Fc domains), serum albumin, serum albumin subdomains, cell surface receptors, T cell receptor (TCR) α chains, T cell receptor β chains, cell surface receptor subdomains, peptides, and peptide tags (e.g., FLAG or myc). The number and location of ADBDs and their respective locations within the adapter can vary. For example, ADBDs can be located at one or all ends of the fusion partner and / or interspersed within the heterologous subunit within the adapter fusion partner. In some embodiments, the adapter comprises two or more ADBDs separated by a heterologous protein (e.g., an antigenic determinant). In some embodiments, the heterologous protein is at least 50, 75, 100, 150, 200, 250, 300, 350, 400, 450, or 500 amino acids in length. In some embodiments, the heterologous protein is 5 to 500, 5 to 400, 10 to 300, 5 to 200, 50 to 100, 5 to 50, 10 to 500, 10 to 400, 10 to 300, 10 to 200, 10 to 100, 10 to 50, 50 to 500, 50 to 400, 50 to 300, 50 to 200, 50 to 100, 50 to 75, 100 to 500, 100 to 400, 100 to 300, 100 to 200, or 100 to 150 amino acids in length.

[0268] In one embodiment, the adapter is bispecific and comprises an ADBD that specifically binds to two different target antigens. In a further embodiment, the bispecific adapter specifically binds to two different target antigens expressed on the surface of two different cell types. In a further embodiment, the bispecific adapter specifically binds to two different target antigens expressed on the surface of a tumor cell. In a further embodiment, the bispecific adapter specifically binds to two different target antigens expressed on the surface of a multiple myeloma cell (e.g., BCMA and CS1). In one embodiment, the bispecific adapter binds to target antigens expressed on different cells. In a further embodiment, the bispecific adapter binds to target antigens expressed on different tumor cells. In another embodiment, the bispecific adapter binds to target antigens expressed on different cells within the tumor vasculature or tumor microenvironment. In one embodiment, the bispecific adapter specifically binds to a cancer cell target and an immune effector cell target. In one embodiment, the bispecific adapter specifically binds to a target expressed on a cancer cell (e.g., CD19) and a target expressed on the surface of a T lymphocyte (e.g., CD3 or CD45). In some embodiments, the bispecific adapter is capable of simultaneously binding to different target antigens. In some embodiments, the bispecific adapter comprises two or more operably linked ADBDs separated by an antigenic determinant. In some embodiments, the antigenic determinant is at least 50, 75, 100, 150, 200, 250, 300, 350, 400, 450, or 500 amino acids in length. In some embodiments, the antigenic determinant is 5-500, 5-400, 10-300, 5-200, 50-100, 5-50, 10-500, 10-400, 10-300, 10-200, 10-100, 10-50, 50-500, 50-400, 50-300, 50-200, 50-100 50-75, 100-500, 100-400, 100-300, 100-200, or 100-150 amino acids in length. In further embodiments, the adapter comprises two or more operably linked ADBDs separated by a BCMA antigenic determinant.In a further embodiment, the adapter comprises two or more operably linked ADBDs separated by a CD45 antigenic determinant, hi a further embodiment, the adapter comprises two or more operably linked ADBDs separated by an AFP p26 antigenic determinant.

[0269] In some embodiments where an adaptor comprises two or more ADBDs, the ADBDs can be any type of ADBD discussed herein (e.g., any of the ADBDs described in Sections III and XI above). For example, the ADBD can be an antibody, an antigen-binding fragment thereof, an scFv, an alternative scaffold binding domain, a D domain, a T cell receptor, or an antigen-binding fragment thereof.

[0270] In some embodiments, an adapter comprises two or more ADBDs, which may be the same or different types of antigen-binding molecules. For example, an adapter may comprise two ADBDs that are D domains. The two D domain ADBDs may be the same or different. An adapter may comprise an ADBD that is a D domain and an ADBD that is an scFv. An adapter may also comprise an ADBD that is a T cell receptor or an antigen-binding fragment thereof, or an ADBD that is an scFv. In some embodiments, an adapter comprises two or more operably linked ADBDs separated by an antigenic determinant. In some embodiments, the antigenic determinant is at least 50, 75, 100, 150, 200, 250, 300, 350, 400, 450, or 500 amino acids in length. In some embodiments, the antigenic determinant is 5-500, 5-400, 10-300, 5-200, 50-100, 5-50, 10-500, 10-400, 10-300, 10-200, 10-100, 10-50, 50-500, 50-400, 50-300, 50-200, 50-100, 50-75, 100-500, 100-400, 100-300, 100-200, or 100-150 amino acids in length. In further embodiments, the adapter comprises two or more operably linked ADBDs separated by a BCMA antigenic determinant. In further embodiments, the adapter comprises two or more operably linked ADBDs separated by a CD45 antigenic determinant. In a further embodiment, the adapter comprises two or more operably linked ADBDs separated by an AFP p26 antigenic determinant.

[0271] In some embodiments, the ADBD of the adaptor is deimmunized.

[0272] In some embodiments, the adapter comprises an ADBD that binds to an antigen target, including an AD of interest, without discernible effect on the function of the target. Alternatively, in some embodiments, the adapter comprises an ADBD that binds to an antigen target, including an AD of interest, and fully or partially inhibits, antagonizes, stimulates, blocks, enhances, stimulates, or interferes with the biological activity of the target. Binding, identified as agonistic or antagonistic, can be determined using, or routinely modified by, assays, bioassays, and / or animal models known in the art for assessing such activity.

[0273] An adapter agonist refers to an adapter that in some way increases or enhances the biological activity of the adapter target or has biological activity equivalent to a known agonist of the adapter target. In another embodiment, an adapter is an antagonist of the target to which it binds. An adapter antagonist refers to an adapter that completely or partially blocks the biological activity of the adapter target or that in some way interferes with the biological activity of the adapter target or has biological activity equivalent to a known antagonist of the adapter target.

[0274] In one embodiment, the adaptor specifically binds to a target that is a serum protein. In one embodiment, the adaptor specifically binds to a serum protein selected from serum albumin (e.g., human serum albumin (HSA)), thyroxine-binding protein, transferrin, fibrinogen, and immunoglobulins (e.g., IgG, IgE, and IgM). Without being bound by theory, it is believed that the attachment of the adaptor to a carrier protein confers improved pharmacodynamic properties to the adaptor, including, but not limited to, improved tumor targeting, tumor penetration, intratumoral spread, and enhanced therapeutic action, compared to an adaptor lacking the protein-binding sequence (see, e.g., WO 01 / 45746, the contents of which are incorporated herein by reference in their entirety).

[0275] In one embodiment, the target of interest specifically bound by the adapter is a disease-associated antigen. The antigen can be an antigen characteristic of cancer, and / or a particular cell type (e.g., a hyperproliferative cell), and / or a pathogen (e.g., a bacterial cell (e.g., tuberculosis, smallpox, and anthrax), a virus (e.g., HIV and H), a parasite (e.g., malaria and leishmaniasis), a fungal infection, a mold, a mycoplasma, a prion antigen, or an antigen associated with a disorder of the immune system.

[0276] In additional embodiments, the target bound by the adapter is a bacterial antigen, a viral antigen, a fungal antigen, a mycoplasmal antigen, a prion antigen, or a parasitic antigen (e.g., one that infects mammals). In one embodiment, the target of the adapter is human papillomavirus, hepatitis B, rabies, Nipah virus, West Nile virus, meningitis virus, or CMV. In additional embodiments, the adapter specifically binds to the pathogen.

[0277] Vc. Adaptor functional domain In some embodiments, the adaptor comprises a first antigenic determinant (AD), a second antigenic determinant-binding domain (ADBD), and further comprises a functional domain that confers one or more additional desirable properties (e.g., improved manufacturing properties) and / or pharmacokinetic properties (e.g., improved half-life). The adaptor's functional domain can be located between the AD and the ADBD. The adaptor can also be located N-terminal to both the AD and the ADBD, or C-terminal to both the AD and the ADBD. In some embodiments, where the adaptor comprises two or more ADs, the adaptor's functional domain can be located between the two or more ADs, N-terminal to the two or more ADs, or C-terminal to the two or more ADs. In some embodiments, where the adaptor comprises two or more ADBDs, the adaptor's functional domain can be located between the two or more ADBDs, N-terminal to the two or more ADBDs, or C-terminal to the two or more ADBDs.

[0278] In some embodiments, the adapter comprises a functional domain selected from Fc or variant Fc (e.g., human Fc or variant Fc domain) or fragment thereof, a serum protein (e.g., human serum albumin) or fragment thereof; an FcRn-binding domain; a serum protein-binding domain; a cytokine, growth factor, hormone, or enzyme; an imaging agent; a labeling agent; and a peptide tag.

[0279] The functional domain of the adapter can be naturally derived or the result of recombinant engineering (e.g., phage display, xeno-mouse, or synthetic techniques). In certain embodiments, the functional domain of the adapter extends half-life, enhances or reduces antibody-dependent cellular cytotoxicity (ADCC) activity, and / or enhances or reduces complement-dependent cytotoxicity (CDC) activity.

[0280] In some embodiments, the adapter comprises a functional domain selected from Fc or variant Fc (e.g., human Fc or variant human Fc domain) or a fragment or derivative thereof, a serum protein (e.g., human serum albumin) or a fragment or derivative thereof (e.g., a serum protein binding domain); an FcRn binding domain; and a serum protein binding domain.

[0281] In one embodiment, the adaptor comprises a functional domain comprising an antibody effector domain or a derivative of an antibody effector domain that confers one or more effector functions to the adaptor, such as the ability to bind to one or more Fc receptors. In some embodiments, the functional domain comprises the CH2 and / or CH3 domains of one or more antibodies having effector functions provided by the CH2 and CH3 domains. In some embodiments, the functional domain comprises a derivative of the CH2 and / or CH3 domains of one or more antibodies having effector functions provided by the CH2 and CH3 domains. Other sequences that can be included in an adaptor to provide effector functions, and are encompassed by the invention, will be apparent to those of skill in the art and can be routinely selected and designed into the adaptors encompassed herein based on the desired effector functions.

[0282] In one embodiment, the adaptor comprises a functional domain that enhances antibody-dependent cellular cytotoxicity (ADCC) conferred by the adaptor (see, e.g., Bruhns et al., Blood 113:3716-3725 (2009); Shields et al., J. Biol. Chem. 276:6591-6604 (2001); Lazar et al., PNAS 103:4005-4010 (2006); Stavenhagen et al., Cancer Res. 67:8882-8890 (2007); Horton et al., Cancer Res. 68:8049-8057 (2008); Zalevsky et al., Blood 113:3735-3743 (2009); Bruckheimer, Neoplasia 11:509-517 (2009); WO 2006 / 020114; Strohl, Curr. Op. Biotechnol. 20:685-691 (2009); and WO 2004 / 074455, each of which is incorporated herein by reference in its entirety. Examples of fragment-engineered modifications of effector function conferred to the Fc portion contained in the functional domain of the adaptor that enhances ADCC include IgG1-S298A, E333A, K334A; IgG1-S239D, I332E; IgG1-S239D, A330L, I332E; IgG1-P247I, A339D or Q; IgG1-D280H, K290S and S298D or V together or K290S alone; IgG1-F243L, R292P, Y300L; IgG1-F243L, R292P, Y300L, P396L; and IgG1-F243L, R292P, Y300L, V305I, P396L (numbering of residues in the Fc region is that of the EU index of Kabat et al., Sequences of proteins of Immunological Interest, 1991 5th ed.)

[0283] Thus, in some embodiments, the adapter comprises a functional domain comprising an antibody fragment that confers a biological or biochemical property of an immunoglobulin to the adapter. In some embodiments, the antibody fragment confers a property selected from the ability to non-covalently dimerize, the ability to localize to tumor sites, and an increased serum half-life compared to the adapter without the antibody fragment. In certain embodiments, the adapter is at least as stable as the corresponding antibody fragment without the adapter. In certain embodiments, the adapter is at least more stable than the corresponding antibody fragment without the adapter. The stability of the adapter protein can be measured using established methods, such as, for example, ELISA techniques. In some embodiments, the adapter is stable in whole blood at 37°C (in vivo or ex vivo) for at least about 10 hours, at least about 15 hours, at least about 20 hours, at least about 24 hours, at least about 25 hours, at least about 30 hours, at least about 35 hours, at least about 40 hours, at least about 45 hours, at least about 48 hours, at least about 50 hours, at least about 55 hours, at least about 60 hours, at least about 65 hours, at least about 70 hours, at least about 72 hours, at least about 75 hours, at least about 80 hours, at least about 85 hours, at least about 90 hours, at least about 95 hours, or at least about 100 hours (including any time in between these amounts). In one embodiment, the adapter comprises an immunoglobulin effector domain or half-life-affecting domain corresponding to an immunoglobulin domain or fragment in which at least a portion of one or more constant region domains has been modified to confer a desired property, such as reduced or increased effector function, ability to non-covalently dimerize, increased ability to localize to tumor sites, shortened serum half-life, or extended serum half-life, compared to an immunoglobulin fragment having a corresponding unmodified immunoglobulin sequence. These constant region domain modifications can be amino acid substitutions, insertions, or deletions.

[0284] In one embodiment, the adapter comprises a functional domain comprising the amino acid sequence of an immunoglobulin effector domain or a derivative of an immunoglobulin effector domain that confers antibody-dependent cellular cytotoxicity (ADCC) to the adapter. In further embodiments, the adapter comprises the sequence of an immunoglobulin effector domain that has been modified to enhance ADCC (e.g., Bruhns, Blood 113:3716-3725 (2009); Shields, J. Biol. Chem. 276:6591-6604 (2001); Lazar, PNAS 103:4005-4010 (2006); Stavenhagen, Cancer Res. 67:8882-8890 (2007); Horton, Cancer Res. 68:8049-8057 (2008); Zalevsky, Blood 113:3735-3743 (2009); Bruckheimer, Neoplasia 11:509-517 (2009); WO 06 / 020114; Strohl, Curr. Op. Biotechnol. 20:685-691 (2009); and WO 04 / 074455, each of which is incorporated herein by reference in its entirety. Examples of immunoglobulin fragment engineering modifications included in the amino acid sequence in the adapter that enhance ADCC include IgG1-S298A, E333A, K334A; IgG1-S239D, I332E; IgG1-S239D, A330L, I332E; IgG1-P247I, A339D or Q; IgG1-D280H, K290S and S298D or V together or K290S alone; IgG1-F243L, R292P, Y300L; IgG1-F243L, R292P, Y300L, P396L; and IgG1-F243L, R292P, Y300L, V305I, P396L (numbering of residues in the Fc region is that of the EU index of Kabat et al., Sequences of proteins of Immunological Included are immunoglobulin effector domain sequences having one or more modifications corresponding to those described in The Journal of Clinical Immunology, Vol. 1, No. 1, pp. 1991-1995, 5th Edition (incorporated herein by reference).

[0285] In a further embodiment, the adaptor comprises a functional domain comprising the amino acid sequence of an immunoglobulin effector domain or a derivative of an immunoglobulin effector domain that confers antibody-dependent cellular phagocytosis (ADCP) to the adaptor. In further embodiments, the adapter comprises the sequence of an immunoglobulin effector domain that has been modified to enhance antibody-dependent cellular phagocytosis (ADCP) (e.g., Shields et al., J. Biol. Chem. 276:6591-6604 (2001); Lazar et al., PNAS 103:4005-4010 (2006); Stavenhagen et al., Cancer Res. 67:8882-8890 (2007); Richards et al., Mol. Cancer Ther. 7:2517-2527 (2008); Horton et al., Cancer Res. 68:8049-8057 (2008); Zalevsky et al., Blood 113:3735-3743 (2009); Bruckheimer et al., Neoplasia 11:509-517 (2009); WO 06 / 020114; Strohl, Curr. Op. Biotechnol. 20:685-691 (2009); and WO 04 / 074455, each of which is incorporated herein by reference in its entirety.Examples of immunoglobulin fragment engineering modifications included in the amino acid sequence load in the adaptor to enhance ADCP include IgG1-S298A, E333A, K334A; IgG1-S239D, I332E; IgG1-S239D, A330L, I332E; IgG1-P247I, A339D or Q; IgG1-D280H, K290S and S298D or V together; IgG1-F243L, R292P, Y300L; IgG1-F243L, R292P, Y300L, P396L; IgG1-F243L, R292P, Y300L, V305I, P396L; and IgG1-G236A, S239D, I332E (residue numbering is that of the EU index of Kabat et al.). Included are immunoglobulin effector domain sequences having one or more modifications corresponding to those in "Proteins of Immunological Interest," 1991, 5th ed. (incorporated herein by reference).

[0286] In further embodiments, the adaptor comprises a functional domain comprising the amino acid sequence of an immunoglobulin effector domain or a derivative of an immunoglobulin effector domain that confers complement-dependent cytotoxicity (CDC) to the adaptor. In further embodiments, the adaptor comprises the sequence of an immunoglobulin effector domain that has been modified to enhance complement-dependent cytotoxicity (CDC) (see, e.g., Idusogie et al., J. Immunol. 166:2571-2575 (2001); Strohl, Curr. Op. Biotechnol. 20:685-691 (2009); and Natsume et al., Cancer Res. 68:3863-3872 (2008) each of which is incorporated herein by reference in its entirety). For example, the adapter can comprise an antibody fragment or domain containing one or more of the following modifications that enhance CDC: IgG1-K326A, E333A; IgG1-K326W, E333S, IgG2-E333S (residue numbering is that of the EU index of Kabat et al. (Sequences of proteins of Immunological Interest, 1991 5th ed.), incorporated herein by reference).

[0287] In a further embodiment, the adapter comprises a functional domain comprising the amino acid sequence of an immunoglobulin effector domain or a derivative of an immunoglobulin effector domain that confers the adapter with the ability to bind to the FcγRIIb receptor. In a further embodiment, the adapter comprises the sequence of an immunoglobulin effector domain that has been modified to enhance inhibitory binding to the FcγRIIb receptor (see, e.g., Chu et al., Mol. Immunol. 45:3926-3933 (2008)). An example of an immunoglobulin fragment engineering modification that enhances inhibitory binding to the FcγRIIb receptor is IgG1-S267E, L328F.

[0288] The half-life of IgG is mediated by pH-dependent binding to its neonatal receptor FcRn. In certain embodiments, the adaptor comprises a functional domain comprising the amino acid sequence of an immunoglobulin effector domain or a derivative of an immunoglobulin effector domain that confers the adaptor with the ability to bind to the neonatal receptor FcRn. In certain embodiments, the adapter comprises a functional domain comprising the sequence of an immunoglobulin FcRn-binding domain that has been modified to enhance binding to FcRn (e.g., Petkova et al., Int. Immunol. 18:1759-1769 (2006); Dall'Acqua et al., J. Immunol. 169:5171-5180 (2002); Oganesyan et al., Mol. Immunol. 46:1750-1755 (2009); Dall'Acqua et al., J. Biol. Chem. 281:23514-23524 (2006), Hinton et al., J. Immunol. 176:346-356 (2006); Datta-Mannan et al., Drug Metab. Dispos. 35:86-94 (2007); Datta-Mannan et al., J. Biol. Chem. 282:1709-1717 (2007); WO 06 / 130834; Strohl, Curr. Op. Biotechnol. 20:685-691 (2009); and Yeung et al., J. Immunol. 182:7663-7671 (2009), the contents of each of which are incorporated herein by reference in their entireties.

[0289] In further embodiments, the adapter comprises a functional domain comprising the sequence of an immunoglobulin effector domain that has been modified to have selective affinity for FcRn at pH 6.0 but no affinity at pH 7.4. For example, the adapter functional domain can comprise an antibody fragment or domain containing one or more of the following half-life-enhancing modifications: IgG1-M252Y, S254T, T256E; IgG1-T250Q, M428L; IgG1-H433K, N434Y; IgG1-N434A; and IgG1-T307A, E380A, N434A (residue numbering is that of the EU index of Kabat et al. (Kabat et al., Sequences of proteins of Immunological Interest, 1991 5th ed.), incorporated herein by reference).

[0290] In another embodiment, the adapter is 238, 239, 246, 248, 249, 252, 254, 255, 256, 258, 265, 267, 268, 269, 270, 272, 276, 278, 280, 283, 285, 286, 289, 290, 292, 293, 294, 295, 296, 298, 301, 303, 305, 307, 309, 312, 315, 320, 322, 324, 326, 327, 329, 330, 331, 332, 333, 334, 335, 337, 338, and a functional domain comprising an amino acid sequence corresponding to an immunoglobulin effector domain that has been modified to include at least one substitution in its sequence corresponding to a position in the Fc region (e.g., Fc gamma) selected from 340, 360, 373, 376, 378, 382, 388, 389, 398, 414, 416, 419, 430, 434, 435, 437, 438, and 439 (numbering of residues in the Fc region is according to the EU numbering system of Kabat et al. (Sequences of proteins of Immunological Interest, 1991 5th ed.; incorporated herein by reference). In certain embodiments, the adapter comprises a functional domain comprising the sequence of an immunoglobulin effector domain derivative in which at least one residue corresponding to position 434 is a residue selected from A, W, Y, F, and H. In another embodiment, the adapter comprises the sequence of an immunoglobulin effector fragment derivative having the following respective substitutions: S298A / E333A / K334A. In an additional embodiment, the adapter comprises an immunoglobulin effector domain derivative having a substitution corresponding to K322A. In another embodiment, the adapter comprises the sequence of an immunoglobulin effector fragment derivative having one or any combination of the following substitutions: K246H, H268D, E283L, S324G, S239D, and I332E. In yet another embodiment, the adapter comprises the sequence of an immunoglobulin effector domain derivative having substitutions corresponding to D265A / N297A.

[0291] In certain embodiments, the adapter comprises a functional domain comprising the sequence of an immunoglobulin effector domain that has been glycoengineered or mutated to enhance effector function using techniques known in the art. For example, inactivation (by point mutation or other means) of the constant region domain sequence contained in the adapter may reduce Fc receptor binding of the circulating adapter, thereby enhancing tumor localization. In other cases, constant region modifications consistent with certain embodiments of the present invention may slow complement binding, thereby shortening serum half-life and reducing nonspecific binding of conjugated cytotoxins. Still other modifications of the constant region may be used to modify disulfide bonds or oligosaccharide moieties that allow for enhanced localization by increasing antigen specificity or antibody flexibility. The resulting physiological profiles, such as tumor localization, biodistribution, and serum half-life, as well as other effects of biochemical modifications, can be readily measured and quantified using known immunological techniques without undue experimentation.

[0292] Adapters as chemical complexes Adapters that facilitate specific binding to target sites can be chemically conjugated to a variety of compounds, such as fluorescent dyes, radioisotopes, chromatographic compositions (e.g., beads, resins, gels, etc.), and chemotherapeutic agents. Adapter conjugates have uses including, but not limited to, purification, diagnostic, analytical, manufacturing, and therapeutic applications.

[0293] The lack of a unique cysteine in the adapter sequence provides an opportunity for the introduction of a unique cysteine for site-specific conjugation purposes.

[0294] In some embodiments, the adapter comprises at least one reactive residue. The reactive residue is useful, for example, as a site for attachment of a conjugate such as a chemotherapeutic agent. The reactive residue can be, for example, a cysteine, a lysine, or another reactive residue. Thus, a cysteine can be added to the N- or C-terminus of the adapter or within the adapter sequence. A cysteine can replace another amino acid in the adapter sequence. Additionally, a lysine can be added to either end of the adapter or within the adapter sequence, and / or a lysine can replace another amino acid in the adapter sequence. In one embodiment, the reactive residue (e.g., cysteine, lysine, etc.) is located in the loop sequence of the ADBD (e.g., Z1 and Z2 in SEQ ID NOs: 22-25 or 26). In one embodiment, the reactive residue is located in a linker located between components of the adapter, e.g., between the ADBD and other components of the adapter fusion protein. The reactive residue (e.g., cysteine, lysine, etc.) can also be located within the adapter sequence. In one embodiment, the adapter comprises at least one, at least two, or at least three reactive residues. In one embodiment, the adaptor comprises at least one, at least two, or at least three cysteine residues.

[0295] Vd. Adapter fabrication The production of adapters useful in practicing the provided methods can be carried out using a variety of standard techniques for chemical synthesis, semi-synthetic methods, and recombinant DNA methods known in the art. Methods for producing adapters, either individually or as part of multidomain fusion proteins, as soluble substances, and as cell-binding proteins, are also provided.

[0296] In some embodiments, the overall adapter generation scheme involves obtaining a reference protein scaffold and identifying multiple residues within the scaffold for modification. Depending on the embodiment, the reference scaffold may include one or more alpha-helical regions or other protein structures with tertiary structure. Once identified, the multiple residues can be modified, for example, by amino acid substitution. In some embodiments, the substitutions are conservative, while in other embodiments, non-conservative substitutions are made. In some embodiments, a naturally occurring amino acid (e.g., one of alanine, arginine, asparagine, aspartic acid, cysteine, glutamine, glutamic acid, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine, or valine) is substituted at the targeted position in the reference scaffold for modification. In certain embodiments, the modification does not include substitution of a cysteine or proline. In certain embodiments, after modifications are made at all desired identified positions, the resulting modified polypeptides (e.g., candidate adapters) can be recombinantly expressed, for example, in plasmids, bacteria, phage, or other vectors (e.g., to expand the number of each modified polypeptide). The modified polypeptides can then be purified and screened to identify those modified polypeptides that specifically bind to a particular target. In some embodiments, certain modified polypeptides exhibit increased binding specificity for a target compared to a reference scaffold, which in some embodiments may exhibit little or no binding to a given target. In further embodiments, depending on the target, the reference scaffold may exhibit some interaction (e.g., non-specific interaction) with the target, but certain modified polypeptides exhibit increased binding specificity for the target by at least about 2-fold, at least about 5-fold, at least about 10-fold, at least about 20-fold, at least about 50-fold, or at least about 100-fold (or more). Optionally, the reference sequence and / or modified polypeptide (e.g., adapter) can be deimmunized. For example, potentially immunogenic residues or motifs can be identified and modified to reduce or eliminate the likelihood of an immune response to the adapter.Additional details regarding various embodiments of adapter production, selection, and isolation are provided in more detail below.

[0297] Recombinant expression of Ve. adaptor In some embodiments, the adapters are "recombinantly produced" (i.e., produced using recombinant DNA technology). Exemplary recombinant methods that can be used to synthesize adapter fusion proteins include, but are not limited to, polymerase chain reaction (PCR)-based synthesis, concatenation, seamless cloning, and recursive directional ligation (RDL) (see, e.g., Meyer et al., Biomacromolecules 3:357-367 (2002), Kurihara et al., Biotechnol. Lett. 27:665-670 (2005), Haider et al., Mol. Pharm. 2:139-150 (2005); and McMillan et al., 32:3643-3646 (1999), the contents of each of which are incorporated herein by reference in their entirety).

[0298] Also provided is a nucleic acid comprising a polynucleotide sequence encoding an adaptor. Such a polynucleotide may optionally further comprise one or more expression control sequences. For example, the polynucleotide may comprise one or more promoters or transcription enhancers, ribosome binding sites, transcription termination signals, and polyadenylation signals as expression control sequences. The polynucleotide may be inserted into any suitable vector, which may be contained in any suitable host cell for expression.

[0299] Expression of a nucleic acid encoding an adapter is typically achieved by operably linking the nucleic acid encoding the adapter to a promoter in an expression vector. Typical expression vectors include transcription and translation terminators, initiation sequences, and promoters useful for regulating expression of the nucleic acid sequence of interest. Expression vectors containing a nucleic acid sequence encoding the adapter along with appropriate transcriptional / translational control signals can be routinely constructed using methods known in the art. These methods include, but are not limited to, in vitro recombinant DNA techniques, synthetic techniques, and in vivo recombination / genetic recombination. Expression of the polynucleotide can be carried out in any suitable expression host known in the art, including, but not limited to, bacterial cells, yeast cells, insect cells, plant cells, or mammalian cells. In one embodiment, the nucleic acid sequence encoding the adapter is operably linked to a suitable promoter sequence such that the nucleic acid sequence is transcribed and / or translated into the adapter in the host. Promoters useful for expression in E. coli include, but are not limited to, the T7 promoter.

[0300] In one embodiment, a vector containing an adapter-encoding nucleic acid is introduced into a host cell (e.g., a phagemid) for expression of the adapter. The vector can remain episomal or be chromosomally integrated, as long as the insert encoding the therapeutic agent can be transcribed. Vectors can be constructed using standard recombinant DNA techniques. Vectors can be plasmids, phages, cosmids, phagemids, viruses, or any other type known in the art that can be used for replication and expression in prokaryotic or eukaryotic cells. Those skilled in the art will understand that such vectors can contain a wide variety of elements (e.g., expression control sequences) known in the art, including a wide variety of transcription signals, e.g., promoters and other sequences that regulate the binding of RNA polymerase to the promoter. Any promoter known or demonstrated to be effective in the cells in which the vector will be expressed could be used to initiate expression of the adapter. Suitable promoters can be inducible (e.g., regulated) or constitutive. Non-limiting examples of suitable promoters include the SV40 early promoter region, promoters contained in the 3' long terminal repeat of Rous sarcoma virus, the HSV-1 (herpes simplex virus-1) thymidine kinase promoter, regulatory sequences of metallothionein genes, and the like, as well as the following animal transcriptional control regions, which exhibit tissue specificity and have been utilized in transgenic animals: the elastase I gene control region, which is active in pancreatic acinar cells; the insulin gene control region, which is active in beta cells of the pancreas; the mouse mammary tumor virus control region, which is active in testicular, breast, lymphoid, and mast cells; the albumin gene control region, which is active in the liver; the alpha fetoprotein gene control region, which is active in the liver; the alpha 1-antitrypsin gene control region, which is active in the liver; the beta globin gene control region, which is active in erythroid cells; the myelin basic protein control region, which is active in oligodendrocyte cells of the brain; the myosin light chain-2 gene control region, which is active in skeletal muscle; and the gonadotropin-releasing hormone gene control region, which is active in the hypothalamus. In a particular embodiment, the promoter is an immunoglobulin gene regulatory region that is active in lymphoid cells.

[0301] In one embodiment, one or more nucleic acids encoding the adapters are expressed under the control of a constitutive promoter or a regulated expression system. Suitable regulated expression systems include, but are not limited to, a tetracycline-regulated expression system, an ecdysone-inducible expression system, a rack switch expression system, a glucocorticoid-inducible expression system, a temperature-inducible promoter system, and a metallothionein metal-inducible expression system. When several different nucleic acids encoding adapters are contained within a host cell system, some nucleic acids may be expressed under the control of a constitutive promoter, while others may be expressed under the control of a regulated promoter. Expression levels may be determined by methods known to those skilled in the art, including Western blot analysis and Northern blot analysis.

[0302] A variety of host-expression vector systems can be utilized to express the nucleic a...

Claims

1. 1. A composition suitable for use in therapy comprising cells expressing a chimeric antigen receptor (CAR) and an adaptor, comprising: (a) the CAR comprises (i) a D domain that binds to a first antigenic determinant (AD), (ii) a transmembrane domain, and (iii) an intracellular domain, and the adaptor comprises (i) the first AD and (ii) an adaptor comprising an ADBD that binds to a second AD on a target cell, wherein the first AD is AFP p26, AFP, CD123, BCMA, or CS1, and the second AD is CS1, BCMA, CD19, CD22, CD45, CD123, HER2, TACI, BAFFR, or PDL1; (b) the CAR comprises (i) an antigenic determinant binding domain (ADBD) that binds to a first antigenic determinant (AD) on a target cell, (ii) a transmembrane domain, and (iii) an intracellular domain, and the adapter comprises an ADBD that binds to (i) the first AD and (ii) a second AD on the target cell; (c) the CAR comprises (i) an antigenic determinant binding domain (ADBD) that binds to a first antigenic determinant (AD) on a target cell, (ii) a transmembrane domain, and (iii) an intracellular domain, and the adapter comprises (i) the first AD and (ii) an ADBD that binds to a second AD on a second target cell; (d) the CAR comprises (i) an antigenic determinant binding domain (ADBD) that is an surrogate scaffold binding domain (ASBD) that binds to a first antigenic determinant (AD), (ii) a transmembrane domain, and (iii) an intracellular domain, and the adapter comprises an ADBD that binds to (i) the first AD and (ii) a second AD on a target cell; (e) the CAR comprises (i) an antigenic determinant binding domain (ADBD) that binds to a first antigenic determinant (AD), (ii) a transmembrane domain, and (iii) an intracellular domain, and the adapter comprises an ADBD that comprises (i) the first AD and (ii) an ASBD that binds to a second AD on a target cell; (f) the CAR comprises (i) an antigenic determinant binding domain (ADBD) that binds to a first antigenic determinant (AD), (ii) a transmembrane domain, and (iii) an intracellular domain, and the adapter comprises (i) the first AD and (ii) a D domain that binds to a second AD on a target cell; (g) the CAR comprises (i) a D domain that binds to a first antigenic determinant (AD), (ii) a transmembrane domain, and (iii) an intracellular domain, and the adapter comprises (i) the first AD and (ii) an ADBD that binds to a second AD on a target cell; (h) The composition, wherein the CAR comprises (i) a first D domain that binds to a first antigenic determinant (AD), (ii) a transmembrane domain, and (iii) an intracellular domain, and the adapter comprises (i) the first AD and (ii) a second D domain that binds to a second AD on a target cell.

2. 2. The composition of claim 1, wherein the CAR is characterized by one or more of the following: (a) the CAR comprises a single chain variable fragment (scFv) ADBD; (b) the CAR comprises an alternative scaffold binding domain (ASBD) ADBD; (c) the CAR comprises a D domain, and optionally the CAR comprises a sequence selected from the group: SEQ ID NOs: 17-26, and 27 or SEQ ID NOs: 44-1078, and 1079; (d) the CAR comprises two ADBDs; (e) the CAR comprises an ASBD and an scFv; (f) the CAR comprises a D domain and an scFv; (g) the CAR comprises two ASBDs; (h) the CAR comprises two D domains; (i) the intracellular domain of the CAR is a signaling domain; (j) the intracellular domain of the CAR comprises a primary signaling domain; (k) the intracellular domain of the CAR comprises a CD3ζ primary signaling domain; (l) the intracellular domain of the CAR further comprises a costimulatory signaling domain; (m) the intracellular domain of the CAR comprises a costimulatory signaling domain selected from CD28, 41BB, CD27, and CD134; (n) the intracellular domain of the CAR comprises a 41BB costimulatory signaling domain; (o) the CAR binds to an antigen selected from CD19, CD22, CD123, BCMA, CS1, HER2, TACI, BAFFR, and PDL1; (p) the CAR binds to BCMA, and optionally the CAR comprises a sequence selected from the group: SEQ ID NOs: 44-338, and 339; (q) the CAR binds to CD123, and optionally the CAR comprises a sequence selected from the group: SEQ ID NOs: 340-77 and 773; (r) the CAR binds to CD19, and optionally the CAR comprises a sequence selected from the group: SEQ ID NOs: 1030-1058 and 1059; (s) the CAR binds to CD22, and optionally the CAR comprises a sequence selected from the group: SEQ ID NOs: 1060-1068 and 1069; (t) the CAR binds to CS1, and optionally the CAR comprises a sequence selected from the group: SEQ ID NOs: 780-794 and 795; (u) the CAR binds to HER2, and optionally the CAR comprises a sequence selected from the group: SEQ ID NOs: 800-839 and 840; (v) the CAR binds to TACI or BAFFR; (w) the CAR binds to PDL1, and optionally the CAR comprises a sequence selected from the group: SEQ ID NOs: 1010-1016, 1074-1078 and 1079; (x) the CAR binds to AFP p26, and optionally the CAR comprises a sequence selected from the group: SEQ ID NOs: 841-983 and 984; (y) the CAR comprises two ADBDs that bind to separate targets; (z) the CAR binds to CD19 and CD123; (aa) the CAR binds to BCMA and CS1; (bb) the CAR binds to CD22 and CD123; (cc) the CAR binds to PDL1 and CD123; (dd) the CAR comprises a first ASBD that binds to CD19 and a second ASBD that binds to CD123; (ee) the CAR comprises a first D domain that binds to CD19 and a second D domain that binds to CD123; (ff) the CAR comprises a D domain that binds to CD19 and an scFv that binds to CD123; (gg) the CAR comprises a first ASBD that binds to BCMA and a second ASBD that binds to CS1; (hh) the CAR comprises a first D domain that binds to BCMA and a second D domain that binds to CS1; (ii) the CAR comprises a D domain that binds to CS1 and an scFv that binds to BCMA; (jj) the CAR comprises a D domain that binds to BCMA and an scFv that binds to CS1; (kk) the CAR comprises a first ASBD that binds to CD22 and a second ASBD that binds to CD123; (ll) the CAR comprises a first D domain that binds to CD22 and a second D domain that binds to CD123; (mm) the CAR comprises a D domain that binds to CD22 and an scFv that binds to CD123; (nn) the CAR comprises a first ASBD that binds to PDL1 and a second ASBD that binds to CD123; (oo) the CAR comprises a first D domain that binds to PDL1 and a second D domain that binds to CD123; (pp) the CAR comprises a D domain that binds to PDL1 and an scFv that binds to CD123; and (qq) The CAR comprises an ASBD that binds to CD19 and an scFv that binds to CD123.

3. 3. The composition of claim 1 or claim 2, wherein the adapter is characterized by one or more of the following: (a) the adapter comprises an AD that is an epitope of AFP p26 or AFP, and optionally comprises amino acid residues of SEQ ID NO: 16 or 1117-1123; (b) the adapter comprises an AD of a tumor antigen, optionally wherein said tumor antigen is selected from the group: BCMA, CD123, CD19, CD22, CS1, HER2, TACI, BAFFR, and PDL1; (c) the adapter comprises an ADBD that is an scFv; (d) the adapter comprises an ADBD that is an ASBD; (e) the adapter comprises a D domain, and optionally the adapter comprises a sequence selected from the group: SEQ ID NOs: 17-26, and 27, or SEQ ID NOs: 44-1078, and 1079; (f) the adapter comprises two ADBDs, and optionally the ADBDs (i) are the same, (ii) bind to the same antigenic determinant, (iii) bind to different ADs of the same antigen, (iv) bind to different antigens on the same cell, or (v) bind to different antigens on different cells; (g) the adapter comprises two ASBDs; (h) the adapter comprises two D domains, and optionally the adapter comprises an amino acid sequence selected from the group: SEQ ID NOs: 44-1079; (i) the adapter comprises an ADBD that is an scFv and an ADBD that is an ASBD; (j) the adapter comprises an ADBD that is an scFv and an ADBD that is a D domain; (k) the adapter comprises an ADBD that binds to a member selected from BCMA, CD123, CD19, CD22, CS1, HER2, TACI, BAFFR, and PDL1; (l) the adapter comprises an ADBD that binds to BCMA, and optionally the ADBD comprises a sequence selected from the group: SEQ ID NOs: 44-338, and 339; (m) the adapter comprises an ADBD that binds to CS1, and optionally the ADBD comprises a sequence selected from SEQ ID NOs: 780-794, and 795; (n) the adapter comprises an ADBD that binds to HER2, and optionally the ADBD comprises a sequence selected from SEQ ID NOs: 800-839, and 840; (o) the adapter comprises an ADBD that binds to CD123, and optionally the ADBD comprises a sequence selected from SEQ ID NOs: 340-772, and 773; (p) the adapter comprises an ADBD that binds to CD19, and optionally the ADBD comprises a sequence selected from SEQ ID NOs: 1030-1058, and 1059; (q) the adapter comprises an ADBD that binds to CD22, and optionally the ADBD comprises a sequence selected from SEQ ID NOs: 1060-1068, and 1069; (r) the adapter comprises an ADBD that binds to TACI; (s) the adapter comprises an ADBD that binds to BAFFR; (t) the adapter comprises an ADBD that binds to PDL1, and optionally the ADBD comprises a sequence selected from SEQ ID NOs: 1010-1016, 1074-1078, and 1079; (u) the adapter is bispecific; (v) the adaptor comprises an ADBD that binds to CD19 and an ADBD that binds to CD123; (w) the adapter comprises an ADBD that binds to BCMA and an ADBD that binds to CS1; (x) the adaptor comprises an ADBD that binds to CD22 and an ADBD that binds to CD123; and (y) the adaptor comprises an ADBD that binds to PDL1 and an ADBD that binds to CD123.

4. The composition according to any one of claims 1 to 3, (a) the target cell is a tumor cell; (b) the target cells are tumor cells selected from acute leukemia, chronic leukemia, polycythemia vera, lymphoma, Hodgkin's disease, non-Hodgkin's lymphoma, multiple myeloma, Waldenstrom's macroglobulinemia, heavy chain disease, myelodysplastic syndrome, hairy cell leukemia, and myelodysplasia; (c) the target cell is multiple myeloma; (d) at least one target cell is a tumor cell, and optionally said tumor cell is selected from acute leukemia, chronic leukemia, polycythemia vera, lymphoma, Hodgkin's disease, non-Hodgkin's lymphoma, multiple myeloma, Waldenstrom's macroglobulinemia, heavy chain disease, myelodysplastic syndrome, hairy cell leukemia, and myelodysplasia; (e) the first and second target cells are tumor cells, and optionally the tumor cells are selected from acute leukemia, chronic leukemia, polycythemia vera, lymphoma, Hodgkin's disease, non-Hodgkin's lymphoma, multiple myeloma, Waldenstrom's macroglobulinemia, heavy chain disease, myelodysplastic syndrome, hairy cell leukemia, and myelodysplasia; (f) the first and second target cells are tumor cells of the same type, and optionally the tumor cells are selected from acute leukemia, chronic leukemia, polycythemia vera, lymphoma, Hodgkin's disease, non-Hodgkin's lymphoma, multiple myeloma, Waldenstrom's macroglobulinemia, heavy chain disease, myelodysplastic syndrome, hairy cell leukemia, and myelodysplasia; (g) the first and second target cells are different types of tumor cells, and optionally the tumor cells are selected from acute leukemia, chronic leukemia, polycythemia vera, lymphoma, Hodgkin's disease, non-Hodgkin's lymphoma, multiple myeloma, Waldenstrom's macroglobulinemia, heavy chain disease, myelodysplastic syndrome, hairy cell leukemia, and myelodysplasia; (h) the target cells are cancer cells selected from breast cancer, prostate cancer, ovarian cancer, pancreatic cancer, colon cancer, and lung cancer; and / or (i) The composition, wherein the target cell is a breast cancer or ovarian cancer.

5. The composition according to any one of claims 1 to 4, (a) the CAR-expressing cell is an immune effector cell; (b) the CAR-expressing cell is a T cell, or (c) the composition, wherein the CAR-expressing cell is a NK cell.

6. A method of killing a target cell comprising contacting said target cell with the composition of any one of claims 1 to 5.

7. 1. A method for killing a target cell, for transmitting an immune response to a target cell, for treating a proliferative disorder, cancer, an autoimmune disease, an infectious disease, or allograft rejection, or for treating a hematological cancer, comprising: (a) contacting a composition comprising target cells with an adaptor, (1) The composition comprising the target cell further comprises a cell expressing a CAR, wherein the CAR comprises (i) an antigenic determinant binding domain (ADBD) that binds to a first AD on the target cell, (ii) a transmembrane domain, and (iii) an intracellular domain; (2) the adapter comprises an ADBD that binds to (i) the first AD and (ii) a second AD on the target cell; (b) contacting a composition comprising a first target cell with an adaptor, (1) The composition comprising the target cells further comprises cells expressing a chimeric antigen receptor (CAR), wherein the CAR comprises (i) an antigenic determinant binding domain (ADBD) that binds to a first AD on the first target cell, (ii) a transmembrane domain, and (iii) an intracellular domain; and (2) the adapter comprises an ADBD that binds to (i) the first AD and (ii) a second AD on a second target cell; (c) contacting a composition comprising target cells with an adaptor, (1) the composition comprising the target cells further comprises cells expressing a chimeric antigen receptor (CAR), the CAR comprising (i) an antigenic determinant binding domain (ADBD) that is an ASBD that binds to a first AD, (ii) a transmembrane domain, and (iii) an intracellular domain; and (2) the adapter comprises an ADBD that binds to (i) the first AD and (ii) a second AD on the target cell; (d) contacting a composition comprising target cells with an adaptor, (1) The composition comprising the target cells further comprises cells expressing a chimeric antigen receptor (CAR), wherein the CAR comprises (i) an antigenic determinant binding domain (ADBD) that binds to a first AD, (ii) a transmembrane domain, and (iii) an intracellular domain; and (2) the adapter comprises an ADBD that is an ASBD that binds to (i) the first AD and (ii) a second AD on the target cell; (e) contacting the composition comprising the target cells with cells expressing a chimeric antigen receptor (CAR), (1) a first antigenic determinant (AD) is present on the target cell; (2) the composition comprising the first target cell further comprises an adaptor comprising an antigenic determinant binding domain (ADBD) that binds to (i) the first AD and (ii) a second AD on the target cell; and (3) the CAR comprises (i) an ADBD that binds to the first AD on the target cell or the adaptor, (ii) a transmembrane domain, and (iii) an intracellular domain; (f) contacting the composition comprising the target cells with cells expressing a chimeric antigen receptor (CAR), (1) a first AD is present on a first target cell; (2) the composition comprising the first target cell further comprises a second target cell and an adaptor comprising an antigenic determinant binding domain (ADBD) that binds to (i) the first AD and (ii) a second AD on the second target cell; and (3) the CAR comprises (i) an ADBD that binds to the first AD on the first target cell or the adaptor, (ii) a transmembrane domain, and (iii) an intracellular domain; (g) contacting the composition comprising the target cells with cells expressing a chimeric antigen receptor (CAR), (1) a first antigenic determinant (AD) is present on the target cell; (2) the composition comprising the target cell further comprises an adaptor comprising (i) an antigenic determinant binding domain (ADBD) that binds to the first AD on the target cell and (ii) a second AD; and (3) the CAR comprises (i) an ADBD that is an ASBD that binds to the second AD, (ii) a transmembrane domain, and (iii) an intracellular domain; (h) contacting the composition comprising the target cells with cells expressing a chimeric antigen receptor (CAR), (1) a first antigenic determinant (AD) is present on the target cell; (2) the composition comprising the target cell further comprises an adaptor comprising (i) an antigenic determinant binding domain (ADBD) that is an ASBD that binds to the first AD on the target cell and (ii) a second AD; and (3) The method, wherein the CAR comprises (i) an ADBD that binds to the second AD, (ii) a transmembrane domain, and (iii) an intracellular domain.

8. Methods for killing target cells, methods for redirecting the killing of target cells in a patient, methods for conveying an immune response to target cells in a patient, methods for inducing an immune response to target cells in a patient, methods for treating a proliferative disorder, cancer, autoimmune disease, infectious disease, or allograft rejection in a patient, methods for redirecting treatment of a proliferative disorder, cancer, autoimmune disease, infectious disease, or allograft rejection in a patient, methods for inducing treatment of a proliferative disorder, cancer, autoimmune disease, infectious disease, or allograft rejection in a patient, methods for treating a hematological cancer in a patient, methods for redirecting treatment of a hematological cancer in a patient, or methods for inducing treatment of a hematological cancer in a patient, (a) administering to the patient an adapter, (1) the patient is treated with cells expressing a chimeric antigen receptor (CAR), the CAR comprising: (i) an antigenic determinant binding domain (ADBD) that binds to a first antigenic determinant (AD) on the target cell; (ii) a transmembrane domain; and (iii) an intracellular domain; and (2) the adapter comprises an ADBD that binds to (i) the first AD and (ii) a second AD on the target cell; (b) administering to the patient an adapter, (1) the patient is treated with cells expressing a chimeric antigen receptor (CAR), the CAR comprising: (i) an antigenic determinant binding domain (ADBD) that binds to a first antigenic determinant (AD) on a first target cell; (ii) a transmembrane domain; and (iii) an intracellular domain; and (2) the adapter comprises an ADBD that binds to (i) the first AD and (ii) a second AD on a second target cell; (c) administering to the patient an adapter, (1) the patient is treated with cells expressing a chimeric antigen receptor (CAR), the CAR comprising: (i) an antigenic determinant binding domain (ADBD), which is an ASBD that binds to a first antigenic determinant (AD); (ii) a transmembrane domain; and (iii) an intracellular domain; and (2) the adapter comprises an ADBD that binds to (i) the first AD and (ii) a second AD on the target cell; (d) administering to the patient an adapter, (1) the patient is treated with cells expressing a chimeric antigen receptor (CAR), the CAR comprising: (i) an antigenic determinant binding domain (ADBD) that binds to a first antigenic determinant (AD); (ii) a transmembrane domain; and (iii) an intracellular domain; and (2) the adapter comprises an ADBD that is an ASBD that binds to (i) the first AD and (ii) a second AD on the target cell; (e) administering to the patient cells expressing a chimeric antigen receptor (CAR), (1) a first antigenic determinant (AD) and a second AD are present on the target cell; (2) the patient has been treated with an adapter comprising an antigenic determinant binding domain (ADBD) that binds to (i) the first AD and (ii) the second AD on the target cell; and (3) the CAR comprises (i) an ADBD that binds to the first AD on the target cell or the adaptor, (ii) a transmembrane domain, and (iii) an intracellular domain; (f) administering to the patient cells expressing a chimeric antigen receptor (CAR), (1) the patient has been treated with an adaptor comprising (i) a first antigenic determinant (AD) and (ii) an antigenic determinant binding domain (ADBD) that binds to a second AD on the target cell; and (2) the CAR comprises (i) an ADBD that binds to the first AD on the adapter, (ii) a transmembrane domain, and (iii) an intracellular domain; (g) administering to the patient cells expressing a chimeric antigen receptor (CAR), (1) a first antigenic determinant (AD) is present on the target cell; (2) the patient has been treated with an adapter comprising (i) an antigenic determinant binding domain (ADBD) that binds to the first AD on the target cell and (ii) a second AD; and (3) the CAR comprises (i) an ADBD that is an ASBD that binds to the second AD, (ii) a transmembrane domain, and (iii) an intracellular domain; (h) administering to the patient cells expressing a chimeric antigen receptor (CAR), (1) a first antigenic determinant (AD) is present on the target cell; (2) the patient has been treated with an adaptor comprising (i) an antigenic determinant binding domain (ADBD) that is an ASBD that binds to the first AD on the target cell and (ii) a second AD; and (3) the CAR comprises (i) an ADBD that binds to the second AD, (ii) a transmembrane domain, and (iii) an intracellular domain; (i) administering to the patient an adapter, (1) the patient is treated with cells expressing a chimeric antigen receptor (CAR), the CAR comprising: (i) an antigenic determinant binding domain (ADBD) that binds to a first antigenic determinant (AD) on the target cell; (ii) a transmembrane domain; and (iii) an intracellular domain; and (2) the adapter comprises an ADBD that binds to (i) the first AD and (ii) a second AD on the target cell; (j) administering to the patient an adapter, (1) the patient is treated with cells expressing a chimeric antigen receptor (CAR), the CAR comprising: (i) an antigenic determinant binding domain (ADBD) that binds to a first antigenic determinant (AD) on a first target cell; (ii) a transmembrane domain; and (iii) an intracellular domain; and (2) the adapter comprises an ADBD that binds to (i) the first AD and (ii) a second AD on a second target cell; (k) administering to the patient an adapter, (1) the patient is treated with cells expressing a chimeric antigen receptor (CAR), the CAR comprising: (i) an antigenic determinant binding domain (ADBD), which is an ASBD that binds to a first antigenic determinant (AD); (ii) a transmembrane domain; and (iii) an intracellular domain; and (2) the adapter comprises an ADBD that binds to (i) the first AD and (ii) a second AD on the target cell; (l) administering to the patient an adapter, (1) the patient is treated with cells expressing a chimeric antigen receptor (CAR), the CAR comprising: (i) an antigenic determinant binding domain (ADBD) that binds to a first antigenic determinant (AD); (ii) a transmembrane domain; and (iii) an intracellular domain; and (2) the adapter comprises an ADBD that is an ASBD that binds to (i) the first AD and (ii) a second AD on the target cell; (m) administering to the patient cells expressing a chimeric antigen receptor (CAR), (1) a first antigenic determinant (AD) and a second AD are present on the target cell; (2) the patient has been treated with an adapter comprising an antigenic determinant binding domain (ADBD) that binds to (i) the first AD and (ii) the second AD on the target cell; and (3) the CAR comprises (i) an ADBD that binds to the first AD on the target cell or the adaptor, (ii) a transmembrane domain, and (iii) an intracellular domain; (n) administering to the patient cells expressing a chimeric antigen receptor (CAR), (1) the patient has been treated with an adaptor comprising (i) a first antigenic determinant (AD) and (ii) an antigenic determinant binding domain (ADBD) that binds to a second AD on the target cell; and (2) the CAR comprises (i) an ADBD that binds to the first AD on the adapter, (ii) a transmembrane domain, and (iii) an intracellular domain; (o) administering to said patient cells expressing a chimeric antigen receptor (CAR), (1) a first antigenic determinant (AD) is present on the target cell; (2) the patient has been treated with an adapter comprising (i) an antigenic determinant binding domain (ADBD) that binds to the first AD on the target cell and (ii) a second AD; and (3) the CAR comprises (i) an ADBD that is an ASBD that binds to the second AD, (ii) a transmembrane domain, and (iii) an intracellular domain; (p) administering to the patient cells expressing a chimeric antigen receptor (CAR), (1) a first antigenic determinant (AD) is present on the target cell; (2) the patient has been treated with an adaptor comprising (i) an antigenic determinant binding domain (ADBD) that is an ASBD that binds to the first AD on the target cell and (ii) a second AD; and (3) The method, wherein the CAR comprises (i) an ADBD that binds to the second AD, (ii) a transmembrane domain, and (iii) an intracellular domain.

9. 9. The method of claim 7 or 8, wherein the proliferative disorder, cancer or hematological cancer is selected from acute leukemia, chronic leukemia, polycythemia vera, lymphoma, Hodgkin's disease, non-Hodgkin's lymphoma, multiple myeloma, Waldenstrom's macroglobulinemia, heavy chain disease, myelodysplastic syndrome, hairy cell leukemia, myelodysplasia, breast cancer, prostate cancer, ovarian cancer, pancreatic cancer, colon cancer, and lung cancer.

10. 10. The method of any one of claims 7 to 9, wherein the CAR is characterized by one or more of the following: (a) the CAR comprises a single chain variable fragment (scFv) ADBD; (b) the CAR comprises an alternative scaffold binding domain (ASBD) ADBD; (c) the CAR comprises a D domain, and optionally the CAR comprises a sequence selected from the group: SEQ ID NOs: 17-26, and 27 or SEQ ID NOs: 44-1078, and 1079; (d) the CAR comprises two ADBDs; (e) the CAR comprises an ASBD and an scFv; (f) the CAR comprises a D domain and an scFv; (g) the CAR comprises two ASBDs; (h) the CAR comprises two D domains; (i) the intracellular domain of the CAR is a signaling domain; (j) the intracellular domain of the CAR comprises a primary signaling domain; (k) the intracellular domain of the CAR comprises a CD3ζ primary signaling domain; (l) the intracellular domain of the CAR further comprises a costimulatory signaling domain; (m) the intracellular domain of the CAR comprises a costimulatory signaling domain selected from CD28, 41BB, CD27, and CD134; (n) the intracellular domain of the CAR comprises a 41BB costimulatory signaling domain; (o) the CAR binds to an antigen selected from CD19, CD22, CD123, BCMA, CS1, HER2, TACI, BAFFR, and PDL1; (p) the CAR binds to BCMA, and optionally the CAR comprises a sequence selected from the group: SEQ ID NOs: 44-338, and 339; (q) the CAR binds to CD123, and optionally the CAR comprises a sequence selected from the group: SEQ ID NOs: 340-77 and 773; (r) the CAR binds to CD19, and optionally the CAR comprises a sequence selected from the group: SEQ ID NOs: 1030-1058 and 1059; (s) the CAR binds to CD22, and optionally the CAR comprises a sequence selected from the group: SEQ ID NOs: 1060-1068 and 1069; (t) the CAR binds to CS1, and optionally the CAR comprises a sequence selected from the group: SEQ ID NOs: 780-794 and 795; (u) the CAR binds to HER2, and optionally the CAR comprises a sequence selected from the group: SEQ ID NOs: 800-839 and 840; (v) the CAR binds to TACI or BAFFR; (w) the CAR binds to PDL1, and optionally the CAR comprises a sequence selected from the group: SEQ ID NOs: 1010-1016, 1074-1078 and 1079; (x) the CAR binds to AFP p26, and optionally the CAR comprises a sequence selected from the group: SEQ ID NOs: 841-983 and 984; (y) the CAR comprises two ADBDs that bind to separate targets; (z) the CAR binds to CD19 and CD123; (aa) the CAR binds to BCMA and CS1; (bb) the CAR binds to CD22 and CD123; (cc) the CAR binds to PDL1 and CD123; (dd) the CAR comprises a first ASBD that binds to CD19 and a second ASBD that binds to CD123; (ee) the CAR comprises a first D domain that binds to CD19 and a second D domain that binds to CD123; (ff) the CAR comprises a D domain that binds to CD19 and an scFv that binds to CD123; (gg) the CAR comprises a first ASBD that binds to BCMA and a second ASBD that binds to CS1; (hh) the CAR comprises a first D domain that binds to BCMA and a second D domain that binds to CS1; (ii) the CAR comprises a D domain that binds to CS1 and an scFv that binds to BCMA; (jj) the CAR comprises a D domain that binds to BCMA and an scFv that binds to CS1; (kk) the CAR comprises a first ASBD that binds to CD22 and a second ASBD that binds to CD123; (ll) the CAR comprises a first D domain that binds to CD22 and a second D domain that binds to CD123; (mm) the CAR comprises a D domain that binds to CD22 and an scFv that binds to CD123; (nn) the CAR comprises a first ASBD that binds to PDL1 and a second ASBD that binds to CD123; (oo) the CAR comprises a first D domain that binds to PDL1 and a second D domain that binds to CD123; (pp) the CAR comprises a D domain that binds to PDL1 and an scFv that binds to CD123; and (qq) The CAR comprises an ASBD that binds to CD19 and an scFv that binds to CD123.

11. 11. The method according to any one of claims 7 to 10, wherein the adapter is characterized by one or more of the following: (a) the adapter comprises an AD that is an epitope of AFP p26 or AFP, and optionally comprises amino acid residues of SEQ ID NO: 16 or 1117-1123; (b) the adapter comprises an AD of a tumor antigen, optionally wherein said tumor antigen is selected from the group: BCMA, CD123, CD19, CD22, CS1, HER2, TACI, BAFFR, and PDL1; (c) the adapter comprises an ADBD that is an scFv; (d) the adapter comprises an ADBD that is an ASBD; (e) the adapter comprises a D domain, and optionally the adapter comprises a sequence selected from the group: SEQ ID NOs: 17-26, and 27, or SEQ ID NOs: 44-1078, and 1079; (f) the adapter comprises two ADBDs, and optionally the ADBDs (i) are the same, (ii) bind to the same antigenic determinant, (iii) bind to different ADs of the same antigen, (iv) bind to different antigens on the same cell, or (v) bind to different antigens on different cells; (g) the adapter comprises two ASBDs; (h) the adapter comprises two D domains, and optionally the adapter comprises an amino acid sequence selected from the group: SEQ ID NOs: 44-1079; (i) the adapter comprises an ADBD that is an scFv and an ADBD that is an ASBD; (j) the adapter comprises an ADBD that is an scFv and an ADBD that is a D domain; (k) the adaptor comprises an ADBD that binds to a member selected from BCMA, CD123, CD19, CD22, CS1, TACI, BAFFR, and PDL1; (l) the adapter comprises an ADBD that binds to BCMA, and optionally the ADBD comprises a sequence selected from the group: SEQ ID NOs: 44-338, and 339; (m) the adapter comprises an ADBD that binds to CS1, and optionally the ADBD comprises a sequence selected from SEQ ID NOs: 780-794, and 795; (n) the adapter comprises an ADBD that binds to HER2, and optionally the ADBD comprises a sequence selected from SEQ ID NOs: 800-839, and 840; (o) the adapter comprises an ADBD that binds to CD123, and optionally the ADBD comprises a sequence selected from SEQ ID NOs: 340-772, and 773; (p) the adapter comprises an ADBD that binds to CD19, and optionally the ADBD comprises a sequence selected from SEQ ID NOs: 1030-1058, and 1059; (q) the adapter comprises an ADBD that binds to CD22, and optionally the ADBD comprises a sequence selected from SEQ ID NOs: 1060-1068, and 1069; (r) the adapter comprises an ADBD that binds to TACI; (s) the adapter comprises an ADBD that binds to BAFFR; (t) the adapter comprises an ADBD that binds to PDL1, and optionally the ADBD comprises a sequence selected from SEQ ID NOs: 1010-1016, 1074-1078, and 1079; (u) the adapter is bispecific; (v) the adaptor comprises an ADBD that binds to CD19 and an ADBD that binds to CD123; (w) the adapter comprises an ADBD that binds to BCMA and an ADBD that binds to CS1; (x) the adaptor comprises an ADBD that binds to CD22 and an ADBD that binds to CD123; and (y) the adaptor comprises an ADBD that binds to PDL1 and an ADBD that binds to CD123.

12. The method according to any one of claims 7 to 11, (a) the target cell is a tumor cell; (b) the target cells are tumor cells selected from acute leukemia, chronic leukemia, polycythemia vera, lymphoma, Hodgkin's disease, non-Hodgkin's lymphoma, multiple myeloma, Waldenstrom's macroglobulinemia, heavy chain disease, myelodysplastic syndrome, hairy cell leukemia, and myelodysplasia; (c) the target cell is multiple myeloma; (d) at least one target cell is a tumor cell, and optionally said tumor cell is selected from acute leukemia, chronic leukemia, polycythemia vera, lymphoma, Hodgkin's disease, non-Hodgkin's lymphoma, multiple myeloma, Waldenstrom's macroglobulinemia, heavy chain disease, myelodysplastic syndrome, hairy cell leukemia, and myelodysplasia; (e) the first and second target cells are tumor cells, and optionally the tumor cells are selected from acute leukemia, chronic leukemia, polycythemia vera, lymphoma, Hodgkin's disease, non-Hodgkin's lymphoma, multiple myeloma, Waldenstrom's macroglobulinemia, heavy chain disease, myelodysplastic syndrome, hairy cell leukemia, and myelodysplasia; (f) the first and second target cells are tumor cells of the same type, and optionally the tumor cells are selected from acute leukemia, chronic leukemia, polycythemia vera, lymphoma, Hodgkin's disease, non-Hodgkin's lymphoma, multiple myeloma, Waldenstrom's macroglobulinemia, heavy chain disease, myelodysplastic syndrome, hairy cell leukemia, and myelodysplasia; (g) the first and second target cells are different types of tumor cells, and optionally the tumor cells are selected from acute leukemia, chronic leukemia, polycythemia vera, lymphoma, Hodgkin's disease, non-Hodgkin's lymphoma, multiple myeloma, Waldenstrom's macroglobulinemia, heavy chain disease, myelodysplastic syndrome, hairy cell leukemia, and myelodysplasia; (j) the target cells are cancer cells selected from breast cancer, prostate cancer, ovarian cancer, pancreatic cancer, colon cancer, and lung cancer; and / or (h) The method, wherein the target cell is a breast cancer or ovarian cancer.

13. The method according to any one of claims 7 to 12, (a) the CAR-expressing cell is an immune effector cell; (b) the CAR-expressing cell is a T cell, or (c) the CAR-expressing cell is a NK cell.

14. 1. A genetically modified human immune effector cell, comprising: a. a chimeric antigen receptor (CAR) comprising: (1) an antigenic determinant binding domain (ADBD) that specifically binds to a human CD45 antigenic determinant (AD), (2) a transmembrane domain, and (3) an intracellular domain; and b. A genetic modification that abolishes expression of human CD45 AD on the genetically modified cells; A genetically modified human immune effector cell, wherein said genetically modified cell is capable of inducing an immune response against cells expressing CD45 AD in an in vitro assay, and wherein said genetically modified cell does not express said CD45 AD.

15. A method for killing a target cell or transmitting an immune response to a target cell, comprising contacting the genetically modified cell of claim 14 with the target cell, wherein the target cell expresses human CD45.

16. A method for inducing an immune response in target cells, treating a proliferative disorder, cancer, autoimmune disease, infectious disease, or allograft rejection in a subject, depleting lymphocytes in a subject, depleting memory T cells in a subject, treating an autoimmune disease or disorder in a subject, or treating a hematological cancer in a subject, or pretreating a subject for transplantation, comprising administering an effective amount of the genetically modified cells of claim 14 to the subject in need thereof.

17. 1. A genetically modified human immune effector cell, comprising: a. a chimeric antigen receptor (CAR) comprising: (1) a first antigenic determinant binding domain (ADBD) that specifically binds to a first antigenic determinant (AD), (2) a transmembrane domain, and (3) an intracellular domain; and b. A genetic modification that abolishes expression of at least one human CD45 AD on the genetically modified cells; A genetically modified human immune effector cell, wherein the first AD is not at least one human CD45 AD, and wherein the genetically modified cell used in combination with the adapter in an in vitro assay is capable of inducing an immune response against CD45 AD-expressing cells, and wherein the adapter comprises a second ADBD that specifically binds to the first AD and at least one human CD45 AD, and wherein the genetically modified cell does not express at least one human CD45 AD.

18. 20. A method of killing a target cell or transmitting an immune response to a target cell, comprising contacting the genetically modified cell of claim 17 with an adaptor and the target cell, wherein the target cell expresses CD45, the adaptor comprises the first AD and a second ADBD that specifically binds to a human CD45 AD, and optionally the target cell is a hematological cancer cell.

19. 1. A method for inducing an immune response in target cells, a method for treating a proliferative disorder, cancer, an autoimmune disease, an infectious disease, or allograft rejection in a subject, a method for lymphodepleting a subject, a method for depleting memory T cells in a subject, a method for treating an autoimmune disease or disorder in a subject, or a method for treating a hematological cancer in a subject, or a method for pre-treating a subject for transplantation, comprising: a. administering to the subject in need thereof a therapeutically effective amount of the genetically modified cell of claim 17 and a therapeutically effective amount of an adaptor comprising the first AD and a second ADBD that specifically binds to a human CD45 AD; b. Administering a therapeutically effective amount of the genetically modified cell of claim 234 to a subject in need thereof, wherein the subject has been administered an adaptor comprising the first AD and a second ADBD that specifically binds to a human CD45 AD; or c) administering to a subject in need thereof a therapeutically effective amount of an adapter comprising the first AD and a second ADBD that specifically binds to a human CD45 AD, wherein the subject has been administered the genetically modified cells of claim 17.

20. 1. A genetically modified human immune effector cell, comprising: a. a chimeric antigen receptor (CAR) comprising: (i) two or more antigenic determinant binding domains (ADBDs), each ADBD comprising a first antigenic determinant (AD), (ii) a transmembrane domain, and (iii) an intracellular domain; and b. A genetic modification that abolishes expression of the first AD on the genetically modified cell; A genetically modified human immune effector cell, wherein said genetically modified cell does not express said first AD.

21. 1. A method for killing target cells or delivering an immune response to a target, comprising: a. contacting the genetically modified cell of claim 20 with the target cell; b. Contacting the genetically modified cell of claim 20 with an adaptor and the target cell, wherein the adaptor comprises an AD recognized by the CAR and a second ADBD; c. contacting the genetically modified cell of claim 20 with an adaptor and the target cell, wherein the adaptor comprises an AD recognized by the CAR and a second ADBD capable of binding to a second AD on the target cell; or d. Contacting the genetically modified cell of claim 20 with an adaptor and a cancer cell, wherein the adaptor comprises an AD recognized by the CAR and a second ADBD capable of binding to a second AD on the cancer cell.

22. 1. A method for inducing an immune response in target cells, a method for treating a proliferative disorder, cancer, an autoimmune disease, an infectious disease, or allograft rejection in a subject, a method for lymphodepleting a subject, a method for depleting memory T cells in a subject, a method for treating an autoimmune disease or disorder in a subject, or a method for treating a hematological cancer in a subject, or a method for pre-treating a subject for transplantation, comprising: a. administering to the subject in need thereof a therapeutically effective amount of the genetically modified cells of claim 20; b. Administering to the subject in need thereof a therapeutically effective amount of the genetically modified cells of claim 20 and a therapeutically effective amount of an adaptor comprising an AD recognized by the CAR and a second ADBD capable of binding to a second AD on the target cell; c. Administering a therapeutically effective amount of the genetically modified cells of claim 20 to the subject in need thereof, wherein the subject comprises an AD recognized by the CAR and a second ADBD capable of binding to a second AD on the target cell; d. A method comprising administering to a subject in need thereof a therapeutically effective amount of an adapter comprising the CAR and an AD recognized by a second ADBD capable of binding to a second AD on the target cell, wherein the subject has been administered the genetically modified cell of claim 20.

23. 1. A genetically modified human immune effector cell, comprising: a. a chimeric antigen receptor (CAR) comprising: (i) a first antigenic determinant binding domain (ADBD) that specifically binds to a first antigenic determinant (AD), (ii) a transmembrane domain, and (iii) an intracellular domain; and b. A genetic modification that abolishes expression of a second AD on the genetically modified cell; A genetically modified human immune effector cell, wherein the genetically modified cell used in combination with an adaptor is capable of inducing an immune response against cells expressing the second AD in an in vitro assay, the adaptor comprising a second ADBD that specifically binds to the first AD and the second AD, and the genetically modified cell does not express the second AD.

24. 1. A genetically modified human immune effector cell, comprising: a. a chimeric antigen receptor (CAR) comprising: (i) two or more antigenic determinant binding domains (ADBDs), each ADBD comprising a first antigenic determinant (AD) that specifically binds to the first antigenic determinant (AD), (ii) a transmembrane domain, and (iii) an intracellular domain; and b. A genetic modification that abolishes expression of a second AD on the genetically modified cell; A genetically modified human immune effector cell, wherein the genetically modified cell used in combination with an adaptor is capable of inducing an immune response against cells expressing the second AD in an in vitro assay, the adaptor comprising a second ADBD that specifically binds to the first AD and the second AD, and the genetically modified cell does not express the second AD.

25. 25. A method of killing a target cell or transmitting an immune response to a target cell, comprising contacting the genetically modified cell of claim 23 or claim 24 with an adaptor and the target cell, wherein the adaptor comprises a second ADBD that specifically binds to the first AD and the second AD, and wherein the second AD is expressed on the target cell, and optionally the target cell is a blood cancer cell.

26. 1. A method for inducing an immune response in target cells, a method for treating a proliferative disorder, cancer, an autoimmune disease, an infectious disease, or allograft rejection in a subject, a method for lymphodepleting a subject, a method for depleting memory T cells in a subject, a method for treating an autoimmune disease or disorder in a subject, or a method for treating a hematological cancer in a subject, or a method for pre-treating a subject for transplantation, comprising: a. administering to a subject in need thereof a therapeutically effective amount of the genetically modified cells of claim 23 or claim 24; b. Administering to the subject in need thereof a therapeutically effective amount of the genetically modified cell of claim 23 or claim 24 and a therapeutically effective amount of an adaptor comprising a second ADBD that specifically binds to the first AD and the second AD, wherein the second AD is expressed on the target cell; c) administering a therapeutically effective amount of the genetically modified cells of claim 23 or claim 24 to the subject in need thereof, wherein the subject has been administered an adaptor comprising the first AD and a second ADBD that specifically binds to the second AD, and wherein the second AD is expressed on the target cell; d. Administering a therapeutically effective amount of an adapter comprising the first AD and a second ADBD that specifically binds to the second AD to a subject in need thereof, wherein the subject has been administered the genetic modification of claim 23 or claim 24, and the second AD is expressed on the target cell.

27. 18. An isolated adaptor polypeptide comprising (1) an antigenic determinant (AD) and (b) one or more antigenic determinant binding domains (ADBDs), wherein at least one ADBD specifically binds to human CD45 AD, and wherein contacting the adaptor with a CD45 AD-expressing target cell in the presence of the genetically modified cell of claim 17 induces an immune response by the genetically modified cell against the target cell in an in vitro assay.

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