Multifunctional immune cell therapy

Abstract

Provided herein are compositions based on multifunctional chimeric antigen receptors (CARs) and their use in directing immune responses to target cells. The compositions have uses including treating hyperproliferative disorders such as cancer. The methods provided generally include the use of CAR cells in combination with adaptors. The adaptors confer the ability to modulate, modify, and / or redirect immune responses mediated by CAR cells in vitro and in vivo. In some embodiments, the CAR cells include genetic modifications to reduce or eliminate the expression of targeted antigenic determinants.

Description

[Technical Field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims priority to U.S. Provisional Patent Application No. 63 / 166,550, filed March 26, 2021, which is incorporated herein in its entirety. [Background technology]

[0002] background The disclosures of International Application Nos. PCT / US2018 / 060902 (Patent Document 1) and PCT / US2018 / 060887 (Patent Document 2), both filed on November 14, 2018, are incorporated herein by reference in their entirety for all purposes.

[0003] Adoptive transfer of genetically modified T cells is a rapidly evolving, innovative treatment for cancer. Chimeric antigen receptor (CAR)-engineered 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 hematologic 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 CAR technology can more fully realize its enormous potential.

[0004] Due to the inability of CAR T cells to address antigen escape and the inherent heterogeneity of cancer or tumor phenotype, current clinical trials using CAR T cells observe a high incidence of relapse within one year. Furthermore, current CAR technologies are not adaptive to such 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 antigen targets expressed over time. In addition to improving the durability and persistence of clinical responses, other significant obstacles to the use of current CAR cell-based technologies include the time required to generate CAR T cells and the suboptimal specificity, efficacy, and safety of CAR cells 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 CAR cell-mediated immune responses in vivo.

[0005] Bone marrow transplantation (BMT) and hematopoietic stem cell transplantation (HSCT) hold the potential to cure any hematological or immunological disease. Czechowicz et al., Blood 128(22):493 (2016). Despite their enormous potential, the clinical use of BMT and HSCT remains significantly limited, in part due to significant safety and toxicity risks associated with current high-dose chemotherapy / irradiation conditioning regimens designed to prepare patients for transplantation and subsequent donor HSC engraftment. The most common toxicities experienced by patients include neutropenia, infections, anemia, mucositis, infertility, organ damage, and secondary malignancies, particularly in the bone marrow compartment. The complete elimination of these toxic conditioning regimens would dramatically improve the safety profile of BMT and HSCT, expanding their potential applications to include more non-malignant hematological disorders, a wide range of autoimmune disorders, and facilitating solid organ transplantation. Therefore, there is a need to develop improved 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] [Patent documents]

[0006] [Patent Document 1] PCT / US2018 / 060902 [Patent Document 2] PCT / US2018 / 060887 [Non-patent literature]

[0007] [Non-Patent Document 1] Czechowicz et al.,Blood 128(22):493(2016) Summary of the Invention

[0008] Quick Overview In some aspects, the present disclosure provides: [1.] A composition comprising: [1.] A method for treating lymphoma or leukemia, comprising administering to a subject in need of treatment for lymphoma or leukemia: a. a therapeutically effective dose of cells expressing a chimeric antigen receptor (CAR), wherein the CAR comprises (i) a D domain that binds to AFP p26 or a variant thereof, (ii) a transmembrane domain, and (iii) an intracellular domain; and b. One or more therapeutically effective doses of an adaptor comprising (i) AFP p26 or a variant thereof and (ii) a D domain that binds to BCMA. The method comprising administering [2.] The method described in [1], wherein the lymphoma or leukemia contains cells that express BCMA. [3.] The method described in [1] or [2], wherein the lymphoma or leukemia is B-cell lymphoma or leukemia. [4.] The method described in [1] or [2], wherein the lymphoma or leukemia is multiple myeloma. [5.] A method for delivering an immune response to multiple myeloma cells, comprising administering to a subject in need thereof an immune response to multiple myeloma cells: a. a therapeutically effective dose of cells expressing a chimeric antigen receptor (CAR), wherein the CAR comprises (i) a D domain that binds to AFP p26 or a variant thereof, (ii) a transmembrane domain, and (iii) an intracellular domain; and b. One or more therapeutically effective doses of an adaptor comprising (i) AFP p26 or a variant thereof and (ii) a D domain that binds to BCMA. The method comprising administering [6.] A method for killing multiple myeloma cells, comprising administering to a subject in need of killing multiple myeloma cells: a. a therapeutically effective dose of cells expressing a chimeric antigen receptor (CAR), wherein the CAR comprises (i) a D domain that binds to AFP p26 or a variant thereof, (ii) a transmembrane domain, and (iii) an intracellular domain; and b. One or more therapeutically effective doses of an adaptor comprising (i) AFP p26 or a variant thereof and (ii) a D domain that binds to BCMA. The method comprising administering [7.] A method for depleting multiple myeloma cells, comprising administering to a subject in need of depletion of multiple myeloma cells: a. a therapeutically effective dose of cells expressing a chimeric antigen receptor (CAR), wherein the CAR comprises (i) a D domain that binds to AFP p26 or a variant thereof, (ii) a transmembrane domain, and (iii) an intracellular domain; and b. One or more therapeutically effective doses of an adaptor comprising (i) AFP p26 or a variant thereof and (ii) a D domain that binds to BCMA. The method comprising administering [8.] The method according to any one of [1] to [7], wherein the cells are immune effector cells. [9.] The method according to any one of [1] to [8], wherein the cells are T cells or NK cells. [10.] The method according to any one of [1] to [9], wherein the D domain that binds to AFP p26 comprises the amino acid sequence of SEQ ID NO:201. [11.] The method described in any one of [1] to

[10] , wherein the CAR comprises the amino acid sequence of SEQ ID NO:1164. [12.] The method described in any one of [1] to

[11] , wherein the D domain that binds to BCMA comprises the amino acid sequence of SEQ ID NO:201. [13.] The method described in any one of [1] to

[12] , wherein the adapter comprises the amino acid sequence of SEQ ID NO:1165. [14.] The method according to any one of [1] to

[13] , wherein the adapter is formulated for intravenous administration. [15.] The method according to any one of [1] to

[13] , wherein the adapter is formulated for subcutaneous administration. [16.] The method according to any one of [1] to

[15] , wherein the cells and the adaptor are administered simultaneously. [17.] The method according to any one of [1] to

[15] , wherein the cells and the adaptor are administered sequentially. [18.] The method according to any one of [1] to

[15] , wherein the cells are administered before the adaptor. [19.] The method according to any one of [1] to

[18] , wherein the cells and the adaptor are administered on the same day. [20.] The method according to any one of [1] to

[18] , wherein the cells and the adaptor are administered on different days. [21.] The method according to

[20] , wherein the cells and the adapter are administered on consecutive days. [22.] The method of

[20] , wherein the cells and the adapter are administered 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, or 7 days apart. [23.] The method according to any one of [1] to

[22] , wherein the cells and the adaptor are administered on different days. [24.] The method described in any one of [1] to

[23] , comprising a step of administering about 20 × 10 6 to about 500 × 10 6 or about 50 × 10 6 to about 200 × 10 6 cells expressing the CAR. [25.] The method described in any one of [1] to

[23] , comprising a step of administering approximately 100 x 106 cells expressing the CAR. [26.] A method according to any one of [1] to

[24] , comprising administering one or more doses of the adapter, the dose being about 1□g / kg to about 10 mg / kg, about 5□g / kg to about 5 mg / kg, about 10□g / kg to about 2 mg / kg, about 20□g / kg to about 1 mg / kg, or about 50□g / kg to about 0.5 mg / kg. [27.] The method described in any one of [1] to

[24] , comprising administering one or more doses of the adapter that are between about 20 □g / kg and about 1 mg / kg. [28.] The method of any one of [1] to

[24] , comprising a step of administering one or more doses of the adapter, the dose being about 0.01 mg / kg, about 0.02 mg / kg, about 0.04 mg / kg, about 0.07 mg / kg, about 0.1 mg / kg, about 0.2 mg / kg, about 0.3 mg / kg, about 0.4 mg / kg, about 0.5 mg / kg, about 0.6 mg / kg, about 0.7 mg / kg, about 1 mg / kg, or about 2 mg / kg. [29.] The method of any one of [1] to

[24] , comprising administering one or more doses of the adapter, the dose being about 0.04 mg / kg, about 0.07 mg / kg, about 0.1 mg / kg, about 0.2 mg / kg, or about 0.3 mg / kg. [30.] The method of any one of [1] to

[24] , comprising a step of administering one or more doses of the adapter, the dose being about 0.4 mg, about 0.8 mg, about 1.6 mg, about 2.8 mg, about 3 mg, about 4 mg, about 8 mg, about 12 mg, about 16 mg, about 20 mg, about 24 mg, about 28 mg, about 40 mg, or about 80 mg. [31.] The method of any one of [1] to

[24] , comprising administering one or more doses of the adapter, the dose being about 1.6 mg, about 2.8 mg, about 3 mg, about 4 mg, about 8 mg, about 12 mg, about 16 mg, or about 20 mg. [32.] The method of any one of [1] to

[31] , comprising administering more than one dose of the adapter, wherein the more than one dose is administered three times a day, twice a day, daily, every two days, twice a week, or weekly. [33.] The method of any one of [1] to

[31] , comprising administering more than one dose of the adapter, wherein the more than one dose is administered daily. [34.] The method of any one of [1] to

[31] , comprising administering more than one dose of the adapter, wherein the more than one dose is administered every two days. [35.] The method of any one of [1] to

[31] , comprising administering more than one dose of the adapter, wherein the more than one dose is administered daily for about 1 to about 4 weeks, followed by administration every other day, twice a week, or weekly. [36.] The method of any one of [1] to

[31] , comprising administering more than one dose of the adapter, wherein the more than one dose is administered daily for about one week, followed by administration every other day. [37.] The method of any one of [1] to

[31] , comprising administering more than one dose of the adapter, wherein the more than one dose is administered daily for about one week, followed by twice weekly administration. [38.] The method of any one of [1] to

[31] , comprising administering more than one dose of the adapter, wherein the more than one dose is administered daily for about one week, followed by weekly administration. [39.] The method according to any one of [1] to

[38] , comprising administering the adapter at different doses. [40.] The method of

[36] , comprising administering one or more loading doses of the adapter of about 0.1 mg / kg to about 10 mg / kg, followed by administering one or more maintenance doses of about 0.01 mg / kg to about 0.1 mg / kg. [41.] The method described in

[39] , comprising administering one or more loading doses of the adapter of about 4 mg to about 200 mg, followed by administering one or more maintenance doses of about 0.4 mg to about 10 mg. [42.] The method of any one of

[40] or

[41] , wherein the one or more loading doses are administered for about 1 day to about 3 weeks. [43.] The method of any one of

[40] or

[41] , wherein the one or more loading doses are administered for about 1 day to about 1 week. [44.] The method of any one of

[40] or

[41] , wherein the one or more loading doses are administered for 1 day, 2 days, 3 days, 4 days, or about 1 week. [45.] A method for treating lymphoma or leukemia, comprising the step of administering to a subject in need of lymphoma or leukemia treatment one or more therapeutically effective doses of an adapter comprising (i) AFP p26 or a variant thereof and (ii) a D domain that binds to BCMA, wherein the subject has previously received a therapeutically effective dose of cells expressing a chimeric antigen receptor (CAR), the CAR comprising (i) a D domain that binds to AFP p26 or a variant thereof, (ii) a transmembrane domain, and (iii) an intracellular domain. [46.] The method described in

[45] , wherein the lymphoma or leukemia contains cells that express BCMA. [47.] The method according to

[45] or

[46] , wherein the lymphoma or leukemia is a B-cell lymphoma or leukemia. [48.] The method according to

[45] or

[46] , wherein the lymphoma or leukemia is multiple myeloma. [49.] A method for delivering an immune response to multiple myeloma cells, comprising the step of administering to a subject in need thereof one or more therapeutically effective doses of an adapter comprising (i) AFP p26 or a variant thereof and (ii) a D domain that binds to BCMA, wherein the subject has previously received a therapeutically effective dose of cells expressing a chimeric antigen receptor (CAR), the CAR comprising (i) a D domain that binds to AFP p26 or a variant thereof, (ii) a transmembrane domain, and (iii) an intracellular domain. [50.] A method for killing multiple myeloma cells, comprising the step of administering to a subject in need of killing multiple myeloma cells one or more therapeutically effective doses of an adapter comprising (i) AFP p26 or a variant thereof and (ii) a D domain that binds to BCMA, wherein the subject has previously received a therapeutically effective dose of cells expressing a chimeric antigen receptor (CAR), the CAR comprising (i) a D domain that binds to AFP p26 or a variant thereof, (ii) a transmembrane domain, and (iii) an intracellular domain. [51.] A method for depleting multiple myeloma cells, comprising the step of administering to a subject in need of depletion of multiple myeloma cells one or more therapeutically effective doses of an adapter comprising (i) AFP p26 or a variant thereof and (ii) a D domain that binds to BCMA, wherein the subject has previously received a therapeutically effective dose of cells expressing a chimeric antigen receptor (CAR), the CAR comprising (i) a D domain that binds to AFP p26 or a variant thereof, (ii) a transmembrane domain, and (iii) an intracellular domain.

[0009] Provided herein are compositions based on multifunctional chimeric antigen receptors (CARs) and their use in directing immune responses to target cells.The compositions have uses including treating hyperproliferative disorders such as cancer.The provided methods generally involve the use of CAR cells in combination with adaptors.The adaptors confer the ability to modulate, modify, and / or redirect the immune response mediated by CAR cells in vitro and in vivo.

[0010] The present disclosure relates to a composition comprising: (a) a cell expressing a chimeric antigen receptor (CAR), wherein the CAR comprises 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.

[0011] In some embodiments, the present disclosure provides a composition comprising: (a) a cell expressing a CAR, wherein the CAR comprises (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 (i) the first AD and (ii) an ADBD that binds to a second AD on the target cell.

[0012] In some embodiments, the present disclosure provides a composition comprising: (a) a cell expressing a CAR, wherein the CAR comprises (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 (i) the first AD and (ii) an ADBD that binds to a second AD on a second target cell.

[0013] In some embodiments, the present disclosure provides a composition comprising: (a) a cell expressing a CAR, wherein the CAR comprises (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 that comprises (i) the first AD and (ii) an ADBD that binds to a second AD on a target cell.

[0014] In some embodiments, the present disclosure provides a composition comprising: (a) a cell expressing a CAR, wherein the CAR comprises (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.

[0015] In some embodiments, the present disclosure provides a composition comprising: (a) a cell expressing a CAR, wherein the CAR comprises (i) an ADBD that binds to a first AD, (ii) a transmembrane domain, and (iii) an intracellular domain; and (b) an adaptor that comprises (i) the first AD and (ii) a D domain that binds to a second AD on a target cell.

[0016] In some embodiments, the present disclosure provides a composition comprising: (a) a cell expressing a CAR, wherein the CAR comprises (i) a 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) an ADBD that binds to a second AD on a target cell.

[0017] In some embodiments, the present disclosure provides a composition comprising: (a) a cell expressing a CAR, wherein the CAR comprises (i) a first D domain that binds to a first AD, (ii) a transmembrane domain, and (iii) an intracellular domain; and (b) an adaptor that comprises (i) the first AD and (ii) a second D domain that binds to a second AD on a target cell.

[0018] In some embodiments, the present disclosure provides a method of killing a target cell, the method 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 the target cell, (ii) a transmembrane domain, and (iii) an intracellular domain, and (b) the adaptor comprises (i) the first AD and (ii) an ADBD that binds to a second AD on the target cell.

[0019] In some embodiments, the present disclosure provides a method of killing one or more target cells, the method 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 the first target cell, (ii) a transmembrane domain, and (iii) an intracellular domain, and (b) the adaptor comprises (i) the first AD and (ii) an ADBD that binds to a second AD on a second target cell.

[0020] In some embodiments, the present disclosure provides a method of killing a target cell, the method 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 (i) the first AD and (ii) an ADBD that binds to a second AD on the target cell.

[0021] In some embodiments, the present disclosure provides a method of killing a target cell, the method 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 (i) the first AD and (ii) an ADBD that is an ASBD that binds to a second AD on the target cell.

[0022] In some embodiments, the present disclosure provides a method of killing a target cell, the method 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 first target cell further comprises an adaptor comprising (i) the first AD and (ii) an ADBD that binds to a second AD on the target cell; and (c) 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.

[0023] In some embodiments, the present disclosure provides a method of killing one or more target cells, the method comprising contacting a composition comprising target cells with cells 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) an ADBD that binds to 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.

[0024] In some embodiments, the present disclosure provides a method of killing a target cell, the method 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 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 the second AD, (ii) a transmembrane domain, and (iii) an intracellular domain.

[0025] In some embodiments, the present disclosure provides a method of killing a target cell, the method 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, the ADBD being an ASBD that binds to the first AD on the target cell, and (ii) a second AD; and (c) the CAR comprises (i) an ADBD that binds to the second AD, (ii) a transmembrane domain, and (iii) an intracellular domain.

[0026] In some embodiments, the present disclosure provides a method of delivering an immune response to a target cell, the method 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 the target cell, (ii) a transmembrane domain, and (iii) an intracellular domain, and (b) the adaptor comprises (i) the first AD and (ii) an ADBD that binds to a second AD on the target cell.

[0027] In some embodiments, the present disclosure provides a method of delivering an immune response to one or more target cells, the method 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 the first target cell, (ii) a transmembrane domain, and (iii) an intracellular domain, and (b) the adaptor comprises (i) the first AD and (ii) an ADBD that binds to a second AD on a second target cell.

[0028] In some embodiments, the present disclosure provides a method of delivering an immune response to a target cell, the method 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, the ADBD being an ASBD that binds to a first AD, (ii) a transmembrane domain, and (iii) an intracellular domain, and (b) the adaptor comprises (i) the first AD and (ii) an ADBD that binds to a second AD on the target cell.

[0029] In some embodiments, the present disclosure provides a method of delivering an immune response to a target cell, the method 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 (i) the first AD and (ii) an ADBD that is an ASBD that binds to a second AD on the target cell.

[0030] In some embodiments, the present disclosure provides a method of delivering an immune response to a target cell, the method comprising contacting a composition comprising a 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 (i) the first AD and (ii) an ADBD that binds to a second AD on the target cell; and (c) 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.

[0031] In some embodiments, the present disclosure provides a method of delivering an immune response to one or more target cells, the method comprising contacting a composition comprising target cells with cells 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) an ADBD that binds to 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.

[0032] In some embodiments, the present disclosure provides a method of delivering an immune response to a target cell, the method comprising contacting a composition comprising a 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 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 the second AD, (ii) a transmembrane domain, and (iii) an intracellular domain.

[0033] In some embodiments, the present disclosure provides a method of delivering an immune response to a target cell, the method comprising contacting a composition comprising a 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 an adaptor comprising (i) an ADBD, the ADBD being an ASBD that binds to the first AD on the target cell, and (ii) a second AD; and (c) the CAR comprises (i) an ADBD that binds to the second AD, (ii) a transmembrane domain, and (iii) an intracellular domain.

[0034] In some embodiments, the present disclosure provides a method of killing a target cell in a patient, comprising administering an adaptor to the patient, 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 on the target cell, (ii) a transmembrane domain, and (iii) an intracellular domain, and (b) the adaptor comprises (i) the first AD and (ii) an ADBD that binds to a second AD on the target cell.

[0035] In some embodiments, the present disclosure provides a method of redirecting target cell killing in a patient, comprising administering an adaptor to the patient, 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 on a first target cell, (ii) a transmembrane domain, and (iii) an intracellular domain, and (b) the adaptor comprises (i) the first AD and (ii) an ADBD that binds to a second AD on a second target cell.

[0036] In some embodiments, the present disclosure provides a method of killing a target cell in a patient, comprising administering an adaptor to the patient, wherein (a) the patient has been treated with 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 (i) the first AD and (ii) an ADBD that binds to a second AD on the target cell.

[0037] In some embodiments, the present disclosure provides a method of killing a target cell in a patient, comprising administering an adaptor to the patient, 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 (i) the first AD and (ii) an ASBD that binds to a second AD on the target cell.

[0038] In some embodiments, the present disclosure provides a method of killing a target cell in a patient, the method 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) the first AD and (ii) an ADBD that binds to the second AD on the target cell, and (c) 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.

[0039] In some embodiments, the present disclosure provides a method of directing targeted cell killing 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 the first AD on the adaptor, (ii) a transmembrane domain, and (iii) an intracellular domain.

[0040] In some embodiments, the present disclosure provides a method of killing 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 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 the second AD, (ii) a transmembrane domain, and (iii) an intracellular domain.

[0041] In some embodiments, the present disclosure provides a method of killing 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, the ADBD being an ASBD that binds to the first AD on the target cell, and (ii) a second AD; and (c) the CAR comprises (i) an ADBD that binds to the second AD, (ii) a transmembrane domain, and (iii) an intracellular domain.

[0042] In some embodiments, the present disclosure provides a method of delivering an immune response to a target cell in a patient, the method comprising administering an adapter to the patient, 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 on the target cell, (ii) a transmembrane domain, and (iii) an intracellular domain, and (b) the adapter comprises (i) the first AD and (ii) an ADBD that binds to a second AD on the target cell.

[0043] In some embodiments, the present disclosure provides a method of redirecting an immune response to a target cell in a patient, comprising administering an adapter to the patient, 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 on a first target cell, (ii) a transmembrane domain, and (iii) an intracellular domain, and (b) the adapter comprises (i) the first AD and (ii) an ADBD that binds to a second AD on a second target cell.

[0044] In some embodiments, the present disclosure provides a method of delivering an immune response to a target cell in a patient, the method comprising administering an adaptor to the patient, wherein (a) the patient has been treated with 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 (i) the first AD and (ii) an ADBD that binds to a second AD on the target cell.

[0045] In some embodiments, the present disclosure provides a method of delivering an immune response to a target cell in a patient, the method comprising administering an adaptor to the patient, 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 (i) the first AD and (ii) an ASBD that binds to a second AD on the target cell.

[0046] In some embodiments, the present disclosure provides a method of delivering 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) the first AD and (ii) an ADBD that binds to the second AD on the target cell, and (c) 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.

[0047] In some embodiments, the present disclosure provides a method of directing an immune response to 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 the first AD on the adaptor, (ii) a transmembrane domain, and (iii) an intracellular domain.

[0048] In some embodiments, the present disclosure provides a method of delivering 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 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 the second AD, (ii) a transmembrane domain, and (iii) an intracellular domain.

[0049] In some embodiments, the present disclosure provides a method of delivering 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 the first AD on the target cell, and (ii) a second AD; and (c) the CAR comprises (i) an ADBD that binds to the second AD, (ii) a transmembrane domain, and (iii) an intracellular domain.

[0050] In some embodiments, the present disclosure provides a method of treating a proliferative disorder, cancer, an autoimmune disease, an infectious disease, or transplant rejection, comprising contacting a composition comprising target cells with cells expressing a CAR, wherein (a) a first AD is present on the target cells; (b) the composition comprising the target cells further comprises an adaptor comprising (i) an ADBD, which is an ASBD that binds to the first AD on the target cells, and (ii) a second AD; and (c) the CAR comprises (i) an ADBD that binds to the second AD, (ii) a transmembrane domain, and (iii) an intracellular domain.

[0051] In some embodiments, the present disclosure provides a method of treating a proliferative disorder, cancer, an autoimmune disease, an infectious disease, or graft rejection, 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 the target cell, (ii) a transmembrane domain, and (iii) an intracellular domain, and (b) the adaptor comprises (i) the first AD and (ii) an ADBD that binds to a second AD on the target cell.

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

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

[0054] In some embodiments, the present disclosure provides a method of treating a proliferative disorder, cancer, an autoimmune disease, an infectious disease, or graft rejection, comprising administering an adapter to said patient, wherein (a) said patient has been treated with a cell expressing a chimeric antigen receptor (CAR), said 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) said adapter comprises an ADBD that is (i) the first AD and (ii) an ASBD that binds to a second AD on the target cell.

[0055] In some embodiments, the present disclosure provides a method of treating a proliferative disorder, cancer, an autoimmune disease, an infectious disease, or graft rejection, 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) the first AD and (ii) an ADBD that binds to the second AD on the target cell; and (c) 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.

[0056] In some embodiments, the present disclosure provides a method for directing the treatment of a proliferative disorder, cancer, an autoimmune disease, an infectious disease, or transplant rejection, comprising administering to said patient cells expressing a CAR, wherein (a) said patient has been treated with an adaptor comprising (i) a first AD and (ii) an ADBD that binds to a second AD on said target cell, and (b) said CAR comprises (i) an ADBD that binds to the first AD on the adaptor, (ii) a transmembrane domain, and (iii) an intracellular domain.

[0057] In some embodiments, the present disclosure provides a method of treating a proliferative disorder, cancer, an autoimmune disease, an infectious disease, or graft rejection, comprising administering to said patient a cell expressing a CAR, wherein (a) a first AD is present on said target cell; (b) said 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) said CAR comprises (i) an ADBD that is an ASBD that binds to said second AD, (ii) a transmembrane domain, and (iii) an intracellular domain.

[0058] In some embodiments, the present disclosure provides a method of treating a proliferative disorder, cancer, an autoimmune disease, an infectious disease, or graft rejection, comprising administering to said patient cells expressing a CAR, wherein (a) a first AD is present on said target cell; (b) said 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) said CAR comprises (i) an ADBD that binds to said second AD, (ii) a transmembrane domain, and (iii) an intracellular domain.

[0059] In some embodiments, the present disclosure provides a method of treating a hematological cancer, the method 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, the ADBD being an ASBD that binds to the first AD on the target cell, and (ii) a second AD; and (c) the CAR comprises (i) an ADBD that binds to the second AD, (ii) a transmembrane domain, and (iii) an intracellular domain.

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

[0061] In some embodiments, the present disclosure provides a method of redirecting treatment of a hematological cancer, comprising administering an adapter 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 a first target cell, (ii) a transmembrane domain, and (iii) an intracellular domain, and (b) the adapter comprises (i) the first AD and (ii) an ADBD that binds to a second AD on a second target cell.

[0062] In some embodiments, the present disclosure provides a method of treating a hematological cancer, comprising administering an adaptor to the patient, wherein (a) the patient has been treated with 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 (i) the first AD and (ii) an ADBD that binds to a second AD on the target cell.

[0063] In some embodiments, the present disclosure provides a method of treating a hematological cancer, comprising administering an adaptor to the patient, wherein (a) the patient has been treated with a cell expressing a CAR, wherein the CAR comprises (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 (i) the first AD and (ii) an ASBD that binds to a second AD on the target cell.

[0064] In some embodiments, the disclosure provides a method of treating a hematological cancer, 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) the first AD and (ii) an ADBD that binds to the second AD on the target cell; and (c) 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.

[0065] In some embodiments, the present disclosure provides a method of directing the treatment of a hematological cancer, comprising administering to the patient a cell 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 the first AD on the adaptor, (ii) a transmembrane domain, and (iii) an intracellular domain.

[0066] In some embodiments, the disclosure provides a method of treating a hematological cancer, 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 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 the second AD, (ii) a transmembrane domain, and (iii) an intracellular domain.

[0067] In some embodiments, the disclosure provides a method of treating a hematological cancer, 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, which is an ASBD that binds to the first AD on the target cell, and (ii) a second AD; and (c) the CAR comprises (i) an ADBD that binds to the second AD, (ii) a transmembrane domain, and (iii) an intracellular domain.

[0068] In some embodiments, the present disclosure provides an engineered human immune effector cell comprising: (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 eliminates expression of said human CD45 AD on said engineered cell, wherein said engineered cell is capable of directing an immune response to cells expressing the CD45 AD in an in vitro assay; and wherein said engineered cell does not express said CD45 AD.

[0069] In some embodiments, the present disclosure provides methods of killing target cells expressing human CD45, comprising contacting the target cells with engineered human immune effector cells, wherein the engineered 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 ablates expression of human CD45 AD on the engineered cells.

[0070] In some embodiments, the present disclosure provides methods of delivering an immune response to target cells expressing human CD45, comprising contacting the target cells with engineered human immune effector cells, wherein the engineered 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 ablates expression of human CD45 AD on the engineered cells.

[0071] In some embodiments, the present disclosure provides methods of treating a proliferative disorder, cancer, an autoimmune disease, an infectious disease, or graft rejection in a subject, the method comprising: administering to a subject in need of treatment for a proliferative disorder, cancer, an autoimmune disease, an infectious disease, or graft rejection a therapeutically effective amount of engineered human immune effector cells, wherein the engineered 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 ablates expression of human CD45 AD on the engineered cells.

[0072] In some embodiments, the present disclosure provides an engineered human immune effector cell 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; and (b) a genetic modification that eliminates expression of at least one human CD45 AD on the engineered cell, wherein the first AD is not the at least one human CD45 AD; wherein the engineered cell, used in combination with an adaptor, is capable of directing an immune response to cells expressing CD45 in an in vitro assay; and wherein the adaptor comprises the first AD and a second ADBD that specifically binds the at least one human CD45 AD; and wherein the engineered cell does not express the at least one human CD45 AD.

[0073] In some embodiments, the present disclosure provides a method of killing a target cell, comprising contacting the target cell with an engineered human immune effector cell and an adaptor, wherein the target cell expresses CD45, and the engineered human immune effector cell comprises (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 eliminates expression of at least one human CD45 AD on the engineered cell, and wherein the adaptor comprises the first AD and a second ADBD that specifically binds to human CD45 AD.

[0074] In some embodiments, the present disclosure provides a method of delivering an immune response to a target cell, the method comprising contacting the target cell with an engineered human immune effector cell and an adaptor, wherein the target cell expresses CD45, and the engineered 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 eliminates expression of at least one human CD45 AD on the engineered cell, and the adaptor comprises the first AD and a second ADBD that specifically binds to human CD45 AD.

[0075] In some embodiments, the present disclosure provides methods for directing an immune response to target cells expressing CD45 in a subject, the method comprising: (a) administering to a subject in need thereof a therapeutically effective amount of engineered human immune effector cells, the engineered human immune effector cells comprising: (i) 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 (ii) a genetic modification that eliminates expression of at least one human CD45 AD on the engineered 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 to the human CD45 AD.

[0076] In some embodiments, the present disclosure provides a method of directing an immune response to target cells expressing CD45 in a subject, the method comprising administering to a subject in need thereof a therapeutically effective amount of engineered human immune effector cells, the engineered human immune effector cells comprising: (i) 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 (ii) a genetic modification that eliminates expression of at least one human CD45 AD on the engineered cells, wherein the subject has previously administered an adaptor comprising the first AD and a second ADBD that specifically binds to the human CD45 AD.

[0077] In some embodiments, the present disclosure provides a method of directing an immune response to target cells expressing CD45 in a subject, the method comprising: administering to a subject in need of directing an immune response to target cells expressing CD45 in the subject 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 previously received engineered 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 ablates expression of at least one human CD45 AD on the engineered cells.

[0078] In some embodiments, the present disclosure provides methods of treating a proliferative disorder, cancer, autoimmune disease, infectious disease, or graft rejection in a subject, the method comprising: (a) administering to a subject in need of treatment a therapeutically effective amount of engineered human immune effector cells, the engineered human immune effector cells comprising: (i) 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 (ii) a genetic modification that eliminates expression of at least one human CD45 AD on the engineered 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 to human CD45 AD.

[0079] In some embodiments, the present disclosure provides a method of treating a proliferative disorder, cancer, autoimmune disease, infectious disease, or graft rejection in a subject, the method comprising administering to a subject in need of treatment a therapeutically effective amount of engineered human immune effector cells, the engineered 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), and (2) a transmembrane domain, and (3) an intracellular domain; and (ii) a genetic modification that eliminates expression of at least one human CD45 AD on the engineered cells, wherein the subject has previously been administered an adaptor comprising the first AD and a second ADBD that specifically binds to human CD45 AD.

[0080] In some embodiments, the present disclosure provides a method of treating a proliferative disorder, cancer, an autoimmune disease, an infectious disease, or graft rejection in a subject, the method comprising: administering a therapeutically effective amount of an adapter comprising a first AD and a second ADBD that specifically binds to a human CD45 AD to a subject in need of treatment for the proliferative disorder, cancer, autoimmune disease, infectious disease, or graft rejection, wherein the subject has previously received engineered 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 ablates expression of at least one human CD45 AD on the engineered cells.

[0081] In some embodiments, the present disclosure provides an engineered human immune effector cell comprising: (a) a chimeric antigen receptor (CAR), wherein the CAR comprises: (i) two or more antigenic determinant binding domains (ADBDs) comprising a first ADBD that binds to a first antigenic determinant (AD); (ii) a transmembrane domain; and (iii) an intracellular domain; and (b) a genetic modification that eliminates expression of the first AD on the engineered cell, such that the engineered cell does not express the first AD.

[0082] In some embodiments, the present disclosure provides a method of killing a target cell, the method comprising contacting the target cell with an engineered human immune effector cell comprising: (a) a chimeric antigen receptor (CAR), wherein the CAR comprises: (i) two or more antigenic determinant binding domains (ADBDs) comprising a first ADBD that binds to a first antigenic determinant (AD); (ii) a transmembrane domain; and (iii) an intracellular domain; and (b) a genetic modification that eliminates expression of the first AD on the engineered cell.

[0083] In some embodiments, the present disclosure provides a method of directing an immune response to target cells in a subject, the method comprising administering to a subject in need thereof a therapeutically effective amount of engineered human immune effector cells, the engineered human immune effector cells comprising: (a) a chimeric antigen receptor (CAR), wherein the CAR comprises: (i) two or more antigenic determinant binding domains (ADBDs) comprising a first ADBD that binds to a first antigenic determinant (AD); (ii) a transmembrane domain; and (iii) an intracellular domain; and (b) a genetic modification that eliminates expression of the first AD on the engineered cells.

[0084] In some embodiments, the present disclosure provides a method of treating a proliferative disorder, cancer, autoimmune disease, infectious disease, or graft rejection in a subject, the method comprising administering to a subject in need of treatment a therapeutically effective amount of engineered human immune effector cells, the engineered human immune effector cells comprising: (a) a chimeric antigen receptor (CAR), wherein the CAR comprises: (i) two or more antigenic determinant binding domains (ADBDs) comprising a first ADBD that binds to a first antigenic determinant (AD); (ii) a transmembrane domain; and (iii) an intracellular domain; and (b) a genetic modification that eliminates expression of the first AD on the engineered cells.

[0085] In some embodiments, the present disclosure provides a method of directing an immune response to target cells in a subject, the method comprising: (a) administering to a subject in need of directing an immune response to target cells in the subject a therapeutically effective amount of engineered human immune effector cells, the engineered human immune effector cells comprising: (i) a chimeric antigen receptor (CAR), the CAR comprising: (1) two or more antigenic determinant binding domains (ADBDs) comprising a first ADBD that binds to a first antigenic determinant (AD), (2) a transmembrane domain, and (3) an intracellular domain; and (ii) a genetic modification that eliminates expression of the first AD on the engineered cells; and (b) administering to the subject 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 cells.

[0086] In some embodiments, the present disclosure provides an engineered human immune effector cell comprising: (a) a chimeric antigen receptor (CAR), wherein the CAR comprises (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 eliminates expression of a second AD on the engineered cell; wherein the engineered cell is used in combination with an adaptor, wherein the engineered cell is capable of directing an immune response to cells expressing the second AD in an in vitro assay, and wherein the adaptor comprises the first AD and a second ADBD that specifically binds to the second AD, and wherein the engineered cell does not express the second AD.

[0087] In some embodiments, the present disclosure provides a method of directing an immune response to target cells in a subject, the method comprising: (a) administering to a subject in need of directing an immune response to target cells in the subject a therapeutically effective amount of engineered human immune effector cells, the engineered human immune effector cells comprising: (i) a chimeric antigen receptor (CAR), the 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 (ii) a genetic modification that eliminates expression of a second AD on the engineered cells; 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, wherein the second AD is expressed on the target cells.

[0088] In some embodiments, the present disclosure provides an engineered human immune effector cell comprising: (a) a chimeric antigen receptor (CAR), wherein the CAR comprises (i) two or more antigenic determinant binding domains (ADBDs) comprising a first 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 eliminates expression of a second AD on the engineered cell; used in combination with an adaptor, wherein the engineered cell is capable of directing an immune response to cells expressing the second AD in an in vitro assay, and wherein the adaptor comprises the first AD and a second ADBD that specifically binds to the second AD, and wherein the engineered cell does not express the second AD.

[0089] In some embodiments, the present disclosure provides a method of directing an immune response to target cells in a subject, the method comprising: (a) administering to a subject in need of directing an immune response to target cells in the subject a therapeutically effective amount of engineered human immune effector cells, the engineered human immune effector cells comprising: (i) a chimeric antigen receptor (CAR), wherein the CAR comprises: (1) two or more antigenic determinant binding domains (ADBDs) comprising a first 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 eliminates expression of a second AD on the engineered cells; 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, wherein the second AD is expressed on the target cells.

[0090] In some aspects, the present disclosure provides: [1.] (a) a cell expressing a chimeric antigen receptor (CAR), wherein 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 (b) an adaptor comprising (i) the first AD and (ii) an ADBD that binds to a second AD on the target cell. A composition comprising: [2.] (a) a cell expressing a chimeric antigen receptor (CAR), wherein 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 (b) an adaptor comprising (i) the first AD and (ii) an ADBD that binds to a second AD on a second target cell. A composition comprising: [3.] (a) a cell expressing a chimeric antigen receptor (CAR), wherein 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 (b) an adaptor comprising (i) the first AD and (ii) an ADBD that binds to a second AD on a target cell. A composition comprising: [4.] (a) a cell expressing a chimeric antigen receptor (CAR), wherein 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 (b) an adaptor comprising (i) the first AD and (ii) an ADBD comprising an ASBD that binds to a second AD on a target cell. A composition comprising: [5.] (a) a cell expressing a chimeric antigen receptor (CAR), wherein 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 (b) an adaptor comprising (i) the first AD and (ii) a D domain that binds to a second AD on a target cell. A composition comprising: [6.] (a) a cell expressing a chimeric antigen receptor (CAR), wherein 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 (b) an adaptor comprising (i) the first AD and (ii) an ADBD that binds to a second AD on a target cell. A composition comprising: [7.] (a) a cell expressing a chimeric antigen receptor (CAR), 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 (b) an adaptor comprising (i) the first AD and (ii) a second D domain that binds to a second AD on a target cell. A composition comprising: [8.] The composition described in any one of [1] to [7], wherein the CAR comprises a single-chain variable fragment (scFv) ADBD. [9.] The composition described in any one of [1] to [7], wherein the CAR comprises an alternative scaffold binding domain (ASBD) ADBD. [10.] The composition described in [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 described in any one of [1] to

[10] , wherein the CAR contains two ADBDs. [12.] The composition described in

[11] , wherein the CAR comprises an ASBD and an scFv. [13.] The composition described in

[11] , wherein the CAR comprises a D domain and an scFv. [14.] The composition described in

[11] , wherein the CAR contains two ASBDs. [15.] The composition described in

[11] , wherein the CAR contains two D domains. [16.] The composition described in any one of [1] to

[15] , wherein the CAR intracellular domain is a signal transduction domain. [17.] The composition described in

[16] , wherein the CAR intracellular domain comprises a primary signaling domain. [18.] The composition described in

[16] , wherein the CAR intracellular domain comprises a CD3ζ primary signaling domain. [19.] The composition described in

[17] or

[18] , wherein the CAR intracellular domain further comprises a costimulatory signaling domain. [20.] The composition described in

[19] , wherein the CAR intracellular domain comprises a costimulatory signaling domain selected from CD28, 41BB, CD27, and CD134. [21.] The composition described in

[20] , wherein the CAR intracellular domain includes a 41BB costimulatory signaling domain. [22.] The composition described in any one 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 described in

[22] , wherein the CAR binds to BCMA, and optionally the CAR comprises a sequence selected from the group: SEQ ID NO: 44 to 338 and 339. [24.] The composition described in

[22] , wherein the CAR binds to CD123, and optionally, the CAR comprises a sequence selected from the group: SEQ ID NO:340-77 and 773. [25.] The composition described in

[22] , wherein the CAR binds to CD19, and optionally, the CAR comprises a sequence selected from the group: SEQ ID NO: 1030 to 1058 and 1059. [26.] The composition described in

[22] , wherein the CAR binds to CD22, and optionally, the CAR comprises a sequence selected from the group: SEQ ID NO: 1060-1068 and 1069. [27.] The composition described in

[22] , wherein the CAR binds to CS1, and optionally, the CAR comprises a sequence selected from the group: SEQ ID NO:780-794 and 795. [28.] The composition described in

[22] , wherein the CAR binds to HER2, and optionally, the CAR comprises a sequence selected from the group: SEQ ID NO:800-839 and 840. [29.] The composition described in

[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 described in any one 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 one of [1] to

[30] , wherein the CAR comprises two ADBDs that bind to different targets. [32.] The composition described in

[31] , wherein the CAR binds to CD19 and CD123. [33.] The composition described in

[31] , wherein the 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 the CAR binds to PDL1 and CD123. [36.] The composition described in

[32] , wherein the CAR comprises a first ASBD that binds to CD19 and a second ASBD that binds to CD123. [37.] The composition described in

[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 described in

[33] , wherein the CAR comprises a first ASBD that binds to BCMA and a second ASBD that binds to CS1. [40.] The composition described in

[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 described in

[33] , wherein the CAR comprises a D domain that binds to CS1 and an scFv that binds to BCMA. [42.] The composition described in

[39] , wherein the CAR comprises a D domain that binds to BCMA and an scFv that binds to CS1. [43.] The composition described in

[34] , wherein the CAR comprises a first ASBD that binds to CD22 and a second ASBD that binds to CD123. [44.] The composition described in

[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 described in

[35] , wherein the CAR comprises a first ASBD that binds to PDL1 and a second ASBD that binds to CD123. [47.] The composition described in

[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 one 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 described in [1] to

[50] , 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. [52.] The composition described in any one of [1] to

[51] , wherein the adaptor comprises an ADBD that is an scFv. [53.] The composition described in any one of [1] to

[51] , wherein the adaptor comprises an ADBD that is an ASBD. [54.] The composition described in

[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 described in any one of [1] to

[54] , wherein the adapter comprises two ADBDs. [56.] The composition described in

[55] , wherein the two ADBDs are (a) identical, (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 described in

[55] or

[56] , wherein the adapter comprises two ASBDs. [58.] The composition described in 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 consisting of SEQ ID NOs: 44 to 1079. [59.] The composition described in any one of

[55] or

[56] , wherein the adapter comprises an ADBD that is an scFv and an ADBD that is an ASBD. [60.] The composition described in

[59] , wherein the adapter comprises an ADBD that is an scFv and an ADBD that is a D domain. [61.] The composition described in any one 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 described in

[61] , wherein the adapter comprises an ADBD that binds to BCMA, and optionally, the ADBD comprises a sequence selected from the group: SEQ ID NO: 44 to 338 and 339. [63.] The composition described in

[61] , wherein the adapter comprises an ADBD that binds to CS1, and optionally, the ADBD comprises a sequence selected from the group: SEQ ID NO:780-794 and 795. [64.] The composition described in

[61] , wherein the adapter comprises an ADBD that binds to CD123, and optionally, the ADBD comprises a sequence selected from the group: SEQ ID NO:340-772 and 773. [65.] The composition described in

[61] , wherein the adapter comprises an ADBD that binds to CD19, and optionally, the ADBD comprises a sequence selected from the group consisting of SEQ ID NOs: 1030 to 1058 and 1059. [66.] The composition described in

[61] , wherein the adapter 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 described in

[61] , wherein the adapter comprises an ADBD that binds to HER2, and optionally, the ADBD comprises a sequence selected from the group: SEQ ID NOs: 800 to 839 and 840. [68.] The composition described in

[61] , wherein the adaptor comprises an ADBD that binds to TACI or BAFFR. [69.] The composition described in

[61] , wherein the adapter 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 described in any one of [1] to

[69] , wherein the adapter is bispecific. [71.] The composition described in

[70] , wherein the adapter comprises an ADBD that binds to CD19 and an ADBD that binds to CD123. [72.] The composition described in

[70] , wherein the adapter comprises an ADBD that binds to BCMA and an ADBD that binds to CS1. [73.] The composition described in

[70] , wherein the adapter comprises an ADBD that binds to CD22 and an ADBD that binds to CD123. [74.] The composition described in

[70] , wherein the adaptor comprises an ADBD that binds to PDL1 and an ADBD that binds to CD123. [75.] The composition described in any one of [1] to

[74] , wherein the target cells are tumor cells. [76.] The composition described in

[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 described in

[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 described in

[79] , wherein the first and second tumor cells are of the same type. [81.] The composition described in

[79] , wherein the first and second tumor cells are of different types. [82.] The composition described in

[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 described in

[82] , wherein the tumor cells are multiple myeloma cells. [84.] The composition described in any one of [1] to

[83] , wherein the cells expressing the CAR are immune effector cells. [85.] The composition described in

[84] , wherein the immune effector cells are T cells. [86.] The composition described in

[84] , wherein the immune effector cells are NK cells. [87.] The composition described in any one of [1] to

[86] , wherein the cells expressing the CAR kill the target cells. [88.] The composition described in any one of [1] to

[87] , wherein binding of the adapter to an antigenic determinant blocks the activity of the antigen, including the AD. [89.] A method for killing target cells, comprising the step of contacting the target cells with the composition described in any one of [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, the 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; (b) the adapter comprises (i) the first AD and (ii) an ADBD that binds to a second AD on the target cell; The method. [91.] A method for 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), the 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; (b) the adapter comprises (i) the first AD and (ii) an ADBD that binds to a second AD on a second target cell; The method. [92.] 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 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; (b) the adapter comprises (i) the first AD and (ii) an ADBD that binds to a second AD on the target cell; The method. [93.] 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 chimeric antigen receptor (CAR), the CAR comprising (i) an antigenic determinant binding domain (ADBD) that binds to a first AD, (ii) a transmembrane domain, and (iii) an intracellular domain; (b) the adapter comprises (i) the first AD and (ii) an ADBD that is an ASBD that binds to a second AD on the target cell; The method. [94.] A method for killing target cells, comprising contacting a composition comprising the 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 a first target cell further comprises an adaptor comprising (i) the first AD and (ii) an antigenic determinant binding domain (ADBD) that binds to a second AD on the target cell; (c) 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; The method. [95.] A method for killing 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 (i) the first AD and (ii) an antigenic determinant binding domain (ADBD) that binds to a second AD on the second target cell; (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; The method. [96.] A method for killing target cells, comprising contacting a composition comprising the 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) 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 the second AD, (ii) a transmembrane domain, and (iii) an intracellular domain; The method. [97.] A method for killing target cells, comprising contacting a composition comprising the 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; (c) the CAR comprises (i) an ADBD that binds to the second AD, (ii) a transmembrane domain, and (iii) an intracellular domain; The method. [98.] A method for delivering 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, the 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; (b) the adapter comprises (i) the first AD and (ii) an ADBD that binds to a second AD on the target cell; The method. [99.] A method for delivering 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), the 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; (b) the adapter comprises (i) the first AD and (ii) an ADBD that binds to a second AD on a second target cell; The method. [100.] A method for delivering 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 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; (b) the adapter comprises (i) the first AD and (ii) an ADBD that binds to a second AD on the target cell; The method. [101.] A method for delivering 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 chimeric antigen receptor (CAR), the CAR comprising (i) an antigenic determinant binding domain (ADBD) that binds to a first AD, (ii) a transmembrane domain, and (iii) an intracellular domain; (b) the adapter comprises (i) the first AD and (ii) an ADBD that is an ASBD that binds to a second AD on the target cell; The method. [102.] A method for delivering an immune response to 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 a first target cell further comprises an adaptor comprising (i) the first AD and (ii) an antigenic determinant binding domain (ADBD) that binds to a second AD on the target cell; (c) 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; The method. [103.] A method for 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 (i) the first AD and (ii) an antigenic determinant binding domain (ADBD) that binds to a second AD on the second target cell; (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; The method. [104.] A method for delivering an immune response to 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; (c) 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; The method. [105.] A method for delivering an immune response to 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; (c) the CAR comprises (i) an ADBD that binds to the second AD, (ii) a transmembrane domain, and (iii) an intracellular domain; The method. [106.] A method of killing target cells in a patient, comprising administering to the patient an adapter; (a) the patient has been treated with a cell 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; (b) the adapter comprises (i) the first AD and (ii) an ADBD that binds to a second AD on the target cell; The method. [107.] A method of redirecting target cell killing in a patient, comprising administering to the patient an adapter; (a) the patient has been treated with a cell 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; (b) the adapter comprises (i) the first AD and (ii) an ADBD that binds to a second AD on a second target cell; The method. [108.] A method of killing target cells in a patient, comprising administering to the patient an adapter; (a) the patient has been treated with a cell 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; (b) the adapter comprises (i) the first AD and (ii) an ADBD that binds to a second AD on the target cell; The method. [109.] A method of killing target cells in a patient, comprising administering to the patient an adapter; (a) the patient has been treated with a cell 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; (b) the adapter comprises (i) the first AD and (ii) an ADBD that is an ASBD that binds to a second AD on the target cell; The method. [110.] 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 the target cell; (b) the patient has been treated with an adaptor comprising (i) the first AD and (ii) an antigenic determinant binding domain (ADBD) that binds to the second AD on the target cell; (c) the CAR comprises (i) an ADBD that binds to the first AD on a target cell or an adaptor, (ii) a transmembrane domain, and (iii) an intracellular domain; The method. [111.] A method of directing target cell killing in a patient, comprising administering to the patient cells expressing a chimeric antigen receptor (CAR), (a) 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; (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; The method. [112.] 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 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; (c) 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; The method. [113.] 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 has been 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 CAR comprises (i) an ADBD that binds to the second AD, (ii) a transmembrane domain, and (iii) an intracellular domain; The method. [114.] A method of delivering an immune response to target cells in a patient, comprising administering an adapter to the patient; (a) the patient has been treated with a cell 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; (b) the adapter comprises (i) the first AD and (ii) an ADBD that binds to a second AD on the target cell; The method. [115.] A method for redirecting an immune response to target cells in a patient, comprising administering to the patient an adapter; (a) the patient has been treated with a cell 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; (b) the adapter comprises (i) the first AD and (ii) an ADBD that binds to a second AD on a second target cell; The method. [116.] A method of delivering an immune response to target cells in a patient, comprising administering an adapter to the patient; (a) the patient has been treated with a cell 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; (b) the adapter comprises (i) the first AD and (ii) an ADBD that binds to a second AD on the target cell; The method. [117.] A method of delivering an immune response to target cells in a patient, comprising administering an adapter to the patient; (a) the patient has been treated with a cell 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; (b) the adapter comprises (i) the first AD and (ii) an ADBD that is an ASBD that binds to a second AD on the target cell; The method. [118.] 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 the target cell; (b) the patient has been treated with an adaptor comprising (i) the first AD and (ii) an antigenic determinant binding domain (ADBD) that binds to the second AD on the target cell; (c) 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; The method. [119.] A method of directing an immune response to target cells in a patient, comprising administering to the patient cells expressing a chimeric antigen receptor (CAR); (a) 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; (b) the CAR comprises (i) an ADBD that binds to the first AD on the adapter, (ii) a transmembrane domain, and (iii) an intracellular domain; The method. [120.] 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 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; (c) 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; The method. [121.] 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 has been 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 CAR comprises (i) an ADBD that binds to the second AD, (ii) a transmembrane domain, and (iii) an intracellular domain; The method. [122.] A method for treating a proliferative disorder, cancer, an autoimmune disease, an infectious disease, or a transplant 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; (c) the CAR comprises (i) an ADBD that binds to the second AD, (ii) a transmembrane domain, and (iii) an intracellular domain; The method. [123.] A method for treating a proliferative disorder, cancer, an autoimmune disease, an infectious disease, or transplant rejection, comprising administering to said patient an adapter; (a) the patient has been treated with a cell 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; (b) the adapter comprises (i) the first AD and (ii) an ADBD that binds to a second AD on the target cell; The method. [124.] A method of redirecting treatment of a proliferative disorder, cancer, autoimmune disease, infectious disease, or transplant rejection, comprising administering to a patient an adapter; (a) the patient has been treated with a cell 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; (b) the adapter comprises (i) the first AD and (ii) an ADBD that binds to a second AD on a second target cell; The method. [125.] A method for treating a proliferative disorder, cancer, an autoimmune disease, an infectious disease, or transplant rejection, comprising administering to a patient an adapter; (a) the patient has been treated with a cell 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; (b) the adapter comprises (i) the first AD and (ii) an ADBD that binds to a second AD on the target cell; The method. [126.] A method for treating a proliferative disorder, cancer, an autoimmune disease, an infectious disease, or transplant rejection, comprising administering to a patient an adapter; (a) the patient has been treated with a cell 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; (b) the adapter comprises (i) the first AD and (ii) an ADBD that is an ASBD that binds to a second AD on the target cell; The method. [127.] A method for treating a proliferative disorder, cancer, an autoimmune disease, an infectious disease, or transplant 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 the target cell; (b) the patient has been treated with an adaptor comprising (i) the first AD and (ii) an antigenic determinant binding domain (ADBD) that binds to the second AD on the target cell; (c) 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; The method. [128.] A method for treating a proliferative disorder, cancer, an autoimmune disease, an infectious disease, or transplant rejection, comprising administering to a patient cells expressing a chimeric antigen receptor (CAR); (a) 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; (b) the CAR comprises (i) an ADBD that binds to the first AD on the adapter, (ii) a transmembrane domain, and (iii) an intracellular domain; The method. [129.] A method of treating a proliferative disorder, cancer, an autoimmune disease, an infectious disease, or transplant 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 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; (c) 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; The method. [130.] A method of treating a proliferative disorder, cancer, an autoimmune disease, an infectious disease, or transplant 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 has been 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 CAR comprises (i) an ADBD that binds to the second AD, (ii) a transmembrane domain, and (iii) an intracellular domain; The method. [131.] The 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. [132.] A method according to any one of

[0122] to

[0130] , wherein the proliferative disorder or cancer is multiple myeloma. [133.] A method of treating 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; (c) the CAR comprises (i) an ADBD that binds to the second AD, (ii) a transmembrane domain, and (iii) an intracellular domain; The method. [134.] A method of treating hematological cancer, comprising administering to a patient an adapter; (a) the patient has been treated with a cell 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; (b) the adapter comprises (i) the first AD and (ii) an ADBD that binds to a second AD on the target cell; The method. [135.] A method of redirecting treatment of a hematological cancer, comprising administering an adapter to a patient; (a) the patient has been treated with a cell 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; (b) the adapter comprises (i) the first AD and (ii) an ADBD that binds to a second AD on a second target cell; The method. [136.] A method of treating hematological cancer, comprising administering to a patient an adapter; (a) the patient has been treated with a cell 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; (b) the adapter comprises (i) the first AD and (ii) an ADBD that binds to a second AD on the target cell; The method. [137.] A method of treating hematological cancer, comprising administering to a patient an adapter; (a) the patient has been treated with a cell 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; (b) the adapter comprises (i) the first AD and (ii) an ADBD that is an ASBD that binds to a second AD on the target cell; The method. [138.] 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 the target cell; (b) the patient has been treated with an adaptor comprising (i) the first AD and (ii) an antigenic determinant binding domain (ADBD) that binds to the second AD on the target cell; (c) 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; The method. [139.] A method for treating a hematological cancer, comprising administering to a patient cells expressing a chimeric antigen receptor (CAR); (a) 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; (b) the CAR comprises (i) an ADBD that binds to the first AD on the adapter, (ii) a transmembrane domain, and (iii) an intracellular domain; The method. [140.] 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 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; (c) 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; The method. [141.] 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 has been 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 CAR comprises (i) an ADBD that binds to the second AD, (ii) a transmembrane domain, and (iii) an intracellular domain; The method. [142.] The 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. [143.] A method according to any one of

[0133] to

[0141] , wherein the blood cancer is multiple myeloma. [144.] A method according to any one of

[90] to

[0131] , wherein the CAR comprises a single-chain variable fragment (scFv) ADBD. [145.] A method according to any one of

[90] to

[0143] , wherein the CAR comprises an alternative scaffold binding domain (ASBD) ADBD. [146.] The method described in

[0145] , 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. [147.] A method according to any one of

[90] to

[0146] , wherein the CAR comprises two ADBDs. [148.] The method described in

[0147] , wherein the CAR comprises an ASBD and an scFv. [149.] The method described in

[0147] , wherein the CAR comprises a D domain and an scFv. [150.] The method described in

[0147] , wherein the CAR comprises two ASBDs. [151.] The method described in

[0147] , wherein the CAR comprises two D domains. [152.] A method according to any one of

[90] to

[0151] , wherein the CAR intracellular domain is a signal transduction domain. [153.] The method described in

[0152] , wherein the CAR intracellular domain comprises a primary signaling domain. [154.] The method described in

[0152] , wherein the CAR intracellular domain comprises a CD3ζ primary signaling domain. [155.] The method described in

[0153] or

[0154] , wherein the CAR intracellular domain further comprises a costimulatory signaling domain. [156.] The method described in

[0155] , wherein the CAR intracellular domain comprises a costimulatory signaling domain selected from CD28, 41BB, CD27, and CD134. [157.] The method described in

[0156] , wherein the CAR intracellular domain comprises a 41BB costimulatory signaling domain. [158.] A method described in 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. [159.] The method described in

[0158] , wherein the CAR binds to BCMA, and optionally the CAR comprises a sequence selected from the group: SEQ ID NO:SEQ ID NO:44-338 and 339. [160.] The method described in

[0158] , wherein the CAR binds to CS1, and optionally the CAR comprises a sequence selected from the group: SEQ ID NO:780-794 and 795. [161.] The method described in

[0158] , wherein the CAR binds to CD123, and optionally the CAR comprises a sequence selected from the group: SEQ ID NO:340-772 and 773. [162.] The method described in

[0158] , wherein the CAR binds to CD19, and optionally, the CAR comprises a sequence selected from the group: SEQ ID NO: 1030-1058 and 1059. [163.] The method described in

[0158] , wherein the CAR binds to CD22, and optionally, the CAR comprises a sequence selected from the group: SEQ ID NO: 1060-1068 and 1069. [164.] The method described in

[0158] , wherein the CAR binds to HER2, and optionally the CAR comprises a sequence selected from the group: SEQ ID NO:800-839 and 840. [165.] The method described in

[0158] , 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. [166.] 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 consisting of SEQ ID NOs: 841 to 983 and 984. [167.] A method according to any one of

[90] to

[0166] , wherein the CAR comprises two ADBDs that bind to different targets. [168.] The method described in

[0167] , wherein the CAR binds to CD19 and CD123. [169.] The method described in

[0167] , wherein the CAR binds to CD22 and CD123. [170.] The method described in

[0167] , wherein the CAR binds to PDL1 and CD123. [171.] The method described in

[0167] , wherein the CAR binds to CS1 and BCMA. [172.] The method described in

[0168] , wherein the CAR comprises a first ASBD that binds to CD19 and a second ASBD that binds to C123. [173.] 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. [174.] The method described in

[0168] , wherein the CAR comprises a D domain that binds to CD19 and an scFv that binds to CD123. [175.] The method described in

[0169] , wherein the CAR comprises a first ASBD that binds to CD22 and a second ASBD that binds to CD123. [176.] 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. [177.] The method described in

[0169] , wherein the CAR comprises a D domain that binds to CD22 and an scFv that binds to CD123. [178.] The method described in

[0170] , wherein the CAR comprises a first ASBD that binds to PDL1 and a second ASBD that binds to CD123. [179.] 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. [180.] The method described in

[0170] , wherein the CAR comprises a D domain that binds to PDL1 and an scFv that binds to CD123. [181.] The method described in

[0168] , wherein the CAR comprises an ASBD that binds to CD19 and an scFv that binds to CD123. [182.] The method described in

[0171] , wherein the CAR comprises a first ASBD that binds to BCMA and a second ASBD that binds to CS1. [183.] 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. [184.] The method described in

[0171] , wherein the CAR comprises a D domain that binds to BCMA and an scFv that binds to CS1. [185.] The method described in

[0171] , wherein the CAR comprises a D domain that binds to CS1 and an scFv that binds to BCMA. [186.] The method of 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. [187.] 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. [188.] The method described in any one of

[90] to

[0187] , wherein the adapter comprises an ADBD that is an scFv. [189.] A method according to any one of

[90] to

[0187] , wherein the adapter comprises an ADBD that is an ASBD. [190.] The method described in

[0189] , 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. [191.] A method according to any one of

[90] to

[0190] , wherein the adapter comprises two ADBDs. [192.] The method described in

[0191] , wherein the two ADBDs are (a) identical, (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. [193.] The method described in

[0191] or

[0192] , wherein the adapter comprises two ASBDs. [194.] 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 consisting of SEQ ID NOs: 44 to 1079. [195.] The method described in

[0191] or

[0192] , wherein the adapter comprises an ADBD that is an scFv and an ADBD that is an ASBD. [196.] The method described in

[0195] , wherein the adapter comprises an ADBD that is an scFv and an ADBD that is a D domain. [197.] A method described in 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. [198.] The method described in

[0197] , wherein the adapter comprises an ADBD that binds to BCMA, and optionally, the ADBD comprises a sequence selected from the group: SEQ ID NO: 44 to 338 and 339. [199.] The method described in

[0197] , wherein the adapter comprises an ADBD that binds to CS1, and optionally, the ADBD comprises a sequence selected from the group: SEQ ID NOs: 780 to 794 and 795. [200.] The method described in

[0197] , wherein the adapter comprises an ADBD that binds to CD123, and optionally, the ADBD comprises a sequence selected from the group consisting of SEQ ID NOs: 340 to 772 and 773. [201.] The method described in

[0197] , wherein the adapter comprises an ADBD that binds to CD19, and optionally, the ADBD comprises a sequence selected from the group consisting of SEQ ID NOs: 1030 to 1058 and 1059. [202.] The method described in

[0197] , wherein the adapter comprises an ADBD that binds to CD22, and optionally, the ADBD comprises a sequence selected from the group: SEQ ID NOs: 1060 to 1068, and 1069. [203.] The method described in

[0197] , wherein the adapter comprises an ADBD that binds to HER2, and optionally, the ADBD comprises a sequence selected from the group consisting of SEQ ID NOs: 800 to 839 and 840. [204.] The method described in

[0197] , wherein the adapter comprises an ADBD that binds to TACI or BAFFR. [205.] The method described in

[0197] , wherein the adapter comprises an ADBD that binds to PDL1, and optionally, the ADBD comprises a sequence selected from the group consisting of SEQ ID NOs: 1010-1016, 1074-1078 and 1079. [206.] The method described in any one of

[90] to

[0205] , wherein the adapter is bispecific. [207.] The method described in

[0206] , wherein the adapter comprises an ADBD that binds to BCMA and an ADBD that binds to CS1. [208.] The method described in

[0206] , wherein the adapter comprises an ADBD that binds to CD19 and an ADBD that binds to CD123. [209.] The method described in

[0206] , wherein the adapter comprises an ADBD that binds to CD22 and an ADBD that binds to CD123. [210.] The method described in

[0206] , wherein the adaptor comprises an ADBD that binds to PDL1 and an ADBD that binds to CD123. [211.] The method according to any one of

[90] to

[0210] , wherein the target cells are tumor cells. [212.] The method described in

[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. [213.] The method described in

[0211] , wherein the tumor cells are multiple myeloma cells. [214.] The method described in

[0211] , wherein at least one target cell is a tumor cell. [215.] The method described in

[0211] , wherein the first and second target cells are tumor cells. [216.] The method described in

[0215] , wherein the first and second tumor cells are of the same type. [217.] The method described in

[0215] , wherein the first and second tumor cells are of different types. [218.] The method described in

[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. [219.] The method described in

[0214] or

[0215] , wherein the tumor cells are multiple myeloma. [220.] A method according to any one of

[90] to

[0219] , wherein the cells expressing the CAR are immune effector cells. [221.] The method described in

[0220] , wherein the immune effector cells are T cells. [222.] The method described in

[0220] , wherein the immune effector cells are NK cells. [223.] A method according to any one of

[90] to

[0222] , wherein cells expressing the CAR kill the target cells. [224.] Engineered 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) a genetic modification that eliminates expression of the human CD45 AD on the engineered cells. Including, the engineered cells are capable of directing an immune response to cells expressing CD45 AD in an in vitro assay, and the engineered cells do not express the CD45 AD. The engineered human immune effector cells. [225.] A method for killing a target cell, comprising contacting the engineered cell described in

[0224] with the target cell, wherein the target cell expresses human CD45. [226.] A method for delivering an immune response to a target cell, comprising contacting the engineered cell described in

[0224] with the target cell, wherein the target cell expresses human CD45. [227.] A method for directing an immune response to target cells in a subject, the method comprising administering a therapeutically effective amount of the engineered cells described in

[0224] to a subject in need of directing an immune response to target cells in the subject. [228.] A method for treating a proliferative disorder, cancer, autoimmune disease, infectious disease, or graft rejection in a subject, comprising administering a therapeutically effective amount of an engineered cell described in

[0224] to a subject in need of treatment for a proliferative disorder, cancer, autoimmune disease, infectious disease, or graft rejection in the subject. [229.] A method for lymphodepleting, comprising administering to a subject in need of lymphodepletion an effective amount of the engineered cells described in

[0224] . [230.] A method for depleting memory T cells, comprising administering to a subject in need of depleting memory T cells an effective amount of the engineered cells described in

[0224] . [231.] A method for treating an autoimmune disease or disorder, comprising administering to a subject in need of treatment for the autoimmune disease or disorder a therapeutically effective amount of an engineered cell described in

[0224] . [232.] A method for pretreating a subject for transplantation, comprising administering to a subject in need of pretreatment for transplantation an effective amount of the engineered cells described in

[0224] . [233.] A method for treating blood cancer, comprising administering to a subject in need of treatment for blood cancer a therapeutically effective amount of an engineered cell described in

[0224] . [234.] Engineered 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) a genetic modification that eliminates expression of at least one human CD45 AD on the engineered cells. Including, the first AD is not at least one human CD45 AD, and the engineered cells used in combination with an adaptor are capable of directing an immune response to cells expressing a CD45 AD in an in vitro assay, the adaptor comprising the first AD and a second ADBD that specifically binds to the at least one human CD45 AD, and the engineered cells do not express the at least one human CD45 AD; The engineered human immune effector cells. [235.] A method for killing a target cell, comprising the step of contacting an engineered cell described in

[0234] with an adaptor and the target cell, wherein the target cell expresses CD45, and the adaptor comprises a first AD and a second ADBD that specifically binds to human CD45 AD. [236.] A method for delivering an immune response to a target cell, comprising contacting an engineered cell described in

[0234] with an adaptor and the target cell, wherein the target cell expresses CD45, and the adaptor comprises a first AD and a second ADBD that specifically binds to human CD45 AD. [237.] A method for treating a hematological cancer, comprising contacting the engineered cells described in

[0234] with an adaptor and cancer cells, wherein the cancer cells express CD45, and the adaptor comprises a first AD and a second ADBD that specifically binds to human CD45 AD. [238.] A method for directing an immune response to target cells in a subject, comprising: (a) administering a therapeutically effective amount of the engineered cells described in

[0234] to a subject in need thereof to direct an immune response against target cells in the subject; (b) administering to the subject a therapeutically effective amount of an adapter comprising a first AD and a second ADBD that specifically binds to a human CD45 AD; The method comprising: [239.] A method for directing an immune response to target cells in a subject, comprising administering a therapeutically effective amount of the engineered cells described in

[0234] to a subject in need of directing an immune response to target cells in the subject, wherein the subject has previously been administered an adapter comprising the first AD and a second ADBD that specifically binds to human CD45 AD. [240.] A method for directing an immune response to 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 previously been administered an engineered cell described in

[0234] . [241.] A method of treating a proliferative disorder, cancer, an autoimmune disease, an infectious disease, or transplant rejection in a subject, comprising: (a) administering a therapeutically effective amount of the engineered cells of

[0234] to a subject in need of treatment for a proliferative disorder, cancer, an autoimmune disease, an infectious disease, or a transplant rejection in the subject; (b) administering to the subject a therapeutically effective amount of an adapter comprising a first AD and a second ADBD that specifically binds to a human CD45 AD; The method comprising: [242.] A method for treating a proliferative disorder, cancer, autoimmune disease, infectious disease, or graft rejection in a subject, comprising administering a therapeutically effective amount of the engineered cells described in

[0234] to a subject in need of treatment for a proliferative disorder, cancer, autoimmune disease, infectious disease, or graft rejection in the subject, wherein the subject has previously been administered an adapter comprising the first AD and a second ADBD that specifically binds to human CD45 AD. [243.] A method for treating a proliferative disorder, cancer, autoimmune disease, infectious disease, or graft rejection in a subject, comprising administering a therapeutically effective amount of an adapter comprising a first AD and a second ADBD that specifically binds to a human CD45 AD to a subject in need of treatment for a proliferative disorder, cancer, autoimmune disease, infectious disease, or graft rejection in the subject, wherein the subject has previously been administered an engineered cell described in

[0234] . [244.] A method for lymphocyte depletion, comprising: (a) administering a therapeutically effective amount of the engineered cells described in

[0234] to a subject in need of lymphocyte depletion; (b) administering to the subject a therapeutically effective amount of an adaptor comprising an adaptor comprising a first AD and a second ADBD that specifically binds to a human CD45 AD; The method comprising: [245.] A method for lymphodepleting, comprising administering a therapeutically effective amount of the engineered cells described in

[0234] to a subject in need of lymphodepletion, wherein the subject has previously been administered an adapter comprising the first AD and a second ADBD that specifically binds to a human CD45 AD. [246.] A method for lymphodepleting, comprising administering to a subject in need of lymphodepletion 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 previously been administered an engineered cell described in

[0234] . [247.] A method for depleting memory T cells, comprising: (a) administering a therapeutically effective amount of the engineered cells described in

[0234] to a subject in need of depleting memory T cells; (b) administering to the subject a therapeutically effective amount of an adapter comprising a first AD and a second ADBD that specifically binds to a human CD45 AD; The method comprising: [248.] A method for depleting memory T cells, comprising administering to a subject in need of depleting memory T cells 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 previously been administered the engineered cells described in

[0234] . [249.] A method for depleting memory T cells, comprising administering to a subject in need of depleting memory T cells 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 previously been administered the engineered cells described in

[0234] . [250.] A method of treating an autoimmune disease or disorder, comprising: (a) administering a therapeutically effective amount of the engineered cells described in

[0234] to a subject in need of treatment for an autoimmune disease or disorder; (b) administering to the subject a therapeutically effective amount of an adapter comprising a first AD and a second ADBD that specifically binds to a human CD45 AD; The method comprising: [251.] A method for treating an autoimmune disease or disorder, comprising administering a therapeutically effective amount of an adapter comprising a first AD and a second ADBD that specifically binds to a human CD45 AD to a subject in need of treatment for the autoimmune disease or disorder, wherein the subject has previously been administered an engineered cell described in

[0234] . [252.] A method for treating an autoimmune disease or disorder, comprising administering a therapeutically effective amount of an adapter comprising a first AD and a second ADBD that specifically binds to a human CD45 AD to a subject in need of treatment for the autoimmune disease or disorder, wherein the subject has previously been administered an engineered cell described in

[0234] . [253.] A method of preparing a subject for transplantation, comprising: (a) administering a therapeutically effective amount of the engineered cells described in

[0234] to a subject in need thereof to condition the subject for transplantation; (b) administering to the subject a therapeutically effective amount of an adapter comprising a first AD and a second ADBD that specifically binds to a human CD45 AD; The method comprising: [254.] A method for pretreating a subject for transplantation, comprising administering a therapeutically effective amount of the engineered cells described in

[0234] to a subject in need of pretreating the subject for transplantation, wherein the subject has previously been administered an adapter comprising the first AD and a second ADBD that specifically binds to a human CD45 AD. [255.] A method for pretreating a subject for transplantation, comprising administering to a subject in need of pretreating the subject for transplantation a therapeutically effective amount of an adaptor comprising a first AD and a second ADBD that specifically binds to a human CD45 AD, wherein the subject has previously received an engineered cell described in

[0234] . [256.] A method for treating hematological cancer, comprising: (a) administering a therapeutically effective amount of the engineered cells described in

[0234] to a subject in need of treatment for hematological cancer; (b) administering to the subject a therapeutically effective amount of an adapter comprising a first AD and a second ADBD that specifically binds to a human CD45 AD; The method comprising: [257.] A method for treating hematological cancer, comprising administering a therapeutically effective amount of the engineered cells described in

[0234] to a subject in need of treatment for hematological cancer, wherein the subject has previously been administered an adapter comprising the first AD and a second ADBD that specifically binds to human CD45 AD. [258.] A method for treating a hematological cancer, comprising administering a therapeutically effective amount of an adapter comprising a first AD and a second ADBD that specifically binds to a human CD45 AD to a subject in need of treatment for the hematological cancer, wherein the subject has previously been administered an engineered cell described in

[0234] . [259.] Engineered human immune effector cells, (a) a chimeric antigen receptor (CAR), the CAR comprising: (i) two or more antigenic determinant binding domains (ADBDs) including a first ADBD that binds to a first antigenic determinant (AD); (ii) a transmembrane domain; and (iii) an intracellular domain; and (b) a genetic modification that eliminates expression of the first AD on the engineered cells, such that the engineered cells do not express the first AD. The engineered human immune effector cell comprising: [260.] A method for killing a target cell, comprising contacting the engineered cell described in

[0259] with the target cell. [261.] A method for delivering an immune response to a target cell, comprising contacting the engineered cell described in

[0259] with the target cell. [262.] A method for directing an immune response to target cells in a subject, the method comprising administering a therapeutically effective amount of the engineered cells described in

[0259] to a subject in need of directing an immune response to target cells in the subject. [263.] A method for treating a proliferative disorder, cancer, autoimmune disease, infectious disease, or graft rejection in a subject, comprising administering a therapeutically effective amount of an engineered cell described in

[0259] to a subject in need of treatment for a proliferative disorder, cancer, autoimmune disease, infectious disease, or graft rejection in the subject. [264.] A method for lymphodepleting, comprising administering to a subject in need of lymphodepletion an effective amount of the engineered cells described in

[0259] . [265.] A method for pretreating a subject for transplantation, comprising administering to a subject in need of pretreatment for transplantation an effective amount of the engineered cells described in

[0259] . [266.] A method for treating blood cancer, comprising administering to a subject in need of treatment for blood cancer a therapeutically effective amount of an engineered cell described in

[0259] . [267.] A method for killing a target cell, comprising the step of contacting an engineered cell described in

[0259] with an adapter and the target cell, wherein the adapter comprises an AD recognized by the CAR and a second ADBD. [268.] A method for delivering an immune response to a target cell, comprising contacting an engineered cell described in

[0259] with an adapter and the target cell, wherein the adapter comprises an AD recognized by the CAR and a second ADBD capable of binding to a second AD on the target cell. [269.] A method for treating hematological cancer, comprising contacting an engineered cell described in

[0259] with an adapter and a cancer cell, wherein the adapter comprises an AD recognized by the CAR and a second ADBD capable of binding to a second AD on the cancer cell. [270.] A method for directing an immune response to target cells in a subject, comprising: (a) administering a therapeutically effective amount of the engineered cells described in

[0259] to a subject in need thereof to direct an immune response to target cells in the subject; (b) administering to the subject a therapeutically effective amount of an adapter comprising an AD recognized by a CAR and a second ADBD capable of binding to a second AD on the target cell; The method comprising: [271.] A method for directing an immune response to a target cell in a subject, comprising administering a therapeutically effective amount of an engineered cell described in

[0259] to a subject in need of directing an immune response to a target cell in the subject, wherein the subject has previously been administered an adapter comprising an AD recognized by a CAR and a second ADBD capable of binding to a second AD on the target cell. [272.] A method for directing an immune response to a target cell in a subject, comprising administering to a subject in need of directing an immune response to a target cell in the subject a therapeutically effective amount of an adapter comprising an AD recognized by a CAR and a second ADBD capable of binding to a second AD on the target cell, wherein the subject has previously been administered an engineered cell described in

[0259] . [273.] A method of treating a proliferative disorder, cancer, an autoimmune disease, an infectious disease, or transplant rejection in a subject, comprising: (a) administering a therapeutically effective amount of the engineered cells of

[0259] to a subject in need of treatment for a proliferative disorder, cancer, an autoimmune disease, an infectious disease, or a transplant rejection in the subject; (b) administering to the subject a therapeutically effective amount of an adapter comprising an AD recognized by the CAR and a second ADBD capable of binding to a second AD on a target cell; The method comprising: [274.] A method for treating a proliferative disorder, cancer, autoimmune disease, infectious disease, or graft rejection in a subject, comprising administering a therapeutically effective amount of an engineered cell described in

[0259] to a subject in need of treatment for a proliferative disorder, cancer, autoimmune disease, infectious disease, or graft rejection in the subject, wherein the subject has previously been administered an adapter comprising an AD recognized by a CAR and a second ADBD capable of binding to a second AD on a target cell. [275.] A method for treating a proliferative disorder, cancer, autoimmune disease, infectious disease, or graft rejection in a subject, comprising administering a therapeutically effective amount of an adapter comprising an AD recognized by a CAR and a second ADBD capable of binding to a second AD on a target cell to a subject in need of treatment for the proliferative disorder, cancer, autoimmune disease, infectious disease, or graft rejection, wherein the subject has previously been administered an engineered cell described in

[0259] . [276.] A method for lymphocyte depletion, comprising: (a) administering a therapeutically effective amount of the engineered cells described in

[0259] to a subject in need of lymphocyte depletion; (b) administering to the subject a therapeutically effective amount of an adapter comprising an AD recognized by the CAR and a second ADBD capable of binding to a second AD on a target cell; The method comprising: [277.] A method for lymphodepleting, comprising administering a therapeutically effective amount of the engineered cells described in

[0259] to a subject in need of lymphodepletion, wherein the subject has previously been administered an adapter comprising an AD recognized by the CAR and a second ADBD capable of binding to a second AD on a target cell. [278.] A method for lymphodepleting, comprising administering to a subject in need of lymphodepletion a therapeutically effective amount of an adapter comprising an AD recognized by a CAR and a second ADBD capable of binding to a second AD on a target cell, wherein the subject has previously been administered an engineered cell described in

[0259] . [279.] A method of preparing a subject for transplantation, comprising: (a) administering a therapeutically effective amount of the engineered cells described in

[0259] to a subject in need thereof to condition the subject for transplantation; (b) administering to the subject a therapeutically effective amount of an adapter comprising an AD recognized by the CAR and a second ADBD capable of binding to a second AD on a target cell; The method comprising: [280.] A method for pretreating a subject for transplantation, comprising administering a therapeutically effective amount of the engineered cells described in

[0259] to a subject in need of pretreatment for transplantation, wherein the subject has previously been administered an adapter comprising an AD recognized by the CAR and a second ADBD capable of binding to a second AD on the target cell. [281.] A method for pretreating a subject for transplantation, comprising administering to a subject in need of pretreatment for transplantation a therapeutically effective amount of an adapter comprising an AD recognized by a CAR and a second ADBD capable of binding to a second AD on a target cell, wherein the subject has previously been administered an engineered cell described in

[0259] . [282.] A method for treating hematological cancer, comprising: (a) administering a therapeutically effective amount of the engineered cells described in

[0259] to a subject in need of treatment for hematological cancer; (b) administering to the subject a therapeutically effective amount of an adapter comprising an AD recognized by the CAR and a second ADBD capable of binding to a second AD on a target cell; The method comprising: [283.] A method for treating hematological cancer, comprising administering a therapeutically effective amount of the engineered cells described in

[0259] to a subject in need of treatment for hematological cancer, wherein the subject has previously been administered an adapter comprising an AD recognized by the CAR and a second ADBD capable of binding to a second AD on a target cell. [284.] A method for treating hematological cancer, comprising administering to a subject in need of treatment for hematological cancer a therapeutically effective amount of an adapter comprising an AD recognized by a CAR and a second ADBD capable of binding to a second AD on a target cell, wherein the subject has previously been administered an engineered cell described in

[0259] . [285.] Engineered human immune effector cells, (a) a chimeric antigen receptor (CAR), the 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 eliminates expression of a second AD on the engineered cells. Including, the engineered cells used in combination with an adaptor are capable of directing an immune response to cells expressing the second AD in an in vitro assay, the adaptor comprising the first AD and a second ADBD that specifically binds to the second AD, and the engineered cells do not express the second AD. The engineered human immune effector cells. [286.] Engineered human immune effector cells, (a) a chimeric antigen receptor (CAR), wherein the CAR comprises: (i) two or more antigenic determinant binding domains (ADBDs), including a first 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 eliminates expression of a second AD on the engineered cells. Including, the engineered cells used in combination with an adaptor are capable of directing an immune response to cells expressing the second AD in an in vitro assay, the adaptor comprising the first AD and a second ADBD that specifically binds to the second AD, and the engineered cells do not express the second AD. The engineered human immune effector cells. [287.] A method for directing an immune response to target cells in a subject, comprising administering a therapeutically effective amount of an engineered cell described in

[0285] or

[0286] to a subject in need of directing an immune response to target cells in the subject. [288.] A method for treating a proliferative disorder, cancer, autoimmune disease, infectious disease, or graft rejection in a subject, comprising administering a therapeutically effective amount of an engineered cell described in

[0285] or

[0286] to a subject in need of treatment for a proliferative disorder, cancer, autoimmune disease, infectious disease, or graft rejection in the subject. [289.] A method for lymphodepleting, comprising administering to a subject in need of lymphodepletion an effective amount of an engineered cell described in

[0285] or

[0286] . [290.] A method for pretreating a subject for transplantation, comprising administering to a subject in need of pretreatment for transplantation an effective amount of an engineered cell described in

[0285] or

[0286] . [291.] A method for treating blood cancer, comprising administering to a subject in need of treatment for blood cancer a therapeutically effective amount of an engineered cell described in

[0285] or

[0286] . [292.] A method for killing a target cell, comprising the step of contacting an engineered cell described in

[0285] or

[0286] with an adaptor and a target cell, wherein the adaptor 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. [293.] A method for delivering an immune response to a target cell, comprising contacting an engineered cell described in

[0285] or

[0286] with an adaptor and a target cell, wherein the adaptor 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. [294.] A method for treating a blood cancer, comprising contacting an engineered cell described in

[0285] or

[0286] with an adaptor and a cancer cell, wherein the adaptor comprises a first AD and a second ADBD that specifically binds to the second AD, and the second AD is expressed on the cancer cell. [295.] A method for directing an immune response to target cells in a subject, comprising: (a) administering a therapeutically effective amount of the engineered cells of

[0285] or

[0286] to a subject in need thereof to direct an immune response to target cells in the subject; (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 second AD, wherein the second AD is expressed on the target cell; The method comprising: [296.] A method for directing an immune response to target cells in a subject, comprising administering a therapeutically effective amount of an engineered cell described in

[0285] or

[0286] to a subject in need of directing an immune response to target cells in the subject, wherein the subject has been administered an adapter comprising a first AD and a second ADBD that specifically binds to a second AD, and the second AD is expressed on the target cells. [297.] A method for directing an immune response to target cells in a subject, comprising the step of administering to a subject in need of directing an immune response to target cells in 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 subject has been administered an engineered cell described in

[0285] or

[0286] , and the second AD is expressed on the target cells. [298.] A method of treating a proliferative disorder, cancer, an autoimmune disease, an infectious disease, or transplant rejection in a subject, comprising: (a) administering a therapeutically effective amount of the engineered cells of

[0285] or

[0286] to a subject in need of treatment for a proliferative disorder, cancer, autoimmune disease, infectious disease, or transplant rejection; (b) administering to the subject a therapeutically effective amount of an adapter comprising a first AD and a second ADBD that specifically binds to a second AD, wherein the second AD is expressed on a target cell associated with the proliferative disorder, cancer, autoimmune disease, infectious disease, or graft rejection; The method comprising: [299.] A method for treating a proliferative disorder, cancer, autoimmune disease, infectious disease, or graft rejection in a subject, comprising administering a therapeutically effective amount of an engineered cell described in

[0285] or

[0286] to a subject in need of treatment for a proliferative disorder, cancer, autoimmune disease, infectious disease, or graft rejection, wherein the subject has been administered an adapter comprising a first AD and a second ADBD that specifically binds to a second AD, and the second AD is expressed on a target cell associated with the proliferative disorder, cancer, autoimmune disease, infectious disease, or graft rejection. [300.] A method for treating a proliferative disorder, cancer, autoimmune disease, infectious disease, or graft rejection in a subject, comprising administering a therapeutically effective amount of an adapter comprising a first AD and a second ADBD that specifically binds to the second AD to a subject in need of treatment for the proliferative disorder, cancer, autoimmune disease, infectious disease, or graft rejection, wherein the subject has previously been administered an engineered 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 graft rejection. [301.] A method for lymphocyte depletion, comprising: (a) administering a therapeutically effective amount of the engineered cells described in

[0285] or

[0286] to a subject in need of lymphocyte depletion; (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 second AD, wherein the second AD is expressed on a lymphocyte target cell; The method comprising: [302.] A method for lymphodepleting, comprising administering a therapeutically effective amount of an engineered cell described in

[0285] or

[0286] to a subject in need of lymphodepletion, wherein the subject has previously been administered an adapter comprising a first AD and a second ADBD that specifically binds to a second AD, and the second AD is expressed on a lymphocyte target cell. [303.] A method for lymphodepleting lymphocytes, comprising administering to a subject in need of lymphodepletion 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 previously received an engineered cell described in

[0285] or

[0286] , and the second AD is expressed on lymphocyte target cells. [304.] A method of preparing a subject for transplantation, comprising: (a) administering a therapeutically effective amount of the engineered cells described in

[0285] or

[0286] to a subject in need of conditioning for transplantation; (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 second AD, wherein the second AD is expressed on a target cell associated with the transplant; The method comprising: [305.] A method for pretreating a subject for transplantation, comprising administering a therapeutically effective amount of an engineered cell described in

[0285] or

[0286] to a subject in need of pretreatment for transplantation, wherein the subject has previously been administered an adapter comprising a first AD and a second ADBD that specifically binds to a second AD, and the second AD is expressed on target cells associated with the transplantation. [306.] A method for pretreating a subject for transplantation, comprising administering to a subject in need of pretreatment for transplantation 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 previously received an engineered cell described in

[0285] or

[0286] , and the second AD is expressed on target cells associated with the transplantation. [307.] A method for treating hematological cancer, comprising: (a) administering a therapeutically effective amount of the engineered cells described in

[0285] or

[0286] to a subject in need of treatment for a hematological cancer; (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 second AD, wherein the second AD is expressed on a target cell associated with the hematological cancer; The method comprising: [308.] A method for treating a hematological cancer, comprising administering a therapeutically effective amount of an engineered cell described in

[0285] or

[0286] to a subject in need of treatment for hematological cancer, wherein the subject has been administered an adapter comprising a first AD and a second ADBD that specifically binds to a second AD, and the second AD is expressed on a target cell associated with the hematological cancer. [309.] A method for treating a hematological cancer, comprising administering a therapeutically effective amount of an adapter comprising a first AD and a second ADBD that specifically binds to the second AD to a subject in need of treatment for the hematological cancer, wherein the subject has been administered an engineered cell described in

[0285] or

[0286] , and the second AD is expressed on a target cell associated with the hematological cancer. [310.] An isolated adaptor polypeptide comprising (1) an antigenic determinant (AD) and (b) one or more antigenic determinant binding domains (ADBDs), at least one ADBD specifically binds to human CD45 AD, and contacting the adapter with a CD45 AD-expressing target cell in the presence of an engineered cell according to claim 234 enables the engineered cell to direct an immune response to the target cell in an in vitro assay. The isolated adaptor polypeptide. [Brief explanation of the drawings]

[0091] [Figure 1]Figures 1A-1D: Using effector cell-to-target cell ratios ranging from 1:4 to 1:64, the cytolytic activity of CD123-specific (cg06) and BCMA-specific (bc-40) CARs was compared to a CAR with no known target specificity (α3D) against a panel of tumors. Briefly, 20,000 T cells expressing bc40, cg06, or α3D CARs 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] Figures 2A-2B: Adapter binding correlates with adapter and CAR binding specificity. 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 (4°C, 20 min), washed, and then stained with anti-HIS PE (clone J095G 46, 4°C, 20 min). Figure 2B shows CAR expression (clone L5) based on FLAG staining compared to mock-transduced Jurkat cells. [Figure 3-1]Figures 3A-3C show the specificity of adapter binding between corresponding CAR:adapter and target:adapter pairs to induce target cell lysis. In Figure 3A, 40,000 CD123+ BCMA-MOLM13-GFP / luciferase cells were incubated for 16 hours with various adapters in the presence or absence of 20,000 T cells transduced with a BCMA-binding CAR (bc40) (1:2 E:T ratio, donor D14-053017, day 7). After 16 hours, cells were washed and luciferase activity was assessed. Percent lysis was assessed based on the percentage of MOLM13-GFP / luciferase T cells cultured in the absence of adapter protein. A control of CD123-specific CAR T cells (cg06) cultured at the same ratio was used as a positive control for lysis. In Figure 3B, 40,000 CD123+ BCMA-MOLM13-GFP / luciferase cells were incubated for 16 hours with the CD123(cg06)-AFP(p26) adaptor in the presence or absence of 20,000 T cells transduced with an AFP-conjugated CAR (Af03 or Af05) (1:2 E:T ratio, donor D16-061317, day 7). After 16 hours, cells were washed and luciferase activity was assessed. Percent lysis was assessed based on wells of MOLM13-GFP / luciferase cultured in the absence of T cells or adaptor protein. A control of CD123-specific CAR T cells (cg06) cultured at the same ratio was used as a positive control for lysis. In Figure 3C, 40,000 BCMA+ NCI H929-GFP / luciferase cells were incubated for 16 hours with the bc40-AFP(p26) adaptor in the presence or absence of 10,000 T cells (1:4 E:T ratio, donor D15-062017, day 8) that were mock-transduced or transduced with an AFP-conjugated CAR (Af03 or Af05). After 16 hours, cells were washed and luciferase activity was assessed. Percent lysis was assessed based on the wells of NCI-H929-GFP / luciferase cultured in the absence of T cells or adaptor protein. [Figure 3-2] See description of Figure 3-1. [Figure 4-1] Figures 4A-4D show that the specificity of the corresponding CAR:adapter and target:adapter adapter binding results in 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 adapters 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). In Figures 4C and 4D, donor D15-062017 T cells transduced with an AFP-binding CAR (Af03 and Af05) were cultured overnight with the cg06-AFP(p26) adapter in the presence or absence of CD123+BCMA-MOLM13 cells (25,000 T cells and target cells). Culture supernatants were collected and evaluated for the production of IL-2 (Fig. 4C) and IFN-γ (Fig. 4D). [Figure 4-2] See description of Figure 4-1. [Figure 5] The specificity of the corresponding CAR:adapter and target:adapter adapter binding drives CAR T cell proliferation. Donor D16-062717 cells transduced with an AFP-conjugated CAR (Af03) were CFSE-labeled (0.5 μM for 10 minutes) and then cultured for 72 hours in the presence of either a CD123-specific or BCMA-specific adapter, with or without mitomycin C-treated CD123-BCMA+ NCI-H929 cells (25,000 cells). At 72 hours, cells were stained for CD3 and then analyzed for absolute CD3+ cell counts by flow cytometry. [Figure 6]Figures 6A and 6B show that adapter binding of the corresponding CAR:adapter and target:adapter leads to signaling by Jurkat NFAT-luciferase reporter cells expressing the CAR. In Figure 6A, 50,000 reporter cells pre-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 assessed for luciferase activity. In Figure 6B, 50,000 reporter cells pre-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 assessed for luciferase activity. [Figure 7] Figures 7A and 7B show that a CD123-specific adaptor bearing a BCMA antigenic determinant can function with either 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 (1:2 E:T ratio, donor D14-062717, day 9) transduced with a nonspecific CAR (α3D), a BCMA-binding D-domain CAR (bc40), or a BCMA-binding scFv CAR (c11D5-3). After 16 hours, cells were washed and luciferase activity was assessed. Percent lysis was assessed based on the percentage of MOLM13-GFP / luciferase cells cultured in the absence of T cells or adaptor protein. The solid lines show the calculated three-parameter nonlinear curves, and the dashed line for c11D5-3 is for illustration purposes only. In Figure 7B, CD123+BCMA-MOLM13-GFP / luciferase cells were cultured with transduced T cells in the absence of adaptor protein in the same experiment as in Figure 7A. [Figure 8]Figures 8A and 8B show that AFP-specific CARs can simultaneously possess CD123 and BCMA binding capabilities through incubation with multiple adaptor proteins. 105 Jurkat NFAT-luciferase transduced with AFP(p26)-binding CAR (af03) were incubated with 0.5 μg total adaptor protein at various BCMA-specific to CD123-specific adaptor ratios (4°C for 20 minutes), washed, and then incubated with CD123-Fc and biotinylated BCMA (0.5 μg each) (4°C for 20 minutes), washed, and then binding was detected with anti-FcA488 and streptavidin-PE. Figure 8A shows flow cytometry analysis of CD123 binding and BCMA binding to their respective target proteins, and Figure 8B provides a comparison of mean fluorescence intensity (MFI) of the A488 MFI (CD123 binding, left axis) and PE MFI (BCMA binding, right axis) flow cytometry data shown in Figure 8A. [Figure 9] Figures 9A and 9B show that dual-binding domain adaptor proteins induce enhanced signaling by CAR-expressing Jurkat NFAT-luciferase reporter cells compared to single-binding domain adaptor proteins. In Figure 9A, 50,000 reporter cells pre-transduced with an AFP (p26 domain)-linked CAR (af03) were cultured for 5 hours in the presence of the CD123-specific Cg06 adaptor (Cg06-p26) or the Cg06 dual adaptor protein (Cg06-p26-Cg06) in the presence of 50,000 CD123+ or CD123-deficient MOLM13 cells, and then assessed for luciferase activity. Using genetic engineering techniques, CD123-deficient cells were generated. In Figure 9B, 50,000 reporter cells pre-transduced with an AFP (p26 domain)-binding 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 adaptor (Bc40-p26) or the Bc40 dual adaptor protein (Bc40-p26-Bc40), and then assessed for luciferase activity. [Figure 10] Figure 1 shows that binding of truncated and full-length p26 to human FcRn is pH dependent. [Figure 11A] Figures 11A-11D show that adaptors containing CS1 (SLAMF7, CRACC, CD319)-specific ADBDs modulate intracellular signaling and killing of CS1-positive tumors. In Figure 11A, the cc02 and cc08 ADBDs exhibited the most potent NFAT signaling when cultured in the presence of af59-CAR-expressing JNL10 cells (Figure 11A) and the CS1-positive tumor cell line MM.1S. Figure 11B shows that the bispecific bc98-p26-cc02 adaptor, which can bind both CS1 and BCMA, was more potent at transducing signals than the monospecific BCMA-binding bc98-p26-α3DQ19E adaptor and the monospecific CS1-binding α3DQ19E-p26-cc02 adaptor. Figures 11C and 11D show that the bispecific bc98-p26-cc02 is an effective adaptor at killing HT929 (high expression of both BCMA and CS1; Figure 11C) and MM.1S (high BCMA, low CS1; Figure 11D). [Figure 11B] See legend to Figure 11A. [Figure 11C] See legend to Figure 11A. [Figure 11D] See legend to Figure 11A. [Figure 12A]Figures 12A-12C show that adapters containing HER2-binding ADBDs induce signaling in Af59-CAR-expressing JNL10 cells cultured with HER2-positive SKBR3 tumors. Figure 12A shows that the eb08 HER2-binding ABDB-containing adapter was the most potent stimulator in this assay. Figure 12B shows that NFAT signaling in JNL10 cells mediated by eb08-containing adapters was greater than that mediated by zHER:4-containing adapters, comparable to that mediated by 9.29-containing adapters, and less than that mediated by G3 and zHER2:342. Figure 12C demonstrates that HER2-binding eb08- or eb04-containing adapters modulate tumor lysis in a dose-dependent manner. [Figure 12B] See legend to Figure 12A. [Figure 12C] See legend to Figure 12A. [Figure 13] T cells transduced with a p26-linked CAR ("ARC-T") in combination with a BCMA-specific adaptor ("SPRX-BCMA") eliminate MM.1S tumor cells in vivo comparably to BCMA-targeting CAR-T cells ("CART-ddBCMA"). Total flux measured by dorsal imaging of NGS mice implanted with MM.1S cells expressing GFP and luciferase after administration of BCMA-targeting CAR-T cells or a combination of T cells transduced with a p26-linked CAR and a BCMA-specific adaptor. [Figure 14] T cells transduced with a p26-binding CAR ("ARC-T") show comparable activity in vivo with intermittent administration of a BCMA-specific adaptor ("SPRX-BCMA") in a disseminated model of BCMA+ B-cell leukemia (NALM6-BCMA). Total flux measured by dorsal imaging of NGS mice transplanted with NALM6-BCMA cells expressing GFP and luciferase after administration of 10 T cells transduced with a p26-binding CAR and a BCMA-specific adaptor. [Figure 15]In vivo residence time of BCMA-specific adaptor ("SPRX-BCMA") on NALM6-BCMA tumor cells in mice after intravenous administration. DETAILED DESCRIPTION OF THE INVENTION

[0092] Detailed Description 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.

[0093] Whenever an embodiment is described herein in terms of "comprising," it is understood that other similar embodiments described in terms of "consisting of" and / or "consisting essentially of" are also provided. However, when used as transitional phrases in the claims, each should be interpreted separately in the appropriate legal and factual context (e.g., "comprising" is considered the more open-ended phrase, while "consisting of" is more exclusive, and "consisting essentially of" achieves a middle ground).

[0094] As used herein, the singular forms "a," "an," and "the" include plural references unless otherwise indicated.

[0095] The term "and / or" as used herein in phrases such as "A and / or B" is intended 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" is intended to encompass 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).

[0096] The term "about," as used herein, when referring to measurable values, such as amounts, temporal durations, and other measurable values ​​known in the art, is meant to encompass variations of ±20%, or in some embodiments ±10%, or in some embodiments ±5%, or in some embodiments ±1%, or in some embodiments ±0.1% from the specified value, since such variations are reasonable for performing the disclosed methods.

[0097] The terms "protein" and "polypeptide" are used interchangeably herein to refer to biological polymers comprising units derived from amino acids linked through peptide bonds, and a protein may be composed of two or more polypeptide chains.

[0098] "Cell surface receptor" refers to molecules and molecular complexes that can receive signals and transmit such signals across the cell membrane of a cell. An example of a cell surface receptor provided herein is an activated integrin receptor, such as an activated αβ integrin receptor on a metastatic cell. As used herein, "cell surface receptor" also includes molecules expressed on the cell surface that contain a CAR that can bind to an antigenic determinant target of interest. The term "receptor" refers to a cell-associated protein that binds to or otherwise interacts with a molecule (e.g., a ligand) and mediates the effect of the ligand on the cell. In some embodiments, the molecule that interacts with the receptor is a biologically active molecule. Membrane-bound cell surface receptors are characterized by a multi-domain structure that includes an extracellular ligand-binding domain, a transmembrane domain, and an intracellular effector domain that is typically involved in signal transduction.

[0099] As used herein, the term "chimeric antigen receptor" or "CAR" or "CARs" refers to an engineered chimeric polypeptide that transfers antigen or target specificity to a cell, such as an immune cell (e.g., a naive T cell, a central memory T cell, an effector memory T cell, a T cell such as an NK cell, an NKT cell, 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, a CAR can activate or inactivate a cytotoxic cell in which it is placed, modulate the anti-tumor activity of the cell, or otherwise modulate the immune response of the cell. In some embodiments, a CAR comprises 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 a natural killer cell receptor (NKR). In some embodiments, the 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, the CAR comprises more than one antigenic determinant binding domain. In some embodiments, the CAR comprises more than one antigenic determinant binding domain that binds to different antigenic determinants of the same antigen, different antigenic determinants on different antigens, or antigenic determinants expressed by different target cells.

[0100] As used herein, the term "immune cell" refers to cells 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.

[0101] 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.

[0102] "Autologous," as the term is used herein, refers to any material that originates from the same individual into whom it is later reintroduced.

[0103] "Allogeneic" as used herein refers to any material derived from a different animal of the same species as the individual into which the material is introduced. Two or more individuals are said to be allogeneic to each other if the genes at one or more loci are not identical. In some aspects, allogeneic materials from individuals of the same species may be genetically sufficiently different to interact antigenically.

[0104] 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 native source, e.g., from blood or PBMCs, as described herein or known in the art. In certain embodiments, the effector cells are human effector cells.

[0105] The term "effector function" refers to a specialized immune function of a differentiated cell. The effector function of a T cell can be, for example, cytolytic activity or helper activity, including secretion of cytokines.

[0106] The term "immune response" as used herein refers to immunity, including but not limited to innate immunity, humoral immunity, cellular immunity, immune, inflammatory response, acquired (adaptive) immunity, autoimmunity, and / or overactive immunity. Indicators of immune response may include the secretion of cytokines by immune cells, expansion of immune cell populations, production of antibodies, degranulation of cytotoxic cells, and killing of target cells. Such indicators can be routinely measured using readily available assays known in the art, such as ELISA or ELISpot.

[0107] As used herein, the term "adapter" refers to a multidomain soluble protein that comprises an antigenic determinant (AD) and an antigenic determinant binding domain (ADBD), where the ADBD binds to a second AD. In addition to an AD and an ADBD, the adapter can comprise additional ADs, additional ADBDs, and / or other additional domains.

[0108] The terms "antibody" and "immunoglobulin," used interchangeably herein, include whole antibodies. Whole antibodies comprise at least two heavy (H) chains and two light (L) chains interconnected by disulfide bonds. Each heavy chain is composed of a heavy chain variable region (abbreviated herein as VH) and a heavy chain constant region. The heavy chain constant region is composed of three domains, CH1, CH2, and CH3. Each light chain is composed of a light chain variable region (abbreviated herein as VL) and a light chain constant region. The light chain constant region is composed of one domain, Cl. The VH and VL regions can be further subdivided into hypervariable regions called complementarity-determining regions (CDRs), interspersed with more conserved regions called framework regions (FWs). Each VH and VL is composed of three CDRs and four FWs, arranged from the amino terminus to the 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 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.

[0109] As used herein, terms such as "antibody fragment" include 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. The antibody fragments described herein may 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, such as nanobodies (either VL or VH)), 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 an effector domain or portion thereof, or other functional region of an antibody, such as a salvage receptor binding epitope or portion thereof.

[0110] 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, linked by a linker peptide. An scFv may contain VL-linker-VH or VH-linker-VL. ScFv antibodies are generally 220-250 amino acids in length and contain a linker that is 10-25 amino acids in length.

[0111] 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 an appropriate immunoglobulin heavy chain constant region 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. It is considered within the level of ordinary skill in the art to select a particular immunoglobulin heavy chain constant region sequence from a given immunoglobulin class and subclass to achieve a particular result. 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 Fcγ or a homologous domain in any of IgA, IgD, IgE, or IgM. Furthermore, it is contemplated that substitution or deletion 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 introducing amino acid substitutions into the upper CH2 region to create an Fc variant with reduced affinity for Fc receptors (Cole, J. Immunol. 159:3613 (1997)).

[0112] "Antibody-dependent cell-mediated cytotoxicity" or "ADCC" refers to a cell-mediated reaction in which nonspecific cytotoxic cells expressing Fc receptors (e.g., natural killer (NK) cells, neutrophils, and macrophages) recognize bound antibodies on target cells, subsequently causing lysis (or other cytotoxic effects) of the target cells. To assess the 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. No. 5,500,362 or U.S. Pat. No. 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, the ADCC activity of a molecule of interest can be assessed in vivo in an animal model, such as that disclosed in Clynes et al., PNAS 95:652-656 (1998).

[0113] The term "antigenic determinant binding domain" or "ADBD," as used herein, refers to a sequence of a polypeptide (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.

[0114] In certain embodiments, the ADBD is a non-antibody-scaffold-based polypeptide sequence sufficient to confer recognition and specific binding to a target antigen 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 antigen 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.

[0115] The terms "specifically bind" or "having selective affinity for" mean that a binding agent, such as an adapter or CAR, reacts or associates with an epitope, protein, or target molecule more frequently, more rapidly, for a longer duration, with higher affinity, or some combination of the above, than with alternative agents, including proteins unrelated to the target epitope. Due to sequence identity between homologous proteins in different species, specific binding can, in some embodiments, involve binding agents that recognize proteins or targets in more than one species. Similarly, due to homology within certain regions of the polypeptide sequences of different proteins, specific binding can involve binding agents that recognize more than one protein or target. It is understood that, in some embodiments, a binding agent that specifically binds a first target may or may not specifically bind a second target. Thus, "specific binding" does not necessarily require (but can include) exclusive binding, e.g., binding to a single target. Thus, a binding agent can, in some embodiments, specifically bind more than one target. In some embodiments, multiple targets may be bound by the same antigen binding site on the binding agent.

[0116] The terms "linker," "spacer," and "hinge" are used interchangeably herein to refer to a peptide or other chemical bond located between two or more originally independent functional domains of an adapter or CAR. For example, a linker can be located between the antigenic determinant domain and the antigenic determinant binding domain of an adapter. Similarly, a linker can be located between two antigenic determinant binding domains or an antigen binding domain and the transmembrane domain of a CAR. Suitable linkers for coupling two or more domains of an adapter are described herein and / or will otherwise be apparent to those skilled in the art.

[0117] As used herein, the term "operably linked" refers to two molecules joined such that each retains at least some level of the functional activity that each molecule has alone (assuming that each molecule has functional activity). In embodiments where one molecule lacks functional activity, a molecule is operably linked to another molecule if the other molecule retains at least some level of that functional activity. Operatively linked can also refer to the linkage of two non-functional molecules. Two molecules can be "operably linked" regardless of whether they are joined directly or indirectly (e.g., via a linker).

[0118] "Target" refers to any molecule or combination of molecules that can be bound by an adapter or CAR, or a component of an adapter or CAR, such as an antigenic determinant binding domain.

[0119] 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.

[0120] Expressions such as "binding affinity for a target," "binding to a target," and similar expressions known in the art refer to the affinity constant, a property of a polypeptide that can be directly measured by determining, for example, the amount of adapter associated and dissociated at a given antigen concentration.To characterize molecular interactions, various methods can be used, including but not limited to, competitive analysis, equilibrium analysis, and microcalorimetry, as well as real-time interaction analysis based on surface plasmon resonance interaction (for example, using a Biacore® instrument).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).

[0121] The terms "antigenic determinant" and "epitope" are used interchangeably herein and refer to a portion of any molecule (e.g., a target of interest or an adapter) that can be recognized and specifically bound by a specific binding factor (e.g., an adapter or CAR). When the molecule being recognized is a polypeptide, epitopes can be formed from adjacent amino acids as well as non-adjacent amino acids juxtaposed by tertiary folding of the protein and / or other chemically active surface groups of the molecule (such as carbohydrates). Epitopes formed from adjacent amino acids are typically retained upon protein denaturation, whereas epitopes formed by tertiary folding are typically lost upon protein denaturation. Epitopes typically contain at least three amino acids, more commonly at least five or eight to ten amino acids, in a unique spatial conformation.

[0122] As used herein, "derived from" refers to a relationship between a first molecule and a second molecule. "Derived from" generally refers to a structural similarity between the first and second molecules and does not imply or imply a limitation to the process or source from which the first molecule is derived from the second molecule. For example, in the case of an intracellular signaling domain derived from the CD3ζ molecule, the intracellular signaling domain retains sufficient CD3ζ structure so that the intracellular signaling domain has the required function, i.e., the ability to generate a signal under appropriate conditions. "Derived from" does not imply or imply a limitation to a particular process for generating the intracellular signaling domain; for example, "derived from" does not mean that one must start from the CD3ζ sequence and delete undesired sequences or impose mutations to arrive at the intracellular signaling domain to provide the intracellular signaling domain.

[0123] The term "naturally occurring," when used in reference to biological materials such as nucleic acid molecules, polypeptides, antigenic determinants, and host cells, refers to something that is found in nature and has not been modified by humans. Conversely, "non-natural" or "synthetic," when used in reference to biological materials, refers to something that is not found in nature and has not been modified by humans.

[0124] As used herein in reference to a reference sequence, a "modification" includes substitutions, deletions, insertions and / or additions in the sequence compared to the corresponding amino acid positions in the reference sequence.

[0125] A "substitution" with respect to a reference sequence refers to the replacement of a particular amino acid residue with a different amino acid residue at the corresponding amino acid position in the reference sequence.

[0126] 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 and include 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)), nonpolar 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, substituting tyrosine for phenylalanine is a conservative substitution. In one embodiment, a conservative substitution in the sequence of an adapter or CAR results in retained specific binding of the adapter or CAR containing the substitution to the target of interest to which the adapter or CAR binds. Methods for identifying conservative and non-conservative substitutions of nucleotides and amino acids 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)).

[0127] A "non-conservative" amino acid substitution is one in which one amino acid residue is replaced with another amino acid residue having a dissimilar side chain. In one embodiment, a non-conservative substitution in the sequence of an adapter or CAR results in retained specific binding of the adapter or CAR containing the substitution to the target of interest to which the adapter or CAR binds.

[0128] 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. Further examples of unnatural amino acids that can be substituted in the 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 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.

[0129] The terms "polynucleotide" and "nucleic acid," used interchangeably herein, refer to polymeric forms of nucleotides of any length, either ribonucleotides or deoxyribonucleotides. 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 non-naturally occurring nucleosides or nucleotides. In some embodiments, the modified mRNA contains 2-thiouridine, pseudouridine, or 1-methylpseudouridine.

[0130] 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 an appropriate 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.

[0131] As used herein, the terms "vector," "cloning vector," and "expression vector" refer to a vehicle that can maintain or amplify a nucleic acid sequence (e.g., an adapter or CAR-encoding sequence) within a host cell (e.g., a cloning vector) or can be introduced into a host cell to transform the host and promote expression (e.g., transcription and translation) of the introduced sequence. Vectors include plasmids, phages, viruses, etc.

[0132] 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, and due to natural, accidental, or deliberate mutations and / or variations, the progeny may not necessarily be completely identical (in morphology or in total DNA complement) to the original parent cell. A host cell includes cells that have been transfected or infected in vivo, in vitro, or ex vivo with a nucleic acid encoding an adapter or CAR. 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.

[0133] As used herein, the terms "pharmaceutically acceptable" or "physiologically tolerable" and grammatical variations thereof are used interchangeably when referring to compositions, carriers, diluents, and reagents and indicate that the substance can be administered to or on a human without producing therapeutically prohibitive undesirable physiological effects, such as nausea, dizziness, stomach upset, and other therapeutically prohibitive undesirable physiological effects known in the art.

[0134] "Parenteral" administration of the immunogenic compositions includes, for example, subcutaneous (sc), intravenous (iv), intramuscular (im), or intrasternal injection or infusion techniques.

[0135] The term "stimulate" or "stimulation" refers to a primary response induced by the binding of a stimulatory molecule to its cognate ligand, thereby mediating a signal transduction event, such as, but not limited to, signaling through an appropriate receptor, e.g., a T or NK receptor.

[0136] "Modulate" or "modulation" means adjusting or controlling the amplitude, frequency, extent, or activity. In another related aspect, such modulation can be positively modulated (e.g., an increase in frequency, extent, or activity) or negatively modulated (e.g., a decrease in frequency, extent, or activity). In some embodiments, modulation in a positive or negative direction is referred to relative to the function of a cell, tissue, or organ prior to administration of the therapeutic agent. In further embodiments, modulation in a positive or negative direction is referred to with respect to a normal, healthy cell, tissue, or organ.

[0137] An "effective amount" of a CAR cell, adaptor, and / or CAR cell / adaptor composition provided herein is an amount sufficient to carry out a specifically stated purpose, such as causing an observable change in the level of one or more biological activities associated with the target to which the CAR cell and / or adaptor binds. In some embodiments, the change increases the level of the target activity. In other embodiments, the change decreases the level of the target activity. An "effective amount" can be determined empirically and routinely, relative to the stated purpose. The term "therapeutically effective amount" refers to an amount of CAR cells and / or adaptors or other therapeutic agents effective to "treat" (e.g., alleviate the symptoms of) a disease or disorder in a subject (mammal). A "prophylactically effective amount" refers to an amount effective, at the dosages and for the duration necessary, to achieve a desired prophylactic result.

[0138] 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, and mice, and other animals. Animals include all vertebrates, e.g., mammals and non-mammals, e.g., 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 ape and monkey species; domestic animals, such as cows, sheep, pigs, goats, and horses; domestic mammals, such as dogs and cats; laboratory animals, including rodents, such as mice, rats, and 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 subjects, as well as embryos and fetuses, regardless of male or female gender, are intended to be included within the scope of this term.

[0139] As used herein, the terms "treat," "treatment," and "treating" refer to both therapeutic treatment and prophylactic or preventative measures, the purpose of which is to prevent or slow (reduce or delay) the symptoms, complications, or biochemical manifestations of a disease, condition, or disorder, to alleviate symptoms, or to prevent or inhibit further development of a disease, condition, or disorder. Treatment can be prophylactic (to prevent or delay the onset of the disease or to prevent the onset of clinical or subclinical symptoms) or therapeutic suppression or alleviation of symptoms after the onset of the pathological symptoms targeted by the disease, condition, or disorder; can prevent pathological symptoms; can pursue or achieve a beneficial outcome; or can reduce the likelihood that an individual will develop a condition, even if treatment is ultimately unsuccessful. Those in need of treatment include those already with the condition, as well as those prone to have the condition or those in whom the condition is to be prevented. Treatment can be with CAR cells, adapters, and / or CAR cell / adapter compositions alone or in combination with an additional therapeutic agent. In some embodiments, the terms "treat," "treatment," and "treating" are used herein to refer to therapeutic treatment and prophylactic or preventative measures, the purpose of which is to prevent or slow (alleviate or delay) symptoms, complications, or biochemical manifestations of a proliferative disorder, or to ameliorate one or more symptoms (preferably one or more discernible symptoms) of a proliferative disorder. In certain embodiments, the terms "treat," "treatment," and "treating" refer to an improvement in at least one measurable physical parameter of a proliferative disorder, e.g., tumor growth, not necessarily discernible by the patient. In other embodiments, the terms "treat," "treatment," and "treating" refer to inhibiting the progression of a proliferative disorder physically, e.g., by stabilization of a discernible symptom, physiologically, e.g., by stabilization of a physical parameter, or both. In other embodiments, the terms "treat," "treatment," and "treating" refer to a reduction or stabilization of tumor size, tumor cell growth or survival, or cancerous cell number.

[0140] "Cancer," "tumor," or "malignancy" are used synonymously and refer to any of a number of diseases characterized by the uncontrolled, abnormal growth of cells, the ability of affected cells to spread (metastasize) locally or via the bloodstream and lymphatic system to other parts of the body, and any of a number of characteristic structural and / or molecular features. As used herein, "tumor" refers to all neoplastic cell growth and proliferation, whether malignant or benign, and all precancerous and cancerous cells and tissues. A "cancerous tumor" or "malignant cell" is understood as a cell that possesses 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, lung, brain, cervical, skin, bone, liver, pancreatic, colorectal, kidney, head and neck, ovarian, hematopoietic (e.g., leukemia), and prostate cancer, and 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 known in the art. Reference to a particular "type" of cancer, tumor, or tumor cell is understood to mean 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 type 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.

[0141] The term "tumor antigen" refers to an antigen common to certain hyperproliferative disorders such as cancer. The terms "tumor antigen" and "cancer antigen" are used interchangeably herein. In some aspects, the antigen is derived from 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 adenocarcinoma, 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 chronic leukemias, including but not limited to B-cell acute lymphoblastic leukemia ("BALL"), T-cell acute lymphoblastic leukemia ("TALL"), acute lymphoblastic leukemia (ALL), acute myeloid leukemia (AML); chronic myeloid leukemia (CML), chronic lymphocytic leukemia (CLL); B-cell prolymphocytic leukemia, blastic plasmacytoid dendritic cell neoplasm, Burkitt's lymphoma, diffuse large B-cell neoplasm and additional blood cancers or conditions including, but not limited to, 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, plasmacytoid dendritic cell neoplasm, Waldenstrom's macroglobulinemia.

[0142] Tumor and cancer antigens can 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 present on other cells in the body. TAAs are antigens found on both tumor cells and some normal cells. TAAs can 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 can be expressed on normal cells during fetal development, when the immune system is immature and unable to respond, or they can be normally present at very low levels on normal cells but at much higher levels on tumor cells. Due to the dynamic nature of tumors, in some instances, tumor cells may express unique antigens at some stages and antigens that are also expressed on non-tumor cells at other stages. Therefore, including a marker as a TAA does not exclude that marker from being considered a TSA. In some embodiments, the TAA and / or TSA containing an antigenic determinant specifically bound by the CAR cells, adaptors and / or CAR cell / adaptor compositions provided herein include, but are not limited to, BCMA, CD19, CD20, CD22, CD30, CD33 / IL3Ra, CD70, CD123, CD171 (L1-CAM), CS1, EGFRvIII, GD2, Lewis YROR1, ringworm, IL13Ra2, cMet, PSMA, free radical α(FR-α), CEA, ErbB2(HER-2 / neu) EGFR(HER), PSCA, PSA, MUC1, MUC16, CD44v6, CD44v6 / 7, CD44v7 / 8 CD55, IL11Ra, EphA2, EGP40, TAG72, CAIX, HMW-MAA(CSPG4), MAGEA4, N KG2D fragment, β-HCG, activator F77, HLA-A2(NY-ESO-1), HMW-MAA, GD3, TCR, MAGE A3, TUESDAY, WT1, L.A., 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;VEG FR2, FAP, FAR, EBVA, HPV vaccine E6, HPV vaccine E7, TSP-180, MAGE4, ​​MAGE5, MAG E6, RAGE, pl85erbB2, pl80erbB3, nm-23Hl, CA19-9, CA72-4, CAM17. 1, NuMa, K-ras, β-linkage, CDK4, Mum-1, p15, p16, 43-9F, α-chain. BCA225, BTA, CA125, CA15-3, CA27.29(BCAA), CA195, CA242, CA50 CAM43, CD68, CO-029, FGF5, G250, HTgp-175, M344, MA50, MG7-Ag, NB / 70K, NY-CO-1, RCAS1, SDCCAG16, M2BP, TAAL6, TLP, and TPS.

[0143] As used herein, the term "CS1" refers to an NK cell receptor that regulates immune function and is also expressed on B cells, T cells, dendritic cells, NK-T cells, and monocytes. 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 differentially spliced ​​isoforms. In some embodiments, a 22-amino acid residue N-terminal signal sequence The amino acid sequence of human CS1 comprising TIFF2024512590000001.tif4128 and an extracellular domain comprising the 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.

[0144] The term "autoimmune disease" as used herein is defined as a disorder caused by an autoimmune response. Autoimmune diseases are the result of an inappropriate and 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.

[0145] As used herein, the term "transduction" refers to the introduction of foreign nucleic acid into a cell using a viral vector. As used herein, "transfection" 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 so that the host cell expresses the introduced nucleic acid to produce a desired substance, such as a protein or enzyme, encoded by the 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 include nonfunctional sequences or sequences without a 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 into a host cell can be from any source, including cells of the same genus or species as the host cell or cells of a different genus or species, or may be non-naturally occurring.

[0146] The term "D domain" refers to the canonical backbone sequence: The term "canonical scaffold" refers to a target-binding polypeptide that shares certain sequences and certain structural features of TIFF2024512590000002.tif10170 (see International Publication Nos. WO 2016 / 164305 and WO 2016 / 164308, each of which is incorporated herein by reference in its entirety). The canonical scaffold is a variant of a non-naturally occurring target-free antiparallel three-helix bundle canonical polypeptide originally engineered as a protein folding exercise (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 canonical scaffold having the amino acid sequence of SEQ ID NO: 1 can specifically bind targets of interest. Thus, D domains, or molecules containing D domains, can specifically (non-randomly) bind to target molecules. Without wishing to be bound by theory, it is believed that when designing a D domain, structural constraints on surface-exposed residues (which may be modified) confer the ability of the surface-exposed residues to specifically bind a target of interest.

[0147] As used herein, "co-expressed" refers to the expression of two or more protein-coding sequences by the same cell or cell population. These coding sequences can be, for example, nucleic acids that each encode a single protein or encode a chimeric protein as a single polypeptide chain.

[0148] As used herein, "antigen loss escape variant" refers to a cell that exhibits reduced expression or loss of expression of a target antigen whose antigenic determinant is targeted by an adapter or CAR provided herein.

[0149] 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 of an antibody or an alternative scaffold binding domain (ASBD) (e.g., a D domain)). The ADs in the adapters and on target cells provided herein can be bound by CARs, as described below.

[0150] In some embodiments, the AD (e.g., in the adapter 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 endogenous to humans.

[0151] In some embodiments, the AD in the adaptor is an AD present on a target cell.

[0152] In some embodiments, the AD (e.g., in the adaptor and / or on the target cell) is an AD present in a transmembrane protein, for example, 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.

[0153] 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.

[0154] 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 of the amino acid residues of SEQ ID NO:5.

[0155] In some embodiments, the AD is an epitope of CD 19. 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 the amino acid residues of SEQ ID NO:3.

[0156] 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 of the amino acid residues of SEQ ID NOs:6-9 or 10.

[0157] 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 of the amino acid residues of SEQ ID NO:41.

[0158] In some embodiments, the AD is an epitope of CD 123. 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 the amino acid residues of SEQ ID NO:11.

[0159] In some embodiments, the AD is an epitope of CD37. 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 the amino acid residues of SEQ ID NO:12 or 13.

[0160] 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 of the amino acid residues of SEQ ID NO: 1139. In further embodiments, the AD is the epitope of CS1 bound by elotuzumab.

[0161] In some embodiments, the AD is an epitope of HER2. 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 the amino acid residues of SEQ ID NO:42.

[0162] In some embodiments, the AD is an epitope of AFP. 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 the amino acid residues of SEQ ID NO:15.

[0163] 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 of the 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 of the amino acid residues of SEQ ID NO: 1117. In further embodiments, the AD comprises SEQ ID NO: 16, 1117, 1118, 1119, 1120, 1121, 1122, or 1123 amino acid residues.

[0164] In some embodiments, the AD (eg, in the adapter and / or on the target cell) is expressed on the surface of an immune effector cell.

[0165] 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.

[0166] 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.

[0167] 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.

[0168] 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.

[0169] 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.

[0170] 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.

[0171] In some embodiments, AD is an epitope of a human intracellular protein. In further embodiments, AD is an epitope of the intracellular portion of a membrane-associated 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, AD is an epitope of the intracellular portion of CD3, CD137, CD279, CD223, CD152, CD28 and VEGFR-2. In some embodiments, AD is an epitope of a human nuclear protein.

[0172] 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 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 an onco-fetal antigen. In some embodiments, the AD is an epitope of 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.

[0173] 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 a 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 other 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.

[0174] In some embodiments, the AD (e.g., in the adapter and / or on the target cell) is an epitope of a hematologic 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.

[0175] In some embodiments, the AD (e.g., in the adapter and / or on the target cell) is an epitope of a solid tumor antigen. In 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.

[0176] In some embodiments, the AD (e.g., in the adaptor and / or on the target cell) is selected from the group: 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 (Plysialic acid) acid), PLAC1, GloboH, NY-BR-1, UPK2, HAVCR1, ADRB3, PANX3, GPR20, LY6K, OR51E2, TARP, LAGE1a, legumain, E7, ETV6-AML, sperm protein 17, XAGE1, Tie2, MAD-CT1, MAD-CT2, Fos-related antigen 1, p53, p53 mutants, prostein, survivin, telomerase, PCTA1 (galectin 8), Ras mutants, hTERT, sarcoma translocation breakpoints, 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, IGLL1, TSP-180, MAGE4, ​​MAGE5, MAGE6, VEGFR1, IGF1R, hepatocyte growth factor receptor, p185E rbB2, p180ErbB-3, nm-23H1, CA19-9, CA72-4, CAM17.1, NuMa, K-ras, β-catenin, CDK4, Mum1 , p15, p16, 43-9F, 5T4, 791Tgp72, β-human chorionic gonadotropin, BCA225, BTAA, CA125, CA15-3, CA27.The epitope is an antigen selected from 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 variant, lectin-reactive AFP, and TLR4.

[0177] 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.

[0178] In some embodiments, the AD (e.g., in the adapter and / or on the target cell) is PDGFRA, VEGFR1, VEGFR3, neuropilin 1 (NRP1), neuropilin 2 (NRP2), betacellulin, PLGF, RET (rearranged during transfection), 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, patched1 (PTCH1), smoothened (SMO), WNT1, WNT2B, WNT3A, WNT4, WNT4A, WNT5A, WNT5B, WNT7B, WNT8A, W NT10A, WNT10B, WNT16B, LKP5, LRP5, LRP6, FZD1, FZD2, FZD4, FZD5, FZD6, FZD7, FZD8, Notch, Notch1, Notch3, Notch4, DLL4, J , Jagged1, Jagged2, Jagged3, TNFRSF1A(TNFR1, p55, p60), TNFRSF1B(TNFR2), TNFRSF6(Fas, CD95), TNFRSF6B(DcR3), T NFRSF7 (CD27), TNFSF9 (41BB ligand), TNFRSF8 (CD30), TNFRSF10A (TRAILR1, DR4), TNFRSF11A (RANK), TNFRSF12 (TWEAKR), TNF RSF19L(KELT), TNFRSF19(TROY), TNFRSF21(DR6), ILIRI, 1L1R2, IL2R, IL5R, IL6R, 1L8R, IL10R, IL12R, IL13R, IL15R, IL18R, IL19R, IL21R, IL23R, XAG1, XAG3, REGIV, FGFR1, FGFR2, FGFR3, ALK, ALK1, ALK7, ALCAM, Axl, TGFb, TGFb2, TGFb3, TGFBR1, 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, HM1.24, LFA1, LYNX, S100A8, S100A9, SCF, von Willebrand factor, Lewis Y6 receptor, CAG250 (CA9), CRYPTO, VLA5, HLADR, MUCl8, mucin CanAg, The epitope is an antigen selected from EGFL7, integrin avb3, integrin α5β, activin B1α, 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.

[0179] 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.

[0180] In some embodiments, the AD (e.g., in the adaptor and / or on the target cell) is an epitope of an immune inhibitory target. In another embodiment, the AD is an epitope of an immune inhibitory target selected from IL1Ra, IL6R, CD26L, CD28, CD80, FcγRIIB. In another embodiment, the AD in the adaptor is an epitope of an immune stimulatory target selected from CD25, CD28, CTLA4, PD1, B7H1 (PDL1), B7H4TGFβ, TNFRSF4 (OX40), TNFRSF5 (CD40), TNFRSF9 (41BB, CD137), TNFRSF14 (HVEM), TNFRSF25 (DR3), and TNFRSF18 (GITR).

[0181] In some embodiments, the AD (e.g., in the adaptor and / or on the target cell) is selected from the group consisting of 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, integrin alpha 4, integrin subunit alpha 5, integrin alpha 6, integrin alpha 7, integrin alpha 8, integrin alpha 9, integrin alpha 10, integrin alpha 11, integrin alpha 12, integrin alpha 13, integrin alpha 14, integrin alpha 15, integrin alpha 16, integrin alpha 17, integrin alpha 18, integrin alpha 19, integrin alpha 20, integrin alpha 21, integrin alpha 22, integrin alpha 23, integrin alpha 24, integrin alpha 25, integrin alpha 26, integrin alpha 27, integrin alpha 28, integrin alpha 29, integrin alpha 30, integrin alpha 31, integrin alpha 32, integrin alpha 33, integrin alpha 34, integrin alpha 35, integrin alpha 36, ​​integrin alpha 37, integrin alpha 38, integrin alpha 39, integrin alpha 41, integrin alpha 42, integrin alpha 43, integrin alpha 44, integrin unit, A4B7 integrin, cathepsin G, TNFRSF3 (LTBR), TNFRSF6 (Fas, CD95), TNFRSF6B (DcR3), TNFRSF8 (CD30), TNFRSF11A (RANK), T NFRSF16(NGFR), TNFRSF19L(RELT), TNFRSF19(TROY), TNFRSF21(DR6), CD14, CD23, CD36, CD36L, CD39, CD91, CD153, CD164 , CD200, CD200R, B71(CD80), B72(CD86), B7h, B7DC(PDL2), ICOS, ICOSL, MHC, CD, B7H2, B7H3, B7x, SLAM, KIM1, SLAMF2, SLA MF3, SLAMF4, SLAMF5, SLAMF6, SLAMF7, TNFRSF1A(TNFR1, p55, p60), TNFRSF1B(TNFR2), TNFRSF7(CD27), TNFRSF12(TWEAKR ), TNFRSF5 (CD40), IL1R, IL2R, IL4Ra, IL5R, IL6RIL15R, IL17R, IL17Rb, IL17RC, IL22RA, IL23R, TSLPR, B7RP1, cKit, GMCSF, GMCSFR, CD2, CD4, CD11a, CD18, CD30, CD40, CD86, CXCR3, CCR2, CCR4, CCR5, CCR8, RhD, IgE, and Rh.

[0182] 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.

[0183] In some embodiments, the AD (e.g., in the adaptor and / or on the target cell) is a target epitope selected from amyloid beta (Aβ), beta amyloid, PLP, ROBO4, ROBO, LINGO, gpIIB, gpIIIa, integrin a2bB3, AOC3, TNFRSF19L (RELT), TNFRSF19 (TROY), and sclerostin.

[0184] The targets listed above and those described elsewhere herein are intended to be exemplary and not limiting.

[0185] 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.

[0186] III. Antigenic Determinant Binding Domain (ADBD) A protein domain that binds to an antigenic determinant (AD) (e.g., as described in Section II) is referred to herein as an "antigenic determinant binding domain" or "ADBD." In some embodiments, the ADBD is sufficient to confer recognition of and specific binding to a target of interest. The ADBDs described herein can be present in an adapter (e.g., as described in Section V) and / or a chimeric antigen receptor (CAR) (e.g., as described in Section VI).

[0187] The target of interest specifically bound by the 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, any of the ADs described herein (e.g., as described in Section II). In some embodiments, the target specifically bound by the ADBD can be any target that has relevance or value for purification, manufacturing, formulation, treatment, diagnosis, or prognosis. In some embodiments, the target of the ADBD can be naturally occurring or synthetic. In some embodiments, the target of the ADBD can be an extracellular component, an intracellular component, a soluble factor (e.g., an enzyme, hormone, cytokine, growth factor, toxin, venom, pollutant, etc.), or a transmembrane protein (e.g., a cell surface receptor).

[0188] In some embodiments, the ADBD (e.g., of the adapter and / or CAR) specifically binds a target of interest on the surface of a target cell. In some embodiments, the ADBD specifically binds a cell surface receptor. In some embodiments, the ADBD specifically binds a target of interest 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., 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.

[0189] 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.

[0190] 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.

[0191] 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.

[0192] In some embodiments, the target of interest bound by the ADBD (e.g., of the adapter and / or CAR) is a human protein. In one embodiment, the ADBD binds to the human protein target of interest and its monkey (e.g., cynomolgus monkey), mouse, rabbit, hamster, and / or rabbit ortholog.

[0193] In another embodiment, the ADBD (for example, of the adaptor and / or CAR) binds to a peptide tag present on the target of interest. Such peptide tags provide a useful means for detecting, monitoring, and / or binding one or more additional moieties to the adaptor. In one embodiment, the ADBD specifically binds to a peptide tag selected from a hexahistidyl (His6) tag, a myc tag, and a FLAG tag. Other peptide tags are described herein or known in the art.

[0194] 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 is bound to a 5×10 -3 M or less, 10 -3 M or less, 5×10 -4 M or less, 10 -4 M or less, 5×10 -5 M or less or 10 -5In a further embodiment, the ADBD binds to the target of interest with a dissociation constant (KD) of 5×10 -6 M or less, 10 -6 M or less, 5×10 -7 M or less, 10 -7 M or less, 5×10 -8 M or less or 10 -8 In a further embodiment, the ADBD binds to the target of interest with a KD of 5×10 or less. -9 M or less, 10 -9 M or less, 5×10 -10 M or less, 10 -10 M or less, 5×10 -11 M or less, 10 -11 M or less, 5×10 -12 M or less, 10 -12 M or less, 5×10 -13 M or less, 10 -13 M or less, 5×10 -14 M or less, 10 -14 M or less, 5×10 -15 M or less or 10 -15 In some embodiments, the ADBDs produced by the methods disclosed herein bind to the target of interest with a KD of no greater than 10. -4 M~10 -5 M, 10 -5 M~10 -6 M, 10 -6 M~10 -7 M, 10 -7 M~10 -8 M, 10 -8 M~10 -9 M, 10 -9 M~10 -10 M, 10 -10 M~10 -11 M, and 10 -11 M~10 -12 M has a dissociation constant selected from

[0195] In one embodiment, the ADBD binds to the target of interest in an active form. In one embodiment, the ADBD reversibly binds to the target of interest in an active form and releases the bound target in an active form. In one embodiment, the ADBD binds to the target of interest in a natural form. In a particular embodiment, the ADBD binds to the target of interest in a natural form.-10 seconds -1 That's it, 5 x 10 -9 seconds -1 That's it, 10 -9 seconds -1 That's it, 5 x 10 -8 seconds -1 That's it, 10 -8 seconds -1 That's it, 5 x 10 -7 seconds -1 That's it, 10 -7 seconds -1 That's it, 5 x 10 -6 seconds -1 That's it, 10 -6 seconds -1 That's it, 5 x 10 -5 seconds -1 That's it, 10 -5 seconds -1 That's it, 5 x 10 -4 seconds -1 That's it, 10 -4 seconds -1 That's it, 5 x 10 -3 seconds -1 That's it, 10 -3 seconds -1 That's it, 5 x 10 -2 seconds -1 That's it, 10 -2 seconds -1 That's it, 5 x 10 -1 seconds -1 or more, or 10 -1 seconds -1 The target of interest is bound with an off-rate or Koff equal to or greater than 0.

[0196] Binding experiments to determine KD and dissociation rates can be performed 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, 0.01% Tween 20], [pH 5.0, 15% ethylene glycol, 0.01% Tween 20], and [pH 9.0, 15% ethylene glycol, 0.01% Tween 20]. The buffers for making these solutions can be readily determined by those skilled in the art and depend largely on the desired pH of the final solution. Low pH solutions (<pH 5.5) can be made, for example, in citrate buffer, glycine-HCl buffer or succinate buffer. High pH solutions can be made, for example, in Tris-HCl, phosphate buffer or sodium bicarbonate buffer. For example, a number of conditions can be used to determine KD and off-rate for the purpose of determining the optimal pH and / or salt concentration.

[0197] In one aspect, the ADBD specifically binds to the target of interest with a KOff in the range of 0.1 to 10 -7 seconds -1 10 -2 to 10 -7 seconds -1 or 0.5×10 -2 to 10 -7 seconds -1 In certain embodiments, the ADBD binds to the target of interest with an off-rate (KOff) of less than 5×10 -2 seconds -1 less than 10 -2 seconds -1 less than 5×10 -3 seconds -1 or less than 10 -3 seconds -1 In a further aspect, the ADBD binds to the target of interest with an off-rate (KOff) of less than 5×10 -4 seconds -1 less than 10 -4 seconds -1 less than 5×10 -5 seconds -1 or less than 10 -5 seconds -1Less than 5 x 10 -6 seconds -1 Less than 10 -6 seconds -1 Less than 5 x 10 -7 seconds -1 Less than or equal to 10 -7 seconds -1 Off-rate (K Off ) to bind to the target of interest.

[0198] 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 the target of interest with a KOn in the range of 10 3 M -1 seconds -1 Larger, 5 x 10 3 M -1 seconds -1 Greater than 10 4 M -1 seconds -1 Greater than, or 5 x 10 4 M -1 seconds -1 In a further embodiment, the ADBD binds to the target of interest with a greater on-rate (KOn). 5 M -1 seconds -1 Bigger, 5Back-10 5 M -1 seconds -1 Greater than 10 6 M -1 seconds -1 Greater than, or 5 x 10 6 M -1 seconds -1 Greater than or 10 7 M -1 seconds -1 Binds to target of interest with a larger KOn.

[0199] In some embodiments, the ADBD (e.g., of the adapter 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.

[0200] IIIa. Antibody-derived antigenic determinant binding domains (ADBDs) In some embodiments, one or more ADBDs (e.g., of an 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, a single domain antibody, for example, one or more of a heavy chain variable domain (VH), a light chain variable domain (VL), and a variable domain (VHH) of human or camelid origin. In some embodiments, the ADBD is derived from the same species in which the adapter or CAR will ultimately be used, for example, for use in humans. It may be beneficial for the adapter and / or CAR to include a human or humanized ADBD. Compositions and techniques for routinely producing such ADBDs are known in the art.

[0201] In some embodiments, the ADBD (e.g., of the adapter and / or CAR) comprises a fragment of an antibody 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, or Fv fragments, scFv antibody fragments, linear antibodies, single domain antibodies such as sdAbs (either VL or VH), camelid VHH domains, and multispecific antibodies formed from antibody fragments.

[0202] 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 short, flexible polypeptide linker, e.g., a linker described herein. scFvs can be routinely prepared according to methods known 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)).

[0203] 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, engineered domains, and single domain scaffolds other than those derived from antibodies. SDAB molecules can be derived from any species, including, but not limited to, mice, humans, camels, llamas, fish, sharks, goats, rabbits, and cows. This term also includes naturally occurring single domain antibody molecules from species other than camelids and sharks.

[0204] 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.

[0205] Antibody humanization is well known in the art and is essentially as described by Winter and coworkers (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 rodent CDRs or CDR sequences for the corresponding sequences of a human antibody, i.e., CDR grafting (EP 239,400; 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.

[0206] IIIb. Alternative scaffold-binding domains In some embodiments, the ADBD (e.g., of an adaptor and / or CAR) is an alternative scaffold-binding domain (ASBD). 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.

[0207] 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 is provided comprising a CAR comprising an ASBD. In some embodiments, the present disclosure provides an adaptor comprising an ADBD that is an ASBD.

[0208] In a further embodiment, the present disclosure provides a composition comprising an adaptor and a CAR, each comprising an ASBD.

[0209] In some embodiments, the binding of the ASBD (e.g., of an adaptor and / or a CAR) to the target AD is mediated by the secondary structure of the binding scaffold, such as an α-helix or a β-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 other embodiments, the ASBD is a Z-domain (Affibody)-based binding domain.

[0210] In some embodiments, the ASBD (e.g., of the adapter and / or CAR) is a D-domain (de novo binding domain)-based AD binding domain. D-domain scaffold-based binding domains generally consist of 70-75 amino acid residues, in which substitutions at up to 20 positions corresponding to structurally constrained, surface-exposed residues in a non-naturally occurring antiparallel three-helix bundle canonical framework (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 International Publication No. 2016164308, the entire contents of which are incorporated herein by reference. In one embodiment, the D-domain comprises an amino acid sequence that differs (e.g., by amino acid modification) from the amino acid sequence of the canonical framework having the sequence of SEQ ID NO: 1 by up to 20 substitutions. 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 a further embodiment, the D domain comprises a sequence selected from the group: SEQ ID NOs: 44-1078 and 1079.

[0211] 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 generally consist of 58 amino acid residues, in which substitutions at up to 13 positions located in the first and second alpha helices of three alpha helices confer target (AD) recognition and binding specificity to 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.

[0212] Further examples of ASBDs that exhibit secondary structure-mediated target binding include DARPins, affilins and armadillo repeat-based binding scaffolds.

[0213] In some embodiments, the ASBD (e.g., of the adaptor and / or CAR) is a DARPin-based AD-binding domain. DARPin-based binding domains generally comprise N-terminal and C-terminal capping repeats (e.g., the sequences TIFF2024512590000003.tif4148 and sequence TIFF2024512590000004.tif4128, the first Gln is at the consensus repeat position X of the preceding repeat 33 DARPins contain two to three repeats of the sequence of SEQ ID NO:30 located between the β-turn and β-turn of the next repeat (corresponding to the β-turn of the next repeat). Each internal repeat consists of 27 framework residues and up to six substituted non-framework residues that form a β-turn followed by two antiparallel helices and a loop connecting to the β-turn of the next repeat. The collective substitutions and structure of DARPins confer target (AD) recognition and binding specificity.

[0214] Table 1. Exemplary secondary structure-based ASBD sequences TIFF2024512590000005.tif128164X = All amino acid residues, including natural and unnatural amino acids Z = an amino acid sequence corresponding to loop 1 (Z1) or loop 2 (Z2) described herein, comprising from about 2 to about 30 natural or unnatural amino acids.

[0215] In some embodiments, the binding specificity of the ASBD (e.g., of an adaptor and / or CAR) for the 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.

[0216] 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 generally adopts a beta-sandwich fold containing seven chains connected by six loops. Substitutions in three surface-exposed loops on one side of the Adnectin domain generate target (AD)-specific binding moieties.

[0217] 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 is composed of eight antiparallel β-strands and generally exhibits a conserved β-barrel structure of 160-180 amino acids. The ligand-binding pocket of anticalin-based binding scaffolds is composed of four loops, each containing up to 24 substitutions, that collectively confer target (AD) recognition and binding specificity.

[0218] In some embodiments, the ASBD (e.g., of the adapter 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 generally 35 amino acids long. The structure of the avimer-based binding domain is maintained by 12 conserved amino acids. Substitution of up to all of the remaining 23 residues of the binding domain confers target (AD) recognition and binding specificity. In some embodiments, the avimer scaffold-based binding domain has the sequence TIFF2024512590000006.tif4128 (wherein 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, the contents of each of which are incorporated by reference in their entirety.

[0219] 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 generally 60-75 amino acids long and is composed of a pair of antiparallel β-sheets connected by two flexible loops. Substitutions / insertions in the loops confer AD target recognition and binding specificity. In some embodiments, a phenomer-based AD-binding domain has the sequence TIFF2024512590000007.tif11159 (X is any amino acid residue, X 13 ~X 21 and X 42 ~X 46 In some embodiments, the phenomer-based AD-binding domain comprises the sequence TIFF2024512590000008.tif11128 (X is any amino acid residue, X 16 ~X 21 is optionally absent) (SEQ ID NO:35).

[0220] In some embodiments, the ASBD (e.g., of the adaptor and / or CAR) is a knottin scaffold-based AD-binding domain. Knottin scaffold-based binding domains correspond to a 30 amino acid protein fold composed of three antiparallel β-strands connected by loops of variable length and multiple disulfide bonds.

[0221] In some embodiments, the ASBD (e.g., of the adapter 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 generally about 60 amino acids long. The hydrophobic core of this ADBD is composed of a twisted two-stranded antiparallel β-sheet and two α-helices stabilized by three pairs of disulfide bonds. 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 TIFF2024512590000009.tif11162X, containing a sequence that is at least 85% identical 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 39 is one of A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y; X 40 is one of G and A. Kunitz scaffold-based binding domains are further described in WO2004063337, the entire contents of which are incorporated herein by reference.

[0222] 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 generally 30-35 amino acids in length. In some embodiments, the WW domain-based AD-binding scaffold has the sequence TIFF2024512590000010.tif4137, including X7, X9, X 12 , X 13 , X 14 , X 18 , X 20 , X22 , X 25 , and X 27 represents any amino acid, and X 14 is arbitrary and non-existent.

[0223] In some embodiments, the WW domain-based AD-binding scaffold comprises the sequence TIFF2024512590000011.tif4133, including X7, X9, X 12 , X 13 , X 17 , X 19 , X 21 , X 24 , and X 26 represents any amino acid, and X 14 is arbitrary and non-existent.

[0224] Table 2. Exemplary loop-based ASBDs TIFF2024512590000012.tif57163X = All amino acid residues

[0225] IV. Linkers A linker is a peptide or other chemical bond positioned between two or more essentially independent functional domains of an adaptor or CAR.

[0226] Suitable linkers for operably linking two or more functional domains of an adaptor in a single amino acid sequence include, but are not limited to, polypeptide linkers such as glycine linkers, serine linkers, mixed glycine / serine linkers, glycine and serine rich linkers or linkers composed mostly of polar polypeptide fragments.

[0227] In one embodiment, the linker is composed of a majority of amino acids selected from glycine, alanine, proline, asparagine, glutamine, and lysine. In one embodiment, one or more linkers in an adaptor or CAR are composed of a majority of amino acids selected from glycine, alanine, proline, asparagine, aspartic acid, threonine, glutamine, and lysine. In one embodiment, one or more linkers in an adaptor or CAR are composed 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 in an adaptor or CAR are composed of a majority of sterically unhindered amino acids. In another embodiment, a linker in which the majority of amino acids are glycine, serine, and / or alanine. In some embodiments, one or more linkers in an adaptor or CAR linker comprise poly(Gly-Ala), and polyalanine, such as polyglycine ((Gly)5 (SEQ ID NO:1099), and (Gly)8 (SEQ ID NO:1100). In some embodiments, the peptide linker contains the sequence Gly-Gly-Gly-Gly-Gly-Thr-Gly-Gly-Gly-Gly-Ser (SEQ ID NO:39). In some embodiments, one or more linkers in an adaptor or CAR comprise the sequence Gly-Gly-Gly-Gly-Asp-Gly-Gly-Gly-Gly-Ser (SEQ ID NO:40).

[0228] 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 contains at least two, at least three, at least four, or at least five ADBDs directly attached to another domain of the adapter or CAR, respectively.

[0229] In another embodiment, the ADBD can be functionally linked to another component of the adaptor or CAR via a linker. The adaptor or CAR can contain a single linker, multiple linkers, or no linker. In one embodiment, the adaptor or CAR comprises an ADBD functionally linked to another component of the adaptor or CAR, respectively, via a linker peptide. In one embodiment, the adaptor or CAR contains at least two, at least three, at least four, or at least five ADBDs functionally linked to another domain of the adaptor or CAR, respectively, via the same linker or different linkers.

[0230] The linker can be of any size or composition, so long as it is capable of operably linking the functional domains of the adaptor or CAR, allowing the functional domains to function (e.g., the ability of an antigenic determinant binding domain to bind to a target of interest). In some embodiments, the linker is about 1 to about 100 amino acids, about 1 to 50 amino acids, about 1 to 20 amino acids, about 1 to 15 amino acids, about 1 to 10 amino acids, about 1 to 5 amino acids, about 2 to 20 amino acids, about 2 to 15 amino acids, about 2 to 10 amino acids, or about 2 to 5 amino acids. It should be apparent that the length, degree of flexibility, and / or other properties of the linker may 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 of interest or one or more other target proteins of interest. When more than one linker is used in an adaptor or CAR, these linkers may be the same or different. In the context and disclosure provided herein, one of skill in the art will be able to routinely determine optimal linker composition and length for the purpose of operably linking the functional domains of an adapter or CAR.

[0231] The linker may 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)-, where s=2 to 20, may be used. These alkyl linkers may be further substituted with any sterically unhindered group, such as lower alkyl (e.g., C1 to C6) lower acyl, halogen (e.g., Cl, Br), CN, NH2, or phenyl. An exemplary non-peptide linker is a PEG linker. In some embodiments, the PEG linker has a molecular weight of about 100 to 5,000 kDa, or about 100 to 500 kDa.

[0232] Suitable linkers for linking adaptor or CAR functional domains by chemical crosslinking include, but are not limited to, homobifunctional chemical crosslinking compounds such as glutaraldehyde, imidoesters such as dimethyl adipimidate (DMA), dimethyl suberimidate (DMS) and dimethyl pimelimidate (DMP), or N-hydroxysuccinimide (NHS) esters such as dithiobis(succinimidyl propionate) (DSP) and dithiobis(sulfosuccinimidyl propionate) (DTSSP). Examples of suitable linkers for linking adaptor or CAR functional domains include, but are not limited to, crosslinkers with one amine-reactive end and a sulfhydryl-reactive moiety at the other end, or with an NHS ester and an SH-reactive group (e.g., maleimide or pyridyl) at one end.

[0233] In further embodiments, one or more of the linkers in the adaptor or CAR are cleavable. Examples of cleavable linkers include, but are not limited to, peptide sequences recognized by various types of proteases (in vitro or in vivo), such as Tev, thrombin, factor Xa, plasmin (blood protease), metalloproteases, cathepsins (e.g., GFLG, etc.), and proteases found in other body compartments.

[0234] In some embodiments, the linker is a "cleavable linker" that facilitates the release of the adaptor functional domain or cytotoxic agent inside or on the cell surface. For example, an acid-labile linker (e.g., hydrazone), a protease-sensitive (e.g., peptidase-sensitive) linker, a photolabile linker, a dimethyl linker, or a disulfide-containing linker (see, for example, Chari, Can.Res.52:127-131(1992); U.S. Patent No. 5,208,020; and U.S. Patent Application Publication No. 20090110753; the contents of each of which are incorporated herein by reference in their entirety) can be used, and it is desirable that the covalent bond between the adaptor or cytotoxic agent is cleaved inside the cell when the composition is internalized into the cell. The terms "intracellularly cleaved" and "intracellular cleavage" refer to a metabolic process or reaction within the cell on an adapter drug conjugate in which the covalent bond, i.e., the linker-mediated connection between the adapter and the cytotoxic agent, is cleaved, resulting in free adapter and / or cytotoxic agent dissociated within the cell.

[0235] In further embodiments, one or more of the linkers in the CAR are cleavable. Examples of cleavable linkers include, but are not limited to, peptide sequences recognized by various types of proteases (in vitro or in vivo), such as Tev, thrombin, factor Xa, plasmin (blood protease), metalloproteases, cathepsins (e.g., GFLG, etc.), and proteases found in other body compartments.

[0236] In some embodiments, short oligopeptide or polypeptide linkers, approximately 1 to 100 amino acids in length, are used to link any of the CAR domains 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 prevent two adjacent domains from sterically interfering with each other.

[0237] In some embodiments, a length of preferably 2 to 10 amino acids forms the link between the transmembrane domain and the cytoplasmic signaling domain of the CAR. In further embodiments, the linker is 10 to 15 amino acids in length, or 15 to 20, or 20 to 30, or 30 to 60, or 60 to 100 amino acids in length (or any range between those listed). In further embodiments, the linker is a glycine-serine doublet sequence. In some embodiments, the ESD corresponds to the human T-cell surface glycoprotein CD8 α-chain ESD region (e.g., amino acid residues 138-182 of the CD8 α-chain; Swiss-Prot Acc. No. P01732). In some embodiments, the ESD corresponds to a CD8 ESD region that has been further modified through amino acid substitutions 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 that confer improved expression function or immunogenicity.

[0238] Linker optimization can be evaluated using techniques described herein and / or known in the art. In some embodiments, the linker does not interfere with the ability of the adapter or CAR to bind to a target antigenic determinant and / or another adapter or CAR functional domain and function properly (e.g., the ability of an effector functional domain in the adapter to induce effector function or the ability of an FcRn-binding domain in the adapter to bind to FcRn).

[0239] V. Adaptor-Soluble Proteins

[0010] Provided herein are multidomain soluble adaptor proteins. The adaptor comprises an antigenic determinant (AD) (e.g., as described in Section II) and an antigenic determinant binding domain (ADBD) (e.g., as described in Section III). The adaptor can further comprise additional ADs, additional ADBDs, and / or other additional domains.

[0240] In the adapters provided herein, the AD may be at the N-terminus of the ADBD. Alternatively, the ADBD may be at the N-terminus of the AD. In some embodiments, the AD and the ADBD are directly fused. In some embodiments, the AD and the ADBD are fused via a linker (a protein linker or a chemical linker) or another protein domain (e.g., a functional domain).

[0241] In some embodiments, the adapter comprises a linker positioned between the ADBD and another functional domain of the adapter. In some embodiments, the linker is positioned between the two ADBDs of the adapter. In some embodiments, the linker is positioned between the AD and ADBD of the adapter. Suitable linkers for linking two or more functional domains of the adapter will be apparent to those skilled in the art and may generally be any linker used in the art to link 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.

[0242] In addition to an AD (or multiple ADs) and an ADBD (or multiple ADBDs), the adapters provided herein can further comprise an additional domain or additional domains, e.g., a domain that confers extended half-life.

[0243] In some embodiments, the adaptor or the ADBD in the adaptor is deimmunized.

[0244] 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 Section VI) expressed on the surface of a cell (e.g., as described in Section VII), e.g., to kill a target cell.

[0245] 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 an adapter comprises two or more ADs, the ADs can be the same or different.

[0246] In the adapters provided herein, the AD can be any AD or combination of ADs (e.g., as described in Section II).

[0247] 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 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 of the amino acid residues of SEQ ID NO:5.

[0248] In some embodiments, the adaptor comprises an extracellular domain of CD123 (e.g., a polypeptide comprising the sequence of SEQ ID NO:11). 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 of the amino acid residues of SEQ ID NO:11.

[0249] In some embodiments, the adapter comprises the extracellular domain of CD19 (e.g., a polypeptide comprising the sequence of SEQ ID NO:2 or 3. In some embodiments, the adapter 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 the amino acid residues of SEQ ID NO:2 or 3.

[0250] 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 of the amino acid residues of SEQ ID NOs:6-9 or 10.

[0251] 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 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 of the amino acid residues of SEQ ID NO:41.

[0252] 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 of the amino acid residues of SEQ ID NO:12 or 13.

[0253] 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 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 of the amino acid residues of SEQ ID NO: 1138.

[0254] In some embodiments, the adapter comprises the extracellular domain of HER2 (e.g., a polypeptide comprising the sequence of SEQ ID NO:42). In some embodiments, the adapter 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 of the amino acid residues of SEQ ID NO:42.

[0255] 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 the amino acid residues of residues 29-766 of SEQ ID NO:1106.

[0256] 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 of the amino acid residues of the extracellular domain of CD26, CD30, CD33, or CD38.

[0257] In some embodiments, the AD is an epitope of AFP. 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 the amino acid residues of SEQ ID NO:15.

[0258] 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 of the 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 of the 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.

[0259] In some embodiments, the adapter comprises a p26 protein (e.g., having the sequence of SEQ ID NO: 16, 1117, 1118, 1119, 1120, 1121, 1122, or 1123). It has been discovered herein that such fusion proteins containing the p26 sequence have surprisingly 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. 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 more than 65 hours, or 1 to 10 hours, 2 to 10 hours, 4 to 10 hours, 6 to 10 hours, or 6 to 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, or 1 to 10 hours, 2 to 10 hours, 4 to 10 hours, 6 to 10 hours, or 6 to 9 hours.

[0260] In some embodiments, the present disclosure provides methods for modifying the in vivo half-life (e.g., in a mouse or a human) of an adaptor comprising a p26 protein (e.g., having the sequence of SEQ ID NOs: 16, 1117, 1118, 1119, 1120, 1121, 1122, or 1123). In some embodiments, the adaptor comprises one or more target-binding DDpp. In some embodiments, the half-life of the adaptor is increased or decreased by substituting or deleting one or more amino acid residues normally found in the human p26 protein, or by inserting one or more amino acid residues not normally found in the human p26 protein. In another embodiment, the p26 sequence of the adaptor is modified through 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 increase or decrease the in vivo half-life of the adaptor. 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 through the deletion of 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 through 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.

[0261] In some embodiments, the AD (e.g., in the adapter 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 endogenous to humans.

[0262] 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 of the amino acid residues of the intracellular protein.

[0263] In some embodiments, the target of interest specifically bound by the ADBD of an adaptor is itself the AD of another adaptor with a different sequence.

[0264] Vb. Antigenic determinant binding domain (ADBD) The adapters provided herein comprise at least one antigenic determinant binding domain (ADBD). In some embodiments, the adapter contains one ADBD. In some embodiments, the adapter contains at least two, three, four, or five, or more than five ADBDs. In some embodiments, the adapter contains one to three, one to four, one to five, or more than five different ADBDs. In some embodiments, the adapter contains at least two, three, four, or five, or more than five different ADBDs. Thus, an adapter can comprise a monomeric ADBD (i.e., containing one antigenic determinant binding domain) or a multimeric ADBD (i.e., containing more than one antigenic determinant binding domain in tandem, optionally operably linked by a linker). In some embodiments, the use of a multimeric adapter provides enhanced (e.g., synergistic) target binding. In further embodiments, the use of a multimeric adapter allows for targeting more than one target using a single adapter construct (e.g., bispecific, trispecific, etc.).

[0265] Multimeric adapters may be homomultimers (i.e., containing two or more of the same ADBDs, optionally connected by a linker) (e.g., homodimers, homotrimers, homotetramers, etc.) or adapter heteromultimers (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 may be determined, at least in part, by the expression system in which the adapter is produced. However, in some embodiments, the fusion protein may contain a multimer 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 (inclusive of numbers and endpoints between the listed ones). Furthermore, the multiple domains of the adapter may contain the same or different ADBDs. In some embodiments, two, three, four, five, or more than five domains are in tandem.

[0266] 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. Linking two or more identical ADBDs that bind to the same target antigen results in a multivalent molecule that offers distinct advantages (e.g., increased binding avidity, target clustering, and receptor activation) over compositions containing only one ADBD for the 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. Linking two or more ADBDs results in a multivalent and multispecific adapter that has the potential to bind more than one target antigen, independently or simultaneously. In some embodiments, the multivalent adapter can simultaneously bind the same target antigen. In some embodiments, the multivalent adapter can simultaneously bind different target antigens. In some embodiments, the 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 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 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 an AFP P26 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 adaptor comprises two or more operably linked ADBDs separated by a CD26, CD30, CD33, or CD38 antigenic determinant.

[0267] The ADBD in the adapters provided herein can bind to any AD (e.g., as described in Section II). 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 ABDB binds to CD45. In some embodiments, the ABDB in the adapters provided herein specifically binds to the AD of human CD26, CD30, CD33, or CD38. Adapters can be "monospecific" or "multispecific." Adapters that are "multispecific" (e.g., bispecific, trispecific, or of greater multispecificity) recognize and bind to two or more different epitopes present on one or more different molecules.

[0268] 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., the 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.

[0269] In some embodiments, the adapter contains 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., the above-mentioned domains). 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. In a further embodiment, the adapter comprises two or more operably linked ADBDs separated by an AFP P26 antigenic determinant.

[0270] In some embodiments, the adapter contains at least two of the same ADBDs (i.e., is multivalent). In some embodiments, the multivalent adapter can simultaneously bind two or more of the same target antigens. In some embodiments, the adapter is multivalent and can simultaneously bind the same target antigen. In some embodiments, the multi-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 adaptor comprises two or more operably linked ADBDs separated by a BCMA antigenic determinant. In further embodiments, the multivalent adaptor 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.

[0271] In some embodiments, the adapter contains at least two ADBDs that bind to different antigens (i.e., is multispecific). In some embodiments, the multispecific adapter can simultaneously bind different target antigens. In some embodiments, the adapter is also multivalent and can simultaneously bind 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.

[0272] In one embodiment, the multispecific adapter contains at least two ADBDs that bind to at least two different epitopes on a single target of interest (i.e., multi-epitopic to the same target antigen). In further embodiments, the multispecific adapter comprises at least one ADBD that specifically binds one epitope on the target of interest and at least one other ADBD that specifically binds 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 antigen 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.

[0273] 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., increased pharmacokinetics, such as increased 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 positions within the adapter can vary. For example, ADBDs can be located at one or all termini of the fusion partner and / or can be interspersed within heterologous subunits 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.

[0274] In one embodiment, the adapter is bispecific and contains 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 tumor cells. In a further embodiment, the bispecific adapter specifically binds to two different target antigens (e.g., BCMA and CS1) expressed on the surface of multiple myeloma cells. 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 cells of a tumor. 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 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 can simultaneously bind 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. In a further embodiment, the adapter comprises two or more operably linked ADBDs separated by an AFP P26 antigenic determinant.

[0275] In some embodiments in which an adaptor comprises more than one ADBD, the ADBD can be any of the types of ADBDs discussed herein (e.g., any ADBD described in Section III 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.

[0276] In some embodiments, when an adapter comprises more than one ADBD, the ADBDs may be the same type of antigen-binding molecule or may be different. For example, an adapter may comprise two ADBDs that are D domains. The two ADBDs that are D domains may be the same or different. An adapter may also 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 its antigen-binding fragment and 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.

[0277] In some embodiments, the ADBD of the adaptor is deimmunized.

[0278] In some embodiments, the adapter comprises an ABBD that binds to an antigen target containing an AD of interest and has no discernible effect on the function of the target. Alternatively, in some embodiments, the adapter comprises an ADBD that binds to an antigen target containing an AD of interest and completely or partially inhibits, antagonizes, agonizes, blocks, increases, stimulates, or interferes with the biological activity of the target. Binding can be identified as agonistic or antagonistic, and can be determined using or routinely modified assays, bioassays, and / or animal models known in the art for evaluating such activity.

[0279] 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 that of a known agonist of the adapter target. In another embodiment, the adapter is an antagonist of the target to which the adapter binds. An adapter agonist refers to an adapter that completely or partially blocks, or in some way interferes with, the biological activity of the adapter target, or has biological activity equivalent to that of a known antagonist or inhibitor of the adapter target.

[0280] In one embodiment, the adaptor specifically binds a target of interest that is a serum protein. In one embodiment, the adaptor specifically binds 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 binding of the adaptor to a carrier protein confers an improved pharmacodynamic profile to the adaptor, including, but not limited to, improved tumor targeting, tumor penetration, intratumoral spread, and enhanced therapeutic activity, compared to an adaptor lacking the carrier protein binding sequence (see, for example, International Publication No. 01 / 45746, the entire contents of which are incorporated herein by reference).

[0281] 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.

[0282] In further embodiments, the target of interest bound by the adapter is a bacterial antigen, a viral antigen, a fungal antigen, a mycoplasma 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 anthrax, hepatitis B, rabies, Nipah virus, West Nile virus, meningitis virus, or CMV. In further embodiments, the adapter specifically binds a pathogen.

[0283] Vc. Adaptor functional domain In some embodiments, the adapter 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) and / or pharmacokinetic properties (e.g., improved half-life). The adapter's functional domain may be located between the AD and the ADBD. The adapter may also be located N-terminally to both the AD and the ADBD, or C-terminally to both the AD and the ADBD. In some embodiments, when the adapter comprises two or more ADs, the adapter's functional domain may be located between the two or more ADs, N-terminally to the two or more ADs, or C-terminally to the two or more ADs. In some embodiments, when the adapter comprises two or more ADBDs, the adapter's functional domain may be located between the two or more ADBDs, N-terminally to the two or more ADBDs, or C-terminally to the two or more ADBDs.

[0284] In some embodiments, the adapter comprises a functional domain selected from Fc or a variant Fc (e.g., a human Fc or a variant Fc domain) or a fragment thereof, a serum protein (e.g., human serum albumin) or a 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.

[0285] The functional domain of an adapter can be naturally derived or can be the result of recombinant engineering (e.g., phage display, xeno-mouse, or synthetic). In some embodiments, the functional domain of an adapter increases half-life, increases or decreases antibody-dependent cellular cytotoxicity (ADCC), and / or increases or decreases complement-dependent cytotoxicity (CDC) activity.

[0286] In some embodiments, the adapter comprises a functional domain selected from an Fc or variant Fc (e.g., a 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) functional domain; an FcRn binding domain; and a serum protein binding domain.

[0287] In one embodiment, the adapter 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 adapter, such as the ability to bind to one or more Fc receptors. In some embodiments, the functional domain comprises one or more CH2 and / or CH3 domains of an antibody having an effector function provided by the CH2 and CH3 domains. In some embodiments, the functional domain comprises one or more derivatives of the CH2 and / or CH3 domains of an antibody having an effector function provided by the CH2 and CH3 domains. Other sequences that can be included in an adapter to provide effector function and are encompassed by the present invention will be apparent to those of skill in the art and can be routinely selected based on the desired effector function and engineered into the adapters encompassed herein.

[0288] In one embodiment, the adapter comprises a functional domain that increases antibody-dependent cellular cytotoxicity (ADCC) conferred by the adapter (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 effector function fragment-engineered modifications that confer portions of the Fc contained in the functional domain of the adaptor that increase ADCC include one or more modifications corresponding to IgG1-S298A, E333A, K334A, IgG1-S239D, I332E; IgG1-S239D, A330L, I332E; IgG1-P247I, A339D or Q; IgG1-D280H, S298D or K290S with or without V; IgG1-F243L, R292P, Y300L; IgG1-F243L, R292P, Y300L, P396L; and IgG1-F243L, R292P, Y300L, V305I, P396L; the numbering of residues in the Fc region is according to the EU index of Kabat et al. (Kabat et al., Sequences of proteins of This is the numbering of the Immunological Interest, 1991 Fifth edition.

[0289] Thus, in some embodiments, the adapter comprises a functional domain comprising an antibody fragment that confers biological or biochemical characteristics of an immunoglobulin to the adapter. In some embodiments, the antibody fragment confers a characteristic 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 some embodiments, the adapter is at least as stable as the corresponding antibody fragment without the adapter. In some embodiments, the adapter is more stable than the corresponding antibody fragment without the adapter. The stability of the adapter protein can be measured using established methods, including, for example, ELISA techniques. In some embodiments, the adapter is stable in whole blood (in vivo or ex vivo) at 37° C. 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 hou...

Claims

1. A chimeric antigen receptor (CAR) that binds to B cell maturation antigen (BCMA) and includes a D domain that includes amino acid residues 21 to 327 of SEQ ID NO:1166.

2. A cell engineered to express the CAR described in claim 1.

3. The cell described in claim 2, wherein the cell is a T cell or a NK cell.

4. A pharmaceutical composition comprising the cells described in claim 2.

5. An isolated nucleic acid encoding the CAR described in claim 1.

6. A vector comprising the isolated nucleic acid described in claim 5.

7. The vector described in claim 6, further comprising a promoter.

8. The vector described in claim 6, wherein the vector is a lentiviral vector.

9. A cell containing the vector described in claim 6.

10. A therapeutically effective dose of cells expressing the CAR described in claim 1.

11. The therapeutically effective dose of cells of claim 10, wherein the therapeutically effective dose comprises about 20x10 6 to about 500x10 6 cells expressing a CAR.

12. The therapeutically effective dose of cells of claim 11, wherein the therapeutically effective dose comprises about 50x10 6 to about 300x10 6 cells expressing a CAR.

13. The therapeutically effective dose of cells of claim 12, wherein the therapeutically effective dose comprises about 50x10 6 to about 200x10 6 cells expressing a CAR.

14. The therapeutically effective dose of cells of claim 13, wherein the therapeutically effective dose comprises about 100x10 6 cells expressing a CAR.

15. A composition comprising a cell expressing a chimeric antigen receptor (CAR) at a dose of 20x10 to 500x10 for use in a method for treating a BCMA associated cancer, the CAR comprising a D domain comprising the amino acid sequence of SEQ ID NO:850, the method comprising: (a) the composition, and (b) at least one dose of an adaptor comprising (i) an amino acid sequence of SEQ ID NO:1118 and (ii) a D domain that binds to BCMA. to a subject in need thereof.

16. The composition described in claim 15, wherein the D domain that binds to BCMA has the amino acid sequence of SEQ ID NO:

201.

17. The composition of claim 15, wherein the dose of cells expressing the chimeric antigen receptor is about 100x10 6 cells.

18. The composition described in claim 15, wherein the dosage of the adapter is 5 ?g / kg to 5 mg / kg.

19. The composition of claim 15, wherein the dosage of the adapter is about 0.2 mg / kg.

20. The composition of claim 15, wherein the dosage of the adapter is about 0.07 mg / kg.

21. The composition of claim 15, wherein the dosage of the adapter is between 2 mg and 25 mg.

22. The composition of claim 15, wherein the dosage of the adapter is about 0.4 mg, 0.8 mg, 1.6 mg, 2.8 mg, 3 mg, 4 mg, 8 mg, 12 mg, 16 mg, 20 mg, 24 mg, 28 mg, 40 mg, or 80 mg.

23. The composition of claim 15, wherein multiple doses of the adaptor are administered.

24. The composition of claim 23, wherein the dose of the adaptor is administered daily.

25. The composition of claim 23, wherein the dose of the adaptor is administered twice a day, three times a day, four times a day, every 12 hours, every 8 hours, every 6 hours, or every 4 hours.

26. The composition of claim 23, wherein the dose of the adaptor is administered every other day, every third day, twice a week, or weekly.

27. The composition of claim 23, wherein the dose of the adaptor is administered for one week to two months.

28. The composition of claim 23, wherein the dose of the adaptor is administered for 1 week, 2 weeks, 3 weeks, 4 weeks, 1 month, 2 months, or 3 months.

29. The composition of claim 23, wherein the dose of the adaptor is administered at a constant frequency over the course of treatment.

30. The composition of claim 23, wherein the dose adaptor is administered at a frequency that decreases over the course of treatment.

31. The composition of claim 15, wherein the at least one dose of the adaptor is administered intravenously.

32. The composition of claim 15, wherein said dose of cells expressing a chimeric antigen receptor and said at least one dose of adaptor are administered on the same day.

33. The composition of claim 15, wherein the BCMA-associated cancer is lymphoma or leukemia.

34. The composition of claim 15, wherein the BCMA-associated cancer is multiple myeloma.

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