Antibody combinations for treatment of cancer in specific patients

Abstract

To provide antibody combinations for treatment of cancer in specific patients.SOLUTION: Described is the combined use of a first antibody molecule that specifically binds to FcγRIIb via the Fab region and binds to an Feγ receptor via the Fc region, and a second antibody molecule that specifically binds to PD-1 via the Fc region and binds to at least one Feγ receptor, in the treatment of cancer in a patient having tumor infiltrating T lymphocytes with a medium or high PD-1 expression, as well as pharmaceutical compositions and kits comprising these two antibody molecules, and methods of treating cancer using these two antibodies. Described is also a diagnostic test for identification of patients benefitting from the treatment described herein.SELECTED DRAWING: Figure 1

Description

[Technical Field]

[0001] The present invention relates to the combination of 1) a first antibody molecule that specifically binds to FcγRIIb via its Fab region and binds to an Fcγ receptor via its Fc region, and 2) a second antibody molecule that specifically binds to PD-1 via its Fc region and binds to at least one Fcγ receptor, in the treatment of cancer in patients with moderate or high expression of PD-1 on CD3-positive tumor-infiltrating lymphocytes (TILs). [Background technology]

[0002] Immunosuppressive checkpoint receptors, such as CTLA-4 or PD-1 (also referred to as PD1), are cell surface receptors that, upon binding to their ligand receptors, such as the B7 family members CD80 and CD86, and PD-L1, respectively, transmit inhibitory signals to the interior of cells, limiting cell activation and proliferation, preventing excessive inflammation, and contributing to the maintenance of self-tolerance. Animals genetically deficient in such inhibitory immune checkpoints are associated with exacerbated inflammatory responses and are unable to develop or maintain tolerance to self, resulting in autoimmune diseases. Antibodies against the immune checkpoint receptors CTLA-4 and PD-1 / PD-L1 have resulted in increased overall survival in patients with a variety of cancers, including multiple solid tumors, such as melanoma, lung cancer, bladder cancer, and head and neck cancer, and such antibodies have been approved by the U.S. Food and Drug Administration (Pardoll, D. M. (2012) Nat Rev Cancer 12(4):252-264; Topalian, S. Let al (2015) Cancer Cell 27(4):450-461; Sharma, P. et al (2017) Cell 168(4):707-723).

[0003] Antibodies against the immunosuppressive checkpoint axis PD-1 / PD-L1 have proven particularly effective in cancer immunotherapy, inducing objective responses (complete and partial responses) in approximately 20% of patients, a significant improvement over standard treatments (Carretero-Gonzalez, A. et al. (2018) Oncotarget 9(9):8706-8715). However, as evidenced by response rates, currently available anti-PD-1 / PD-L1 antibodies are effective in only a minority of patients. Furthermore, a proportion of patients who initially respond eventually develop resistance and are no longer able to benefit from treatment. Therefore, mechanisms of nonresponsiveness and resistance to PD-1 / PD-L1 antibodies are clinically significant. Identifying and overcoming mechanisms of resistance to PD-1 / PD-L1 antibodies represents a major challenge and opportunity for improving cancer patient survival with this clinically important drug class.

[0004] Furthermore, it is generally accepted that predictive biomarkers to identify patients most likely to respond to anti-PD-1 / PD-L1 checkpoint blockade are an important strategy for identifying patients likely to respond to treatment. Conversely, it is equally important to prevent unnecessary treatment of patients with these drugs, which are often associated with significant, sometimes fatal, tolerability issues. Given their high costs, these therapies place an even greater burden on payers and healthcare systems. Examples of existing predictive biomarkers with clinical significance for anti-PD-1 / PD-L1 antibody therapy include microsatellite instability (MSI) (Le, DT et al (2015) N Engl J Med 372(26):2509-2520; Le, DT et al (2017) Science 357(6349):409-413), tumor mutation burden (Gubin, MM et al (2014) Nature 515(7528):577-581; Snyder, A., et al (2014) N Engl J Med 371(23):2189-2199; Tran, E. et al (2014) Science 344(6184):641-645, Tran, E. et al (2015) Science 350(6266):1387-1390), and tumor PD-L1 expression (Gibney, Weiner et al. 2016, Topalian, Taube et al. 2016), tumor mutation burden, and tumor PD-L1 expression.

[0005] Fc gamma receptors (FcγRs) are membrane proteins found on the cell surface of immune effector cells, such as monocytes, macrophages, dendritic cells, neutrophils, mast cells, basophils, eosinophils, and natural killer cells, as well as B lymphocytes. Their name derives from their binding specificity to the Fc region of antibodies. Fc receptors are found on the cell membrane, also known as the plasma membrane or cytoplasmic membrane. FcγRs can be subdivided into activating FcγRs and inhibitory FcγRs, which coordinately regulate cell activation through the binding of clustered immunoglobulin G Fc and transmit activating or inhibitory signals to cells via intracellular ITAM or ITIM motifs, respectively. FcγR binding of clustered immunoglobulins or immune complexes mediates antibody internalization into cells, potentially leading to antibody-mediated phagocytosis, antibody-dependent cell-mediated cytotoxicity, or antigen presentation or cross-presentation. FcγRs are also known to mediate or enhance cross-linking of antibody-bound cell surface receptors. Such cross-linking is required for some antibodies (Li, F. et al (2011) Science 333(6045):1030-1034; White, AL et al (2011) J Immunol 187(4):1754-1763), but not all (Richman, LP et al (2014) Oncoimmunology 3:e28610) antibodies to activate signaling in target cells, and may or may not be required to achieve a therapeutic effect.

[0006] In humans, FcγRIIb (CD32b) is an inhibitory Fcγ receptor, while FcγRI (CD64), FcγRIIa (CD32a), FcγRIIc (CD32c), and FcγRIIIa (CD16a) are activating Fcγ receptors. FcγRIIIb is a GPI-linked receptor expressed on neutrophils that lacks ITAM motifs and is thought to act as a decoy receptor that counteracts activating FcγR signaling (Treffers, L. Wet et al. (2018) Front Immunol 9:3124). In mice, the activating receptors are FcγRI, FcγRIII, and FcγRIV.

[0007] It is known that antibodies can regulate immune cell activity through interaction with Fcγ receptor. Specifically, how antibody immune complexes regulate the activation of immune cells is determined by the relative involvement of activating Fcγ receptors and inhibitory Fcγ receptors. Different antibody isotypes bind to activating Fcγ receptors and inhibitory Fcγ receptors with different affinities, resulting in different A:I ratios (activation:inhibition ratios) (Nimmerjahn et al.; Science. 2005 Dec 2; 310(5753): 1510-2).

[0008] Antibodies can inhibit, block, and / or downregulate effector cell function by binding to inhibitory Fcγ receptors via their Fc domains. Antibodies can stimulate cell activation by aggregating antibody-targeted signaling receptors on target cells via binding to inhibitory FcγRs via their Fc domains (Li, F. et al (2011) Science 333(6045):1030-1034; White, A. L. et al (2011) J Immunol 187(4):1754-1763; White, A. L. et al (2011) J Immunol 187(4):1754-1763 White, A. L. et al (2014) J Immunol 193(4):1828-1835).

[0009] By binding to activating Fcγ receptors, antibodies can activate effector cell function, thereby inducing mechanisms such as antibody-dependent cellular cytotoxicity (ADCC), antibody-dependent cellular phagocytosis (ADCP), cytokine release, and / or antibody-dependent endocytosis, and in the case of neutrophils, NETosis (i.e., activation and release of NETs (neutrophil extracellular traps)). Antibodies that bind to activating Fcγ receptors can also cause an increase in certain activation markers, such as CD40, MHCII, CD38, CD80, and / or CD86.

[0010] Consistent with the coordinate regulation of antibody-induced effector cell responses by activating and inhibitory Fcγ receptors, activating Fcγ receptors have been shown to promote tumor cell depletion and the therapeutic activity of antibodies that directly target tumors. Preclinical and clinical studies have demonstrated that the anti-tumor activity of antibodies that directly target tumors, i.e., antibodies whose therapeutic activity involves direct binding and killing of tumor cells, such as anti-CD20, anti-Her2, and anti-EGFR antibodies, is enhanced in patients carrying alleles that exhibit higher affinity for activating Fcγ receptors (Cartron, G. et al (2002) Blood 99(3):754-758; Musolino, A., et al (2008) J Clin Oncol 26(11):1789-1796; Zhang, W. et al (2007) J Clin Oncol 25(24):3712-3718), and antibody isotypes and formats that exhibit stronger binding to activating Fcγ receptors compared to inhibitory Fcγ receptors (high A:I ratio) (Goede, V. et al (2014) N Engl J Med 370(12):1101-1110). Conversely, the therapeutic activity of antibodies that directly target tumors is enhanced in animals lacking inhibitory FcγRIIB (Clynes, RA et al (2000) Nat Med 6(4):443-446) or, as more recently shown by some of the present inventors, when inhibitory FcγRIIB is blocked by antagonistic anti-FcγRIIB antibodies (Roghanian, A. et al (2015) Cancer Cell 27(4):473-488).

[0011] Emerging preclinical and clinical data indicate that Fcγ receptors also regulate the efficacy of immune-modulating antibodies, including immune checkpoint inhibitors targeting CTLA-4, PD-1, and PD-L1. In humans and mice, there is evidence that the therapeutic activity of anti-CTLA-4 antibodies is enhanced by the engagement of activating Fcγ receptors: melanoma patients harboring high-affinity alleles of the FcγRIIIa gene showed improved survival in response to ipilimumab compared with patients expressing low-affinity FcγRIIIa alleles. Furthermore, in mice humanized for activating and inhibitory receptors, therapeutic efficacy was shown to be FcγR-dependent and enhanced with antibody isotypes with a high A:I ratio (Arce-Vargas, F. et al. (2018) Cancer Cell 33(4):649-663 e644).

[0012] These findings prompted us to investigate the ability of FcγRIIB-blocking antibodies to enhance the activity of anti-CTLA-4 antibodies. Two different types of FcγRIIB-blocking antibodies have been previously generated and described by some of the present inventors: a human IgG1 with enhanced binding to both activating and inhibitory human FcγRs, and an Fc-engineered variant that exhibits significantly impaired Fc domain-mediated FcγR binding (Roghanian, A., et al. (2015) Cancer Cell 27(4):473-488). These two different types of anti-FcγRIIB antibodies have been shown to be equally antagonistic in blocking CD20 internalization and FcγRIIB signaling in B cells, and both improved anti-CD20 mAb-mediated B cell depletion in transgenic animals expressing human FcγRIIB and human CD20.

[0013] In International Patent Application No. PCT / EP2019 / 050566, we demonstrated that only anti-FcγRIIB antibodies lacking an Fc region or whose Fc region exhibited reduced or impaired binding to FcγRs could enhance the therapeutic activity of anti-CTLA-4 antibodies. In different mouse experimental models of solid tumors, combination treatment with an anti-FcγRIIB antibody with impaired Fc:FcγR binding, but not with an anti-FcγRIIB antibody with enhanced Fc:FcγR binding, enhanced the therapeutic activity of anti-CTLA-4. In an in vivo model of humanized PBMCs, the clinically relevant anti-CTLA-4 antibody ipilimumab enhanced Treg depletion. Similar enhancing effects on depletion and / or therapeutic efficacy of antibodies specific for IL-2R (CD25) and PD-L1 were observed after combination treatment with an anti-FcγRIIB antibody with impaired Fc:FcγR binding, indicating that the enhancing effects were not restricted to specific targets or cell types.

[0014] Although preclinical studies have demonstrated the role of Fcγ receptors in regulating the therapeutic activity of anti-PD-1 / PD-L1 antibodies, the individual roles of activating and inhibitory Fcγ receptors remain unclear. Dahan et al. reported that murine antibodies against PD-L1 benefit from FcγR engagement for their antitumor activity, whereas the activity of anti-PD-1 antibodies is impaired by FcγR engagement (Dahan, R. et al. (2015) Cancer Cell 28(3):285-295). In particular, anti-PD-1 antibody isotypes with a high A:I ratio (mIgG2a), i.e., strong engagement of activating Fcγ receptors relative to inhibitory Fcγ receptors, exhibited lower therapeutic activity compared with mIgG1 isotypes with a lower A:I ratio (i.e., relatively strong engagement of inhibitory Fcγ receptors), and compared with anti-PD-1 mutant antibodies defective in Fc:FcγR binding.

[0015] Using the clinically relevant human anti-PD-1 IgG4 antibody nivolumab and a claimed surrogate rat IgG2a antibody with a similar Fcγ receptor binding profile, Arlauckas and colleagues similarly found that anti-PD-1 mutant antibodies with impaired Fc:FcγR binding (deglycosylated) improved therapeutic activity compared with wild-type human IgG4 and rat IgG2a anti-PD-1 counterparts with high Fc:FcγR binding capacity (Arlauckas, SP et al (2017) Sci Transl Med 9 (389)). However, in contrast to Dahan et al., the authors used blocking antibodies against individual Fcγ receptors to identify activating mouse Fcγ receptor III and inhibitory mouse Fcγ receptor II as underlying the reduced efficacy of anti-PD-1 antibodies. As a result, it is not clear from the prior art the relative importance of (individual) activating and inhibitory Fcγ receptors underlying the reduced efficacy of anti-PD-1 antibodies, how they limit the efficacy of clinically relevant human anti-PD-1 antibodies, or which activating or inhibitory FcγRs need to be blocked to enhance anti-PD-1 antibody activity. Summary of the Invention

[0016] Disclosed herein is a method for producing a medicament for the treatment of a pulmonary arthritis. for use in the treatment of cancer in patients with tumor-infiltrating T lymphocytes with intermediate or high PD-1 expression, a first antibody molecule that specifically binds to FcγRIIb via the Fab region and binds to an Fcγ receptor via the Fc region; It is a combination with a second antibody molecule that specifically binds to PD-1 via the Fc region and binds to at least one Fcγ receptor.

[0017] for use in the treatment of cancer in patients with tumor-infiltrating T lymphocytes with intermediate or high PD-1 expression, (i) a first antibody molecule that specifically binds to FcγRIIb via its Fab region and binds to an Fcγ receptor via its Fc region; (ii) a second antibody molecule that specifically binds to PD-1 via its Fc region and binds to at least one Fcγ receptor.

[0018] 1. A kit for use in treating cancer in a patient having tumor-infiltrating T lymphocytes with moderate or high PD-1 expression, comprising: (i) a first antibody molecule that specifically binds to FcγRIIb via its Fab region and binds to an Fcγ receptor via its Fc region; (ii) a second antibody molecule that specifically binds to PD-1 via its Fc region and binds to at least one Fcγ receptor.

[0019] In the manufacture of a drug for use in treating cancer in patients with tumor-infiltrating T lymphocytes with moderate or high PD-1 expression, (i) a first antibody molecule that specifically binds to FcγRIIb via its Fab region and binds to an Fcγ receptor via its Fc region; and (ii) Further disclosed herein is the use of a second antibody molecule that specifically binds to PD-1 via its Fc region and binds to at least one Fcγ receptor.

[0020] 1. A method for treating cancer in a patient having tumor-infiltrating T lymphocytes with intermediate or high PD-1 expression, comprising: (i) a first antibody molecule that specifically binds to FcγRIIb via its Fab region and binds to an Fcγ receptor via its Fc region; (ii) a second antibody molecule that specifically binds to PD-1 via its Fc region and binds to at least one Fcγ receptor.

[0021] The patient, (i) a first antibody molecule that specifically binds to FcγRIIb via its Fab region and binds to an Fcγ receptor via its Fc region; (ii) a diagnostic test to determine whether a patient would benefit from combination therapy with a second antibody molecule that specifically binds PD-1 via its Fc region and binds to at least one Fcγ receptor, Further disclosed herein is a diagnostic test in which the test involves determining PD-1 expression on tumor-infiltrating T lymphocytes of a patient, wherein moderate or high PD-1 expression indicates that the patient will benefit from the combination therapy. Correspondingly, a lack of moderate or high PD-1 expression on T lymphocytes, while low PD-1 expression, suggests that the patient will not benefit from the combination therapy. DETAILED DESCRIPTION OF THE INVENTION

[0022] Here, we demonstrate that only anti-FcγRIIB with high Fc:FcγR binding capacity, but not anti-FcγRIIB with impaired Fc:FcγR binding, enhances the therapeutic efficacy of anti-PD-1 antibodies in vivo and prevents phagocytosis of clinically relevant human anti-PD-1 antibodies in PD-1-high expressing T cells in vitro. This finding is novel and unexpected, as previous studies on the role of FcγRs in anti-PD-1 therapy have demonstrated either a broader role for activating FcγRs compared with inhibitory FcγRs (Dahan, R. et al. (2015) Cancer Cell 28(3):285-295) or the individual activating (FcγRIII) and inhibitory FcγRIIB (Arlauckas, SP et al. (2017) Sci Transl Med 9(389)) FcγRIIB as underlying impaired anti-PD-1 antibody activity.

[0023] The present invention is even more surprising in light of some of our previous findings with antibodies against other immune checkpoints, including anti-CTLA-4, where only anti-FcγRIIB with impaired Fc:FcγR binding, but not anti-FcγRIIB with enhanced Fc:FcγR binding, enhances therapeutic activity.

[0024] Thus, the present invention provides a method for the treatment of cancer in patients with tumor-infiltrating T lymphocytes with intermediate or high PD-1 expression. (i) an antibody molecule (referred to herein as a first antibody molecule) that specifically binds to FcγRIIb via its Fab region and binds to an Fcγ receptor via its Fc region; (ii) in combination with an antibody molecule (referred to herein as the second antibody molecule) that specifically binds to PD-1 via its Fc region and binds to at least one Fcγ receptor.

[0025] This combination is intended for use in treating cancer, such as solid tumors, in patients with cancer, with the aim of improving the therapeutic efficacy of antibody molecules that specifically bind to PD-1, i.e., anti-PD-1 antibodies, by reducing binding to FcγR, including FcγRIIB.

[0026] The antibody molecule of the present invention, i.e., the first antibody, that specifically binds to FcγRIIb binds or interacts with this Fcγ receptor via the Fab region of the antibody, i.e., via the antigen-binding region on the antibody that binds to the antigen, which is composed of one constant region and one variable region of the heavy and light chains, respectively. In particular, it binds to FcγRIIb present on immune effector cells, particularly FcγRIIb present on the surface of immune effector cells.

[0027] In addition to the above, the antibody molecule according to the present invention that specifically binds to FcγRIIb, i.e., the first antibody molecule, also binds to activating Fcγ receptors via interactions between the Fc region and the Fc receptor, as is known (Bruhns, P. et al (2009) Blood 113(16):3716-3725) and has been extensively characterized for antibodies of the human IgG1 isotype.

[0028] This has at least the following therapeutically important consequences: both activating and inhibitory Fcγ receptors are blocked in an anti-FcγRIIB antibody-dependent (Fab- and Fc-) manner, preventing macrophage phagocytosis or anti-PD-1 antibody-mediated elimination of anti-tumor T cells coated with anti-PD-1 antibodies from immune effector cells expressing other Fcγ receptors (coated in this context means that the anti-PD-1 antibody is bound to the cells). This mechanism may further involve the inhibition of macrophage FcγR-dependent transfer of PD-1 antibodies from T cells to FcγR-expressing effector cells, such as macrophages, as previously described (Arlauckas, SP et al (2017) Sci Transl Med 9 (389)).

[0029] Immune effector cells expressing Fc gamma receptors herein primarily refer to innate effector cells, specifically including macrophages, neutrophils, monocytes, natural killer (NK) cells, basophils, eosinophils, mast cells, and platelets. Cytotoxic T cells and memory T cells do not normally express FcγR, but may do so under certain circumstances. In some embodiments, the immune effector cells are innate immune effector cells. In some embodiments, the immune effector cells are macrophages.

[0030] The antibody molecule that specifically binds to or interacts with PD-1, i.e., the second antibody molecule, has an Fc region that binds to or interacts with an activating Fcγ receptor, allowing antibody-PD-1 antibody-dependent FcγR effector cell-dependent elimination of anti-tumor T cells coated with the anti-PD-1 antibody. The immune cells that the anti-PD-1 antibody molecule binds are immune cells that confer important anti-tumor activity, such as CD8+ or CD4+ T cells.

[0031] As a result, any anti-PD-1 mutant antibody, including those of the human IgG4, IgG1, IgG2, and IgG3 isotypes, whose Fc region binds to or interacts with activating Fcγ receptors to an extent that results in depletion of FcγR-expressing effector cells of PD-1-expressing anti-tumor T cells, can be combined with an anti-FcγRIIB antibody with high Fc:FcγR binding ability, i.e., an antibody molecule that specifically binds FcγRIIb via the Fab region and binds to Fcγ receptors via the Fc region.

[0032] The second antibody is an anti-PD-1 antibody. PD-1 (programmed cell death protein 1), also known as CD279, is an immune checkpoint, i.e., a checkpoint protein on immune cells. It promotes apoptosis of antigen-specific T cells in lymph nodes and reduces apoptosis of regulatory T cells. PD-1 inhibitors, such as nivolumab (OPDIVO®), pembrolizumab (KEYTRUDA®), and cemiplimab (LIBTAYO®), are used in cancer treatment to activate the immune system to attack tumors. Monoclonal antibodies targeting either PD-1 or PD-L1 can block the binding of PD-1 to PD-L1, potentially enhancing the immune response against cancer cells.

[0033] Anti-PD-1 antibodies bind to PD-1 expressed on intratumoral T cells.

[0034] Patients who will benefit from treatment according to the present invention are those who are eligible for anti-PD-1 therapy according to the standard criteria for approved anti-PD-1 antibody-containing regimens and who have tumor-infiltrating T lymphocytes with intermediate or high PD-1 expression (i.e., tumor-infiltrating CD3+ lymphocytes). Patients eligible for anti-PD-1 therapy include those with melanoma; lung cancer, including small cell lung cancer (SCLC) and non-small cell lung cancer (NSCLC), including non-squamous NSCLC and squamous NSCLC, and including metastatic NSCLC; head and neck cancer, including head and neck squamous cell carcinoma (HNSCC); Hodgkin lymphoma; primary mediastinal B-cell lymphoma (PMBCL); bladder cancer, including advanced urothelial carcinoma; colorectal cancer, including cancers that are highly instable (MSI-H) and / or mismatch repair deficient (dMMR); gastric cancer, including advanced gastric cancer and gastric or gastroesophageal junction (GEJ) adenocarcinoma; cervical cancer; liver cancer, including hepatocellular carcinoma; Merkel cell carcinoma (MCC); kidney cancer, including renal cell carcinoma (RCC) and cutaneous squamous cell carcinoma (CSCC), including locally advanced CSCC in patients who are not candidates for curative surgery or curative radiation therapy. As known to those skilled in the art, the number of indications that can be treated is rapidly expanding with new clinical trials, new anti-PD-1 antibodies, and new combinations.

[0035] In the studies leading to the present invention, it was observed that when an anti-PD-1 antibody is administered to T cells with moderate or high expression of PD-1, this can lead to phagocytosis of anti-PD-1 antibody-coated T cells, particularly CD8-positive T cells. In the studies leading to the present invention, it was further discovered that phagocytosis can be blocked in T cells with moderate or high expression of PD-1 by administering an antibody molecule that specifically binds to FcγRIIb via its Fab region and Fcγ receptor via its Fc region together with an anti-PD-1 antibody. The in vivo relevance of the in vitro assay used for the above discovery, which will be further described in the Examples below, was confirmed in two different solid cancer experimental models involving immunocompetent mice, and it was observed that combined treatment with anti-FcγRIIB with high Fc:FcγR binding activity and anti-PD-1 antibody significantly improved survival compared with monotherapy with anti-PD-1. This is also shown in more detail in the Examples below. Further supporting the in vivo relevance of the in vitro assay, PD-1 expression levels on intratumoral in vivo T cells were similar in the two settings. PD-1 expression on intratumoral T cells in vivo ranged from approximately 20,000 to 80,000 PD-1 molecules per cell, and in vitro PD-1 expression levels ranged from moderate (15,500 to 78,000 PD-1 molecules) to highly expressing (65,000 to 391,000 PD-1 molecules per cell) human T cells, both of which were sensitive to blockade of human anti-PD-1-mediated phagocytosis by the highly potent anti-FcγRIIB Fc:FcγR binding.

[0036] Thus, in the context of the present invention, patients who may benefit from the treatment described herein are those with at least 10% of tumor-infiltrating T lymphocytes with intermediate or high expression of PD-1.

[0037] As used herein, moderate or high expression refers to expression of ≥ 15,500 PD-1 molecules per cell in at least 10% of tumor-infiltrating T lymphocytes. As further described below, the absolute number of PD-1 expression may vary depending on which anti-PD-1 antibody and / or which method is used to measure PD-1 expression. Thus, as used herein, moderate or high expression of ≥ 15,500 PD-1 molecules per cell is measured using the methods described herein and / or the anti-human PD-1 antibody EH12.2H7 (available from BioLegend).

[0038] Similar to in vivo settings, intratumoral T cells of individual patients exhibit heterogeneous PD-1 expression. Individual patients may have different cell populations with different expression of PD-1, for example, some populations with low expression and some populations with intermediate or high expression. In some embodiments, at least 15% of the patient's tumor-infiltrating CD3+ T lymphocytes have intermediate or high PD-1 expression. In some embodiments, at least 20% of the patient's tumor-infiltrating CD3+ T lymphocytes have intermediate or high PD-1 expression. In some embodiments, at least 25% of the patient's tumor-infiltrating CD3+ T lymphocytes have intermediate or high PD-1 expression. In some embodiments, at least 30% of the patient's tumor-infiltrating CD3+ T lymphocytes have intermediate or high PD-1 expression. In some embodiments, at least 35% of the patient's tumor-infiltrating CD3+ T lymphocytes have intermediate or high PD-1 expression. In some embodiments, at least 40% of the patient's tumor-infiltrating CD3+ T lymphocytes have moderate or high PD-1 expression. In some embodiments, at least 45% of the patient's tumor-infiltrating CD3+ T lymphocytes have moderate or high PD-1 expression. In some embodiments, at least 50% of the patient's tumor-infiltrating CD3+ T lymphocytes have moderate or high PD-1 expression. In some embodiments, at least 55% of the patient's tumor-infiltrating CD3+ T lymphocytes have moderate or high PD-1 expression. In some embodiments, at least 60% of the patient's tumor-infiltrating CD3+ T lymphocytes have moderate or high PD-1 expression. In some embodiments, at least 65% of the patient's tumor-infiltrating CD3+ T lymphocytes have moderate or high PD-1 expression. In some embodiments, at least 70% of the patient's tumor-infiltrating CD3+ T lymphocytes have moderate or high PD-1 expression. In some embodiments, at least 75% of the patient's tumor-infiltrating CD3+ T lymphocytes have intermediate or high PD-1 expression, hi some embodiments, at least 80% of the patient's tumor-infiltrating CD3+ T lymphocytes have intermediate or high PD-1 expression.In some embodiments, at least 85% of the patient's tumor-infiltrating CD3+ T lymphocytes have intermediate or high PD-1 expression, hi some embodiments, at least 90% of the patient's tumor-infiltrating CD3+ T lymphocytes have intermediate or high PD-1 expression.

[0039] In some embodiments, it is the patient's tumor-infiltrating CD3-positive and CD8-positive (CD3+CD8+) T lymphocytes that have intermediate or high PD-1 expression.

[0040] In some embodiments, at least 10% of the patient's tumor-infiltrating CD3+CD8+ T lymphocytes have intermediate or high PD-1 expression. In some embodiments, at least 15% of the patient's tumor-infiltrating CD3+CD8+ T lymphocytes have intermediate or high PD-1 expression. In some embodiments, at least 20% of the patient's tumor-infiltrating CD3+CD8+ T lymphocytes have intermediate or high PD-1 expression. In some embodiments, at least 25% of the patient's tumor-infiltrating CD3+CD8+ T lymphocytes have intermediate or high PD-1 expression. In some embodiments, at least 30% of the patient's tumor-infiltrating CD3+CD8+ T lymphocytes have intermediate or high PD-1 expression. In some embodiments, at least 35% of the patient's tumor-infiltrating CD3+CD8+ T lymphocytes have intermediate or high PD-1 expression. In some embodiments, at least 40% of the patient's tumor-infiltrating CD3+CD8+ T lymphocytes have intermediate or high PD-1 expression. In some embodiments, at least 45% of the patient's tumor-infiltrating CD3+CD8+ T lymphocytes have intermediate or high PD-1 expression. In some embodiments, at least 50% of the patient's tumor-infiltrating CD3+CD8+ T lymphocytes have intermediate or high PD-1 expression. In some embodiments, at least 55% of the patient's tumor-infiltrating CD3+CD8+ T lymphocytes have intermediate or high PD-1 expression. In some embodiments, at least 60% of the patient's tumor-infiltrating CD3+CD8+ T lymphocytes have intermediate or high PD-1 expression. In some embodiments, at least 65% of the patient's tumor-infiltrating CD3+CD8+ T lymphocytes have intermediate or high PD-1 expression. In some embodiments, at least 70% of the patient's tumor-infiltrating CD3+CD8+ T lymphocytes have intermediate or high PD-1 expression. In some embodiments, at least 75% of the patient's tumor-infiltrating CD3+CD8+ T lymphocytes have intermediate or high PD-1 expression. In some embodiments, at least 80% of the patient's tumor-infiltrating CD3+CD8+ T lymphocytes have intermediate or high PD-1 expression.In some embodiments, at least 85% of the patient's tumor-infiltrating CD3+CD8+ T lymphocytes have intermediate or high PD-1 expression, hi some embodiments, at least 90% of the patient's tumor-infiltrating CD3+CD8+ T lymphocytes have intermediate or high PD-1 expression.

[0041] PD-1 expression on tumor cells in individual patients can be measured using cells or tissues from tumor biopsies. More specifically, absolute expression levels on T cells can be quantified using flow cytometry and bead-based antibody and cellular epitope quantification kits described herein or equivalent. Alternatively, semiquantitative analysis can be performed by immunohistochemistry using tumor tissue biopsies, and anti-PD-1 staining of patient biopsies can be compared to staining of tissues or cytospun cells expressing defined and determined PD-1 levels associated with sensitivity (≥15,500 PD-1 molecules per cell) or insensitivity (<15,500 PD-1 molecules per cell) to anti-FcγRIIB-mediated enhancement of anti-PD-1 antibody activity. Importantly, if the method for determining human T cell PD-1 expression differs from the quantification method described herein or uses a different anti-PD-1 antibody clone or fluorescent label, the assay should be compared and validated, for example, with the method described in detail in the Examples below, particularly Example 1 with reference to Figure 1. Generally, as illustrated in Example 1 with reference to Figure 1, one method for quantifying PD-1 expression is to use a fluorochrome-labeled antibody at a defined ratio, e.g., and preferably in some embodiments, an antibody labeled with phycoerythrin (PE) at a 1:1 ratio. This allows for the generation of a standard curve using beads containing a defined number of fluorochrome (e.g., PE) molecules to determine the number of antibody molecules bound to the cells. The cells to be tested are incubated with a labeled anti-PD1 antibody (e.g., BioLegend's anti-human PD-1 antibody EH12.2H7) and analyzed using a FACS machine configured to run beads and cells under the same settings. For example, a standard curve can be generated by plotting Log molecules per bead versus Log fluorescence, and then fluorochrome-labeled anti-PD1 antibody-stained cells are run and the Log mean fluorescence intensity (MFI) is used to calculate the number of bound antibody.

[0042] As mentioned above, the absolute numbers may vary depending on which anti-PD1 antibody is used in the measurement.

[0043] In addition to specifically binding to PD-1 on immune cells, the second antibody molecule binds to at least one Fcγ receptor via its Fc region. In some embodiments, the second antibody molecule binds to at least one activating Fcγ receptor via its Fc region. The second antibody may be capable of binding to an activating Fcγ receptor, such as an activating Fcγ receptor present on an immune effector cell, via its Fc region. To enable binding to an activating Fcγ receptor, the Fc region of the second antibody may, at least in some embodiments, be glycosylated at position 297. Carbohydrate residues at this position are useful for binding to the Fcγ receptor. In some embodiments, these residues are preferably biantennary carbohydrates containing GlnNAc, mannose, along with terminal galactose residues and sialic acid. This should comprise the CH2 portion of the Fc molecule.

[0044] The present invention further relates to a diagnostic test that can be used to identify patients who will benefit from the treatment described herein, i.e., a combination treatment of (i) a first antibody molecule that specifically binds to FcγRIIb via its Fab region and to an Fcγ receptor via its Fc region, and (ii) a second antibody molecule that specifically binds to PD-1 via its Fc region and to at least one Fcγ receptor. Based on in vivo PD-1 expression levels associated with improved efficacy of anti-FcγRIIB-mediated anti-PD-1 antibodies, as well as in vitro phagocytosis assays incorporating therapeutically relevant human anti-PD-1 antibodies and human T cells and macrophages, the inventors have determined that specific T cell PD-1 receptor expression levels are associated with and required for enhanced efficacy of anti-FcγRIIB-mediated anti-PD-1 therapy. A diagnostic test according to the present invention is based on this finding and therefore involves measuring PD-1 expression on tumor cells in a sample, such as cells or tissue from a tumor biopsy obtained from a patient. As described above, absolute or semi-quantitative analysis of PD-1 expression levels on T cells can be used. In some embodiments, the diagnostic test is based on the use of the anti-PD1 antibody EH12.2H7 to measure PD-1 expression. Expression of at least 15,500 PD-1 molecules per T lymphocyte predicts that the patient is likely to benefit from a combination treatment according to the present invention, as further described above and in Example 1.

[0045] Antibodies are known to those skilled in the art of immunology and molecular biology. Typically, an antibody comprises two heavy chains (H) and two light chains (L). This complete antibody molecule is sometimes referred to herein as a full-size antibody or full-length antibody. The heavy chain of an antibody comprises one variable region (VH) and three constant regions (CH1, CH2, CH3), and the light chain of an antibody molecule comprises one variable region (VL) and one constant region (CL). The variable regions (sometimes referred to as F VThe variable regions (collectively referred to as CDRs) bind to the antibody's target, or antigen. Each variable region contains three loops called complementarity-determining regions (CDRs), which are involved in target binding. The constant regions are not directly involved in binding the antibody to the antigen, but exhibit various effector functions. Depending on the amino acid sequence of the constant regions of their heavy chains, antibodies or immunoglobulins can be assigned to different classes. There are five major classes of immunoglobulins: IgA, IgD, IgE, IgG, and IgM; in humans, some of these are further divided into subclasses (isotypes), e.g., IgG1, IgG2, IgG3, IgG4; IgA1, and IgA2.

[0046] Another part of an antibody is the Fc region (also known as the fragment crystallizable domain), which contains two constant domains in each of the antibody's heavy chains. As mentioned above, the Fc region is involved in the interaction between the antibody and the Fc receptor.

[0047] As used herein, the term antibody molecule includes full-length or full-size antibodies, as well as functional fragments of full-length antibodies and derivatives of such antibody molecules.

[0048] A functional fragment of a full-size antibody has the same antigen-binding characteristics as the corresponding full-size antibody, and contains either the same variable domain (i.e., VH sequence and VL sequence) and / or the same CDR sequence as the corresponding full-size antibody. Having the same antigen-binding characteristics as the corresponding full-size antibody means that the functional fragment binds to the same epitope on the target as the full-size antibody. Such a functional fragment may correspond to the Fv portion of a full-size antibody. Alternatively, such a fragment may be Fab, also referred to as F(ab), which is a monovalent antigen-binding fragment that does not contain the Fc portion, or F(ab')2, which is a bivalent antigen-binding fragment containing two antigen-binding Fab portions linked together by disulfide bonds, or a monovalent variant of F(ab'), i.e., F(ab')2. Such a fragment may also be a single-chain variable fragment (scFv).

[0049] Functional fragments do not always contain all six CDRs of the corresponding full-size antibody.It is understood that molecules containing three or fewer CDR regions (sometimes only a single CDR or a portion thereof) can retain the antigen binding activity of the antibody derived from that CDR.For example, Gao et al., 1994, J.Biol.Chem.,269:32389-93, has described that the entire VL chain (comprising all three CDRs) has high affinity for its substrate.

[0050] Molecules containing two CDR regions are described, for example, in Vaughan & Sollazzo 2001, Combinatorial Chemistry & High Throughput Screening, 4:417-430. On page 418 (right column -3 (Our Strategy for Design)), a minibody containing only H1 and H2 CDR hypervariable regions interspersed within framework regions is described. Minibodies are described as being capable of binding to targets. Pessi et al., 1993, Nature, 362:367-9, and Bianchi et al., 1994, J. Mol. Biol., 236:649-59, referenced by Vaughan & Sollazzo, provide more detailed descriptions of H1 and H2 minibodies and their properties. Qiu et al., 2007, Nature Biotechnology, 25:921-9, demonstrate that molecules consisting of two linked CDRs are capable of binding to antigens. Quiocho 1993, Nature, 362:293-4 provides an overview of "minibody" technology. Ladner 2007, Nature Biotechnology, 25:875-7 observes that molecules containing two CDRs can retain antigen-binding activity.

[0051] Antibody molecules containing a single CDR region are described, for example, in Laune et al., 1997, JBC, 272:30937-44, where it was demonstrated that a series of hexapeptides derived from CDRs exhibited antigen-binding activity, and it was noted that synthetic peptides of a complete single CDR exhibited strong binding activity. Monnet et al., 1999, JBC, 274:3789-96, demonstrated that various 12-mer peptides and related framework regions had antigen-binding activity, and stated that a CDR3-like peptide alone could bind to the antigen. Heap et al., 2005, J. Gen. Virol., 86:1791-1800, reported that "microantibodies" (molecules containing a single CDR) could bind to antigens, and demonstrated that a cyclic peptide from an anti-HIV antibody had antigen-binding activity and function. Nicaise et al., 2004, Protein Science, 13:1882-91, show that a single CDR can confer antigen binding activity and affinity for its lysozyme antigen.

[0052] Thus, antibody molecules having five, four, three or fewer CDRs can retain the antigen-binding properties of the full-length antibody from which they are derived.

[0053] An antibody molecule may be a derivative of a full-length antibody or a fragment of such an antibody, where the derivative has the same antigen-binding characteristics as the corresponding full-size antibody, meaning that it binds to the same epitope on the target as the full-size antibody.

[0054] Thus, as used herein, the term "antibody molecule" includes all types of antibody molecules, including monoclonal antibodies, polyclonal antibodies, synthetic antibodies, recombinantly produced antibodies, multispecific antibodies, bispecific antibodies, human antibodies, human-derived antibodies, humanized antibodies, chimeric antibodies, single-chain antibodies, single-chain Fvs (scFvs), Fab fragments, F(ab') fragments, F(ab') fragments, disulfide-linked Fvs (sdFvs), antibody heavy chains, antibody light chains, antibody heavy chain homodimers, antibody light chain homodimers, antibody heavy chain heterodimers, antibody light chain heterodimers, antigen-binding functional fragments of such homodimers and heterodimers, as well as functional fragments and derivatives thereof.

[0055] Furthermore, as used herein, the term "antibody molecule" includes all classes of antibody molecules and functional fragments, including IgG, IgG1, IgG2, IgG3, IgG4, IgA, IgM, IgD, and IgE, unless otherwise specified.

[0056] In some embodiments, the first antibody is human IgG1. Those skilled in the art will understand that mouse IgG2a and human IgG1 can bind to block activating Fc gamma receptors, thereby preventing anti-PD-1 antibody-mediated engagement of activating Fc gamma receptors on immune effector cells and subsequent elimination of anti-PD-1 antibody-coated effector T cells, for example, by ADCP or ADCC. Thus, in embodiments in which mouse IgG2a is the preferred isotype for deletion in mice, human IgG1 is the preferred isotype for deletion in humans in such embodiments.

[0057] In some embodiments, the first antibody is a human IgG1. In other embodiments, the first antibody is a human IgG4, IgG3, or IgG2. In other embodiments, the first antibody is a human IgG antibody that has been Fc-engineered to enhance binding to Fc gamma receptors. In some embodiments, the human IgG antibody is Fc-engineered to improve binding to one or several activating Fc gamma receptors and / or engineered to improve relative binding to activating receptors over inhibitory Fc gamma receptors. In some embodiments, the anti-Fc gamma RIIB antibody is an Fc-engineered human IgG antibody. Examples of such engineered antibody variants include non-fucosylated antibodies with selectively improved antibody binding to FcγRIIIA, and antibodies engineered by directed, mutational, or other means amino acid substitutions that result in improved binding to one or several activating Fcγ receptors compared to the inhibitory FcγRIIB (Richards et al. 2008. 'Optimization of antibody binding to FcgammaRIIa enhances macrophage phagocytosis of tumor cells', Mol Cancer Ther, 7:2517-27; Lazar et al. 2006. 'Engineered antibody Fc variants with enhanced effector function', Proc Natl Acad Sci USA, 103:4005-10). In some embodiments, the human IgG antibody engineered to improve binding to activating Fc gamma receptors may be a human IgG antibody having two mutations, S239D and I332E, or three mutations, S239D, I332E, and A330L, and / or a G236A mutation, in its Fc portion. In some embodiments, the human IgG antibody engineered to improve binding to activating Fc gamma receptors may be a non-fucosylated human IgG antibody.

[0058] In some embodiments, the second antibody is a human IgG4, which is the isotype of the currently FDA-approved anti-PD-1 antibodies nivolumab, pembrolizumab, and ceplizumab. Those skilled in the art will understand that some mouse antibody isotypes can bind to both activating and inhibitory Fc gamma receptors. Importantly, those skilled in the art will understand that the rat IgG2a isotype, which binds to mouse activating and inhibitory Fc gamma receptors, is known to closely mimic the human IgG4 isotype, which binds to human activating and inhibitory Fc gamma receptors (Arlauckas, SP et al (2017) Sci Transl Med 9 (389)). Those skilled in the art will further understand that, in addition to human isotypes IgG1 and IgG4, human IgG3 and IgG2 antibodies can productively bind to human FcγR (Sanders, LA et al (1995) Infect Immun 63(1):73-81) and can mediate antibody-dependent T cell depletion, for example, by ADCP and ADCC, following activation of activating Fc gamma receptors with immune cells (Arce Vargas, F. et al (2018) Cancer Cell 33(4):649-663e644). Consequently, in some embodiments, the second antibody can be a human IgG1 or IgG2 or IgG3 antibody.

[0059] As outlined above, different types and forms of antibody molecules are encompassed by the present invention and would be known to one skilled in the art of immunology. It is known that antibodies used for therapeutic purposes are often modified with additional moieties that modify the properties of the antibody molecule.

[0060] Thus, antibody molecules of the invention or used in accordance with the invention (e.g., monoclonal and / or polyclonal and / or bispecific antibody molecules) may comprise a detectable moiety and / or a cytotoxic moiety.

[0061] A "detectable moiety" includes one or more from the group consisting of an enzyme, a radioactive atom, a fluorescent moiety, a chemiluminescent moiety, and a bioluminescent moiety. The detectable moiety allows for the visualization of the antibody molecule in vitro, and / or in vivo, and / or ex vivo.

[0062] "Cytotoxic moieties" include radioactive moieties and / or enzymes, where the enzymes are caspases and / or toxins, where the toxins are bacterial toxins or venoms, and where the cytotoxic moieties are capable of inducing cell lysis.

[0063] Additionally, antibody molecules may be in isolated and / or purified form and / or may be PEGylated, which is the process of adding polyethylene glycol polymers to a molecule such as an antibody molecule or derivative to modify its behavior, e.g., increase its hydrodynamic size and prevent renal clearance, thereby increasing its half-life.

[0064] As discussed above, the CDRs of an antibody bind to the antibody target. The amino acid assignments for each CDR described herein are as defined by Kabat EA et al. 1991, "Sequences of Proteins of Immunological Interest," Fifth Edition, NIH Publication No. 91-3242, pp. xv-xvii.

[0065] As those skilled in the art will recognize, there are other methods for assigning amino acids to each CDR, such as the International ImMunoGeneTics information system (IMGT®) (http: / / www.imgt.org / and Lefranc and Lefranc "The Immunoglobulin Facts Book" published by Academic Press, 2001).

[0066] In a further embodiment, the antibody molecule of the present invention or used in accordance with the present invention is an antibody molecule that is capable of competing with the specific antibodies provided herein for binding to a specific target, e.g., an antibody molecule comprising any of the amino acid sequences set out in SEQ ID NOs: 1 to 194.

[0067] By "capable of competing" is meant that the competing antibody is capable of at least partially inhibiting or otherwise interfering with the binding of an antibody molecule as defined herein to a particular target.

[0068] For example, such a competitor antibody molecule may be capable of inhibiting binding of an antibody molecule described herein by at least about 10%, e.g., at least about 20%, or at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, at least about 95%, or about 100% and / or may be capable of inhibiting the binding ability of an antibody described herein to prevent or reduce binding to a specific target by at least about 10%, e.g., at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, at least about 95%, or about 100%.

[0069] Competitive binding can be determined by methods known to those skilled in the art, such as enzyme-linked immunosorbent assay (ELISA).

[0070] ELISA assay can be used to evaluate epitope-modifying or blocking antibodies. Additional suitable methods for identifying competitive antibodies are disclosed in Antibodies: A Laboratory Manual, Harlow & Lane (see, for example, pages 567-569, 574-576, 583, and 590-612, 1988, CSHL, NY, ISBN 0-87969-314-2), which is incorporated herein by reference.

[0071] It is well known that antibodies specifically bind to or interact with a defined target molecule or antigen, and that this means that the antibody binds preferentially and selectively to that target over non-target molecules.

[0072] Targets of antibodies according to the invention, or for use in accordance with the invention, are expressed on the surface of cells, i.e., they are cell surface antigens that comprise the epitope (alternatively known in this context as a cell surface epitope) for the antibody. Cell surface antigen and epitope are terms readily understood by those skilled in the art of immunology or cell biology.

[0073] The term "cell surface antigen" includes cases where the cell surface antigen is exposed on the extracellular side of the cell membrane, but only transiently exposed on the extracellular side of the cell membrane. "Transiently exposed" includes cases where the cell surface antigen is internalized into the cell or released from the extracellular side of the cell membrane into the extracellular space. Cell surface antigens can be released from the extracellular side of the cell membrane by cleavage, which can be mediated by proteases.

[0074] Also, cell surface antigens may bind to cell membranes, but may only be transiently associated with the cell membrane. "Transiently associated" includes cases where cell surface antigens are released from the extracellular side of the cell membrane into the extracellular space. Cell surface antigens may be released from the extracellular side of the cell membrane by cleavage, which may be mediated by proteases.

[0075] Furthermore, the cell surface antigen may be an epitope present on a peptide, or a polypeptide, or a carbohydrate, or an oligosaccharide chain, or a lipid, and / or a protein, or a glycoprotein, or a lipoprotein.

[0076] Methods for assessing protein binding are known to those skilled in the art of biochemistry and immunology.Those skilled in the art will understand that these methods can be used to assess the binding of antibody to target and / or the binding of antibody Fc region to Fc receptor, and the relative strength, specificity, or inhibition, prevention, or reduction of their interactions.Examples of methods that can be used to assess protein binding include, for example, immunoassay, BIAcore, Western blot, radioimmunoassay (RIA), and enzyme-linked immunosorbent assay (ELISA) (for a discussion of antibody specificity, see Fundamental Immunology Second Edition, Raven Press, New York at pages 332-336 (1989)).

[0077] Thus, a "specifically binding antibody molecule" or a "target-specific antibody molecule" includes cases where the antibody molecule specifically binds to a target but does not bind to a non-target, or binds to the non-target weaker than it does to the target (e.g., with lower affinity).

[0078] It also includes the meaning that the antibody specifically binds to a target at least 2 times stronger, or at least 5 times stronger, or at least 10 times stronger, or at least 20 times stronger, or at least 50 times stronger, or at least 100 times stronger, or at least 200 times stronger, or at least 500 times stronger, or at least about 1000 times stronger to a target than to a non-target.

[0079] In addition, the antibody targets at least about 10 -1 K d , or at least about 10 -2 K d , or at least about 10 -3 K d , or at least about 10 -4 K d , or at least about 10 -5 K d , or at least about 10 -6 K d, or at least about 10 -7 K d , or at least about 10 -8 K d , or at least about 10 -9 K d , or at least about 10 -10 K d , or at least about 10 -11 K d , or at least about 10 -12 K d , or at least about 10 -13 K d , or at least about 10 -14 K d , or at least about 10 -15 K d K d By binding, it is meant that the antibody specifically binds to the target.

[0080] In some embodiments, the antibody molecule that specifically binds to FcγRIIb is a human antibody.

[0081] In some embodiments, antibody molecules that specifically bind to FcγRIIb are human-derived antibodies, i.e., human-derived antibodies that have been modified as described herein.

[0082] In some embodiments, antibody molecules that specifically bind to FcγRIIb are humanized antibodies, i.e., antibodies of non-human origin that have been modified to increase their similarity to human antibodies. The humanized antibody may be, for example, a murine antibody or a llama antibody.

[0083] In some embodiments, an antibody molecule that specifically binds to FcγRIIb comprises the following constant regions (CH and CL): IgG1-CH [SEQ ID NO: 1] ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGV EVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK IgG1-CL [SEQ ID NO: 2] QPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECS

[0084] In some embodiments, an antibody molecule that specifically binds to FcγRIIb comprises the sequence of one or more of the following clones: Antibody clone: ​​1A01 1A01-VH [SEQ ID NO: 3] EVQLLESGGGLVQPGGSLRLSCAASGFTFSDYYMNWIRQTPGKGLEWVSLIGWDGGSTYYADSVKGRFTISRDNSENTLYLQMNSLRAEDTAVYYCARAYSGYELDYWGQGTLVTVSS 1A01-VL [SEQ ID NO: 27] QSVLTQPPSASGTPGQRVTISSCSGSSSNIGNNAVNWYQQLPGTAPKLLIYDNNNRPSGVPDRFSGSKSGTSASLAISGLRSEDEADYYCAAWDDSLNASIFGGGTKLTVLG CDR area CDRH1:DYYMN [SEQ ID NO:51] CDRH2:LIGWDGGSTYYADSVKG [SEQ ID NO: 52] CDRH3:AYSGYELDY [SEQ ID NO: 53] CDRL1:SGSSSNIGNNAVN [SEQ ID NO: 54] CDRL2:DNNNRPS [SEQ ID NO: 55] CDRL3:AAWDDSLNASI [SEQ ID NO: 56] Antibody clone: ​​1B07 1B07-VH [SEQ ID NO: 4] EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYGMHWVRQAPGKGLEWVAFTRYDGSNKYYADSVRGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARENIDAFDVWGQGTLVTVSS 1B07-VL [SEQ ID NO: 28] QSVLTQPPSASGTPGQRVTISSCSGSSSNIGNNAVNWYQQLPGTAPKLLIYDNQQRPSGVPDRFSGSKSGTSASLAISGLRSEDEADYYCEAWDDRLFGPVFGGGTKLTVLG CDR area CDRH1:SYGMH [SEQ ID NO: 57] CDRH2:FTRYDGSNKYYADSVRG [SEQ ID NO: 58] CDRH3:ENIDAFDV [SEQ ID NO: 59] CDRL1:SGSSSNIGNNAVN [SEQ ID NO: 60] CDRL2:DNQQRPS [SEQ ID NO: 61] CDRL3: WDDRLFGPV [SEQ ID NO: 62] Antibody clone: ​​1C04 1C04-VH [SEQ ID NO: 5] EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYAMSWVRQAPGKGLEWVSSISDSGAGRYYADSVEGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARTHDSGELLDAFDIWGQGTLVTVSS 1C04-VL [SEQ ID NO: 29] QSVLTQPPSASGTPGQRVTISCSGSSSNIGSNHVLWYQQLPGTAPKLLIYGNSNRPSGVPDRFSGSKSGTSASLAISGLRSEDEADYYCAAWDDSLNGWVFGGGTKLTVLG CDR area CDRH1:SYAMS [SEQ ID NO: 63] CDRH2:SISDSGAGRYYADSVEG [SEQ ID NO: 64] CDRH3:THDSGELLDAFDI [SEQ ID NO: 65] CDRL1: SGSSSNIGSNHVL [SEQ ID NO: 66] CDRL2: GNSNRPS [SEQ ID NO: 67] CDRL3:AAWDDSLNGWV [SEQ ID NO: 68] Antibody clone: ​​1E05 1E05-VH [SEQ ID NO: 6] EVQLLESGGGLVQPGGSLRLSCAASGFTFSTYAMNWVRQVPGKGLEWVAVISYDGSNKNYVDSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARNFDNSGYAIPDAFDIWGQGTLVTVSS 1E05-VL [SEQ ID NO: 30] QSVLTQPPSASGTPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYDNNSRPSGVPDRFSGSKSGTSASLAISGLRSEDEADYYCAAWDDSLGGPVFGGGTKLTVLG CDR area CDRH1:TYAMN [SEQ ID NO: 69] CDRH2: VISYDGSNKNYVDSVKG [SEQ ID NO: 70] CDRH3:NFDNSGYAIPDAFDI [SEQ ID NO: 71] CDRL1:TGSSSNIGAGYDVH [SEQ ID NO: 72] CDRL2:DNNSRPS [SEQ ID NO: 73] CDRL3:AAWDDSLGGPV [SEQ ID NO: 74] Antibody clone: ​​2A09 2A09-VH [SEQ ID NO: 7] EVQLLESGGGLVQPGGSLRLSCAASGFTFSNAWMSWVRQAPGKGLEWVAYISRDADITHYPASVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCTTGFDYAGDDAFDIWGQGTLVTVSS 2A09-VL [SEQ ID NO: 31] QSVLTQPPSASGTPGQRVTISSCSGSSSNIGSNAVNWYQQLPGTAPKLLIYGNSDRPSGVPDRFSGSKSGTSASLAISGLRSEDEADYYCAAWDDSLNGRWVFGGGTKLTVLG CDR area CDRH1:NAWMS [SEQ ID NO: 75] CDRH2: YISRDADITHYPASVKG [SEQ ID NO: 76] CDRH3:GFDYAGDDAFDI [SEQ ID NO: 77] CDRL1:SGSSSNIGSNAVN [SEQ ID NO: 78] CDRL2: GNSDRPS [SEQ ID NO: 79] CDRL3:AAWDDSLNGRWV [SEQ ID NO: 80] Antibody clone: ​​2B08 2B08-VH [SEQ ID NO: 8] EVQLLESGGGLVQPGGSLRLSCAASGFTFSDYYMSWVRQAPGKGLEWVALIGHDGNNKYYLDSLEGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARATDSGYDLLYWGQGTLVTVSS 2B08-VL [SEQ ID NO: 32] QSVLTQPPSASGTPGQRVTISSCSGSSSNIGNNAVNWYQQLPGTAPKLLIYYDDLLPSGVPDRFSGSKSGTSASLAISGLRSEDEADYYCTTWDDSLSGVVFGGGTKLTVLG CDR area CDRH1:DYYMS [SEQ ID NO:81] CDRH2: LIGHDGNNKYYLDSLEG [SEQ ID NO: 82] CDRH3:ATDSGYDLLY [SEQ ID NO: 83] CDRL1:SGSSSNIGNNAVN [SEQ ID NO: 84] CDRL2: YDDLLPS [SEQ ID NO: 85] CDRL3:TTWDDSLSGVV [SEQ ID NO: 86] Antibody clone: ​​2E8-VH 2E8-VH [SEQ ID NO: 9] EVQLLESGGGLVQPGGSLRLSCAASGFTFSDYYMSWIRQAPGKGLEWVSAIGFSDDNTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAGGDGSGWSFWGQGTLVTVSS 2E8-VL [SEQ ID NO: 33] QSVLTQPPSASGTPGQRVTISCSGSSSNIGNNAVNWYQQLPGTAPKLLIYDNNKRPSGVPDRFSGSKSGTSASLAISGLRSEDEADYYCATWDDSLRGWVFGGGTKLTVLG CDR area CDRH1:DYYMS [SEQ ID NO:87] CDRH2:AIGFSDDNTYYADSVKG [SEQ ID NO: 88] CDRH3: GDGSGWSF [SEQ ID NO: 89] CDRL1:SGSSSNIGNNAVN [SEQ ID NO: 90] CDRL2:DNNKRPS [SEQ ID NO: 91] CDRL3:ATWDDSLRGWV [SEQ ID NO: 92] Antibody clone: ​​5C04 5C04-VH [SEQ ID NO: 10] EVQLLESGGGLVQPGGSLRLSCAASGFTFSNYGMHWVRQAPGKGLEWVAVISYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAREWRDAFDIWGQGTLVTVSS 5C04-VL [SEQ ID NO: 34] QSVLTQPPSASGTPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYSDNQRPSGVPDRFSGSKSGTSASLAISGLRSEDEADYYCAAWDDSLSGSWVFGGGTKLTVLG CDR area CDRH1:NYGMH [SEQ ID NO: 93] CDRH2:VISYDGSNKYYADSVKG [SEQ ID NO: 94] CDRH3:WRDAFDI [SEQ ID NO: 95] CDRL1:TGSSSNIGAGYDVH [SEQ ID NO: 96] CDRL2:SDNQRPS [SEQ ID NO:97] CDRL3:AAWDDSLSGSWV [SEQ ID NO: 98] Antibody clone: ​​5C05 5C05-VH [SEQ ID NO: 11] EVQLLESGGGLVQPGGSLRLSCAASGFTFSTYGMHVRQAPGKGLEWVAVISYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARENFDAFDVWGQGTLVTVSS 5C05-VL [SEQ ID NO: 35] QSVLTQPPSASGTPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYSNSQRPSGVPDRFSGSKSGTSASLAISGLRSEDEADYYCAAWDDSLNGQVVFGGGTKLTVLG CDR area CDRH1:TYGMH [SEQ ID NO: 99] CDRH2:VISYDGSNKYYADSVKG [SEQ ID NO: 100] CDRH3:ENFDAFDV [SEQ ID NO: 101] CDRL1:TGSSSNIGAGYDVH [SEQ ID NO: 102] CDRL2: SNSQRPS [SEQ ID NO: 103] CDRL3:AAWDDSLNGQVV [SEQ ID NO: 104] Antibody clone: ​​5D07 5D07-VH [SEQ ID NO: 12] EVQLLESGGGLVQPGGSLRLSCAASGFTFSTYGMHVRQAPGKGLEWVAVIAYDGSKKDYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAREYRDAFDIWGQGTLVTVSS 5D07-VL [SEQ ID NO: 36] QSVLTQPPSASGTPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYGNSNRPSGVPDRFSGSKSGTTASLAISGLRSEDEADYYCAAWDDSVSGWMFGGGTKLTVLG CDR area CDRH1:TYGMH [SEQ ID NO: 105] CDRH2: VIAYDGSKKDYADSVKG [SEQ ID NO: 106] CDRH3:EYRDAFDI [SEQ ID NO: 107] CDRL1:TGSSSNIGAGYDVH [SEQ ID NO: 108] CDRL2: GNSNRPS [SEQ ID NO: 109] CDRL3:AAWDDSVSGWM [SEQ ID NO: 110] Antibody clone: ​​5E12 5E12-VH [SEQ ID NO: 13] EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYGMHWVRQAPGKGLEWVAVISYDGINKDYADSMKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARERKDAFDIWGQGTLVTVSS 5E12-VL [SEQ ID NO: 37] QSVLTQPPSASGTPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYSNNQRPSGVPDRFSGSKSGTSASLAISGLRSEDEADYYCATWDDSLNGLVFGGGTKLTVLG CDR area CDRH1:SYGMH [SEQ ID NO: 111] CDRH2: VISYDGINKDYADSMKG [SEQ ID NO: 112] CDRH3:ERKDAFDI [SEQ ID NO: 113] CDRL1:TGSSSNIGAGYDVH [SEQ ID NO: 114] CDRL2: SNNQRPS [SEQ ID NO: 115] CDRL3:ATWDDSLNGLV [SEQ ID NO: 116] Antibody clone: ​​5G08 5G08-VH [SEQ ID NO: 14] EVQLLESGGGLVQPGGSLRLSCAASGFTFNNYGMHWVRQAPGKGLEWVAVISYDGSNRYYADSVKGRFTMSRDNSKNTLYLQMNSLRAEDTAVYYCARDRWNGMDVWGQGTLVTVSS 5G08-VL [SEQ ID NO: 38] QSVLTQPPSASGTPGQRVTISCSGSSSNIGAGYDVHWYQQLPGTAPKLLIYANNQRPSGVPDRFSGSKSGTSASLAISGLRSEDEADYYCAAWDDSLNGPWVFGGGTKLTVLG CDR area CDRH1:NYGMH [SEQ ID NO: 117] CDRH2:VISYDGSNRYYADSVKG [SEQ ID NO: 118] CDRH3:DRWNGMDV [SEQ ID NO: 119] CDRL1:SGSSSNIGAGYDVH [SEQ ID NO: 120] CDRL2:ANNQRPS [SEQ ID NO: 121] CDRL3:AAWDDSLNGPWV [SEQ ID NO: 122] Antibody clone: ​​5H06 5H06-VH [SEQ ID NO: 15] EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYGMHWVRQAPGKGLEWVAVISYDGSDTAYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDHSVIGAFDIWGQGTLVTVSS 5H06-VL [SEQ ID NO: 39] QSVLTQPPSASGTPGQRVTISCSGSSSNIGSNTVNWYQQLPGTAPKLLIYDNNKRPSGVPDRFSGSKSGTSASLAISGLRSEDEADYYCSSYAGSNNVVFGGGTKLTVLG CDR area CDRH1:SYGMH [SEQ ID NO: 123] CDRH2: VISYDGSDTAYADSVKG [SEQ ID NO: 124] CDRH3:DHSVIGAFDI [SEQ ID NO: 125] CDRL1:SGSSSNIGSNTVN [SEQ ID NO: 126] CDRL2:DNNKRPS [SEQ ID NO: 127] CDRL3: SSYAGSNNVV [SEQ ID NO: 128] Antibody clone: ​​6A09 6A09-VH [SEQ ID NO: 16] EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYGMHWVRQAPGKGLEWVAVTSYDGNTKYYANSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAREDCGGDCFDYWGQGTLVTVSS 6A09-VL [SEQ ID NO: 40] QSVLTQPPSASGTPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYGNSNRPSGVPDRFSGSKSGTSASLAISGLRSEDEADYYCAAWDDSLNEGVFGGGTKLTVLG CDR area CDRH1:SYGMH [SEQ ID NO: 129] CDRH2:VTSYDGNTKYYANSVKG [SEQ ID NO: 130] CDRH3: EDCGGDCFDY [SEQ ID NO: 131] CDRL1:TGSSSNIGAGYDVH [SEQ ID NO: 132] CDRL2: GNSNRPS [SEQ ID NO: 133] CDRL3:AAWDDSLNEGV [SEQ ID NO: 134] Antibody clone: ​​6B01 6B01-VH [SEQ ID NO: 17] EVQLLESGGGLVQPGGSLRLSCAASGFTFSNYGMHWVRQAPGKGLEWVAVISYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDQLGEAFDIWGQGTLVTVSS 6B01-VL [SEQ ID NO: 41] QSVLTQPPSASGTPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYDNNKRPSGVPDRFSGSKSGTSASLAISGLRSEDEADYYCATWDDSLSGPVFGGGTKLTVLG CDR area CDRH1:NYGMH [SEQ ID NO: 135] CDRH2:VISYDGSNKYYADSVKG [SEQ ID NO: 136] CDRH3:DQLGEAFDI [SEQ ID NO: 137] CDRL1:TGSSSNIGAGYDVH [SEQ ID NO: 138] CDRL2:DNNKRPS [SEQ ID NO: 139] CDRL3:ATWDDSLSGPV [SEQ ID NO: 140] Antibody clone: ​​6C11 6C11-VH [SEQ ID NO: 18] EVQLLESGGGLVQPGGSLRLSCAASGFTFDDYGMSWVRQAPGKGLEWVSAISGSGSSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAGGDIDYFDYWGQGTLsVTVSS 6C11-VL [SEQ ID NO: 42] QSVLTQPPSASGTPGQRVTISCTGSSSNFGAGYDVHWYQQLPGTAPKLLIYENNKRPSGVPDRFSGSKSGTSASLAISGLRSEDEADYYCAAWDDSLNGPVFGGGTKLTVLG CDR area CDRH1:DYGMS [SEQ ID NO: 141] CDRH2:AISGSGSSTYYADSVKG [SEQ ID NO: 142] CDRH3: GDIDYFDY [SEQ ID NO: 143] CDRL1:TGSSSNFGAGYDVH [SEQ ID NO: 144] CDRL2:ENNKRPS [SEQ ID NO: 145] CDRL3:AAWDDSLNGPV [SEQ ID NO: 146] Antibody clone: ​​6C12 6C12-VH [SEQ ID NO: 19] EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYGMHWVRQAPGKGLEWVAVISYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARERRDAFDIWGQGTLVTVSS 6C12-VL [SEQ ID NO: 43] QSVLTQPPSASGTPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYSDNQRPSGVPDRFSGSKSGTSASLAISGLRSEDEADYYCATWDSDTPVFGGGTKLTVLG CDR area CDRH1:SYGMH [SEQ ID NO: 147] CDRH2:VISYDGSNKYYADSVKG [SEQ ID NO: 148] CDRH3:ERRDAFDI [SEQ ID NO: 149] CDRL1:TGSSSNIGAGYDVH [SEQ ID NO: 150] CDRL2:SDNQRPS [SEQ ID NO: 151] CDRL3:ATWDSDTPV [SEQ ID NO: 152] Antibody clone: ​​6D01 6D01-VH [SEQ ID NO: 20] EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYGMHWVRQAPGKGLEWVAVISYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAMYYCARDHSAAGYFDYWGQGTLVTVSS 6D01-VL [SEQ ID NO: 44] QSVLTQPPSASGTPGQRVTISSCSGSSSNIGSNTVNWYQQLPGTAPKLLIYGNSIRPSGGPDRFSGSKSGTSASLAISGLRSEDEADYYCASWDDSLSSPVFGGGTKLTVLG CDR area CDRH1:SYGMH [SEQ ID NO: 153] CDRH2:VISYDGSNKYYADSVKG [SEQ ID NO: 154] CDRH3:DHSAAGYFDY [SEQ ID NO: 155] CDRL1:SGSSSNIGSNTVN [SEQ ID NO: 156] CDRL2: GNSIRPS [SEQ ID NO: 157] CDRL3:ASWDDSLSSPV [SEQ ID NO: 158] Antibody clone: ​​6G03 6G03-VH [SEQ ID NO: 21] EVQLLESGGGLVQPGGSLRLSCAASGFTFGSYGMHWVRQAPGKGLEWVSGISWDSAIIDYAGSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKDEAAAGAFDIWGQGTLVTVSS 6G03-VL [SEQ ID NO: 45] QSVLTQPPSASGTPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYGNTDRPSGVPDRFSGSKSGTSASLAISGLRSEDEADYYCAAWDDSLSGPVVFGGGTKLTVLG CDR area CDRH1:SYGMH [SEQ ID NO: 159] CDRH2:GISWDSAIIDYAGSVKG [SEQ ID NO: 160] CDRH3:DEAAAGAFDI [SEQ ID NO: 161] CDRL1:TGSSSNIGAGYDVH [SEQ ID NO: 162] CDRL2: GNTDRPS [SEQ ID NO: 163] CDRL3:AAWDDSLSGPVV [SEQ ID NO: 164] Antibody clone: ​​6G08 6G08-VH [SEQ ID NO: 22] EVQLLESGGGLVQPGGSLRLSCAASGFTLSSYGISWVRQAPGKGLEWVSGISGSGGNTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCASSVGAYANDAFDIWGQGTLVTVSS 6G08-VL [SEQ ID NO: 46] QSVLTQPPSASGTPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYGDTNRPSGVPDRFSGSKSGTSASLAISGLRSEDEADYYCAAWDDSLNGPVFGGGTKLTVLG CDR area CDRH1:SYGIS [SEQ ID NO: 165] CDRH2:GISGSGGNTYYADSVKG [SEQ ID NO: 166] CDRH3:SVGAYANDAFDI [SEQ ID NO: 167] CDRL1:TGSSSNIGAGYDVH [SEQ ID NO: 168] CDRL2: GDTNRPS [SEQ ID NO: 169] CDRL3:AAWDDSLNGPV [SEQ ID NO: 170] Antibody clone: ​​6G11 6G11-VH [SEQ ID NO: 23] EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYGMHWVRQAPGKGLEWMAVISYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARELYDAFDIWGQGTLVTVSS 6G11-VL [SEQ ID NO: 47] QSVLTQPPSASGTPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYADDHRPSGVPDRFSGSKSGTSASLAISGLRSEDEADYYCASWDDSQRAVIFGGGTKLTVLG CDR area CDRH1:SYGMH [SEQ ID NO: 171] CDRH2:VISYDGSNKYYADSVKG [SEQ ID NO: 172] CDRH3:ELYDAFDI [SEQ ID NO: 173] CDRL1:TGSSSNIGAGYDVH [SEQ ID NO: 174] CDRL2: ADDHRPS [SEQ ID NO: 175] CDRL3:ASWDDSQRAVI [SEQ ID NO: 176] Antibody clone: ​​6H08 6H08-VH [SEQ ID NO: 24] EVQLLESGGGLVQPGGSLRLSCAASGFTFNNYGMHWVRQAPGKGLEWVAVISYDGSNKYYADSVKGRFTISKDNSKNTLYLQMNSLRAEDTAVYYCAREYKDAFDIWGQGTLVTVSS 6H08-VL [SEQ ID NO: 48] QSVLTQPPSASGTPGQRVTISCTGSSSNIGSNTVNWYQQLPGTAPKLLIYDNNKRPSGVPDRFSGSKSGTSASLAISGLRSEDEADYYCQAWGTGIRVFGGGTKLTVLG CDR area CDRH1:NYGMH [SEQ ID NO: 177] CDRH2:VISYDGSNKYYADSVKG [SEQ ID NO: 178] CDRH3:EYKDAFDI [SEQ ID NO: 179] CDRL1:TGSSSNIGSNTVN [SEQ ID NO: 180] CDRL2:DNNKRPS [SEQ ID NO: 181] CDRL3: QAWGTGIRV [SEQ ID NO: 182] Antibody clone: ​​7C07 7C07-VH [SEQ ID NO: 25] EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYGMHWVRQAPGKGLEWVAVISYDGSNKYYADSVKGRFTISRDNSQNTLYLQMNSLRAEDTAVYYCAREFGYIILDYWGQGTLVTVSS 7C07-VL [SEQ ID NO: 49] QSVLTQPPSASGTPGQRVTISSCSGSSSNIGSNTVNWYQQLPGTAPKLLIYRDYERPSGVPDRFSGSKSGTSASLAISGLRSEDEADYYCMAWDDSLSGVVFGGGTKLTVLG CDR area CDRH1:SYGMH [SEQ ID NO: 183] CDRH2:VISYDGSNKYYADSVKG [SEQ ID NO: 184] CDRH3:EFGYIILDY [SEQ ID NO: 185] CDRL1:SGSSSNIGSNTVN [SEQ ID NO: 186] CDRL2:RDYERPS [SEQ ID NO: 187] CDRL3:MAWDDSLSGVV [SEQ ID NO: 188] Antibody clone: ​​4B02 4B02-VH [SEQ ID NO: 26] EVQLLESGGGLVQPGGSLRLSCAASGFTFSNHGMHWVRQAPGKGLEWVAVISYDGTNKYYADSVRGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARETWDAFDVWGQGTLVTVSS 4B02-VL [SEQ ID NO: 50] QSVLTQPPSASGTPGQRVTISCSGSSSNIGSNNANWYQQLPGTAPKLLIYDNNKRPSGVPDRFSGSKSGTSASLAISGLRSEDEADYYCQAWDSSTVVFGGGTKLTVLG CDR area CDRH1:NHGMH [SEQ ID NO: 189] CDRH2:VISYDGTNKYYADSVRG [SEQ ID NO: 190] CDRH3:ETWDAFDV [SEQ ID NO: 191] CDRL1:SGSSSNIGSNNAN [SEQ ID NO: 192] CDRL2:DNNKRPS [SEQ ID NO: 193] CDRL3:QAWDSSTVV [SEQ ID NO: 194]

[0085] In some embodiments, and sometimes preferred embodiments, antibody molecules that specifically bind to FcγRIIb comprise the following CDR regions: SEQ ID NO: 171 (CDRH1), SEQ ID NO: 172 (CDRH2), SEQ ID NO: 173 (CDRH3), SEQ ID NO: 174 (CDRL1), SEQ ID NO: 175 (CDRL2) and SEQ ID NO: 176 (CDRL3), i.e., the CDR regions of clone 6G11.

[0086] In some embodiments, and sometimes preferred embodiments, an antibody molecule that specifically binds to FcγRIIb comprises the following constant regions: SEQ ID NO: 1 (CH) and SEQ ID NO: 2 (CL), and the following variable regions: SEQ ID NO: 23 (VL) and SEQ ID NO: 47 (VH), i.e., the constant and variable regions of clone 6G11.

[0087] In some embodiments, the PD-1 molecule is a human or human-derived antibody molecule. In some such embodiments, the human or human-derived antibody molecule is an IgG antibody. In some such embodiments, the human or human-derived antibody molecule is an IgG4.

[0088] An anti-PD-1 antibody molecule is an antibody molecule that specifically binds to PD-1.

[0089] In some embodiments, the anti-PD-1 antibody molecule blocks the binding of PD-L1 and / or PD-L2 to PD-1 and may then be considered a PD-1 antagonist.

[0090] In some embodiments, the anti-PD-1 antibody molecule is a humanized antibody molecule.

[0091] In some embodiments, the anti-PD-1 antibody molecule is a chimeric antibody.

[0092] As mentioned above, the PD-1 antibody must have the ability to bind to FcγR.

[0093] In some embodiments, the anti-PD-1 antibody molecule is selected from the group consisting of nivolumab (OPDIVO®), pembrolizumab (KEYTRUDA®), and cemiplimab (LIBTAYO®).

[0094] In some embodiments, the antibody molecule that specifically binds FcγRIIb and the anti-PD-1 antibody molecule are administered to a patient simultaneously, meaning that they are administered either at the same time or separately within a very close time period of each other.

[0095] In some embodiments, the antibody molecule that specifically binds FcγRIIb is administered to the patient before the administration of the anti-PD-1 antibody molecule. Such sequential administration can be achieved by temporally separating the two antibodies. Alternatively, or in combination with the first option, sequential administration can be achieved by spatially separating the two antibody molecules, such as by administering the antibody molecule that specifically binds FcγRIIb, e.g., intratumorally, so that the antibody molecule reaches the cancer before the anti-PD-1 antibody molecule, and then administering the antibody molecule that specifically binds FcγRIIb, e.g., by systemic administration, so that the antibody molecule reaches the cancer after the antibody molecule that specifically binds FcγRIIb.

[0096] In some embodiments, the anti-PD-1 antibody molecule is administered to a patient prior to administration of the antibody molecule that specifically binds to FcγRIIb. As above, such sequential administration can be achieved by temporal separation of the two antibodies and / or spatial separation of the two antibody molecules. In the case of spatial administration, the anti-PD-1 antibody molecule is administered by a method such as intratumoral administration so that it reaches the cancer prior to the antibody molecule that specifically binds to FcγRIIb, and then administered by a method such as systemic administration so that it reaches the cancer after the PD-1 antibody molecule.

[0097] For example, it will be known to those skilled in the art of medicine that drugs can be modified with different additives to alter the rate at which they are absorbed by the body, and can be modified in different forms to allow, for example, specific routes of administration to the body.

[0098] Thus, the compositions of the present invention, and / or antibodies, and / or drugs include those combined with excipients and / or pharmaceutically acceptable carriers and / or pharmaceutically acceptable diluents and / or adjuvants.

[0099] The compositions, and / or antibodies, and / or drugs of the invention may be suitable for parenteral administration, including aqueous and / or non-aqueous sterile injection solutions, which may contain antioxidants, and / or buffers, and / or bacteriostats, and / or solutes that render the formulation isotonic with the blood of the intended recipient, and / or aqueous and / or non-aqueous sterile suspensions that may contain suspending agents and / or thickening agents. The compositions, and / or antibodies, and / or drugs of the invention may be presented in unit-dose or multi-dose containers, for example, sealed ampoules and vials, and may be stored in a freeze-dried (i.e., lyophilized) condition requiring only the addition of a sterile liquid carrier, for example, water for injections, immediately prior to use.

[0100] Extemporaneous injection solutions and suspensions may be prepared from sterile powders and / or granules and / or tablets of the kind previously described.

[0101] For parenteral administration to human patients, daily dosage levels of antibody molecules that specifically bind to FcγRIIb and / or anti-PD-1 antibody molecules typically range from 1 mg to 20 mg / kg (patient body weight), or in some cases up to 100 mg / kg, administered in single or divided doses. Lower doses may be used under special circumstances, e.g., in combination with chronic administration. In any event, the physician will determine the actual dosage that will be most suitable for an individual patient, which will vary with the age, weight, and response of the particular patient. The dosages described above are exemplary of the average case. Of course, there may be individual cases in which higher or lower dosage ranges are merited, and these are within the scope of this invention.

[0102] Typically, the compositions and / or medicaments of the present invention will contain antibody molecules that specifically bind to FcγRIIb and / or anti-PD-1 antibodies at a concentration of approximately 2 mg / mL to 150 mg / mL, or approximately 2 mg / mL to 200 mg / mL. In a preferred embodiment, the compositions and / or medicaments of the present invention will contain antibody molecules that specifically bind to FcγRIIb and / or anti-PD-1 antibody molecules at a concentration of 10 mg / mL.

[0103] Generally, in humans, oral or parenteral administration of the compositions, antibodies, agents, and / or drugs of the present invention is the preferred and most convenient route. For veterinary use, the compositions, antibodies, agents, and / or drugs of the present invention are administered in a suitably acceptable formulation in accordance with normal veterinary practice, and the veterinarian will determine the dosage regimen and route of administration that will be most appropriate for a particular animal. Accordingly, the present invention provides pharmaceutical formulations comprising antibodies and / or drugs of the present invention in an amount effective to treat various conditions (described above and further below). Preferably, the compositions, antibodies, agents, and / or drugs are adapted for delivery by a route selected from the group consisting of intravenous (IV), subcutaneous (SC), intramuscular (IM), or intratumoral.

[0104] The present invention also includes compositions, antibodies, pharmaceutical agents, and / or drugs, comprising pharmaceutically acceptable acid or base addition salts of the polypeptide binding moieties of the present invention. The acids used to prepare pharmaceutically acceptable acid addition salts of the aforementioned base compounds useful in the present invention are, among others, those that form non-toxic acid addition salts, i.e., salts containing pharmacologically acceptable anions, such as hydrochloride, hydrobromide, hydroiodide, nitrate, sulfate, bisulfate, phosphate, acid phosphate, acetate, lactate, citrate, acid citrate, tartrate, bitartrate, succinate, maleate, fumarate, gluconate, saccharate, benzoate, methanesulfonate, ethanesulfonate, benzenesulfonate, p-toluenesulfonate, and pamoate [i.e., 1,1'-methylene-bis-(2-hydroxy-3-naphthoate)] salts. Pharmaceutically acceptable base addition salts may also be used to produce pharmaceutically acceptable salt forms of the agents of the present invention. Chemical bases that can be used as reagents to prepare pharmaceutically acceptable base salts of the present agents, which are acidic in nature, are those that form non-toxic base salts with such compounds. Such non-toxic base salts include, but are not limited to, those derived from pharmacologically acceptable cations such as alkali metal cations (e.g., potassium and sodium) and alkaline earth metal cations (e.g., calcium and magnesium), ammonium or water-soluble amine addition salts, such as N-methylglucamine (meglumine), and lower alkanolammonium, as well as other pharmaceutically acceptable organic amine base salts. The agents and / or polypeptide binding moieties of the present invention may be lyophilized for storage and reconstituted with a suitable carrier prior to use. Any suitable lyophilization method (e.g., spray drying, cake drying) and / or reconstitution technique may be used. Those skilled in the art will understand that lyophilization and reconstitution may lead to varying degrees of loss of antibody activity (e.g., in conventional immunoglobulins, IgM antibodies tend to have greater loss of activity than IgG antibodies), and that usage levels may need to be adjusted upward to compensate.In one embodiment, a lyophilized (freeze-dried) polypeptide binding moiety loses less than about 20%, or less than about 25%, or less than about 30%, or less than about 35%, or less than about 40%, or less than about 45%, or less than about 50% of its activity (before lyophilization) when rehydrated.

[0105] A combination of an antibody molecule that specifically binds to FcγRIIb and an anti-PD-1 antibody molecule can be used in the treatment of cancer.

[0106] As used herein, the term "patient" refers to an animal, including a human, that has been diagnosed with an FcγRIIb-negative cancer, or that is considered likely to be an FcγRIIb-negative cancer and / or that exhibits symptoms of such a cancer.

[0107] The patient may be a mammal or a non-mammal. Preferably, the patient is a human, or a mammal such as a horse, cow, sheep, pig, camel, dog, or cat. Most preferably, the mammalian patient is a human.

[0108] "Exhibiting" includes when a subject exhibits cancer symptoms and / or cancer diagnostic markers and / or when cancer symptoms and / or cancer diagnostic markers can be measured and / or assessed and / or quantified.

[0109] It will be readily apparent to those skilled in the art of medicine what cancer symptoms and cancer diagnostic markers are, and how to measure and / or assess and / or quantify whether there is a reduction or increase in the severity of cancer symptoms or whether there is a reduction or increase in cancer diagnostic markers, and how cancer symptoms and / or cancer diagnostic markers can be used to form a prognosis for cancer.

[0110] Cancer treatments are often administered as a course of treatment, i.e., the therapeutic agents are administered over a period of time. The length of the course of treatment depends on several factors, including the type of therapeutic agent being administered, the type of cancer being treated, the severity of the cancer being treated, and the age and health of the patient, among other factors.

[0111] "Currently undergoing treatment" includes when a patient is currently undergoing a course of treatment and / or is currently receiving a therapeutic agent and / or is currently receiving a course of therapeutic agent.

[0112] Patients treated according to the present invention have cancer characterized by PD-1 positive tumors.

[0113] In some embodiments, the cancer being treated is a solid tumor.

[0114] In some embodiments, the solid cancer being treated is one whose treatment normally consists of or includes immunotherapy with an anti-PD-1 antibody.

[0115] In some embodiments, the cancer to be treated is selected from the group consisting of melanoma; lung cancer, including small cell lung cancer (SCLC) and non-small cell lung cancer (NSCLC), including non-squamous NSCLC and squamous NSCLC, including metastatic NSCLC; head and neck cancer, including head and neck squamous cell carcinoma (HNSCC); Hodgkin's lymphoma; primary mediastinal B-cell lymphoma (PMBCL); bladder cancer, including advanced urothelial carcinoma; colorectal cancer, including cancers that are highly instable (MSI-H) and / or mismatch repair deficient (dMMR); gastric cancer, including advanced gastric cancer and gastric or gastroesophageal junction (GEJ) adenocarcinoma; cervical cancer; liver cancer, including hepatocellular carcinoma; Merkel cell carcinoma (MCC); kidney cancer, including renal cell carcinoma (RCC) and cutaneous squamous cell carcinoma (CSCC), including locally advanced CSCC in patients who are not candidates for curative surgery or curative radiation therapy. Those skilled in the art will be aware that the number and variety of indications associated with anti-PD-1 immunotherapy are rapidly expanding.

[0116] In some embodiments, the cancer is a refractory cancer. In some such embodiments, a refractory cancer is a cancer that is already known to be resistant to treatment with an anti-PD-1 antibody at the start of treatment. This resistance may be manifested by the patient not responding to treatment at all, or by some progression of the cancer despite treatment. In some embodiments, a refractory cancer is a cancer that becomes resistant to an anti-PD-1 antibody during treatment with the antibody, meaning that the patient stops responding to treatment or shows a reduced response to treatment. In some embodiments, a refractory cancer shows resistance after successful treatment with an anti-PD-1 antibody or at the end stage, meaning that if the cancer recurs, the anti-PD-1 antibody is ineffective or less effective.

[0117] All of the above-mentioned cancers are well known, and the symptoms and cancer diagnostic markers are well described, as are the therapeutic agents used to treat those cancers. Therefore, the symptoms, cancer diagnostic markers and therapeutic agents used to treat the above-mentioned types of cancers will be known to those skilled in the art of medicine.

[0118] The clinical definition of the diagnosis, prognosis, and progression of most cancers is based on a specific classification known as staging. These staging systems collate many different cancer diagnostic markers and cancer symptoms to provide an overview of the diagnosis, prognosis, and / or progression of cancer. Those skilled in the art of oncology will know how to use staging systems to assess the diagnosis, prognosis, and / or progression of cancer, and which cancer diagnostic markers and cancer symptoms to use for this purpose.

[0119] "Cancer staging" includes the Rai staging system, including stage 0, stage I, stage II, stage III, and stage IV, and / or the Binet staging system, including stage A, stage B, and stage C, and / or the Ann Arbour staging system, including stage I, stage II, stage III, and stage IV.

[0120] It is known that cancer can cause abnormalities in cell morphology. These abnormalities often occur reproducibly in certain cancers, which means that examining these changes in morphology (also known as histological examination) can be used to diagnose or prognose cancer. Techniques for visualizing and preparing samples for examining cell morphology are known in the art, such as, for example, light microscopy or confocal microscopy.

[0121] "Histological examination" includes the presence of small mature lymphocytes, and / or the presence of small mature lymphocytes with narrow cytoplasmic borders, the presence of small mature lymphocytes with dense nuclei lacking discernible nucleoli, and / or the presence of small mature lymphocytes with dense nuclei lacking discernible nucleoli, and / or the presence of atypical cells, and / or cleaved cells, and / or prolymphocytes.

[0122] It is known that cancer is the result of the mutation of cellular DNA, which can lead to cell death avoidance or uncontrollable proliferation.Therefore, the examination of these mutations (also known as cytogenetic examination) can be a useful tool for assessing the diagnosis and / or prognosis of cancer.An example of this is the deletion of chromosome position 13q14.1, which is characteristic of chronic lymphocytic leukemia.Techniques for examining mutations in cells are known in the art, such as fluorescence in situ hybridization (FISH).

[0123] "Cytogenetic testing" includes testing of DNA in cells, specifically chromosomes. Cytogenetic testing can be used to identify DNA alterations that may be associated with the presence of refractory and / or recurrent cancer. These include deletion of the long arm of chromosome 13, and / or deletion of chromosome location 13q14.1, and / or trisomy of chromosome 12, and / or deletion of the long arm of chromosome 12, and / or deletion of the long arm of chromosome 11, and / or deletion of 11q, and / or deletion of the long arm of chromosome 6, and / or deletion of 6q, and / or deletion of the short arm of chromosome 17, and / or deletion of 17p, and / or t(11:14) translocation, and and / or (q13:q32) translocations, and / or antigen gene receptor rearrangements, and / or BCL2 rearrangements, and / or BCL6 rearrangements, and / or t(14:18) translocations, and / or t(11:14) translocations, and / or (q13:q32) translocations, and / or (3:v) translocations, and / or (8:14) translocations, and / or (8:v) translocations, and / or t(11:14) and (q13:q32) translocations.

[0124] Cancer patients are known to exhibit certain physical symptoms, which are often the result of the burden that cancer places on the body.These symptoms often recur with the same cancer, and can therefore be diagnostic and / or prognostic and / or progression characteristics of the disease.Those skilled in the art of medicine will understand which physical symptoms are associated with which cancer, and how the evaluation of these body systems can be correlated with the diagnosis and / or prognosis and / or progression of the disease."Physical symptoms" include hepatomegaly and / or splenomegaly. [Brief explanation of the drawings]

[0125] In the examples below, reference is made to the following figures:

[0126] [Figure 1]PD-1 expression in PD-1-transfected human Jurkat T cells is shown. PD-1-transfected Jurkat cells were categorized into low, moderate, and high PD-1 expressing cells. After expansion, PD-1 expression was quantified in the three different subsets, and the number of PD-1 molecules / cell is shown in Figure 1A (low), Figure 1B (moderate), and Figure 1C (high). [Figure 2A-B] BI-1206 (6G11 WT) inhibits PD-1-mediated phagocytosis of moderately and highly expressing cells, but not of lowly expressing cells. Figure 2A shows an example of phagocytosed Jurkat cells. FL4 on the y-axis indicates CD14+ macrophages, and FL1 on the x-axis indicates CFSE-labeled Jurkat cells. Thus, the circled upper right quadrant shows double-positive CD14+CFSE+ cells, which are phagocytosed Jurkat cells. This example shows the phagocytosis of high PD-1 expressing Jurkat cells. Figure 2B shows the phagocytosis of medium PD-1 expressing Jurkat cells. [Figure 2C-D] Figure 2C shows high-expressing Jurkat cells. Values ​​are normalized to isotype opsonization (set to zero%) and anti-CD3 opsonization (OKT3 hIgG1, set to 100%). This figure shows that BI-1206 (represented in the figure as 6G11 WT) inhibits nivolumab-mediated phagocytosis at all concentrations tested. Furthermore, this figure demonstrates that the 6G11 antibody requires an intact Fc portion to inhibit phagocytosis, as disruption of FcγR binding caused by mutation of position 297 from the amino acid asparagine (N) to the amino acid glutamine (Q) (i.e., the antibody represented herein as 6G11NQ) reduces its ability to inhibit nivolumab-mediated phagocytosis. This figure shows two experiments with moderately expressing cells and three experiments with highly expressing cells. Figure 2D shows that low-expressing cells do not exhibit nivolumab-mediated phagocytosis. [Figure 3A]We demonstrate that anti-FcγRIIB with enhanced Fc:FcγR binding (AT-130-2 mIgG2a and mIgG1), but not anti-FcγRIIB with impaired Fc:FcγR binding (AT-130-2 mIgG1 NA), enhances the therapeutic efficacy and in vivo survival of anti-PD-1 antibodies. Mice bearing CT26 (Figures A and B) or MC38 (Figures 3C and 3D) tumors were treated three times (on days 8, 12, and 15 after subcutaneous inoculation of 5 x 10 tumor cells in 100 μl PBS) with 200 μg of anti-PD-1 (clone 29F.1A12; Bioxcell) antibody alone or in combination with 200 μg of the indicated anti-FcγRIIB antibody variant or isotype control (WR17). For the first treatment, AT130-2 was administered 6 hours before the anti-PD-1 antibody. For subsequent treatments, both antibodies were administered together. All injections were given intraperitoneally in 200 μl of PBS. Tumors were considered terminal when they reached an area of ​​400 mm for CT26 and 225 mm for MC38. Graphs show tumor growth (Figures 3A and 3C) and survival (Figures 3B and 3D) of animals (**P<0.01; log-rank test). Experiments were performed on 8- to 14-week-old female mice. [Figure 3B]We demonstrate that anti-FcγRIIB with enhanced Fc:FcγR binding (AT-130-2 mIgG2a and mIgG1), but not anti-FcγRIIB with impaired Fc:FcγR binding (AT-130-2 mIgG1 NA), enhances the therapeutic efficacy and in vivo survival of anti-PD-1 antibodies. Mice bearing CT26 (Figures A and B) or MC38 (Figures 3C and 3D) tumors were treated three times (on days 8, 12, and 15 after subcutaneous inoculation of 5 x 10 tumor cells in 100 μl PBS) with 200 μg of anti-PD-1 (clone 29F.1A12; Bioxcell) antibody alone or in combination with 200 μg of the indicated anti-FcγRIIB antibody variant or isotype control (WR17). For the first treatment, AT130-2 was administered 6 hours before the anti-PD-1 antibody. For subsequent treatments, both antibodies were administered together. All injections were given intraperitoneally in 200 μl of PBS. Tumors were considered terminal when they reached an area of ​​400 mm for CT26 and 225 mm for MC38. Graphs show tumor growth (Figures 3A and 3C) and survival (Figures 3B and 3D) of animals (**P<0.01; log-rank test). Experiments were performed on 8- to 14-week-old female mice. [Figure 3C]We demonstrate that anti-FcγRIIB with enhanced Fc:FcγR binding (AT-130-2 mIgG2a and mIgG1), but not anti-FcγRIIB with impaired Fc:FcγR binding (AT-130-2 mIgG1 NA), enhances the therapeutic efficacy and in vivo survival of anti-PD-1 antibodies. Mice bearing CT26 (Figures A and B) or MC38 (Figures 3C and 3D) tumors were treated three times (on days 8, 12, and 15 after subcutaneous inoculation of 5 x 10 tumor cells in 100 μl PBS) with 200 μg of anti-PD-1 (clone 29F.1A12; Bioxcell) antibody alone or in combination with 200 μg of the indicated anti-FcγRIIB antibody variant or isotype control (WR17). For the first treatment, AT130-2 was administered 6 hours before the anti-PD-1 antibody. For subsequent treatments, both antibodies were administered together. All injections were given intraperitoneally in 200 μl of PBS. Tumors were considered terminal when they reached an area of ​​400 mm for CT26 and 225 mm for MC38. Graphs show tumor growth (Figures 3A and 3C) and survival (Figures 3B and 3D) of animals (**P<0.01; log-rank test). Experiments were performed on 8- to 14-week-old female mice. [Figure 3D]We demonstrate that anti-FcγRIIB with enhanced Fc:FcγR binding (AT-130-2 mIgG2a and mIgG1), but not anti-FcγRIIB with impaired Fc:FcγR binding (AT-130-2 mIgG1 NA), enhances the therapeutic efficacy and in vivo survival of anti-PD-1 antibodies. Mice bearing CT26 (Figures A and B) or MC38 (Figures 3C and 3D) tumors were treated three times (on days 8, 12, and 15 after subcutaneous inoculation of 5 x 10 tumor cells in 100 μl PBS) with 200 μg of anti-PD-1 (clone 29F.1A12; Bioxcell) antibody alone or in combination with 200 μg of the indicated anti-FcγRIIB antibody variant or isotype control (WR17). For the first treatment, AT130-2 was administered 6 hours before the anti-PD-1 antibody. For subsequent treatments, both antibodies were administered together. All injections were given intraperitoneally in 200 μl of PBS. Tumors were considered terminal when they reached an area of ​​400 mm for CT26 and 225 mm for MC38. Graphs show tumor growth (Figures 3A and 3C) and survival (Figures 3B and 3D) of animals (**P<0.01; log-rank test). Experiments were performed on 8- to 14-week-old female mice. [Figure 4] Figure 1 shows PD-1 expression on immune cells from tumor-bearing mice. Immune cells from mouse tumors were quantified for PD-1 expression. Mice were injected with MC38 cells, and tumors were harvested approximately 20 days later. Cells were stained for different T cell subsets, and PD-1 expression on CD8+ T cells was analyzed by FACS. Mean fluorescence intensity values ​​for PD-1 on cells were correlated with values ​​from Quantum™ Simply Cellular® beads stained with the same anti-PD-1 antibody to determine the number of receptors per cell. [Figure 5] Jurkat cells expressing different levels of PD-1 are shown: low, intermediate, and high. PD-1 expression was defined using a saturating concentration of Alexa Fluor 647 human anti-human PD-1 (pembrolizumab). [Figure 6]PD-1 expression in "Jurkat PD-1 medium expressing cells" is shown. The gate indicates the full width at half maximum gate used to define the lower end of "medium-high" expression of PD-1 in tumor samples. [Figure 7] The gating strategy used to define PD-1 expression in human tumor samples is shown. CD45+ events were first defined (A), followed by live cells (B), and then CD3+ (C) or CD3+CD8+ (D). PD-1 high expression gates were set on the CD3+ (E) and CD3+CD8+ populations (F), respectively. PD-1 "high expression" gates were defined based on PD-1-transfected Jurkat cells, with the lower edge set according to the lower edge of the full-width half-maximum gate for PD-1-moderately expressing Jurkat cells. (G) and (H) show FMOs of Alexa Fluor 647 human anti-human PD-1 (pembrolizumab) in the CD3+ and CD3+CD8+ populations, respectively. [Figure 8A] A table summarizing data for each patient from whom a tumor sample was obtained is shown, including patient characteristics such as PD-1 expression and predicted response. [Figure 8B] A table summarizing data for each patient from whom a tumor sample was obtained is shown, including patient characteristics such as PD-1 expression and predicted response. [Figure 9] The percentage of CD3+ and CD3+CD8+ lymphocytes with moderate to high PD-1 expression is shown. The dotted line defines 10%. Letters (F, G, H, etc.) correspond to patient IDs in the table in Figure 8. [Example]

[0127] Certain non-limiting examples illustrating certain aspects of the present invention will now be described, which should be read in conjunction with the brief description of the figures provided above.

[0128] Example 1 Transfection of Jurkat cells For transfection of Jurkat cells, cells were cultured in RPMI-1640 medium containing 10% fetal bovine serum, FCS (Sigma), L-glutamine (Life Technologies), sodium pyruvate (Life Technologies), and Pen-Strep (Life Technologies). The day before transfection, cells were cultured at 0.5 × 10 6 To transfect the cells, 1 × 10 cells were split into 1 × 10 cells / mL and cultured overnight. 6 The cells were centrifuged at 90xG for 10 minutes and then resuspended in 100µL of Nucleofector solution (Amaxa® Cell Line Nucleofector® Kit V, Lonza) supplemented with 2µg of DNA (HPD-1 in pcDNA3). The mixture was then transferred to a Nucleofector cuvette. The cuvette was placed in a Nucleofector II machine and nucleofected using program X-005. After 10 minutes of incubation at room temperature (approximately 18-22°C), 500mL of medium was added to the cuvette and transferred to a 12-well plate containing 1mL of medium. Geneticin was added at 1mg / mL 48 hours after transfection to select transfected cells. After 10-14 days, positive cells were purified into low, medium, and high PD-1 expressors by FACS sorting using a FACSAria II machine. The transfected cells were then maintained in medium containing 1 mg / mL Geneticin.

[0129] PD-1 quantification The basic principle of quantification using this set of beads is based on the fact that phycoerythrin (PE) labels antibodies in a 1:1 ratio. Therefore, by creating a standard curve using beads containing a defined number of PE molecules, the number of antibody molecules bound to cells can be determined.

[0130] Jurkat cells were stained with PE-labeled anti-PD1 antibody (EH12.2H7, BioLegend) or isotype control in FACS buffer (PBS containing 2% FCS) for 30 minutes at 4°C and then washed with FACS buffer. One tube of Quantibrite™ beads (PE Phycoerythrin Quantification Kit, BD Biosciences (Cat. No. 340495)) was resuspended in 500 μl of PBS.

[0131] The FACS machine was then configured to run Quantibrite™ beads and Jurkat cells under the same settings. Beads were run until 10,000 events were collected, from which a standard curve was generated by plotting Log molecules per bead (lot-specific information in the kit) versus Log MFI (fluorescence intensity) for the four delivered populations of Quantibrite™ beads, as described in the BD Biosciences for the PE Phycoerythrin Fluorescence Quantitation Kit. This standard curve was then used to calculate the number of molecules on the cell lines by converting MFI to molecule count. Given that the antibody was used at a saturating concentration and that the antibody binding to PD-1 molecules per cell was 1:1, the number of bound antibody per cell corresponds to the number of PD-1 molecules present per cell.

[0132] Jurkat cells stained with PD1-PE were then run by FACS, and the logarithmic mean fluorescence intensity (MFI) of the sample of interest was used to calculate the number of bound antibodies.

[0133] The results are shown in Figure 1.

[0134] From Figure 1, it is clear that the cell population with low expression contains some cells (approximately 2%) with moderate expression, but it is a negligible portion of the population, and from the phagocytosis experiments presented below (and shown in Figure 2), it is clear that this small portion does not affect phagocytosis. Similarly, it is clear that the cell population with moderate expression contains some cells with low expression, but again, this small cell fraction does not affect the outcome of phagocytosis, as shown below.

[0135] The low-expressing subset (Figure 1A) averaged 3,249 PD-1 molecules / cell, with a lower 5% cutoff of 1,253 PD-1 molecules / cell and an upper 5% cutoff of 10,643 PD-1 molecules / cell. The medium-expressing subset (Figure 1B) averaged 32,951 PD-1 molecules / cell, with a lower 5% cutoff of 15,498 PD-1 molecules / cell and an upper 5% cutoff of 77,822 PD-1 molecules / cell. The high-expressing subset (Figure 1C) averaged 165,968 PD-1 molecules / cell, with a lower 5% cutoff of 65,406 PD-1 molecules / cell and an upper 5% cutoff of 390,946 PD-1 molecules / cell. Upper and lower 5% cutoff values ​​are provided to minimize overlap between the different subsets. A lower 5% cutoff value of 15,498, i.e., approximately 15,500 PD-1 molecules / cell as measured above, is used herein to define the lower limit of moderate or high expression.

[0136] phagocytosis Human PBMCs isolated from leukocyte cones obtained from the National Blood Service, Southampton, were incubated in RPMI medium (Life Technologies) containing glutamine, pyruvate, PenStrep, and 1% heat-inactivated human serum from Sigma Heat for 2 hours to allow monocyte attachment. The medium was then replaced with RPMI medium containing glutamine, pyruvate, PenStrep, and 10% FCS (Sigma). After 24 hours, MCSF (produced at the University of Southampton) was added. Macrophages were induced over 7 days with two medium changes (containing MCSF). Macrophages were then harvested by removing the medium, adding 2 mL of PBS, placing on ice for 15 minutes, and gently scraping. The macrophages were then reseeded into 96-well plates for 2 hours. Macrophages were pretreated with anti-hFcγRIIb mAb (6G11WT or 6G11NQ) for 45 minutes at 2x the final concentration, followed by the addition of CFSE (Molecular Probes)-labeled Jurkat cells opsonized with nivolumab (hIgG4) for 15 minutes at 2x the final concentration. Cells were co-cultured at 37°C for 1 hour, then stained with anti-CD14 (BD Bioscience) by incubating in FACS buffer at 4°C for 30 minutes and washing, and then read on a FACS machine.

[0137] The results are shown in Figure 2.

[0138] Example 2. Quantification of PD-1 in immune cells from tumor-bearing mice The number of PD-1 receptors on immune cells from mouse tumors was determined using Quantum™ Simply Cellular® beads (Bangs Laboratories, Inc.). Briefly, beads were stained with a rat anti-PD-1 antibody (clone 29F.1A12, BioLegend) to generate a standard curve. Cell samples were then read against the curve to determine expression.

[0139] Quantification was performed on cells from tumor-bearing mice. They were bred and maintained in a local facility according to the guidelines of the MAU. Female C57 / BL6 mice, 6–8 weeks old, were supplied by Taconic (Bomholt, Denmark) and maintained in a local animal facility. MC38 cells (ATCC) were grown in Glutamax-buffered RPMI supplemented with 10% FBS. When cells became semi-confluent, they were detached with trypsin and cultured at 10 × 10 6 Resuspended in sterile PBS at 1 x 10 cells / mL. 6 A volume of 100 μl of cell suspension, equivalent to 100 cells / mouse, was injected subcutaneously. Tumors were allowed to grow for approximately 20 days before harvest. CD8+ T cell subsets were identified by FACS using CD45, CD3, CD4, and CD8 markers (all from BD Biosciences). PD-1 expression on different T cell subsets was quantified using a commercially available rat anti-PD-1 antibody (clone 29F.1A12) along with the corresponding isotype control (BioLegend). The results are shown in Figure 4.

[0140] Example 3: In vivo effect of combination with anti-PD-1 PD-1 expression in mouse cells corresponds to the "medium-to-high" expression level in transfected Jurkat cells (Example 1, Figure 1). In phagocytosis assays, BI-1206 was shown to significantly reduce the level of phagocytosis of these "medium-to-high" PD-1-expressing cells (Example 1, Figure 2). This data, coupled with the improved therapeutic antitumor effect seen when anti-PD-1 was combined with anti-FcγRIIb (BI-1206 mouse surrogate) in the in vivo MC38 model (Figure 3), suggests improved therapeutic efficacy of anti-PD-1 in combination with BI-1206 in patients with moderate or high PD-1 expression, i.e., PD-1 expression of 15,500 PD-1 molecules / cell or greater.

[0141] Example 4 Quantification of PD-1 expression on human T cells Dissociated viable frozen tumor samples (see table in Figure 8) were purchased from Discovery Life Sciences. Cells were thawed and washed with phosphate-buffered saline (PBS) before staining with the following antibody mixture: Alexa Fluor 700 mouse anti-human CD45 (clone HI30, BD 560566), BV605 mouse anti-human CD8 (clone SK1, BD 564116), PerCP-Cy5.5 mouse anti-human CD3 (clone UCHT1, BD 560835), and Alexa Fluor 647 human anti-human PD-1 (pembrolizumab (KEYTRUDA), clinical grade, lot number 8SNL80406, Merck Sharp & Dohme Limited). Fixable Viability Dye eFluor 780 was also included in the antibody staining mixture (Invitrogen, 65-0865-14). Staining was performed in BD Horizon Brilliant Stain Buffer (BD 563794). Anti-human PD-1 was conjugated in-house with Alexa Fluor 647 and used at a receptor-saturating concentration (5.5 μg / mL) as indicated in previous titration experiments. The remaining antibodies were used at concentrations recommended by the manufacturer. Cells were incubated with the antibodies for 20 minutes, then washed and resuspended in PBS before acquisition using a BD FACSAria II. Analysis was performed using FlowJo software. PD-1 expression analysis in PD-1-transfected Jurkat cells was performed in a similar manner, except that the staining mixture contained only Alexa Fluor 647 human anti-human PD-1 and Fixable Viability Dye eFluor 780. The results are shown in Figure 5. In tumor samples, PD-1 expression was defined within the CD3+ and CD3+CD8+ populations, respectively, pre-gated against live CD45+ cells (Figure 7). The gate for PD-1 high expression was defined based on PD-1 transfected Jurkat cells, and the lower end was set according to the lower end of the full-width at half-maximum gate for PD-1 medium-expressing Jurkat cells (Fig. 6 ).

[0142] Figure 9 shows the percentage of CD3+ lymphocytes and CD3+CD8+ lymphocytes with moderate to high PD-1 expression in individual tumor samples from different patients. The dotted line at 10% expression is included because patients with 10% of their T cells expressing PD-1 at moderate or high levels would be expected to benefit from anti-FcγRIIb in combination with anti-PD-1.

Claims

1. for use in treating cancer in patients with tumor-infiltrating T lymphocytes with moderate or high PD-1 expression; a first antibody molecule that specifically binds to FcγRIIb via its Fab region and to an Fcγ receptor via its Fc region; In combination with a second antibody molecule that specifically binds to PD-1 via its Fc region and binds to at least one Fcγ receptor.

2. for use in treating cancer in patients with tumor-infiltrating T lymphocytes with moderate or high PD-1 expression; (i) a first antibody molecule that specifically binds to FcγRIIb via its Fab region and binds to an Fcγ receptor via its Fc region; (ii) a second antibody molecule that specifically binds to PD-1 via the Fc region and binds to at least one Fcγ receptor.

3. 1. A kit for use in treating cancer in a patient having tumor-infiltrating T lymphocytes with moderate or high PD-1 expression, comprising: (i) a first antibody molecule that specifically binds to FcγRIIb via its Fab region and binds to an Fcγ receptor via its Fc region; (ii) a second antibody molecule that specifically binds to PD-1 via its Fc region and binds to at least one Fcγ receptor.

4. In the manufacture of a medicament for use in treating cancer in patients with tumor-infiltrating T lymphocytes with moderate or high PD-1 expression, (i) a first antibody molecule that specifically binds to FcγRIIb via its Fab region and to an Fcγ receptor via its Fc region; and (ii) The use of a second antibody molecule that specifically binds to PD-1 via its Fc region and binds to at least one Fcγ receptor.

5. 1. A method for treating cancer in a patient having tumor-infiltrating T lymphocytes with intermediate or high PD-1 expression, comprising: (i) a first antibody molecule that specifically binds to FcγRIIb via its Fab region and binds to an Fcγ receptor via its Fc region; (ii) a second antibody molecule that specifically binds to PD-1 via its Fc region and binds to at least one Fcγ receptor.

6. The patient, (i) a first antibody molecule that specifically binds to FcγRIIb via its Fab region and binds to an Fcγ receptor via its Fc region; (ii) a diagnostic test to determine whether a patient would benefit from combination therapy with a second antibody molecule that specifically binds PD-1 via its Fc region and binds to at least one Fcγ receptor, comprising: A diagnostic test, wherein the test comprises determining PD-1 expression on tumor-infiltrating T lymphocytes of said patient, wherein moderate or high PD-1 expression indicates that said patient will benefit from combination therapy.

7. 7. The combination for use according to claim 1, the pharmaceutical composition for use according to claim 2, the kit according to claim 3, the use according to claim 4, the method according to claim 5, or the diagnostic test according to claim 6, wherein the patient's tumor-infiltrating CD3-positive T lymphocytes show intermediate or high PD-1 expression.

8. 8. The combination for use, pharmaceutical composition for use, kit, use, method, or diagnostic test of claim 7, wherein at least 10% of the patient's tumor-infiltrating CD3-positive T lymphocytes have intermediate or high PD-1 expression.

9. The combination for use, pharmaceutical composition for use, kit, use, method, or diagnostic test according to claim 7 or 8, wherein the patient's tumor-infiltrating CD3-positive CD8-positive T lymphocytes have intermediate or high PD-1 expression.

10. 10. The combination for use, pharmaceutical composition for use, kit, use, method, or diagnostic test according to claim 9, wherein at least 10% of the patient's tumor-infiltrating CD3+CD8+ T lymphocytes have moderate or high PD-1 expression.

11. 11. The combination for use, pharmaceutical composition for use, kit, use, method, or diagnostic test according to any one of claims 1 to 10, wherein moderate or high PD-1 expression is defined as at least 10% of tumor-infiltrating T lymphocytes in a sample from a patient having expression of at least 15,500 PD-1 molecules per T lymphocyte.

12. 12. The combination for use, pharmaceutical composition for use, kit, use, method, or diagnostic test according to claim 11, wherein moderate or high PD-1 expression is measured using the anti-PD1 antibody EH12.2H7.

13. The combination for use, pharmaceutical composition for use, kit, use, method or diagnostic test according to any one of claims 1 to 12, wherein the cancer is a solid cancer.

14. 14. The combination for use, pharmaceutical composition for use, kit, use, method, or diagnostic test of claim 13, wherein the solid cancer is selected from the group consisting of melanoma, lung cancer, head and neck cancer, Hodgkin's lymphoma, primary mediastinal B-cell lymphoma (PMBCL), bladder cancer, colorectal cancer, gastric cancer, cervical cancer, liver cancer, Merkel cell carcinoma, renal cancer, and cutaneous squamous cell carcinoma.

15. 15. The combination for use, pharmaceutical composition for use, kit, use, method, or diagnostic test according to claim 13 or 14, wherein the cancer is refractory.

16. The combination for use, pharmaceutical composition for use, kit, use, method, or diagnostic test according to any one of claims 1 to 15, wherein the first antibody molecule and / or the second antibody molecule is selected from the group consisting of a human antibody molecule, a humanized antibody molecule, and an antibody molecule of human origin.

17. 17. The combination for use, pharmaceutical composition for use, kit, use, method or diagnostic test according to any one of claims 1 to 16, wherein said first antibody molecule and / or said second antibody molecule is a monoclonal antibody molecule or a monoclonal-derived antibody molecule.

18. 18. The combination for use, pharmaceutical composition for use, kit, use, method, or diagnostic test according to any one of claims 1 to 17, wherein the first antibody molecule and / or the second antibody molecule is selected from the group consisting of full-size antibodies, chimeric antibodies, single-chain antibodies, and antigen-binding fragments thereof that retain the ability to bind to an Fc receptor via their Fc region.

19. The combination for use, pharmaceutical composition for use, kit, use, method, or diagnostic test according to any one of claims 1 to 18, wherein the first antibody molecule and / or the second antibody molecule is a human IgG antibody, a humanized IgG antibody molecule, or an IgG antibody molecule of human origin.

20. 20. The combination for use, pharmaceutical composition for use, kit, use, method, or diagnostic test according to claim 19, wherein said first antibody molecule is an IgG1 antibody molecule.

21. 21. The combination for use, pharmaceutical composition for use, kit, use, method, or diagnostic test according to claim 19 or 20, wherein said second antibody molecule is an IgG4 antibody molecule.

22. 22. The combination for use, pharmaceutical composition for use, kit, use, method or diagnostic test according to any one of claims 1 to 21, wherein said first antibody molecule and / or said second antibody molecule has been engineered to improve binding to an activating Fc gamma receptor.

23. said first antibody molecule comprising the following CDRs: (i) SEQ ID NO: 51 and SEQ ID NO: 52 and SEQ ID NO: 53, or (ii) SEQ ID NO: 57 and SEQ ID NO: 58 and SEQ ID NO: 59, or (iii) SEQ ID NO: 63, SEQ ID NO: 64, and SEQ ID NO: 65, or (iv) SEQ ID NO: 69 and SEQ ID NO: 70 and SEQ ID NO: 71, or (v) SEQ ID NO: 75 and SEQ ID NO: 76 and SEQ ID NO: 77, or (vi) SEQ ID NO: 81 and SEQ ID NO: 82 and SEQ ID NO: 83, or (vii) SEQ ID NO: 87 and SEQ ID NO: 88 and SEQ ID NO: 89, or (viii) SEQ ID NO: 93 and SEQ ID NO: 94 and SEQ ID NO: 95, or (ix) SEQ ID NO: 99 and SEQ ID NO: 100 and SEQ ID NO: 101, or (x) SEQ ID NO: 105 and SEQ ID NO: 106 and SEQ ID NO: 107, or (xi) SEQ ID NO: 111 and SEQ ID NO: 112 and SEQ ID NO: 113, or (xii) SEQ ID NO: 117 and SEQ ID NO: 118 and SEQ ID NO: 119, or (xiii) SEQ ID NO: 123, SEQ ID NO: 124, and SEQ ID NO: 125, or (xiv) SEQ ID NO: 129 and SEQ ID NO: 130 and SEQ ID NO: 131, or (xv) SEQ ID NO: 135, SEQ ID NO: 136, and SEQ ID NO: 137, or (xvi) SEQ ID NO: 141 and SEQ ID NO: 142 and SEQ ID NO: 143, or (xvii) SEQ ID NO: 147 and SEQ ID NO: 148 and SEQ ID NO: 149, or (xviii) SEQ ID NO: 153 and SEQ ID NO: 154 and SEQ ID NO: 155, or (xix) SEQ ID NO: 159 and SEQ ID NO: 160 and SEQ ID NO: 161, or (xx) SEQ ID NO: 165, SEQ ID NO: 166, and SEQ ID NO: 167, or (xxi) SEQ ID NO: 171 and SEQ ID NO: 172 and SEQ ID NO: 173, or (xxii) SEQ ID NO: 177 and SEQ ID NO: 178 and SEQ ID NO: 179, or (xxiii) SEQ ID NO: 183 and SEQ ID NO: 184 and SEQ ID NO: 185, or (xxiv) The combination for use, pharmaceutical composition for use, kit, use, method, or diagnostic test according to any one of claims 1 to 22, comprising a variable heavy chain (VH) comprising SEQ ID NO: 189 and SEQ ID NO: 190 and SEQ ID NO:

191.

24. said first antibody molecule comprising the following CDRs: (i) SEQ ID NO: 54 and SEQ ID NO: 55 and SEQ ID NO: 56, or (ii) SEQ ID NO: 60 and SEQ ID NO: 61 and SEQ ID NO: 62, or (iii) SEQ ID NO: 66, SEQ ID NO: 67, and SEQ ID NO: 68, or (iv) SEQ ID NO: 72 and SEQ ID NO: 73 and SEQ ID NO: 74, or (v) SEQ ID NO: 78 and SEQ ID NO: 79 and SEQ ID NO: 80, or (vi) SEQ ID NO: 84 and SEQ ID NO: 85 and SEQ ID NO: 86, or (vii) SEQ ID NO: 90 and SEQ ID NO: 91 and SEQ ID NO: 92, or (viii) SEQ ID NO: 96 and SEQ ID NO: 97 and SEQ ID NO: 98, or (ix) SEQ ID NO: 102 and SEQ ID NO: 103 and SEQ ID NO: 104, or (x) SEQ ID NO: 108 and SEQ ID NO: 109 and SEQ ID NO: 110, or (xi) SEQ ID NO: 114 and SEQ ID NO: 115 and SEQ ID NO: 116, or (xii) SEQ ID NO: 120 and SEQ ID NO: 121 and SEQ ID NO: 122, or (xiii) SEQ ID NO: 126, SEQ ID NO: 127, and SEQ ID NO: 128, or (xiv) SEQ ID NO: 132 and SEQ ID NO: 133 and SEQ ID NO: 134, or (xv) SEQ ID NO: 138 and SEQ ID NO: 139 and SEQ ID NO: 140, or (xvi) SEQ ID NO: 144 and SEQ ID NO: 145 and SEQ ID NO: 146, or (xvii) SEQ ID NO: 150 and SEQ ID NO: 151 and SEQ ID NO: 152, or (xviii) SEQ ID NO: 156 and SEQ ID NO: 157 and SEQ ID NO: 158, or (xix) SEQ ID NO: 162 and SEQ ID NO: 163 and SEQ ID NO: 164, or (xx) SEQ ID NO: 168 and SEQ ID NO: 169 and SEQ ID NO: 170, or (xxi) SEQ ID NO: 174 and SEQ ID NO: 175 and SEQ ID NO: 176, or (xxii) SEQ ID NO: 180 and SEQ ID NO: 181 and SEQ ID NO: 182, or (xxiii) SEQ ID NO: 186 and SEQ ID NO: 187 and SEQ ID NO: 188, or (xxiv) The combination for use, pharmaceutical composition for use, kit, use, method, or diagnostic test according to any one of claims 1 to 23, comprising a variable light chain (VL) comprising SEQ ID NO: 192 and SEQ ID NO: 193 and SEQ ID NO:

194.

25. 25. The combination for use, pharmaceutical composition for use, kit, use, method, or diagnostic test of any one of claims 1 to 24, wherein said first antibody molecule comprises a variable heavy (VH) amino acid sequence selected from the group consisting of SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:13, SEQ ID NO:14, SEQ ID NO:15, SEQ ID NO:16, SEQ ID NO:17, SEQ ID NO:18, SEQ ID NO:19, SEQ ID NO:20, SEQ ID NO:21, SEQ ID NO:22, SEQ ID NO:23, SEQ ID NO:24, SEQ ID NO:25, and SEQ ID NO:

26.

26. 26. The combination for use, pharmaceutical composition for use, kit, use, method, or diagnostic test of any one of claims 1 to 25, wherein said first antibody molecule comprises a variable light (VL) amino acid sequence selected from the group consisting of SEQ ID NO:27, SEQ ID NO:28, SEQ ID NO:29, SEQ ID NO:30, SEQ ID NO:31, SEQ ID NO:32, SEQ ID NO:33, SEQ ID NO:34, SEQ ID NO:35, SEQ ID NO:36, SEQ ID NO:37, SEQ ID NO:38, SEQ ID NO:39, SEQ ID NO:40, SEQ ID NO:41, SEQ ID NO:42, SEQ ID NO:43, SEQ ID NO:44, SEQ ID NO:45, SEQ ID NO:46, SEQ ID NO:47, SEQ ID NO:48, SEQ ID NO:49, and SEQ ID NO:

50.

27. the first antibody molecule having the following CDR amino acid sequences: (i) SEQ ID NO: 51 and SEQ ID NO: 52 and SEQ ID NO: 53 and SEQ ID NO: 54 and SEQ ID NO: 55 and SEQ ID NO: 56, or (ii) SEQ ID NO: 57 and SEQ ID NO: 58 and SEQ ID NO: 59 and SEQ ID NO: 60 and SEQ ID NO: 61 and SEQ ID NO: 62, or (iii) SEQ ID NO: 63 and SEQ ID NO: 64 and SEQ ID NO: 65 and SEQ ID NO: 66 and SEQ ID NO: 67 and SEQ ID NO: 68, or (iv) SEQ ID NO: 69 and SEQ ID NO: 70 and SEQ ID NO: 71 and SEQ ID NO: 72 and SEQ ID NO: 73 and SEQ ID NO: 74, or (v) SEQ ID NO: 75 and SEQ ID NO: 76 and SEQ ID NO: 77 and SEQ ID NO: 78 and SEQ ID NO: 79 and SEQ ID NO: 80, or (vi) SEQ ID NO: 81 and SEQ ID NO: 82 and SEQ ID NO: 83 and SEQ ID NO: 84 and SEQ ID NO: 85 and SEQ ID NO: 86, or (vii) SEQ ID NO: 87 and SEQ ID NO: 88 and SEQ ID NO: 89 and SEQ ID NO: 90 and SEQ ID NO: 91 and SEQ ID NO: 92, or (viii) SEQ ID NO:93 and SEQ ID NO:94 and SEQ ID NO:95 and SEQ ID NO:96 and SEQ ID NO:97 and SEQ ID NO:98, or (ix) SEQ ID NO: 99 and SEQ ID NO: 100 and SEQ ID NO: 101 and SEQ ID NO: 102 and SEQ ID NO: 103 and SEQ ID NO: 104, or (x) SEQ ID NO: 105 and SEQ ID NO: 106 and SEQ ID NO: 107 and SEQ ID NO: 108 and SEQ ID NO: 109 and SEQ ID NO: 110, or (xi) SEQ ID NO: 111 and SEQ ID NO: 112 and SEQ ID NO: 113 and SEQ ID NO: 114 and SEQ ID NO: 115 and SEQ ID NO: 116, or (xii) SEQ ID NO: 117 and SEQ ID NO: 118 and SEQ ID NO: 119 and SEQ ID NO: 120 and SEQ ID NO: 121 and SEQ ID NO: 122, or (xiii) SEQ ID NO: 123 and SEQ ID NO: 124 and SEQ ID NO: 125 and SEQ ID NO: 126 and SEQ ID NO: 127 and SEQ ID NO: 128, or (xiv) SEQ ID NO: 129 and SEQ ID NO: 130 and SEQ ID NO: 131 and SEQ ID NO: 132 and SEQ ID NO: 133 and SEQ ID NO: 134, or (xv) SEQ ID NO: 135 and SEQ ID NO: 136 and SEQ ID NO: 137 and SEQ ID NO: 138 and SEQ ID NO: 139 and SEQ ID NO: 140, or (xvi) SEQ ID NO: 141 and SEQ ID NO: 142 and SEQ ID NO: 143 and SEQ ID NO: 144 and SEQ ID NO: 145 and SEQ ID NO: 146, or (xvii) SEQ ID NO: 147 and SEQ ID NO: 148 and SEQ ID NO: 149 and SEQ ID NO: 150 and SEQ ID NO: 151 and SEQ ID NO: 152, or (xviii) SEQ ID NO: 153 and SEQ ID NO: 154 and SEQ ID NO: 155 and SEQ ID NO: 156 and SEQ ID NO: 157 and SEQ ID NO: 158, or (xix) SEQ ID NO: 159 and SEQ ID NO: 160 and SEQ ID NO: 161 and SEQ ID NO: 162 and SEQ ID NO: 163 and SEQ ID NO: 164, or (xx) SEQ ID NO: 165 and SEQ ID NO: 166 and SEQ ID NO: 167 and SEQ ID NO: 168 and SEQ ID NO: 169 and SEQ ID NO: 170, or (xxi) SEQ ID NO: 171 and SEQ ID NO: 172 and SEQ ID NO: 173 and SEQ ID NO: 174 and SEQ ID NO: 175 and SEQ ID NO: 176, or (xxii) SEQ ID NO: 177 and SEQ ID NO: 178 and SEQ ID NO: 179 and SEQ ID NO: 180 and SEQ ID NO: 181 and SEQ ID NO: 182, or (xxiii) SEQ ID NO: 183 and SEQ ID NO: 184 and SEQ ID NO: 185 and SEQ ID NO: 186 and SEQ ID NO: 187 and SEQ ID NO: 188, or (xxiv) The combination for use, pharmaceutical composition for use, kit, use, method, or diagnostic test according to any one of claims 1 to 26, comprising SEQ ID NO: 189 and SEQ ID NO: 190 and SEQ ID NO: 191 and SEQ ID NO: 192 and SEQ ID NO: 193 and SEQ ID NO:

194.

28. the first antibody molecule having the following amino acid sequence: (i) SEQ ID NO: 3 and SEQ ID NO: 27, or (ii) SEQ ID NO: 4 and SEQ ID NO: 28, or (iii) SEQ ID NO: 5 and SEQ ID NO: 29, or (iv) SEQ ID NO: 6 and SEQ ID NO: 30, or (v) SEQ ID NO: 7 and SEQ ID NO: 31, or (vi) SEQ ID NO: 8 and SEQ ID NO: 32, or (vii) SEQ ID NO: 9 and SEQ ID NO: 33, or (viii) SEQ ID NO: 10 and SEQ ID NO: 34, or (ix) SEQ ID NO: 11 and SEQ ID NO: 35, or (x) SEQ ID NO: 12 and SEQ ID NO: 36, or (xi) SEQ ID NO: 13 and SEQ ID NO: 37, or (xii) SEQ ID NO: 14 and SEQ ID NO: 38, or (xiii) SEQ ID NO: 15 and SEQ ID NO: 39, or (xiv) SEQ ID NO: 16 and SEQ ID NO: 40, or (xv) SEQ ID NO: 17 and SEQ ID NO: 41, or (xvi) SEQ ID NO: 18 and SEQ ID NO: 42, or (xvii) SEQ ID NO: 19 and SEQ ID NO: 43, or (xviii) SEQ ID NO: 20 and SEQ ID NO: 44, or (xix) SEQ ID NO: 21 and SEQ ID NO: 45, or (xx) SEQ ID NO: 22 and SEQ ID NO: 46, or (xxi) SEQ ID NO: 23 and SEQ ID NO: 47, or (xxii) SEQ ID NO: 24 and SEQ ID NO: 48, or (xxiii) SEQ ID NO: 25 and SEQ ID NO: 49, or (xxiv) A combination for use, pharmaceutical composition for use, kit, use, method, or diagnostic test according to any one of claims 1 to 27, comprising SEQ ID NO: 26 and SEQ ID NO:

50.

29. 29. The combination for use, pharmaceutical composition for use, kit, use, method or diagnostic test according to any one of claims 1 to 22, wherein said first antibody molecule is an antibody molecule capable of competing for binding to FcyRIIb with an antibody molecule according to any one of claims 23 to 28.

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