Antibody combinations for treating cancer in specific patients

Abstract

The combined use of a first antibody molecule that specifically binds to Fc [gamma] RIIb through its Fab region and binds to Fc [gamma] receptor through its Fc region and a second antibody molecule that specifically binds to PD-1 and binds to at least one Fc [gamma] receptor through its Fc region in the treatment of cancer in a patient having tumor-infiltrating T lymphocytes having medium or high PD-1 expression is described, as well as pharmaceutical compositions and kits comprising these two antibody molecules, as well as methods of using these two antibodies to treat cancer. A diagnostic test for identifying patients who benefit from the treatment described herein is also described.

Description

[0001] This application is a divisional application of the Chinese national phase of application No. 202080051736.5, filed on July 17, 2020, entitled “Combination of antibodies for treating cancer in specific patients”. TECHNICAL FIELD

[0002] The present invention relates to the combined use of 1) a first antibody molecule that specifically binds FcyRIIb through its Fab region and binds an Fcy receptor through its Fc region, and 2) a second antibody molecule that specifically binds PD-1 and binds at least one Fcy receptor through its Fc region, in the treatment of cancer in patients having intermediate or high PD-1 expression on CD3-positive tumor infiltrating lymphocytes (TILs). BACKGROUND

[0003] Immune inhibitory checkpoint receptors, such as CTLA-4 or PD-1 (also denoted PD1), are cell surface receptors that, upon binding to their ligand receptors (e.g. CD80 and CD86, members of the B7 family, and PD-L1, respectively), transmit inhibitory signals into the cell, limiting cell activation and proliferation, preventing excessive inflammation and contributing to the maintenance of self-tolerance. Animals with genetic defects in such inhibitory immune checkpoints are associated with exacerbated inflammatory responses, fail to develop or maintain tolerance to self, leading to autoimmune diseases. Antibodies to the immune checkpoint receptors CTLA-4 and PD-1 / PD-L1 improve overall survival in patients with various cancers, including a variety of solid cancer types, such as melanoma, lung cancer, bladder cancer and head and neck cancer, and such antibodies have been approved by the US Food and Drug Administration (Pardoll, D. M. (2012) Nat Rev Cancer 12(4): 252-264; Topalian, S. L. et al. (2015) Cancer Cell 27(4): 450-461; Sharma, P. et al. (2017) Cell 168(4): 707-723).

[0004] Antibodies to the immune inhibitory checkpoint axis PD-1 / PD-L1 have proven particularly effective in cancer immunotherapy, eliciting objective responses (complete and partial responses) in ~20% of patients - a significant improvement over the standard of care (Carretero-Gonzalez, A. et al. (2018) Oncotarget 9(9): 8706-8715). However, as the response rates demonstrate, currently available anti-PD-1 / PD-L1 antibodies are only active in a minority of patients. In addition, a fraction of patients that initially respond will eventually develop resistance and no longer benefit from the treatment. Thus, the mechanisms of non-responsiveness and resistance to PD-1 / PD-L1 antibodies is a clinically important question. Identifying and overcoming mechanisms of resistance to PD-1 / PD-L1 antibodies is a major challenge and opportunity to improve cancer patient survival for this clinically important drug class.

[0005] Furthermore, it is generally accepted that predictive biomarkers for identifying patients most likely to respond to anti-PD-1 / PD-L1 checkpoint blockade are an important strategy to identify patients likely to respond to therapy. Conversely, it is also important to prevent unnecessary treatment of patients with these drugs that are often associated with serious, occasionally fatal, tolerability issues. These therapies further place a huge burden on payers and healthcare systems due to their high cost. Examples of existing predictive biomarkers of clinical significance for anti-PD-1 / PD-L1 antibody therapy are Micro Satellite Instability (MSI) (Le, D. T. et al. (2015) N Engl J Med 372(26): 2509-2520; Le, D. T. et al. (2017) Science 357 (6349): 409-413), tumor mutational burden (Gubin, M. M. 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 mutational burden and tumor PD-L1 expression.

[0006] Fc gamma receptors (FcyR) are membrane proteins found on the cell surface of immune effector cells, including monocytes, macrophages, dendritic cells, neutrophils, mast cells, basophils, eosinophils, and natural killer cells, as well as B lymphocytes. The name is derived from its binding specificity for the Fc region of antibodies. Fc receptors are found on the cell membrane - the cell membrane is otherwise known as the plasma membrane or cytoplasmic membrane. FcyR can be subdivided into activating FcyR and inhibitory FcyR, known to coordinately regulate cell activation by the binding of clustered immunoglobulin G Fc's and transmit activating or inhibitory signals into the cell through intracellular ITAM or ITIM motifs, respectively. FcyR binding of clustered immunoglobulins or immune complexes can mediate antibody internalization into the cell and can lead to antibody-mediated phagocytosis, antibody-dependent cell-mediated cytotoxicity, or antigen presentation or cross-presentation. FcyR are known to also mediate or enhance cross-linking of antibody-bound cell surface receptors. Such cross-linking is known to be required for some (Li, F. et al. (2011) Science 333(6045): 1030-1034; White, A. L. et al. (2011) J Immunol 187(4): 1754-1763) but not all (Richman, L. P. et al. (2014) Oncoimmunology 3: e28610) antibodies in the ability to activate signaling in targeted cells, and can or can not be required to achieve a therapeutic effect.

[0007] In humans, FcyRIIb (CD32b) is an inhibitory Fcy receptor, while FcyRI (CD64), FcyRIIa (CD32a), FcyRIIc (CD32c), and FcyRIIIa (CD16a) are activating Fcy receptors. FcygRIIIb is a GPI-linked receptor expressed on neutrophils, lacks an ITAM motif, and is thought to act as a decoy receptor to offset activating FcyR signaling (Treffers, L. W. et al. (2018) Front Immunol 9: 3124). In mice, the activating receptors are FcyRI, FcyRIII, and FcyRIV.

[0008] It is well known that antibodies can modulate immune cell activity through interactions with Fcy receptors. Specifically, how antibody immune complexes modulate immune cell activation is determined by the relative engagement of its activating and inhibitory Fcy receptors. Different antibody isotypes bind to activating and inhibitory Fcy receptors with different affinities, resulting in different A:I ratios (activating: inhibitory ratios) (Nimmerjahn et al; Science. 2005 Dec 2; 310(5753): 1510-2).

[0009] By virtue of binding to inhibitory Fcy receptors with their Fc domains, antibodies can inhibit, block, and / or downregulate effector cell function. By virtue of binding to inhibitory FcyR with their Fc domains, antibodies can stimulate cell activation through clustering of the signaling receptors targeted by the antibody on the target cell (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 J Immunol 187(4): 1754-1763 White, A. L. et al. (2014) Blood 193(4): 1828-1835).

[0010] By virtue of binding to activating Fcy receptors, antibodies can activate effector cell function and thereby trigger 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 NET neutrophil extracellular traps). Binding of antibodies to activating Fcy receptors can also result in an increase in certain activation markers such as CD40, MHCII, CD38, CD80, and / or CD86.

[0011] In line with the synergistic regulation of antibody-induced effector cell responses by activating and inhibitory Fcy receptors, activating Fcy receptors have been shown to promote tumor cell depletion and therapeutic activity of tumor-directed antibodies. Preclinical and clinical studies have shown that the anti-tumor activity of tumor-directed antibodies (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 higher in individuals carrying higher affinity alleles for activating Fcy receptors (Cartron, G. et al. (2002) J Clin Oncol 99 (3): 754-758; Musolino, A. et al. (2008) J Clin OncolN Engl J Med 26 (11): 1789-1796; Zhang, W. et al. (2007) Nat Med Cancer Cell 25 (24): 3712-3718) and with antibody isotypes and formats that exhibit stronger binding to activating Fcy receptors compared to inhibitory Fcy receptors (high A:I ratio) (Goede, V. et al. (2014) Cancer Cell 370 (12): 1101-1110) in patients. Conversely, the therapeutic activity of tumor- directed antibodies is enhanced in animals lacking the inhibitory FcyRIIB (Clynes, R. A. et al. (2000) Cancer Cell 6 (4): 443-446) or when the inhibitory FcyRIIB is blocked by an antagonistic anti-FcyRIIB antibody as recently shown by some inventors of the present invention (Roghanian, A. et al. (2015) Cancer Cell 27 (4): 473-488).

[0012] Emerging preclinical and clinical data indicate that Fcy receptors also control the efficacy of immunomodulatory antibodies, including immune checkpoint inhibitory antibodies targeting CTLA-4, PD-1 / PD-L1. In humans and mice, there is evidence that the therapeutic activity of anti-CTLA-4 antibodies is promoted by engagement of activating Fcy receptors; melanoma patients carrying the high-affinity allele of the FcyRIIIa gene responded to ipilimumab and showed improved survival compared to patients expressing the lower-affinity FcyRIIIa allele. Moreover, in mice humanized for activating and inhibitory receptors, therapeutic efficacy was shown to be FcyR-dependent and enhanced by antibody isotypes with high A:I ratio (Arce Vargas, F. et al. (2018) Sci Transl Med 33 (4): 649-663 e644).

[0013] These findings prompted us to investigate the ability of FcyRIIB blocking antibodies to enhance the activity of anti-CTLA-4 antibodies. Some inventors of the present invention have previously generated and disclosed two different types of FcyRIIB blocking antibodies (Roghanian, A. et al. (2015) Cancer Cell27 (4): 473-488); proficient binding to both activating and inhibitory human FcyRs, as well as Fc engineered variants showing severely impaired binding to FcyRs through their Fc domains. Both different types of anti-FcyRIIB antibodies showed the same antagonistic effect in blocking CD20 internalization and FcyRIIB signaling in B cells and both improved anti-CD20 mAb-mediated B cell depletion in transgenic animals expressing human FcyRIIB and human CD20.

[0014] In International Patent Application No. PCT / EP2019 / 050566, we demonstrated that only anti-FcyRIIB antibodies lacking the Fc region or showing reduced or impaired binding to FcyRs of the Fc region can enhance the therapeutic activity of anti-CTLA-4 antibodies. In different mouse experimental models of solid cancers, anti-FcyRIIB antibodies with impaired Fc:FcyR binding, but not anti-FcyRIIB antibodies with proficient Fc:FcyR binding, improved the therapeutic activity of anti-CTLA-4 and demonstrated an enhanced Treg depletion of the clinically relevant anti-CTLA-4 antibody ipilimumab in a humanized PBMC in vivo model. Similar enhancement of depletion and / or therapeutic efficacy of antibodies specific for IL-2R (CD25) and PD-L1 was observed upon combination treatment with anti-FcyRIIB antibodies with impaired Fc:FcyR binding, indicating that the enhancement is not limited to a specific target or cell type.

[0015] Preclinical studies have indicated a role of Fcy receptors in controlling the therapeutic activity of anti-PD-1 / PD-L1 antibodies, but the individual role of activating and inhibitory Fcy receptors was not clear. Dahan et al. reported that the anti-tumor activity of a mouse antibody against PD-L1 benefits from FcyR engagement, but in contrast, the activity of an anti-PD-1 antibody is impaired by FcyR engagement (Dahan, R. et al. (2015) Sci Transl Med 28 (3): 285-295). Notably, the anti-PD-1 antibody isotype with a high A:I ratio (i.e. offering strong engagement of activating Fcy receptors compared to inhibitory Fcy receptors) showed lower therapeutic activity compared to the mIgGl isotype with a lower A:I ratio (i.e. stronger engagement of inhibitory Fcy receptors) and compared to an anti-PD-1 variant antibody with Fc:FcyR binding defects.

[0016] Using the clinically relevant human anti-PD-1 IgG4 antibody nivolumab and an alternative rat IgG2a antibody with the claimed similar Fcy receptor binding characteristics, Arlauckas and colleagues similarly found that anti-PD-1 variant antibodies with impaired Fc:FcyR binding (de-glycosylated) had improved therapeutic activity compared to their Fc:FcyR good wild-type human IgG4 and rat IgG2a anti-PD-1 counterparts (Arlauckas, S. P. et al. (2017) Blood Sci Transl Med 9 (389)). However, in contrast to Dahan et al., using blocking antibodies against individual Fcy receptors, the authors identified the activating mouse Fcy receptor III and the inhibitory mouse Fcy receptor II as potential causes for the reduced efficacy of anti-PD-1 antibodies. Thus, the relative importance of (individual) activating and inhibitory Fcy receptors as potential causes for 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 FcyR should be blocked to enhance anti-PD-1 antibody activity, is not clear from the prior art. SUMMARY

[0017] Disclosed herein is a combination of:

[0018] a first antibody molecule that specifically binds FcyRIIb via its Fab region and binds a Fcy receptor via its Fc region, and

[0019] a second antibody molecule that specifically binds PD-1 and binds at least one Fcy receptor via its Fc region;

[0020] for use in the treatment of cancer in a patient having tumor infiltrating T lymphocytes with medium or high PD-1 expression.

[0021] Also disclosed herein is a pharmaceutical composition comprising:

[0022] (i) a first antibody molecule that specifically binds FcyRIIb via its Fab region and binds a Fcy receptor via its Fc region, and

[0023] (ii) a second antibody molecule that specifically binds PD-1 and binds at least one Fcy receptor via its Fc region;

[0024] for use in the treatment of cancer in a patient having tumor infiltrating T lymphocytes with medium or high PD-1 expression.

[0025] Also disclosed herein is a kit for use in the treatment of cancer in a patient having tumor infiltrating T lymphocytes with medium or high PD-1 expression, comprising:

[0026] (i) a first antibody molecule which specifically binds FcyRIIb through its Fab region and which binds an Fcy receptor through its Fc region, and

[0027] (ii) a second antibody molecule which specifically binds PD-1 and which binds at least one Fcy receptor through its Fc region.

[0028] Also disclosed herein is the use of:

[0029] (i) a first antibody molecule which specifically binds FcyRIIb through its Fab region and which binds an Fcy receptor through its Fc region, and

[0030] (ii) a second antibody molecule which specifically binds PD-1 and which binds at least one Fcy receptor through its Fc region;

[0031] for the manufacture of a medicament for the treatment of cancer in a patient having tumor infiltrating T lymphocytes with medium or high PD-1 expression.

[0032] Also disclosed herein is a method for the treatment of cancer in a patient having tumor infiltrating T lymphocytes with medium or high PD-1 expression, comprising administering:

[0033] (i) a first antibody molecule which specifically binds FcyRIIb through its Fab region and which binds an Fcy receptor through its Fc region, and

[0034] (ii) a second antibody molecule which specifically binds PD-1 and which binds at least one Fcy receptor through its Fc region.

[0035] Also disclosed herein is a diagnostic test for determining whether a patient would benefit from a combination treatment using:

[0036] (i) a first antibody molecule which specifically binds FcyRIIb through its Fab region and which binds an Fcy receptor through its Fc region, and

[0037] (ii) a second antibody molecule which specifically binds PD-1 and which binds at least one Fcy receptor,

[0038] The test comprises determining PD-1 expression on tumor infiltrating T lymphocytes of the patient, wherein medium or high PD-1 expression indicates that the patient would benefit from the combination treatment. Accordingly, a lack of medium or high PD-1 expression on T lymphocytes but rather low PD-1 expression indicates that the patient would not benefit from the combination treatment. DETAILED DESCRIPTION

[0039] Here, we demonstrate that only anti-FcyRIIB antibodies with good Fc:FcyR binding but not impaired Fc:FcyR binding enhance the in vivo therapeutic efficacy of anti-PD-1 antibodies and prevent in vitro phagocytosis of PD-1 high expressing T cells by a clinically relevant human anti-PD-1 antibody. This finding is novel and unexpected as the above referenced studies on the role of FcyRs in anti-PD-1 therapy suggest a broad role of activating FcyRs compared to inhibitory FcyRs (Dahan, R. et al. (2015) Figure 1 28 (3): 285-295), or the separate activating (FcyRIII) and inhibitory FcyRIIB (Arlauckas, S. P. et al. (2017) Figure 1 9(389)) as potential causes for impaired anti-PD-1 antibody activity.

[0040] In view of some earlier findings by the inventors related to antibodies against other immune checkpoints, including anti-CTLA-4 in particular, where only anti-FcyRIIB antibodies with impaired Fc:FcyR binding but not good Fc:FcyR binding enhanced therapeutic activity, the present invention is even more surprising.

[0041] Thus, the present invention relates to the combined use of:

[0042] (i) an antibody molecule that specifically binds FcyRIIb by its Fab region and that binds a Fcy receptor by its Fc region (denoted herein as first antibody molecule), and

[0043] (ii) an antibody molecule that specifically binds PD-1 and that binds at least one Fcy receptor by its Fc region (denoted herein as second antibody molecule)

[0044] for the treatment of cancer in a patient having tumor infiltrating T lymphocytes with medium or high PD-1 expression.

[0045] This combination is intended for the treatment of cancer in a patient, such as a solid cancer, with the aim to improve the therapeutic efficacy of an antibody molecule that specifically binds PD-1 (i.e. an anti-PD-1 antibody) by reducing binding to FcyRs, including FcyRIIB.

[0046] The antibody molecule specifically binding to FcyRIIb according to the present application, i.e. the first antibody, binds or interacts with this Fcy 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 and one variable domain of each of the heavy and light chain. In particular, it binds to FcyRIIb present on immune effector cells, e.g. macrophages, and in particular to FcyRIIb present on the surface of immune effector cells.

[0047] In addition to the above, the antibody molecule specifically binding to FcyRIIb according to the present application, i.e. the first antibody molecule, also binds to activating Fcy receptors via interaction between the Fc region and the Fc receptor, as known and extensively characterized for antibodies of the human IgGl isotype (Bruhns, P. et al. (2009) Sci Transl Med 113 (16): 3716-3725).

[0048] This has at least the following therapeutically important consequences: The activating and inhibitory Fcy receptors are blocked in an anti-FcyRIIB antibody-dependent (Fab and Fc) manner, preventing anti-PD-1 antibody-mediated elimination of anti-tumor T cells coated with anti-PD-1 antibody by macrophages or other immune effector cells expressing Fcy receptors (coating in this context means that the anti-PD-1 antibody has bound to the cell). The mechanism can also involve inhibition of macrophage FcyR-dependent transfer of PD-1 antibody from T cells to FcyR-expressing effector cells, e.g. macrophages, as previously described (Arlauckas, S. P. et al. (2017) Infect Immun 9 (389)).

[0049] The immune effector cells expressing Fcy receptors herein primarily refer to innate effector cells and specifically include macrophages, neutrophils, monocytes, natural killer (NK) cells, basophils, eosinophils, mast cells, and platelets. Cytotoxic T cells and memory T cells generally do not express FcyR, but can do so in specific cases. In some embodiments, the immune effector cell is an innate immune effector cell. In some embodiments, the immune effector cell is a macrophage.

[0050] The antibody molecule specifically binding to PD-1, i.e. the second antibody molecule, has an Fc region that binds or interacts with activating Fcy receptors, allowing antibody PD-1 antibody-dependent FcyR effector cell-dependent elimination of anti-tumor T cells coated with anti-PD-1 antibody. The immune cells to which the anti-PD-1 antibody molecule binds are immune cells that confer key anti-tumor activity, such as CD8+ or CD4+ T cells.

[0051] Thus, any anti-PD-1 variant antibody, including those of human IgG4, IgGl, IgG2 and IgG3 isotypes, whose Fc region binds to or interacts with an activating Fcy receptor to the extent that it results in the elimination of FcyR-expressing effector cells of anti-tumor T cells expressing PD-1, can be combined with an anti-FcyRIIB antibody that binds well to Fc:FcyR, i.e. with an antibody molecule that specifically binds FcyRIIb through its Fab region and binds Fcy receptors through its Fc region.

[0052] 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 therapy to activate the immune system to attack tumors. Monoclonal antibodies targeting PD-1 or PD-L1 can block the binding of PD-1 to PD-L1, which can enhance the immune response against cancer cells.

[0053] The anti-PD-1 antibody binds to PD-1 expressed on T cells within a tumor.

[0054] A patient who benefits from the treatment according to the present application is a patient who is eligible for anti-PD-1 therapy according to the criteria of an approved regimen containing an anti-PD-1 antibody and in addition has tumor-infiltrating T lymphocytes, i.e. tumor-infiltrating CD3+ lymphocytes, with medium or high PD-1 expression. Patients eligible for anti-PD-1 therapy include patients 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 cancer that is highly unstable (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 amenable to curative surgery or definitive radiation. As the skilled person knows, the number of indications that can be treated expands rapidly with new trials, new anti-PD-1 antibodies and new combinations.

[0055] In the work leading to the present application it was observed that when anti-PD-1 antibodies are administered to T cells having intermediate or high PD-1 expression this can lead to phagocytosis of the anti-PD-1 antibody-coated T cells, in particular CD8-positive T cells. In the work leading to the present application it was further discovered that by administering antibody molecules that specifically bind FcyRIIb via their Fab region and bind Fcy receptors via their Fc region together with anti-PD-1 antibodies, this phagocytosis can be blocked for T cells having intermediate or high PD-1 expression. The in vitro relevance of the above findings (further explained in the examples below) was confirmed in two different solid cancer experimental models comprising immune-competent mice, in which a significant increase in survival was observed using combination therapy with anti-FcyRIIB and anti-PD-1 antibodies that bind FcyR well compared to monotherapy with anti-PD-1. This is also shown in more detail in the examples below. Further supporting the in vitro relevance of the in vivo, the in vivo T cell PD-1 expression levels in the tumor were similar in both settings; the in vivo T cell PD-1 expression in the tumor ranged from ~20,000 to 80,000 PD-1 molecules per cell, spanning the in vitro expression levels of PD-1 of PD-1 cultured (15,500 to 78,000 PD-1 molecules) and highly expressing (65,000 to 391,000 PD-1 molecules per cell) human T cells, both of which are sensitive to Fc:FcyR binding good anti-FcyRIIB blockade of human anti-PD-1 mediated phagocytosis.

[0056] Thus, in the context of the present application, a patient that can benefit from the treatment described herein is a patient having at least 10% of the tumor infiltrating T lymphocytes having intermediate or high PD-1 expression.

[0057] In the present context, intermediate or high expression means that for at least 10% of the tumor infiltrating T lymphocytes, each cell expresses > 15,500 PD-1 molecules. As further explained below, the absolute number of PD-1 expression can vary depending on which anti-PD-1 antibody is used and / or which method is used to measure PD-1 expression. Thus, intermediate or high expression as used herein is measured using the methods described herein and / or using the anti-human PD-1 antibody EH12.2H7 (available from BioLegend) as equal to or higher than 15,500 PD-1 molecules per cell.

[0058] Similar to the in vivo environment, individual patient's intratumoral T cells will exhibit heterogeneous PD-1 expression. An individual patient can have different cell populations with varying PD-1 expression, such as one population with low expression and one population with moderate or high expression. In some embodiments, at least 15% of the patient's tumor infiltrating CD3+ T lymphocytes have moderate or high PD-1 expression. In some embodiments, at least 20% of the patient's tumor infiltrating CD3+ T lymphocytes have moderate or high PD-1 expression. In some embodiments, at least 25% of the patient's tumor infiltrating CD3+ T lymphocytes have moderate or high PD-1 expression. In some embodiments, at least 30% of the patient's tumor infiltrating CD3+ T lymphocytes have moderate or high PD-1 expression. In some embodiments, at least 35% of the patient's tumor infiltrating CD3+ T lymphocytes have moderate 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 moderate or high PD-1 expression. In some embodiments, at least 80% of the patient's tumor infiltrating CD3+ T lymphocytes have moderate or high PD-1 expression. In some embodiments, at least 85% of the patient's tumor infiltrating CD3+ T lymphocytes have moderate or high PD-1 expression. In some embodiments, at least 90% of the patient's tumor infiltrating CD3+ T lymphocytes have moderate or high PD-1 expression.

[0059] In some embodiments, the patient's tumor infiltrating CD3 positive and CD8 positive (CD3+CD8+) T lymphocytes have moderate or high PD-1 expression.

[0060] In some embodiments, at least 10% of the patient's tumor infiltrating CD3+CD8+ T lymphocytes have medium or high PD-1 expression. In some embodiments, at least 15% of the patient's tumor infiltrating CD3+CD8+ T lymphocytes have medium or high PD-1 expression. In some embodiments, at least 20% of the patient's tumor infiltrating CD3+CD8+ T lymphocytes have medium or high PD-1 expression. In some embodiments, at least 25% of the patient's tumor infiltrating CD3+CD8+ T lymphocytes have medium or high PD-1 expression. In some embodiments, at least 30% of the patient's tumor infiltrating CD3+CD8+ T lymphocytes have medium or high PD-1 expression. In some embodiments, at least 35% of the patient's tumor infiltrating CD3+CD8+ T lymphocytes have medium or high PD-1 expression. In some embodiments, at least 40% of the patient's tumor infiltrating CD3+CD8+ T lymphocytes have medium or high PD-1 expression. In some embodiments, at least 45% of the patient's tumor infiltrating CD3+CD8+ T lymphocytes have medium or high PD-1 expression. In some embodiments, at least 50% of the patient's tumor infiltrating CD3+CD8+ T lymphocytes have medium or high PD-1 expression. In some embodiments, at least 55% of the patient's tumor infiltrating CD3+CD8+ T lymphocytes have medium or high PD-1 expression. In some embodiments, at least 60% of the patient's tumor infiltrating CD3+CD8+ T lymphocytes have medium or high PD-1 expression. In some embodiments, at least 65% of the patient's tumor infiltrating CD3+CD8+ T lymphocytes have medium or high PD-1 expression. In some embodiments, at least 70% of the patient's tumor infiltrating CD3+CD8+ T lymphocytes have medium or high PD-1 expression. In some embodiments, at least 75% of the patient's tumor infiltrating CD3+CD8+ T lymphocytes have medium or high PD-1 expression. In some embodiments, at least 80% of the patient's tumor infiltrating CD3+CD8+ T lymphocytes have medium or high PD-1 expression. In some embodiments, at least 85% of the patient's tumor infiltrating CD3+CD8+ T lymphocytes have medium or high PD-1 expression. In some embodiments, at least 90% of the patient's tumor infiltrating CD3+CD8+ T lymphocytes have medium or high PD-1 expression.

[0061] PD-1 expression on tumor cells of an individual patient can be measured using cells or tissue derived from a tumor biopsy. More specifically, absolute T cell expression levels can be quantified using flow cytometry and bead-based antibody and cell epitope quantitation kits described herein or equivalent. Alternatively, tumor tissue biopsies can be used to perform semi-quantitative analysis by immunohistochemistry, comparing anti-PD-1 staining of patient biopsies to staining of cells of tissue or cell cytoslide, indicating 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-FcyRIIB-mediated enhancement of activity of anti-PD-1 antibodies. Importantly, if the method of determining human T cell PD-1 expression differs from the quantitative method described herein, or uses a different anti-PD-1 antibody clone or fluorescent label, the assay needs to be compared and validated, for example against the methods detailed in Example 1 (especially with reference to the Cancer Cell In Example 1). In general, a method of quantifying PD-1 expression, as exemplified in Example 1 with reference to 1A01-VH antibodies labeled with fluorescent dyes at defined ratios; for example - and as preferred in some embodiments - using antibodies labeled with phycoerythrin (PE) at a ratio of 1 : 1. Thus, using beads with a defined number of fluorescent dye (e.g. PE) molecules to generate a standard curve, the number of antibody molecules bound to the cell can be determined. Cells to be tested are incubated with labeled anti-PD1 antibody, such as anti-human PD-1 antibody EH12.2H7 from Biolegend, and analyzed using a FAC machine set in such a way that beads and cells can be run in the same environment. A standard curve is generated, for example by plotting Log number of molecules per bead against Log fluorescence, then cells stained with fluorescent dye-labeled anti-PD1 antibody are run, and the number of antibody bound is calculated using Log mean fluorescence intensity (MFI).

[0062] As mentioned above, the absolute number can vary depending on the anti-PD1 antibody used for the measurement.

[0063] In addition to specifically binding to PD-1 on immune cells, the second antibody molecule also binds to at least one Fey receptor through its Fc region. In some embodiments, the second antibody molecule binds to at least one activating Fey receptor through its Fc region. The second antibody can be capable of binding an activating Fey receptor, such as an activating Fey receptor present on an immune effector cell, through its Fc region. To be capable of binding an activating Fey receptor, at least in some embodiments, the Fc region of the second antibody can be glycosylated at position 297. The carbohydrate residue in this position facilitates binding to the Fey receptor. In some embodiments, it is preferred that these residues are biantennary, containing GlnNAc, mannose, sialic acid with terminal galactose residues, and a CH2 portion of the Fc molecule.

[0064] The present application also relates to a diagnostic test that can be used to identify patients who would benefit from the treatment described herein, i.e. the combination of (i) a first antibody molecule that specifically binds FcyRIIb through its Fab region and binds a Fcy receptor through its Fc region, and (ii) a second antibody molecule that specifically binds PD-1 and binds at least one Fcy receptor through its Fc region. Based on the in vivo PD-1 expression levels associated with anti-FcyRIIB-mediated enhancement of anti-PD-1 antibody efficacy, and in vitro phagocytosis assays incorporating the human anti-PD-1 antibody relevant to the treatment, as well as human T cells and macrophages, the inventors have determined that certain T cell PD-1 receptor expression levels are associated with and necessary for anti-FcyRIIB-mediated enhancement of anti-PD-1 treatment efficacy. The diagnostic test according to the present application is based on this finding, and thus comprises measuring PD-1 expression on tumor cells in a sample, such as cells or tissue derived 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 for measuring PD-1 expression; at least 15,500 PD-1 molecules expressed per T lymphocyte is predictive of a patient who can benefit from the combination treatment according to the present application, as further described above and in Example 1.

[0065] Antibodies are well known to those skilled in the art of immunology and molecular biology. Typically, an antibody comprises two heavy (H) chains and two light (L) chains. In this document, this complete antibody molecule is sometimes referred to as a full-size or full-length antibody. The heavy chain of an antibody comprises one variable domain (VH) and three constant domains (CH1, CH2 and CH3), and the light chain of an antibody comprises one variable domain (VL) and one constant domain (CL). The variable domains (sometimes collectively referred to as Fv) are primarily responsible for binding to an antigen. The constant domains are primarily responsible for the effector functions of antibodies. VThe variable domain includes three loops, known as complementarity determining regions (CDRs), which are responsible for target binding. The constant domains are not directly involved in the binding of antibody to antigen, but exhibit various effector functions. Depending on the amino acid sequence of the heavy chain constant region of an antibody or immunoglobulin, it can be assigned to different classes. There are five major classes of immunoglobulins: IgA, IgD, IgE, IgG, and IgM, and in humans, several of these classes are further divided into subclasses (isotypes), e.g., IgGl, IgG2, IgG3, and IgG4, IgAl and IgA2.

[0066] Another part of an antibody is the Fc region (otherwise known as the fragment crystallizable domain), which includes two of the constant domains in each of the heavy chains of the antibody. As mentioned above, the Fc region is responsible for the interaction between the antibody and an Fc receptor.

[0067] As used herein, the term antibody molecule encompasses full-length or full-size antibodies as well as functional fragments of full-length antibodies and derivatives of such antibody molecules.

[0068] A functional fragment of a full-size antibody has the same antigen binding characteristics as the corresponding full-size antibody and includes the same variable domains (i.e., VH and VL sequences) and / or the same CDR sequences 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 can correspond to the Fv portion of the full-size antibody. Alternatively, such a fragment can be a Fab, also denoted as a monovalent antigen binding fragment F(ab) or a divalent antigen binding fragment F(ab')2, which does not contain the Fc portion, which contains two antigen binding Fab portions linked together by disulfide bonds or F(ab') (i.e., a monovalent variant of F(ab')2). Such a fragment can also be a single chain variable fragment (scFv).

[0069] A functional fragment does not always contain all six CDRs of the corresponding full-size antibody. It will be appreciated that a molecule containing three or fewer CDR regions, in some cases even just a single CDR or a portion thereof, is capable of retaining the antigen binding activity of the antibody from which the CDR(s) is derived. For example, in Gao et al., 1994, J. Biol. Chem., 269:32389-93, it is described that the entire VL chain, including all three CDRs, has high affinity for its substrate.

[0070] Molecules containing two CDR regions are described, for example, in Vaughan and Sollazzo 2001, Combinatorial Chemistry & High Throughput Screening, 4: 417-430. On page 418 (right column - 3 "Our Strategy for Design"), minibodies containing only the H1 and H2 CDR hypervariable regions interspersed within framework regions are described. The minibodies are described as being able to bind to a target. Vaughan and Sollazzo cite Pessi et al. 1993, Nature, 362: 367-9 and Bianchi et al. 1994, J. Mol. Biol., 236: 649-59, and these papers describe H1 and H2 minibodies and their properties in more detail. In Qiu et al. 2007, Nature Biotechnology, 25: 921-9, molecules consisting of two linked CDRs are shown to be able to bind antigen. Quiocho 1993, Nature, 362: 293-4 provides a summary of the "minibody" technology. Ladner 2007, Nature Biotechnology, 25: 875-7 states that molecules containing two CDRs are able to retain antigen binding activity.

[0071] Antibody molecules containing a single CDR region are described, for example, in Laune et al. 1997, J. Biol. Chem., 272: 30937-44, where a series of hexapeptides derived from CDRs are shown to display antigen binding activity, and it is noted that synthetic peptides of the entire single CDRs display strong binding activity. In Monnet et al. 1999, J. Biol. Chem., 274: 3789-96, a series of 12-mer peptides and related framework regions are shown to have antigen binding activity, and it is stated that CDR3-like peptides alone are able to bind antigen. In Heap et al. 2005, J. Gen. Virol., 86: 1791-1800, "minibodies" (molecules containing a single CDR) are reported to be able to bind antigen, and it is shown that a cyclic peptide from an anti-HIV antibody has antigen binding activity and function. In Nicaise et al. 2004, Protein Science, 13: 1882-91, it is shown that a single CDR can confer antigen binding activity and affinity for its lysozyme antigen.

[0072] Accordingly, 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.

[0073] The antibody molecule can also be a derivative of a full-length antibody or a fragment of such antibody. When a derivative is used, the derivative should have the same antigen binding characteristics as the corresponding full-length antibody, in the sense that the derivative binds to the same epitope on the target as the full-length antibody.

[0074] Accordingly, as used herein, the term "antibody molecule" includes all types of antibody molecules and functional fragments and derivatives thereof, 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 Fv (scFv), Fab fragments, F(ab')2 fragments, F(ab') fragments, disulfide-linked Fv (sdFv), antibody heavy chains, antibody light chains, homodimers of antibody heavy chains, homodimers of antibody light chains, heterodimers of antibody heavy chains, heterodimers of antibody light chains, antigen-binding functional fragments of such homodimers and heterodimers.

[0075] Further, as used herein, the term "antibody molecule" includes all classes of antibody molecules and functional fragments, including: IgG, IgGl, IgG2, IgG3, IgG4, IgA, IgM, IgD, and IgE, unless otherwise specified.

[0076] In some embodiments, the first antibody is human IgGl. The skilled artisan will recognize that mouse IgG2a and human IgGl engage and are able to block activated Fc gamma receptors, thereby preventing the engagement of an anti-PD-1 antibody-mediated activation of Fc gamma receptors on immune effector cells and the subsequent elimination of the anti-PD-1 antibody-coated effector T cells by, for example, ADCP or ADCC. As such, in embodiments where mouse IgG2a is the preferred isotype for deletion in mice, human IgGl is the preferred isotype for deletion in humans in such embodiments.

[0077] In some embodiments, the first antibody is a human IgGl. 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 is Fc engineered to enhance binding to Fcy receptors. In some embodiments, the human IgG antibody is Fc engineered to improve binding to one or several activating Fcy receptors, and / or engineered to improve the relative binding of activating Fcy receptors compared to inhibitory Fcy receptors. In some embodiments, the anti-FcyRIIB antibody is a Fc engineered human IgG antibody. Examples of such engineered antibody variants include afucosylated antibodies whose binding to FcyRIIIA is selectively improved, and antibodies engineered by directed, mutational or other means to bear amino acid substitutions that result in improved binding to one or several activating Fcy receptors compared to inhibitory FcyRIIB (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 U S A, 103: 4005-10). In some embodiments, the human IgG antibody engineered to improve binding to activating Fcy receptors can be a human IgG antibody bearing two mutations S239D and I332E or three mutations S239D, I332E and A330L and / or G236A mutations in its Fc portion. In some embodiments, the human IgG antibody engineered to improve binding to activating Fcy receptors can be an afucosylated human IgG antibody.

[0078] In some embodiments, the second antibody is a human IgG4, i.e. the isotype of the FDA currently approved anti-PD-1 antibodies nivolumab, pembrolizumab and ceplizumab. The skilled person will recognize that several murine antibody isotypes are capable of binding activating and inhibitory Fcy receptors. Importantly, the skilled person will know that the rat IgG2a isotype, which is known to bind to mouse activating and inhibitory Fcy receptors, is very similar to the human IgG4 isotype, which binds to human activating and inhibitory Fcy receptors (Arlauckas, S. P. et al. (2017) 1A01-VL 9 (389)). The skilled person will further know that in addition to the human isotypes IgG1 and IgG4, human IgG3 and IgG2 antibodies can effectively engage human FcyRs (Sanders, L. A. et al. (1995) CDR regions 63 (1): 73-81), and mediate antibody-dependent T cell depletion after activation of immune cells carrying activating Fcy receptors by, e.g., ADCP and ADCC (Arce Vargas, F. et al. (2018) 1B07-VH 33 (4): 649-663 e644). Thus, in some embodiments, the second antibody can be a human IgG1 or IgG2 or IgG3 antibody.

[0079] As mentioned above, the present application encompasses different types and forms of antibody molecules and are known to the person skilled in the art of immunology. It is well known that antibodies for therapeutic purposes are often modified with additional components that modify the properties of the antibody molecule.

[0080] Thus, it is encompassed that the antibody molecule of the present application or the antibody molecule for use according to the present application (e.g. monoclonal antibody molecule and / or polyclonal antibody molecule and / or bispecific antibody molecule) comprises a detectable moiety and / or a cytotoxic moiety.

[0081] A “detectable moiety” comprises one or more from the group comprising: an enzyme; a radioactive atom; a fluorescent moiety; a chemiluminescent moiety; a bioluminescent moiety. The detectable moiety allows the visualization of the antibody molecule in vitro and / or in vivo and / or ex vivo.

[0082] A “cytotoxic moiety” comprises a radioactive moiety and / or an enzyme, wherein the enzyme is a caspase and / or a toxin, wherein the toxin is a bacterial toxin or venom; wherein the cytotoxic moiety is capable of inducing cell lysis.

[0083] Further included are antibody molecules which can be in isolated form and / or in purified form, and / or which can be pegylated. Pegylation is a method of adding a polyethylene glycol polymer to a molecule, such as an antibody molecule or derivative, to modify its behaviour, for example to prolong its half-life by increasing its hydrodynamic size, thereby preventing renal clearance.

[0084] As discussed above, the CDRs of an antibody bind to the antibody target. The assignment of amino acids to each CDR described herein is in accordance with the definition according to: Kabat EA et al., 1991, “Sequences of Proteins of Immunological Interest” Fifth Edition, NIH Publication No. 91-3242, pages xv-xvii.

[0085] As the skilled person will appreciate, there are other methods for assigning amino acids to each CDR. For example, the International ImMunoGeneTics information system (IMGT®) (http: / / www.imgt.org / and Lefranc and Lefranc “The Immunoglobulin FactsBook”, published by Academic Press, 2001).

[0086] In another embodiment, the antibody molecule of the application or for use according to the application is an antibody molecule which is capable of competing with the specific antibodies provided herein, for example an antibody molecule comprising any of the sequences included in the amino acid sequences set out in, for example, SEQ ID NOs: 1-194 for binding to the specific target.

[0087] “Capable of competing” means 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 the specific target.

[0088] For example, such a competing antibody molecule can be capable of inhibiting the 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%, about 100%) and / or inhibiting the ability of an antibody described herein to prevent or reduce binding to the 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%).

[0089] Competitive binding can be determined by methods well known to those skilled in the art, such as enzyme-linked immunosorbent assay (ELISA).

[0090] Epitope-modifying or blocking antibodies can be evaluated using an ELISA assay. Additional methods suitable for identifying competing antibodies are disclosed in Antibodies: A Laboratory Manual, Harlow and Lane, incorporated herein by reference (see, e.g., pages 567 to 569, 574 to 576, 583, and 590 to 612, 1988, CSHL, NY, ISBN 0-87969-314-2).

[0091] It is well known that antibodies specifically bind or interact with defined target molecules or antigens, and this means that the antibody preferentially and selectively binds its target rather than molecules that are not the target.

[0092] The target of the antibody according to the present application, or the target of the antibody for use according to the present application, is expressed on the surface of a cell, i.e. the target is a cell surface antigen, which will comprise an epitope for the antibody (also referred to in this context as a cell surface epitope). Cell surface antigens and epitopes are terms readily understood by the skilled person in the field of immunology or cell biology.

[0093] A“cell surface antigen” comprises: the cell surface antigen is exposed to the extracellular side of the cell membrane, but can be exposed to the extracellular side of the cell membrane only transiently. “Transiently exposed” comprises: the cell surface antigen can be internalized into the cell, or released from the extracellular side of the cell membrane into the extracellular space. The cell surface antigen can be released from the extracellular side of the cell membrane by lysis, which can be mediated by proteases.

[0094] Also included: the cell surface antigen can be attached to the cell membrane, but can be associated with the cell membrane only transiently. “Transiently associated” comprises: the cell surface antigen can be released from the extracellular side of the cell membrane into the extracellular space. The cell surface antigen can be released from the extracellular side of the cell membrane by lysis, which can be mediated by proteases.

[0095] Further included: the cell surface antigen can be a peptide, or a polypeptide, or a carbohydrate, or an oligosaccharide chain or a lipid; and / or an epitope present on a protein, glycoprotein or lipoprotein.

[0096] Methods of assessing protein binding are known to those of skill in biochemistry and immunology. Those of skill will understand that those methods can be used to assess binding of an antibody to a target and / or binding of the Fc region of an antibody to an Fc receptor; and relative strength or specificity, or inhibition or prevention or reduction in those interactions. Examples of methods that can be used to assess protein binding are, for example, immunoassays, BIAcore, Western blots, radioimmunoassays (RIA), and enzyme-linked immunoabsorbent assays (ELISA) (see Fundamental Immunology, Second Edition, Raven Press, New York, pages 332-336 (1989) for a discussion of antibody specificity).

[0097] Thus, an "antibody molecule that specifically binds" or "a target-specific antibody molecule" includes an antibody molecule that specifically binds to a target and does not bind to a non-target, or binds to a non-target less strongly (e.g., with lower affinity) than to the target.

[0098] Also included is the implication that the antibody specifically binds to a target at least twice as strongly, or at least five times as strongly, or at least 10 times as strongly, or at least 20 times as strongly, or at least 50 times as strongly, or at least 100 times as strongly, or at least 200 times as strongly, or at least 500 times as strongly, or at least about 1000 times as strongly, as the antibody specifically binds to a non-target.

[0099] Also included is the implication that if an antibody binds to a target with a Kd of at least about 10 -1 K d of at least about 10 -2 K d of at least about 10 -3 K d of at least about 10 -4 K d of at least about 10 -5 K d of at least about 10 -6 K d of at least about 10 -7 K d of at least about 10 -8 K d of at least about 10 -9 K d of at least about 10 -10 K d of at least about 10 -11 K d of at least about 10 -12 Kd of at least about 10 -13 K d of at least about 10 -14 K dor at least about 10 -15 K d K d The antibody specifically binds to a target if the antibody binds to the target with a K

[0100] In some embodiments, the antibody molecule that specifically binds FcyRIIb is a human antibody.

[0101] In some embodiments, the antibody molecule that specifically binds FcyRIIb is a human-derived antibody, i.e., an antibody originally derived from a human that has been modified as described herein.

[0102] In some embodiments, the antibody molecule that specifically binds FcyRIIb is a humanized antibody, i.e., an antibody originally derived from a non-human that has been modified to increase its similarity to a human antibody. The humanized antibody can be, for example, a murine antibody or a lama antibody.

[0103] In some embodiments, the antibody molecule that specifically binds FcyRIIb includes the following constant regions (CH and CL):

[0104] IgGl-CH [SEQ ID NO: 1]

[0105] ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK

[0106] IgGl-CL [SEQ ID NO: 2]

[0107] QPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECS

[0108] In some embodiments, the antibody molecule that specifically binds FcyRIIb comprises one or more sequences from the following clones:

[0109] Antibody Clone: 1A01

[0110] 1B07-VL [SEQ ID NO: 3]

[0111] EVQLLESGGGLVQPGGSLRLSCAASGFTFSDYYMNWIRQTPGKGLEWVSLIGWDGGSTYYADSVKGRFTISRDNSENTLYLQMNSLRAEDTAVYYCARAYSGYELDYWGQGTLVTVSS

[0112] CDR regions [SEQ ID NO: 27]

[0113] QSVLTQPPSASGTPGQRVTISCSGSSSNIGNNAVNWYQQLPGTAPKLLIYDNNNRPSGVPDRFSGSKSGTSASLAISGLRSEDEADYYCAAWDDSLNASIFGGGTKLTVLG

[0114] 1C04-VH

[0115] CDRH1: DYYMN [ SEQ ID NO: 51]

[0116] CDRH2: LIGWDGGSTYYADSVKG [ SEQ ID NO: 52]

[0117] CDRH3: AYSGYELDY [ SEQ ID NO: 53]

[0118] CDRL1: SGSSSNIGNNAVN [ SEQ ID NO: 54]

[0119] CDRL2: DNNNRPS [ SEQ ID NO: 55]

[0120] CDRL3: AAWDDSLNASI [ SEQ ID NO: 56]

[0121] Antibody Clone: 1B07

[0122] 1C04-VL[SEQ ID NO: 4]

[0123] EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYGMHWVRQAPGKGLEWVAFTRYDGSNKYYADSVRGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARENIDAFDVWGQGTLVTVSS

[0124] CDR regions [SEQ ID NO: 28]

[0125] QSVLTQPPSASGTPGQRVTISCSGSSSNIGNNAVNWYQQLPGTAPKLLIYDNQQRPSGVPDRFSGSKSGTSASLAISGLRSEDEADYYCEAWDDRLFGPVFGGGTKLTVLG

[0126] 1E05-VH

[0127] CDRH1: SYGMH [ SEQ ID NO: 57]

[0128] CDRH2: FTRYDGSNKYYADSVRG [ SEQ ID NO: 58]

[0129] CDRH3: ENIDAFDV [ SEQ ID NO: 59]

[0130] CDRL1: SGSSSNIGNNAVN [ SEQ ID NO: 60]

[0131] CDRL2: DNQQRPS [ SEQ ID NO: 61]

[0132] CDRL3: WDDRLFGPV [ SEQ ID NO: 62]

[0133] Antibody Clone: 1C04

[0134] 1E05-VL [SEQ ID NO: 5]

[0135] EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYAMSWVRQAPGKGLEWVSSISDSGAGRYY ADSVEGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARTHDSGELLDAFDIWGQGTLVTVSS

[0136] CDR regions [SEQ ID NO: 29]

[0137] QSVLTQPPSASGTPGQRVTISCSGSSSNIGSNHVLWYQQLPGTAPKLLIYGNSNRPSGVPDRFSGSKSGTSASLAISGLRSEDEADYYCAAWDDSLNGWVFGGGTKLTVLG

[0138] 2A09-VH

[0139] CDRH1: SYAMS [ SEQ ID NO: 63]

[0140] CDRH2: SISDSGAGRYYADSVEG [ SEQ ID NO: 64]

[0141] CDRH3: THDSGELLDAFDI [ SEQ ID NO: 65]

[0142] CDRL1: SGSSSNIGSNHVL [ SEQ ID NO: 66]

[0143] CDRL2: GNSNRPS [ SEQ ID NO: 67]

[0144] CDRL3: AAWDDSLNGWV [ SEQ ID NO: 68]

[0145] Antibody Clone: 1E05

[0146] 2A09-VL [SEQ ID NO: 6]

[0147] EVQLLESGGGLVQPGGSLRLSCAASGFTFSTYAMNWVRQVPGKGLEWVAVISYDGSNKNYVDSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARNFDNSGYAIPDAFDIWGQGTLVTVSS

[0148] CDR regions [SEQ ID NO: 30]

[0149] QSVLTQPPSASGTPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYDNNSRPSGVPDRFSGSKSGTSASLAISGLRSEDEADYYCAAWDDSLGGPVFGGGTKLTVLG

[0150] 2B08-VH

[0151] CDRH1: TYAMN [ [SEQ ID NO: 69]

[0152] CDRH2: VISYDGSNKNYVDSVKG [ [SEQ ID NO: 70]

[0153] CDRH3: NFDNSGYAIPDAFDI [ [SEQ ID NO: 71]

[0154] CDRL1: TGSSSNIGAGYDVH [ [SEQ ID NO: 72]

[0155] CDRL2: DNNSRPS [ [SEQ ID NO: 73]

[0156] CDRL3: AAWDDSLGGPV [ [SEQ ID NO: 74]

[0157] Antibody Clone: 2A09

[0158] 2B08-VL [SEQ ID NO: 7]

[0159] EVQLLESGGGLVQPGGSLRLSCAASGFTFSNAWMSWVRQAPGKGLEWVAYISRDADITHYPASVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCTTGFDYAGDDAFDIWGQGTLVTVSS

[0160] CDR regions [SEQ ID NO: 31]

[0161] QSVLTQPPSASGTPGQRVTISCSGSSSNIGSNAVNWYQQLPGTAPKLLIYGNSDRPSGVPDRFSGSKSGTSASLAISGLRSEDEADYYCAAWDDSLNGRWVFGGGTKLTVLG

[0162] 2E8-VH

[0163] CDRH1 : NAWMS [ SEQ ID NO: 75]

[0164] CDRH2: YISRDADITHYPASVKG [ SEQ ID NO: 76]

[0165] CDRH3: GFDYAGDDAFDI [ SEQ ID NO: 77]

[0166] CDRL1 : SGSSSNIGSNAVN [ SEQ ID NO: 78]

[0167] CDRL2: GNSDRPS [ SEQ ID NO: 79]

[0168] CDRL3: AAWDDSLNGRWV [ SEQ ID NO: 80]

[0169] Antibody Clone: 2B08

[0170] 2E8-VL [SEQ ID NO: 8]

[0171] EVQLLESGGGLVQPGGSLRLSCAASGFTFSDYYMSWVRQAPGKGLEWVALIGHDGNNKYYLDSLEGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARATDSGYDLLYWGQGTLVTVSS

[0172] CDR regions [SEQ ID NO: 32]

[0173] QSVLTQPPSASGTPGQRVTISCSGSSSNIGNNAVNWYQQLPGTAPKLLIYYDDLLPSGVPDRFSGSKSGTSASLAISGLRSEDEADYYCTTWDDSLSGVVFGGGTKLTVLG

[0174] 5C04-VH

[0175] CDRH1 : DYYMS [ [SEQ ID NO: 81 ]

[0176] CDRH2: LIGHDGNNKYYLDSLEG [ [SEQ ID NO: 82]

[0177] CDRH3: ATDSGYDLLY [ [SEQ ID NO: 83]

[0178] CDRL1 : SGSSSNIGNNAVN [ [SEQ ID NO: 84]

[0179] CDRL2: YDDLLPS [ [SEQ ID NO: 85]

[0180] CDRL3: TTWDDSLSGVV [ [SEQ ID NO: 86]

[0181] Antibody Clone: 2E8-VH

[0182] 5C04-VL [SEQ ID NO: 9]

[0183] EVQLLESGGGLVQPGGSLRLSCAASGFTFSDYYMSWIRQAPGKGLEWVSAIGFSDDNTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAGGDGSGWSFWGQGTLVTVSS

[0184] CDR regions [SEQ ID NO: 33]

[0185] QSVLTQPPSASGTPGQRVTISCSGSSSNIGNNAVNWYQQLPGTAPKLLIYDNNKRPSGVPDRFSGSKSGTSASLAISGLRSEDEADYYCATWDDSLRGWVFGGGTKLTVLG

[0186] 5C05-VH

[0187] CDRH1: DYYMS [ SEQ ID NO: 87]

[0188] CDRH2: AIGFSDDNTYYADSVKG [ SEQ ID NO: 88]

[0189] CDRH3: GDGSGWSF [ SEQ ID NO: 89]

[0190] CDRL1: SGSSSNIGNNAVN [ SEQ ID NO: 90]

[0191] CDRL2: DNNKRPS [ SEQ ID NO: 91]

[0192] CDRL3: ATWDDSLRGWV [ SEQ ID NO: 92]

[0193] Antibody Clone: 5C04

[0194] 5C05-VL [SEQ ID NO: 10]

[0195] EVQLLESGGGLVQPGGSLRLSCAASGFTFSNYGMHWVRQAPGKGLEWVAVISYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAREWRDAFDIWGQGTLVTVSS

[0196] CDR regions [ SEQ ID NO: 34]

[0197] QSVLTQPPSASGTPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYSDNQRPSGVPDRFSGSKSGTSASLAISGLRSEDEADYYCAAWDDSLSGSWVFGGGTKLTVLG

[0198] 5D07-VH

[0199] CDRH1: NYGMH [ SEQ ID NO: 93]

[0200] CDRH2: VISYDGSNKYYADSVKG [ SEQ ID NO: 94]

[0201] CDRH3: WRDAFDI [ SEQ ID NO: 95]

[0202] CDRL1: TGSSSNIGAGYDVH [ SEQ ID NO: 96]

[0203] CDRL2: SDNQRPS [ SEQ ID NO: 97]

[0204] CDRL3: AAWDDSLSGSWV [ SEQ ID NO: 98]

[0205] Antibody Clone: 5C05

[0206] 5D07-VL [SEQ ID NO: 11]

[0207] EVQLLESGGGLVQPGGSLRLSCAASGFTFSTYGMHWVRQAPGKGLEWVAVISYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARENFDAFDVWGQGTLVTVSS

[0208] CDR regions [ SEQ ID NO: 35]

[0209] QSVLTQPPSASGTPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYSNSQRPSGVPDRFSGSKSGTSASLAISGLRSEDEADYYCAAWDDSLNGQVVFGGGTKLTVLG

[0210] 5E12-VH

[0211] CDRH1: TYGMH [ SEQ ID NO: 99]

[0212] CDRH2: VISYDGSNKYYADSVKG [ SEQ ID NO: 100]

[0213] CDRH3: ENFDAFDV [ SEQ ID NO: 101]

[0214] CDRL1: TGSSSNIGAGYDVH [ SEQ ID NO: 102]

[0215] CDRL2: SNSQRPS [ SEQ ID NO: 103]

[0216] CDRL3: AAWDDSLNGQVV [ SEQ ID NO: 104]

[0217] Antibody Clone: 5D07

[0218] 5E12-VL [SEQ ID NO: 12]

[0219] EVQLLESGGGLVQPGGSLRLSCAASGFTFSTYGMHWVRQAPGKGLEWVAVIAYDGSKKDYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAREYRDAFDIWGQGTLVTVSS

[0220] CDR regions [ SEQ ID NO: 36]

[0221] QSVLTQPPSASGTPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYGNSNRPSGVPDRFSGSKSGTTASLAISGLRSEDEADYYCAAWDDSVSGWMFGGGTKLTVLG

[0222] 5G08-VH

[0223] CDRH1: TYGMH [ SEQ ID NO: 105]

[0224] CDRH2: VIAYDGSKKDYADSVKG [ SEQ ID NO: 106]

[0225] CDRH3: EYRDAFDI [ SEQ ID NO: 107]

[0226] CDRL1: TGSSSNIGAGYDVH [ SEQ ID NO: 108]

[0227] CDRL2: GNSNRPS [ SEQ ID NO: 109]

[0228] CDRL3: AAWDDSVSGWM [ SEQ ID NO: 110]

[0229] Antibody Clone: 5E12

[0230] 5G08-VL [SEQ ID NO: 13]

[0231] EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYGMHWVRQAPGKGLEWVAVISYDGINKDYADSMKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARERKDAFDIWGQGTLVTVSS

[0232] CDR regions [ SEQ ID NO: 37]

[0233] QSVLTQPPSASGTPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYSNNQRPSGVPDRFSGSKSGTSASLAISGLRSEDEADYYCATWDDSLNGLVFGGGTKLTVLG

[0234] 5H06-VH

[0235] CDRH1: SYGMH [ SEQ ID NO: 111]

[0236] CDRH2: VISYDGINKDYADSMKG [ SEQ ID NO: 112]

[0237] CDRH3: ERKDAFDI [ SEQ ID NO: 113]

[0238] CDRL1: TGSSSNIGAGYDVH [ SEQ ID NO: 114]

[0239] CDRL2: SNNQRPS [ SEQ ID NO: 115]

[0240] CDRL3: ATWDDSLNGLV [ SEQ ID NO: 116]

[0241] Antibody Clone: 5G08

[0242] 5H06-VL [SEQ ID NO: 14]

[0243] EVQLLESGGGLVQPGGSLRLSCAASGFTFNNYGMHWVRQAPGKGLEWVAVISYDGSNRYYADSVKGRFTMSRDNSKNTLYLQMNSLRAEDTAVYYCARDRWNGMDVWGQGTLVTVSS

[0244] CDR regions [ SEQ ID NO: 38]

[0245] QSVLTQPPSASGTPGQRVTISCSGSSSNIGAGYDVHWYQQLPGTAPKLLIYANNQRPSGVPDRFSGSKSGTSASLAISGLRSEDEADYYCAAWDDSLNGPWVFGGGTKLTVLG

[0246] 6A09-VH

[0247] CDRH1: NYGMH [ SEQ ID NO: 117]

[0248] CDRH2: VISYDGSNRYYADSVKG [ SEQ ID NO: 118]

[0249] CDRH3: DRWNGMDV [ SEQ ID NO: 119]

[0250] CDRL1: SGSSSNIGAGYDVH [ SEQ ID NO: 120]

[0251] CDRL2: ANNQRPS [ SEQ ID NO: 121]

[0252] CDRL3: AAWDDSLNGPWV [ SEQ ID NO: 122]

[0253] Antibody Clone: 5H06

[0254] 6A09-VL [SEQ ID NO: 15]

[0255] EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYGMHWVRQAPGKGLEWVAVISYDGSDTAYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDHSVIGAFDIWGQGTLVTVSS

[0256] CDR regions [ SEQ ID NO: 39]

[0257] QSVLTQPPSASGTPGQRVTISCSGSSSNIGSNTVNWYQQLPGTAPKLLIYDNNKRPSGVPDRFSGSKSGTSASLAISGLRSEDEADYYCSSYAGSNNVVFGGGTKLTVLG

[0258] 6B01-VH

[0259] CDRH1: SYGMH [ SEQ ID NO: 123]

[0260] CDRH2: VISYDGSDTAYADSVKG [ SEQ ID NO: 124]

[0261] CDRH3: DHSVIGAFDI [ SEQ ID NO: 125]

[0262] CDRL1: SGSSSNIGSNTVN [ SEQ ID NO: 126]

[0263] CDRL2: DNNKRPS [ SEQ ID NO: 127]

[0264] CDRL3: SSYAGSNNVV [ SEQ ID NO: 128]

[0265] Antibody Clone: 6A09

[0266] 6B01-VL [SEQ ID NO: 16]

[0267] EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYGMHWVRQAPGKGLEWVAVTSYDGNTKYYANSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAREDCGGDCFDYWGQGTLVTVSS

[0268] CDR regions [ SEQ ID NO: 40]

[0269] QSVLTQPPSASGTPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYGNSNRPSGVPDRFSGSKSGTSASLAISGLRSEDEADYYCAAWDDSLNEGVFGGGTKLTVLG

[0270] 6C11-VH

[0271] CDRH1: SYGMH [ SEQ ID NO: 129]

[0272] CDRH2: VTSYDGNTKYYANSVKG [ SEQ ID NO: 130]

[0273] CDRH3: EDCGGDCFDY [ SEQ ID NO: 131]

[0274] CDRL1: TGSSSNIGAGYDVH [ SEQ ID NO: 132]

[0275] CDRL2: GNSNRPS [ SEQ ID NO: 133]

[0276] CDRL3: AAWDDSLNEGV [ SEQ ID NO: 134]

[0277] Antibody Clone: 6B01

[0278] 6C11-VL [SEQ ID NO: 17]

[0279] EVQLLESGGGLVQPGGSLRLSCAASGFTFSNYGMHWVRQAPGKGLEWVAVISYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDQLGEAFDIWGQGTLVTVSS

[0280] CDR regions [ SEQ ID NO: 41]

[0281] QSVLTQPPSASGTPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYDNNKRPSGVPDRFSGSKSGTSASLAISGLRSEDEADYYCATWDDSLSGPVFGGGTKLTVLG

[0282] 6C12-VH

[0283] CDRH1: NYGMH [ SEQ ID NO: 135]

[0284] CDRH2: VISYDGSNKYYADSVKG [ SEQ ID NO: 136]

[0285] CDRH3: DQLGEAFDI [ SEQ ID NO: 137]

[0286] CDRL1: TGSSSNIGAGYDVH [ SEQ ID NO: 138]

[0287] CDRL2: DNNKRPS [ SEQ ID NO: 139]

[0288] CDRL3: ATWDDSLSGPV [ SEQ ID NO: 140]

[0289] Antibody Clone: 6C11

[0290] 6C12-VL [SEQ ID NO: 18]

[0291] EVQLLESGGGLVQPGGSLRLSCAASGFTFDDYGMSWVRQAPGKGLEWVSAISGSGSSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAGGDIDYFDYWGQGTLsVTVSS

[0292] CDR regions [ SEQ ID NO: 42]

[0293] QSVLTQPPSASGTPGQRVTISCTGSSSNFGAGYDVHWYQQLPGTAPKLLIYENNKRPSGVPDRFSGSKSGTSASLAISGLRSEDEADYYCAAWDDSLNGPVFGGGTKLTVLG

[0294] 6D01-VH

[0295] CDRH1: DYGMS [ SEQ ID NO: 141]

[0296] CDRH2: AISGSGSSTYYADSVKG [ SEQ ID NO: 142]

[0297] CDRH3: GDIDYFDY [ SEQ ID NO: 143]

[0298] CDRL1: TGSSSNFGAGYDVH [ SEQ ID NO: 144]

[0299] CDRL2: ENNKRPS [ SEQ ID NO: 145]

[0300] CDRL3: AAWDDSLNGPV [ SEQ ID NO: 146]

[0301] Antibody Clone: 6C12

[0302] 6D01-VL [SEQ ID NO: 19]

[0303] EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYGMHWVRQAPGKGLEWVAVISYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARERRDAFDIWGQGTLVTVSS

[0304] CDR regions [ SEQ ID NO: 43]

[0305] QSVLTQPPSASGTPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYSDNQRPSGVPDRFSGSKSGTSASLAISGLRSEDEADYYCATWDSDTPVFGGGTKLTVLG

[0306] 6G03-VH

[0307] CDRH1: SYGMH [ SEQ ID NO: 147]

[0308] CDRH2: VISYDGSNKYYADSVKG [ SEQ ID NO: 148]

[0309] CDRH3: ERRDAFDI [ SEQ ID NO: 149]

[0310] CDRL1: TGSSSNIGAGYDVH [ SEQ ID NO: 150]

[0311] CDRL2: SDNQRPS [ SEQ ID NO: 151]

[0312] CDRL3: ATWDSDTPV [ SEQ ID NO: 152]

[0313] Antibody Clone: 6D01

[0314] 6G03-VL [SEQ ID NO: 20]

[0315] EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYGMHWVRQAPGKGLEWVAVISYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAMYYCARDHSAAGYFDYWGQGTLVTVSS

[0316] CDR regions [ SEQ ID NO: 44]

[0317] QSVLTQPPSASGTPGQRVTISCSGSSSNIGSNTVNWYQQLPGTAPKLLIYGNSIRPSGGPDRFSGSKSGTSASLAISGLRSEDEADYYCASWDDSLSSPVFGGGTKLTVLG

[0318] 6G08-VH

[0319] CDRH1: SYGMH [ SEQ ID NO: 153]

[0320] CDRH2: VISYDGSNKYYADSVKG [ SEQ ID NO: 154]

[0321] CDRH3: DHSAAGYFDY [ SEQ ID NO: 155]

[0322] CDRL1: SGSSSNIGSNTVN [ SEQ ID NO: 156]

[0323] CDRL2: GNSIRPS [ SEQ ID NO: 157]

[0324] CDRL3: ASWDDSLSSPV [ SEQ ID NO: 158]

[0325] Antibody Clone: 6G03

[0326] 6G08-VL [SEQ ID NO: 21]

[0327] EVQLLESGGGLVQPGGSLRLSCAASGFTFGSYGMHWVRQAPGKGLEWVSGISWDSAIIDYAGSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKDEAAAGAFDIWGQGTLVTVSS

[0328] CDR regions [ SEQ ID NO: 45]

[0329] QSVLTQPPSASGTPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYGNTDRPSGVPDRFSGSKSGTSASLAISGLRSEDEADYYCAAWDDSLSGPVVFGGGTKLTVLG

[0330] 6G11-VH

[0331] CDRH1: SYGMH [ SEQ ID NO: 159]

[0332] CDRH2: GISWDSAIIDYAGSVKG [ SEQ ID NO: 160]

[0333] CDRH3: DEAAAGAFDI [ SEQ ID NO: 161]

[0334] CDRL1: TGSSSNIGAGYDVH [ SEQ ID NO: 162]

[0335] CDRL2: GNTDRPS [ SEQ ID NO: 163]

[0336] CDRL3: AAWDDSLSGPVV [ SEQ ID NO: 164]

[0337] Antibody Clone: 6G08

[0338] 6G11-VL [SEQ ID NO: 22]

[0339] EVQLLESGGGLVQPGGSLRLSCAASGFTLSSYGISWVRQAPGKGLEWVSGISGSGGNTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCASSVGAYANDAFDIWGQGTLVTVSS

[0340] CDR regions [ SEQ ID NO: 46]

[0341] QSVLTQPPSASGTPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYGDTNRPSGVPDRFSGSKSGTSASLAISGLRSEDEADYYCAAWDDSLNGPVFGGGTKLTVLG

[0342] 6H08-VH

[0343] CDRH1: SYGIS [ SEQ ID NO: 165]

[0344] CDRH2: GISGSGGNTYYADSVKG [ SEQ ID NO: 166]

[0345] CDRH3: SVGAYANDAFDI [ SEQ ID NO: 167]

[0346] CDRL1: TGSSSNIGAGYDVH [ SEQ ID NO: 168]

[0347] CDRL2: GDTNRPS [ SEQ ID NO: 169]

[0348] CDRL3: AAWDDSLNGPV [ SEQ ID NO: 170]

[0349] Antibody Clone: 6G11

[0350] 6H08-VL [SEQ ID NO: 23]

[0351] EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYGMHWVRQAPGKGLEWMAVISYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARELYDAFDIWGQGTLVTVSS

[0352] CDR regions [ SEQ ID NO: 47]

[0353] QSVLTQPPSASGTPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIY ADDHRPSGVPDRFSGSKSGTSASLAISGLRSEDEADYYCASWDDSQRAVIFGGGTKLTVLG

[0354] 7C07-VH

[0355] CDRH1: SYGMH [ SEQ ID NO: 171]

[0356] CDRH2: VISYDGSNKYYADSVKG [ SEQ ID NO: 172]

[0357] CDRH3: ELYDAFDI [ SEQ ID NO: 173]

[0358] CDRL1: TGSSSNIGAGYDVH [ SEQ ID NO: 174]

[0359] CDRL2: ADDHRPS [ SEQ ID NO: 175]

[0360] CDRL3: ASWDDSQRAVI [ SEQ ID NO: 176]

[0361] Antibody Clone: 6H08

[0362] 7C07-VL [SEQ ID NO: 24]

[0363] EVQLLESGGGLVQPGGSLRLSCAASGFTFNNYGMHWVRQAPGKGLEWVAVISYDGSNKYYADSVKGRFTISKDNSKNTLYLQMNSLRAEDTAVYYCAREYKDAFDIWGQGTLVTVSS

[0364] CDR regions [ SEQ ID NO: 48]

[0365] QSVLTQPPSASGTPGQRVTISCTGSSSNIGSNTVNWYQQLPGTAPKLLIYDNNKRPSGVPDRFSGSKSGTSASLAISGLRSEDEADYYCQAWGTGIRVFGGGTKLTVLG

[0366] 4B02-VH

[0367] CDRH1: NYGMH [ SEQ ID NO: 177]

[0368] CDRH2: VISYDGSNKYYADSVKG [ SEQ ID NO: 178]

[0369] CDRH3: EYKDAFDI [ SEQ ID NO: 179]

[0370] CDRL1: TGSSSNIGSNTVN [ SEQ ID NO: 180]

[0371] CDRL2: DNNKRPS [ SEQ ID NO: 181]

[0372] CDRL3: QAWGTGIRV [ SEQ ID NO: 182]

[0373] Antibody Clone: 7C07

[0374] 4B02-VL [SEQ ID NO: 25]

[0375] EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYGMHWVRQAPGKGLEWVAVISYDGSNKYYADSVKGRFTISRDNSQNTLYLQMNSLRAEDTAVYYCAREFGYIILDYWGQGTLVTVSS

[0376] CDR regions [ SEQ ID NO: 49]

[0377] QSVLTQPPSASGTPGQRVTISCSGSSSNIGSNTVNWYQQLPGTAPKLLIYRDYERPSGVPDRFSGSKSGTSASLAISGLRSEDEADYYCMAWDDSLSGVVFGGGTKLTVLG

[0378] Figure 1

[0379] CDRH1: SYGMH [ SEQ ID NO: 183]

[0380] CDRH2: VISYDGSNKYYADSVKG [ SEQ ID NO: 184]

[0381] CDRH3: EFGYIILDY [ SEQ ID NO: 185]

[0382] CDRL1: SGSSSNIGSNTVN [ SEQ ID NO: 186]

[0383] CDRL2: RDYERPS [ SEQ ID NO: 187]

[0384] CDRL3: MAWDDSLSGVV [ SEQ ID NO: 188]

[0385] Antibody Clone: 4B02

[0386] Figure 1 [SEQ ID NO: 26]

[0387] EVQLLESGGGLVQPGGSLRLSCAASGFTFSNHGMHWVRQAPGKGLEWVAVISYDGTNKYYADSVRGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARETWDAFDVWGQGTLVTVSS

[0388] Figure 1 [ SEQ ID NO: 50]

[0389] QSVLTQPPSASGTPGQRVTISCSGSSSNIGSNNANWYQQLPGTAPKLLIYDNNKRPSGVPDRFSGSKSGTSASLAISGLRSEDEADYYCQAWDSSTVVFGGGTKLTVLG

[0390] Figure 1

[0391] CDRH1: NHGMH [ SEQ ID NO: 189]

[0392] CDRH2: VISYDGTNKYYADSVRG [ SEQ ID NO: 190]

[0393] CDRH3: ETWDAFDV [ SEQ ID NO: 191]

[0394] CDRL1: SGSSSNIGSNNAN [ SEQ ID NO: 192]

[0395] CDRL2: DNNKRPS [ SEQ ID NO: 193]

[0396] CDRL3: QAWDSSTVV [ SEQ ID NO: 194]

[0397] In some embodiments, sometimes preferred embodiments, the antibody molecule that specifically binds FcyRIIb comprises 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.

[0398] In some embodiments, sometimes preferred embodiments, the antibody molecule that specifically binds FcyRIIb 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.

[0399] In some embodiments, the anti-PD-1 antibody molecule is a human antibody molecule or an antibody molecule of human origin. In some such embodiments, the human antibody molecule or antibody molecule of human origin is an IgG antibody. In some such embodiments, the human antibody molecule or antibody molecule of human origin is an IgG4.

[0400] The anti-PD-1 antibody molecule is an antibody molecule that specifically binds to PD-1.

[0401] In some embodiments, the anti-PD-1 antibody molecule blocks the binding of PD-L1 and / or PD-L2 to PD-1, and can thus be considered a PD-1 antagonist.

[0402] In some embodiments, the anti-PD-1 antibody molecule is a humanized antibody molecule.

[0403] In some embodiments, the anti-PD-1 antibody molecule is a chimeric antibody.

[0404] As mentioned above, the anti-PD-1 antibody must have the ability to engage FcyR.

[0405] In some embodiments, the anti-PD-1 antibody molecule is selected from the group consisting of nivolumab (OPDIVO®), pembrolizumab (KEYTRUDA®), and cemiplimab (LIBTAYO®).

[0406] In some embodiments, the antibody molecule that specifically binds FcyRIIb and the anti-PD-1 antibody molecule are administered to the patient simultaneously, meaning that the two antibody molecules are administered together at one point in time or are administered separately in time very close to each other.

[0407] In some embodiments, the antibody molecule that specifically binds FcyRIIb is administered to the patient prior to the anti-PD-1 antibody molecule. This sequential administration can be achieved by temporarily separating the two antibodies. Alternatively, or in combination with the first option, the sequential administration can also be achieved by spatially separating the two antibody molecules; administering the antibody molecule that specifically binds FcyRIIb in a manner such as intratumorally, so that it reaches the cancer before the anti-PD-1 antibody molecule, and then administering the anti-PD-1 antibody molecule in a manner such as systemically, so that it reaches the cancer after the antibody molecule that specifically binds FcyRIIb.

[0408] In some embodiments, the patient is administered an anti-PD-1 antibody molecule prior to administration of the antibody molecule that specifically binds FcyRIIb. This sequential administration can be achieved by temporally separating the two antibodies and / or by spatially separating the two antibody molecules, similar to the above. For spatial administration, the anti-PD-1 antibody molecule is administered in a manner (such as intratumorally) so that it reaches the cancer prior to the antibody molecule that specifically binds FcyRIIb, which is then administered in a manner (such as systemically) so that it reaches the cancer after the anti-PD-1 antibody molecule.

[0409] Those skilled in the medical arts will appreciate that drugs can be modified with different additives, for example to alter the rate at which the body absorbs the drug; and drugs can be modified in different forms, for example to allow a particular route of administration to the body.

[0410] Thus, the compositions and / or antibodies and / or medicaments of the application can be combined with excipients and / or pharmaceutically acceptable carriers and / or pharmaceutically acceptable diluents and / or adjuvants.

[0411] Also included are compositions and / or antibodies and / or medicaments of the application that are suitable for parenteral administration, including aqueous and / or non-aqueous sterile injection solutions that can contain anti-oxidants 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 can include suspending agents and / or thickening agents. The compositions and / or antibodies and / or medicaments and / or medicaments of the application can be presented in unit-dose or multi-dose containers, for example sealed ampoules and vials, and can be stored in a freeze-dried (i.e., lyophilized) condition, requiring only the addition of the sterile liquid carrier, for example water for injections, immediately prior to use.

[0412] Extemporaneous injection solutions and suspensions can be prepared from sterile powders and / or granules and / or tablets of the type described above.

[0413] For parenteral administration to human patients, the daily dosage level of the antibody molecule that specifically binds FcyRIIb and / or the anti-PD-1 antibody molecule administered in single or divided doses will generally be 1 mg / kg to 20 mg / kg of patient body weight, or in some cases even up to 100 mg / kg. In special cases, for example in conjunction with long-term administration, lower dosages can be used. In any event, the physician in charge will determine the actual dosage that will be most appropriate for any individual patient, and this will vary with the age, weight and response of the particular patient. The above dosages are exemplary of the average case. There can, of course, be individual instances where higher or lower dosages are merited, and such are within the scope of the application.

[0414] Generally, the compositions and / or medicaments of the present application will contain an antibody molecule that specifically binds FcyRIIb and / or an anti-PD-1 antibody in a concentration of between about 2 mg / ml and 150 mg / ml or between about 2 mg / ml and 200 mg / ml. In a preferred embodiment, the medicaments and / or compositions of the present application will contain an antibody molecule that specifically binds FcyRIIb and / or an anti-PD-1 antibody molecule in a concentration of 10 mg / ml.

[0415] Generally, oral or parenteral administration of the compositions and / or antibodies and / or agents and / or medicaments of the present application are the most convenient preferred routes in humans. For veterinary uses, the compositions and / or antibodies and / or agents and / or medicaments of the present application are administered as appropriate acceptable formulations in accordance with normal veterinary practice, and the veterinarian will determine the most appropriate dosage regimen and route of administration for a particular animal. Thus, the present application provides a pharmaceutical formulation comprising an amount of an antibody and / or agent of the present application effective to treat various disorders (as described above and further below). Preferably, the compositions and / or antibodies and / or agents and / or medicaments are suitable for delivery by a route selected from the group comprising: intravenous (IV); subcutaneous (SC); intramuscular (IM) or intratumoral.

[0416] The present application also includes compositions and / or antibodies and / or agents and / or medicaments comprising a pharmaceutically acceptable acid addition salt or base addition salt of a polypeptide binding moiety of the present application. The acids used to prepare the pharmaceutically acceptable acid addition salts of the above-mentioned base compounds useful in the present application are those which 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., l,l'-methylene-bis-(2-hydroxy-3-naphthoate)] salts, and the like. Pharmaceutically acceptable base salts can also be used to provide the agents according to the present application in pharmaceutically acceptable salt form. The chemical bases which can be used as reagents to prepare the pharmaceutically acceptable base salts of the agents of the present application which are acidic in nature are those which form non-toxic base salts with such compounds. Such non-toxic base salts include, but are not limited to, those derived from such 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 other base salts of non-toxic organic amines, and the like. Agents and / or polypeptide binding moieties of the present application can be lyophilized for storage and reconstituted in a suitable carrier prior to use. Any suitable lyophilization method (e.g., spray drying, cake drying) and / or reconstitution technique can be employed. Those of skill in the art will appreciate that lyophilization and reconstitution can result in varying degrees of loss of antibody activity (e.g., IgM antibodies tend to have greater loss of activity than IgG antibodies for conventional immunoglobulins), and use levels can have to be adjusted upward to compensate. In one embodiment, a lyophilized (freeze-dried) polypeptide binding moiety loses no more than about 20%, or no more than about 25%, or no more than about 30%, or no more than about 35%, or no more than about 40%, or no more than about 45%, or no more than about 50% of its activity (prior to lyophilization) when rehydrated.

[0417] The combination of an antibody molecule that specifically binds FcyRIIb and an anti-PD-1 antibody molecule can be used to treat cancer.

[0418] The term "patient" as used herein refers to an animal, including a human, that has been diagnosed with an FcyRIIb-negative cancer or diagnosed with a cancer that is believed to be likely an FcyRIIb-negative cancer and / or exhibits symptoms of such a cancer.

[0419] It is included that the patient can be a mammal or a non-mammal. Preferably, the patient is a human or a mammal, such as a horse, or a cow, or a sheep, or a pig, or a camel, or a dog or a cat. Most preferably, the mammal patient is a human.

[0420] "Manifestation" includes that the subject exhibits a cancer symptom and / or a cancer diagnostic marker, and / or that the cancer symptom and / or cancer diagnostic marker can be measured and / or assessed and / or quantified.

[0421] It will be readily understood by those skilled in the art that what the cancer symptoms and cancer diagnostic markers will be and how the severity of the cancer symptoms and / or cancer diagnostic markers are reduced or increased, or the cancer diagnostic markers are reduced or increased, and how those cancer symptoms and / or cancer diagnostic markers can be used to form a prognosis of the cancer.

[0422] Cancer treatment is typically administered in a treatment regimen, that is, the therapeutic agent is administered over a period of time. The length of time of the treatment regimen will depend on many factors, which can include the type of therapeutic agent administered, the type of cancer being treated, the severity of the cancer being treated, and the age and health of the patient, among other reasons.

[0423] "During treatment" includes that the patient is currently undergoing a treatment regimen and / or is receiving a therapeutic agent, and / or is receiving a series of therapeutic agents.

[0424] The patient to be treated according to the present application has a cancer characterized by a PD-1 positive tumor.

[0425] In some embodiments, the cancer to be treated is a solid cancer.

[0426] In some embodiments, the solid cancer to be treated is a cancer whose treatment typically consists of or includes immunotherapy using an anti-PD-1 antibody.

[0427] 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, 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 cancer that is high-grade instability (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 amenable to curative surgery or definitive radiation. Those skilled in the art will be aware that the number and type of indications associated with anti-PD-1 immunotherapy is rapidly expanding.

[0428] In some embodiments, the cancer is a refractory cancer. In some such embodiments, the refractory cancer is a cancer that is found to be resistant to treatment with an anti-PD-1 antibody at the start of treatment. Such resistance can be evidenced by the patient having no response at all to the treatment or some progression of the cancer despite the treatment. In some embodiments, the 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 the treatment or shows a reduced response to the treatment. In some embodiments, the refractory cancer is resistant after or at the last stage of successful treatment with an anti-PD-1 antibody, meaning that the anti-PD-1 antibody will have no or reduced effect if the cancer recurs.

[0429] Each of the above cancers is well known and the symptoms and cancer diagnostic markers are well described as are the therapeutic agents used to treat those cancers. Thus, the symptoms, cancer diagnostic markers and therapeutic agents used to treat the above-mentioned cancer types will be known to those skilled in the medical arts.

[0430] The clinical definition of diagnosis, prognosis and progression of a large number of cancers relies on certain classifications known as staging. Those staging systems are used to collate a number of different cancer diagnostic markers and cancer symptoms to provide an overview of the diagnosis and / or prognosis and / or progression of the cancer. Those skilled in oncology will understand how to use the staging systems to assess the diagnosis and / or prognosis and / or progression of a cancer and which cancer diagnostic markers and cancer symptoms should be used for the assessment.

[0431] "Cancer staging" includes: Rai staging, which includes Stage 0, Stage I, Stage II, Stage III, and Stage IV; and / or Binet staging, which includes Stage A, Stage B, and Stage C; and / or Ann Arbor staging, which includes Stage I, Stage II, Stage III, and Stage IV.

[0432] It is known that cancer can cause abnormalities in cell morphology. These abnormalities often reproducibly occur in certain cancers, which means that examining these changes in morphology (otherwise known as histological examination) can be used in the diagnosis or prognosis of cancer. Techniques for visualising samples to examine cell morphology and for preparing samples for visualisation are well known in the art; for example, light microscopy or confocal microscopy.

[0433] "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 narrow cytoplasmic borders and dense nuclei lacking discernible nucleoli; and / or the presence of atypical cells and / or fragmented cells and / or prolymphocytes.

[0434] It is well known that cancer is the result of mutations in cell DNA, which can cause the cell to avoid cell death or to proliferate uncontrollably. Therefore, examining 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 location 13q14.1, which is characteristic of chronic lymphocytic leukaemia. Techniques for examining mutations in cells are well known in the art; for example, fluorescence in situ hybridisation (FISH).

[0435] "Cytogenetic examination" includes examination of cells and specifically DNA in chromosomes. Cytogenetic examination can be used to identify changes in DNA that can be associated with the presence of refractory cancer and / or recurrent cancer. Such changes can include: deletion in the long arm of chromosome 13; and / or deletion of chromosome location 13ql4.1; and / or trisomy of chromosome 12; and / or deletion in the long arm of chromosome 12; and / or deletion in the long arm of chromosome 11; and / or 11q deletion; and / or deletion in the long arm of chromosome 6; and / or 6q deletion; and / or deletion in the short arm of chromosome 17; and / or 17p deletion; and / or t(11:14) translocation; and / or (q13:q32) translocation; and / or antigen gene receptor rearrangement; and / or BCL2 rearrangement; and / or BCL6 rearrangement; and / or t(14:18) translocation; and / or t(11:14) translocation; and / or (q13:q32) translocation; and / or (3:v) translocation; and / or (8:14) translocation; and / or (8:v) translocation; and / or t(11:14) and (q13:q32) translocation.

[0436] It is known that patients with cancer exhibit certain physical symptoms, which are typically caused by the burden of the cancer on the body. Those symptoms are typically recurring in the same cancer and can therefore be characteristic of the diagnosis and / or prognosis and / or progression of the disease. The skilled person in the medical field will understand which physical symptoms are associated with which cancer and how those physical systems are relevant to the diagnosis and / or prognosis and / or progression of the disease. "Physical symptoms" include hepatomegaly and / or splenomegaly. BRIEF DESCRIPTION OF DRAWINGS

[0437] In the examples hereinafter reference is made to the following drawings:

[0438] Figure 2 PD-1 expression on human Jurkat T cells transfected with PD-1 is shown. PD-1 transfected Jurkat cells were sorted into low, intermediate and high PD-1 expressing cells. After expansion, PD-1 expression was quantified in the three different subpopulations and the number of PD-1 molecules / cell is shown in Figure 2 A (low), Figure 2 B (intermediate) and Figure 2 C (high).

[0439] Figure 2 BI-1206 (6G11 WT) is shown to inhibit PD-1 mediated phagocytosis in intermediate and high expressing cells but not in low expressing cells. Figure 3A shows an example of phagocytosed Jurkat cells. FL4 on the y-axis depicts CD14+ macrophages and FL1 on the x-axis depicts CFSE-labeled Jurkat cells. Therefore, the circled upper right quadrant shows double-positive CD14+ CFSE+ cells, which are phagocytosed Jurkat cells. This example illustrates the phagocytic activity of PD-1-highly expressed Jurkat cells. Figure 3 B shows the phagocytic activity of Jurkat cells expressing PD-1 and Figure 3 C shows Jurkat cells with high expression. The values ​​are normalized for allotype opsonization (set to 0%) and anti-CD3 opsonization (OKT3 hIgG1, set to 100%). This figure shows that BI-1206 (denoted as 6G11 WT in the figure) inhibited nivolumab-mediated phagocytosis at all tested concentrations. Furthermore, this figure shows that the 6G11 antibody requires the intact Fc moiety to inhibit phagocytosis because disruption of FcγR binding caused by inducing a mutation at position 297 from amino acid asparagine (N) to amino acid glutamine (Q) (i.e., the antibody is denoted as 6G11NQ here) reduces its ability to inhibit nivolumab-mediated phagocytosis. This figure shows two experiments for moderately expressing cells and three experiments for highly expressing cells. Figure 3 D indicates that nivolumab-mediated phagocytosis is absent in cells with low expression.

[0440] Figure 3 This demonstrates that Fc:FcγR binds well to anti-FcγRIIB (AT-130-2 mIgG2a and mIgG1), rather than Fc:FcγR binding poorly to anti-FcγRIIB (AT-130-2 mIgG1 NA), enhancing the therapeutic efficacy and survival rate of anti-PD-1 antibodies in vivo. CT26 (Figures A and B) or MC38 ( Figure 3 C and Figure 3 D) Tumor-bearing mice were treated three times with 200 μg of anti-PD-1 (clone 29F.1A12; Bioxcell) antibody, either alone or in combination with 200 μg of the indicated anti-FcγRIIB antibody variant or isotype control (WR17) (subcutaneously injected in 100 μl PBS at a dose of 5 × 10⁻⁶). 5 (Days 8, 12, and 15 after tumor cell count). For the first treatment, AT130-2 was administered 6 hours before the anti-PD1 antibody. For subsequent treatments, both antibodies were administered together. All injections were intraperitoneal, in 200 μl PBS. When the tumor reached 400 mm... 2 (For CT26) or 225 mm 2 (For MC38) the area is considered terminal. The graph shows the tumor growth in the animal (Figure 4 A and Figure 5 C) and survival rate Figure 6 B and Figure 7 D). ( P < 0.01 (log-rank test). The experiment was conducted in female mice aged 8–14 weeks.

[0441] Figure 8 This study illustrates PD-1 expression on immune cells from tumor-bearing mice. PD-1 expression was quantified on immune cells derived from mouse tumors. Mice were injected with MC38 cells, and tumors were collected ~20 days later. Cells were stained against different T cell subsets, and PD-1 expression on CD8+ T cells was analyzed by FACS. The mean fluorescence intensity of PD-1 on cells was compared with that from Quantum™ Simply Cellular cells stained with the same anti-PD-1 antibody. ® The value of the bead is correlated to determine the number of receptors per cell.

[0442] Figure 9 Jurkat cells expressing different levels of PD-1 are shown: low PD-1, intermediate PD-1, and high PD-1. PD-1 expression was defined using a saturation concentration of Alexa Fluor 647 human anti-human PD-1 (pembrolizumab).

[0443] Figure 8 This shows PD-1 expression on cells in “Jurkat PD-1”. The gates show the full width / half height gates used to define “medium-high” PD-1 expression on tumor samples.

[0444] Example The gating strategy for defining PD-1 expression on human tumor samples is demonstrated. First, CD45+ events are defined (A), then live cells (B), followed by CD3+ (C) or CD3+CD8+ (D). PD-1 high gates are set in the CD3+ (E) and CD3+CD8+ populations (F), respectively. The PD-1 "high" gate is defined based on PD-1 transfected Jurkat cells, and the lower end is set according to the lower end of the full-width / half-height gate in Jurkat cells with PD-1. (G) and (H) show the FMO of Alexa Fluor 647 human anti-human PD-1 (pembrolizumab) in the CD3+ and CD3+CD8+ populations, respectively.

[0445] Figure 1 A table summarizing data for each patient from whom tumor samples were obtained is shown, including patient characteristics, including PD-1 expression and predicted response.

[0446] Figure 1 The percentage of CD3+ and CD3+CD8+ lymphocytes with medium-high expression of PD-1 is shown. The dotted line defines 10%. The letters (F, G, H, etc.) correspond to Figure 2 Patient ID in table.

[0447] Figure 1

[0448] Specific non-limiting examples embodying certain aspects of the application will now be described. These examples should be read in conjunction with the description of the figures provided hereinabove.

[0449] Example 1

[0450] Transfection of Jurkat cells

[0451] For transfection of Jurkat cells, cells were cultured in RPMI-1640 medium containing 10% fetal calf serum, FCS (Sigma), L-glutamine (Life Technologies), sodium pyruvate (Life Technologies) and Pen-Strep (Life Technologies). One day before transfection, cells were split to 0.5 x 10 6 cells / ml and cultured overnight. To transfect the cells, 1 x 10 6 cells were centrifuged at 90 x G for 10 min and then resuspended in 100 μΙ of nucleofector solution (Amaxa ® Cell Line Nucleofector® Kit V, Lonza) to which 2 μg of DNA (hPD-1 in pcDNA3) was added. The mixture was then transferred to a nucleofector cuvette. The cuvette was placed in a nucleofector II machine and nucleofection was performed with program X-005. After incubation for 10 min at room temperature (approximately 18-22°C), 500 μΙ of medium was added to the cuvette and transferred to a 12-well plate containing 1 ml of medium. To select the transfected cells, geneticin was added at 1 mg / ml 48 hours after transfection. After 10-14 days, positive cells were purified as low, medium and high PD-1 expressing cells by FACS sorting on a FACSAria II machine. Then, the transfected cells were maintained in medium containing 1 mg / ml geneticin.

[0452] Quantification of PD-1

[0453] The basic principle behind using these beads for quantification is based on the fact that phycoerythrin (PE) labels antibodies at a 1:1 ratio. Therefore, by generating a standard curve using beads with a defined number of PE molecules, the number of antibody molecules binding to cells can be determined.

[0454] Jurkat cells were stained with PE-labeled anti-PD1 antibody (EH12.2H7, Baijin Biotechnology) or an allotype control in FACS buffer (PBS containing 2% FCS) at 4°C for 30 minutes, and then washed in FACS buffer. One tube of Quantibrite™ beads (PE phycoerythrin quantitative kit, BD Bioscience (catalog number 340495)) was resuspended in 500 µl of PBS.

[0455] Next, the FAC machine was set up in a manner that allowed Quantibrite™ beads and Jurkat cells to be run in the same environment. The beads were run until 10,000 events were collected, and then a standard curve was generated according to BD Biosciences' description for the PE phycoerythrin quantitative PCR kit, i.e., by using four Quantibrite samples provided. TM The LogMFI (fluorescence intensity) of the bead cluster was plotted against the Log of the number of molecules per bead (batch-specific information in the kit). The number of molecules on the cell line was then calculated using this standard curve by converting its MFI to a molecular count. Given that the antibody was used at a saturation concentration and that each cell binds to PD-1 molecules 1:1, the number of antibodies bound per cell corresponds to the number of PD-1 molecules present in each cell.

[0456] Then, the PD1-PE stained Jurkat cells were run on FACS and the number of bound antibodies was calculated using the log mean fluorescence intensity (MFI) of the sample of interest.

[0457] The results are shown in Figure 1 middle.

[0458] from Figure 1 It is clear that the cell population with low expression includes some cells with moderate expression (approximately 2%); however, that is a negligible portion of the population, as clearly seen in the phagocytosis experiments described below (and in...). Figure 2 (As demonstrated in the previous text), this small subset does not affect phagocytosis. Similarly, it can be clearly seen that the cell population with moderate expression includes some cells with low expression, but again, this small subset of cells does not affect the phagocytic outcome, as shown below.

[0459] Low expression subgroup ( Figure 4A) with a mean of 3,249 PD-1 molecules / cell with a bottom 5% cutoff of 1,253 PD-1 molecules / cell and a top 5% cutoff of 10,643 PD-1 molecules / cell. Intermediate expression subpopulation Figure 1 B) with a mean of 32,951 PD-1 molecules / cell with a bottom 5% cutoff of 15,498 PD-1 molecules / cell and a top 5% cutoff of 77,822 PD-1 molecules / cell. High expression subpopulation Figure 2 C) with a mean of 165,968 PD-1 molecules / cell with a bottom 5% cutoff of 65,406 PD-1 molecules / cell and a top 5% cutoff of 390,946 PD-1 molecules / cell. The top and bottom 5% cutoffs are provided to reduce overlap between the different subpopulations. The bottom 5% cutoff of 15,498, or approximately 15,500 PD-1 molecules / cell as measured above, is used herein to define the lower limit of intermediate or high expression.

[0460] phagocytosis

[0461] Human PBMC isolated from leukocyte cones obtained from the National Blood Service in Southampton, UK were incubated in plates with RPMI media (Life Technologies) containing glutamine, pyruvate, PenStrep and 1% heat inactivated human serum from Sigma. Incubated for 2 hours to allow monocytes to adhere. Media was then replaced with RPMI media containing glutamine, pyruvate, PenStrep and +10% FCS (Sigma). MCSF (prepared at the University of Southampton) was added after 24 hours. Macrophages were derived over 7 days with 2 media changes (including MCSF). Thereafter, macrophages were harvested by removing media, adding 2ml PBS and placing on ice for 15 minutes, then gently scraped off. Macrophages were then replated in 96 well plates for 2 hours. Macrophages were pre-treated with anti-hFcyRIIb mAb (6G11 WT or 6G11 NQ) at 2x final concentration for 45 minutes, then CFSE (Molecular Probes) labelled Jurkat cells opsonised with Nivolumab (hlgG4) at 2x final concentration for 15 minutes were added. Cells were co-cultured for 1 hour at 37°C, then stained with anti-CD14 (BD Biosciences) by incubating in FACS buffer for 30 min at 4°C, then washed and then read in a FACS machine.

[0462] Results are shown in Figure 3 ​

[0463] Example 2 - Quantification of PD-1 on immune cells from tumor-bearing mice

[0464] Quantum™ Simply Cellular ® beads (Bangs Laboratories, Inc.) were used to determine the number of PD-1 receptors on immune cells from mouse tumors. Briefly, beads were stained with rat anti-PD-1 antibody (clone 29F.1A12, Biolegend) to create a standard curve. Cell samples were then read against the curve to determine expression.

[0465] Cells from tumor-bearing mice were quantified. Mice were housed and maintained in a local facility in accordance with home office guidelines. Female C57 / BL6 mice, six to eight weeks of age, 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% FCS. When cells were semi-confluent, they were detached with trypsin and resuspended in sterile PBS at 10 x 10 6 cells / ml. Mice were injected subcutaneously (s.c.) with 100 μΐ of cell suspension, which corresponds to 1 x 10 6 cells / mouse. Tumors were allowed to grow for ~20 days, then collected. 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 commercial rat anti-PD-1 antibody (clone 29F.1A12) with corresponding isotype control (Biolegend). Results are shown in Figure 8

[0466] Example 3 - In vivo combination effect with anti-PD-1

[0467] PD-1 expression on mouse cells corresponds to the expression level between "medium and high" on transfected Jurkat cells (Example 1, Figure 5 ). In phagocytosis assays, BI-1206 was shown to significantly reduce the level of phagocytosis of these "medium and high" PD-1 expressing cells (Example 1, Figure 7 ). This data is combined with the improved therapeutic anti-tumor effect observed when combining anti-PD-1 with anti-FcyRIIb (BI-1206 mouse surrogate) in vivo in the MC38 model Figure 6 ​), indicating an improved therapeutic effect of the combination of anti-PD-1 and BI-1206 in patients with medium or high PD-1 expression (i.e. PD-1 expression equal to or higher than 15,500 PD-1 molecules / cell).

[0468] Example 4 - Quantification of PD-1 expression on human T cells

[0469] Frozen tumor samples (see Table in Figure 9 ) were purchased from Discovery Life Sciences. Cells were thawed and washed in phosphate buffered saline (PBS) and then stained with a cocktail of the following antibodies: 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), Alexa Fluor 647 human anti-human PD-1 (pembrolizumab (KEYTRUDA), clinical grade, lot 8SNL80406, Merck Sharp & Dohme Limited). Fixable Viability Dye eFluor 780 was also included in the antibody staining cocktail (Invitrogen, 65-0865-14). Staining was performed in BD Horizon Brilliant Staining Buffer (BD 563794). Anti-human PD-1 was conjugated to Alexa Fluor 647 in-house and used at the receptor saturating concentration (5.5 µg / ml) as indicated by pre-titration experiments. The remaining antibodies were used at the manufacturer’s recommended concentration. Cells were incubated with antibodies for 20 minutes, followed by washing and resuspension in PBS, before acquisition using a BD FACSAria II. Analysis was performed using FlowJo software. PD-1 expression analysis on PD-1 transfected Jurkat cells was performed in a similar manner, but only including Alexa Fluor 647 human anti-human PD-1 and Fixable Viability Dye eFluor 780 in the staining cocktail. Results are shown in ​ In tumor samples, PD-1 expression was defined in the CD3+and CD3+CD8+populations, respectively, pre-gated on live CD45+cells ( ​ ). The PD-1 high gate was defined based on PD-1 transfected Jurkat cells and set at the lower end of the full-width / half-height gate for PD-1 on Jurkat cells (lower end of ​ ).

[0470] ​ The percentage of CD3+ lymphocytes and CD3+CD8+ lymphocytes with medium-high expression of PD-1 in each tumor sample obtained from different patients is shown. It is expected that patients with 10% of T cells expressing PD-1 at medium or high levels will benefit from the combination of anti-FcyRIIb with anti-PD-1, and therefore a dotted line at 10% expression is included.

Claims

1. The use of a combination of a first antibody molecule and a second antibody molecule in the manufacture of a drug for treating cancer in patients with tumor-infiltrating T lymphocytes exhibiting intermediate or high PD-1 expression: (i) wherein the first antibody molecule specifically binds to FcγRIIb via its Fab region and to the Fcγ receptor via its Fc region, and (ii) wherein the second antibody molecule specifically binds to PD-1 and binds to at least one Fcγ receptor through its Fc region.

2. The use according to claim 1, wherein the patient’s tumor-infiltrating CD3-positive T lymphocytes have moderate or high PD-1 expression.

3. The use according to claim 2, wherein at least 10% of the tumor-infiltrating CD3-positive T lymphocytes of the patient have moderate or high PD-1 expression.

4. The use according to claim 2 or 3, wherein the patient’s tumor-infiltrating CD3-positive, CD8-positive T lymphocytes have moderate or high PD-1 expression.

5. The use according to claim 4, wherein at least 10% of the tumor-infiltrating CD3-positive, CD8-positive T lymphocytes of the patient have moderate or high PD-1 expression.

6. The use according to any one of claims 1-5, wherein intermediate or high PD-1 expression is defined as at least 10% of the tumor-infiltrating T lymphocytes in the sample from the patient having expression of at least 15,500 PD-1 molecules per T lymphocyte.

7. The use according to claim 6, wherein the anti-PD1 antibody EH12.2H7 is used to measure intermediate or high PD-1 expression.

8. The use according to any one of claims 1-7, wherein the cancer is a solid cancer.

9. The use according to claim 8, wherein the solid cancer is selected from the group consisting of: melanoma, lung cancer, head and neck cancer, Hodgkin lymphoma, primary mediastinal B-cell lymphoma (PMBCL), bladder cancer, colorectal cancer, gastric cancer, cervical cancer, liver cancer, Merkel cell carcinoma, kidney cancer, and squamous cell carcinoma of the skin.

10. The use according to claim 8 or 9, wherein the cancer is refractory.

11. The use according to any one of claims 1-10, wherein the first antibody molecule and / or the second antibody molecule is selected from the group consisting of human antibody molecules, humanized antibody molecules and human-derived antibody molecules.

12. The use according to any one of claims 1-11, wherein the first antibody molecule and / or the second antibody molecule is a monoclonal antibody molecule or an antibody molecule of monoclonal origin.

13. The use according to any one of claims 1-12, 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 Fc receptors through their Fc regions.

14. The use according to any one of claims 1-13, wherein the first antibody molecule and / or the second antibody molecule is a human IgG antibody, a humanized IgG antibody molecule, or a human-derived IgG antibody molecule.

15. The use according to claim 14, wherein the first antibody molecule is an IgG1 antibody molecule.

16. The use according to claim 14 or 15, wherein the second antibody molecule is an IgG4 antibody molecule.

17. The use according to any one of claims 1-16, wherein the first antibody molecule and / or the second antibody molecule has been engineered to improve binding to the activated Fcγ receptor.

18. The use according to any one of claims 1-17, wherein the first antibody molecule comprises a variable heavy chain (VH), the variable heavy chain 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 and 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 and 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 and 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 and 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)SEQ ID NO: 189 and SEQ ID NO: 190 and SEQ ID NO:

191.

19. The use according to any one of claims 1-18, wherein the first antibody molecule comprises a variable light chain (VL), the variable light chain 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 and 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 and 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)SEQ ID NO: 192 and SEQ ID NO: 193 and SEQ ID NO:

194.

20. The use according to any one of claims 1-10, wherein the first antibody molecule comprises a variable heavy chain (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.

21. The use according to any one of claims 1-20, wherein the first antibody molecule comprises a variable light chain (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.

22. The use according to any one of claims 1-21, wherein the first antibody molecule comprises the following CDR amino acid sequence: (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)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.

23. The use according to any one of claims 1-22, wherein the first antibody molecule comprises 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) SEQ ID NO: 26 and SEQ ID NO:

50.

24. The use according to any one of claims 1-17, wherein the first antibody molecule is an antibody molecule capable of competitively binding FcyRIIb with an antibody molecule as defined in any one of claims 18-23.

25. The uses of the following items: (i) The first antibody molecule, which specifically binds to FcγRIIb via its Fab region and to the Fcγ receptor via its Fc region, and (ii) A second antibody molecule that specifically binds to PD-1 and binds to at least one Fcγ receptor through its Fc region. It is used to manufacture drugs for treating cancer in patients with tumor-infiltrating T lymphocytes that have intermediate or high PD-1 expression.

26. A diagnostic test for determining whether a patient will benefit from a combination of treatments including: (i) The first antibody molecule, which specifically binds to FcγRIIb via its Fab region and to the Fcγ receptor via its Fc region, and (ii) A second antibody molecule that specifically binds to PD-1 and binds to at least one Fcγ receptor through its Fc region. The test includes determining PD-1 expression on the patient's tumor-infiltrating T lymphocytes, where moderate or high PD-1 expression indicates that the patient will benefit from combination therapy.

27. A method for transfecting Jurkat cells, comprising the following steps: Jurkat cells were cultured in RPMI-1640 medium containing 10% fetal bovine serum, L-glutamine, sodium pyruvate, and Pen-Strep. The day before transfection, the cells were divided into 0.5 × 10⁶ cells. 6 1 / ml and incubate overnight; To transfect cells, use 1 × 10 6 The cells were centrifuged at 90×G for 10 minutes and then resuspended in 100 µl of nuclear transfection solution with 2 µg DNA (hPD-1 in pcDNA3) added. The mixture was then transferred to a small tank in the nuclear transfection apparatus; Place the small trough in the nuclear transfection instrument II and perform nuclear transfection using program X-005; After incubating at room temperature for 10 min, add 500 ml of culture medium to the small trough and transfer it to a 12-well plate containing 1 ml of culture medium; To select transfected cells, genimycin was added at 1 mg / ml 48 hours post-transfection; After 10-14 days, positive cells were purified into low, medium, and high PD-1 expression cells using FACS sorting on a FACSAria II machine; and The transfected cells were maintained in a culture medium containing 1 mg / ml of genimycin.

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