Anti-MS4A4A antibodies and methods of use thereof

Abstract

The present disclosure generally relates to compositions including antibodies, such as monoclonal antibodies, humanized antibodies, and antibody fragments, that specifically bind to MS4A4A polypeptides, such as mammalian MS4A4A or human MS4A4A, and the use of such compositions in preventing, reducing risk, or treating individuals in need thereof.

Description

[0001] Submit sequence listing as ASCII text file

[0002] The following, submitted as an ASCII text file, is incorporated herein by reference in its entirety: Sequence Listing in Computer Readable Form (CRF) (File Name: 4503_008PC06_Seqlisting_ST25.TXT; Creation Date: July 28, 2020; Size: 289,448 bytes). Technical Field

[0003] The present disclosure relates to anti-MS4A4A antibodies and therapeutic uses of such antibodies. Background Art

[0004] The transmembrane 4 domain subfamily A (MS4A) gene cluster is present on chromosome 11q12 and includes 18 genes. The MS4A gene family encodes membrane proteins that generally have a four-span topology (Ishibashi et al., 2001, Gene, 265:87-93; Liang and Tedder, 2001, Genomics, 72:119-127; Efthymiou and Goate, 2017, Molecular Neurodegeneration, 12:43). The transmembrane domain is interconnected by an intracellular loop and two extracellular loops, with both the N and C termini located in the cytoplasm. Most MS4A proteins share amino acid sequence homology (20-30% similarity) with MS4A1 (CD20), with the highest degree of sequence identity being present in the first three transmembrane domains. The highly conserved motifs across different MS4A proteins within these transmembrane regions indicate that the transmembrane domain plays an important general role in MS4A protein function. The regions that vary most among MS4A proteins are found within their N- and C-terminal cytoplasmic domains and a putative second extracellular loop (Ishibashi et al., 2001, Gene, 265:87-93), suggesting that these regions confer unique functional properties.

[0005] Despite this diversity, the MS4A domain has some shared elements. For example, a notable feature conserved in the MS4A proteins (except MS4A8B and MS4A12) is the conservation of two cysteine ​​residues that can form disulfide bonds in the putative second extracellular loop. The N- and C-terminal domains of the MS4A protein are also rich in proline residues, but their functional significance remains to be elucidated (Hulett et al., 2001, Genomics, 72: 119-127). However, the proline-rich region is generally involved in various cellular processes, such as cytoskeletal rearrangement, transcription initiation, signal transduction cascades, and association with the SH3 domain as part of the linking system to promote protein-protein interactions (Kay et al., 2000, FASEB J, 14: 231-241).

[0006] The MS4A family of proteins is relatively uncharacterized functionally, with some important exceptions: MS4A1 (CD20) is expressed exclusively in B lymphocytes, where it has signaling functions through the B cell antigen receptor and calcium influx. CD20 is the target of immunotherapeutic antibodies used to deplete pathogenic B cells in chronic lymphocytic leukemia, lymphomas, autoimmune diseases, and solid organ transplantation. MS4A2 (FcεRβ) is the signaling subunit of the high-affinity IgE receptor (FcεRI) and the low-affinity IgG receptor (FcεRIII) on mast cells, which play a key role in allergic and anaphylactic reactions. MS4A2 is an ITAM domain protein that amplifies signals through the four-protein high-affinity IgE receptor complex. MS4A3 (Htm4) is expressed on the intracellular membranes of lymphoid and myeloid cells and functions as an adaptor protein in cell cycle regulation.

[0007] Although most MS4A family members have not been characterized, reports indicate that MS4A proteins act as chemical sensors and chemical receptors for a variety of exogenous and endogenous ligands, including fatty acids, peptides, and sulfated steroids, and are involved in mediating calcium influx, regulating endocytosis, trafficking, and can serve as adaptors for signal transduction complexes (Cruse et al., 2015, Mol Biol Cell, 26:1711-1727; Greer et al., 2016, Cell, 165:1734-1748; Eon Kuek et al., 2016, Cell, 165:1734-1748; Koslowski et al., 2008, Cancer Res, 68:3458-3466; Bubien et al., 1993; J Cell Biol, 121:1121-1132).

[0008] Certain MS4A genes have been genetically associated with various conditions and diseases, particularly neurodegenerative conditions. For example, genome-wide significant association analysis has identified the MS4A gene cluster on chromosome 11q12 as one of the most significant Alzheimer's disease loci. One gene of particular interest is MS4A4A (Lambert et al., 2013, Nat Genet, 45:1452-1458; Hollingworth et al., 2011, Nat Genet, 43:429-435; Naj et al., 2011, Nat Genet, 43:436-441).

[0009] Therefore, there is a need for therapies that target MS4A4A, including antibodies that specifically bind to MS4A4A, and / or therapies that can modulate (e.g., inhibit or reduce; activate or enhance) MS4A4A activity, such as by reducing or increasing MS4A4A protein levels or activity, to treat various diseases, disorders, and conditions associated with MS4A4A activity.

[0010] All references cited herein, including patent applications and publications, are hereby incorporated by reference in their entirety. Summary of the Invention

[0011] The present disclosure generally relates to anti-MS4A4A antibodies and methods of using such antibodies. The methods provided herein can be used to prevent, reduce the risk of, or treat an individual suffering from a neurodegenerative disease, disorder, or condition. In some embodiments, the present disclosure provides methods for preventing, reducing the risk of, or treating an individual suffering from a neurodegenerative disease, disorder, or condition selected from Alzheimer's disease, late-onset Alzheimer's disease, dementia, and cognitive impairment, comprising administering to the individual in need thereof a therapeutically effective amount of an anti-MS4A4A antibody. In some embodiments, the present disclosure provides methods for preventing, reducing the risk of, or treating an individual suffering from a disease, disorder, or condition associated with increased expression or activity of MS4A4A, comprising administering to the individual in need thereof a therapeutically effective amount of an anti-MS4A4A antibody.

[0012] In one aspect, the present disclosure relates to an isolated antibody that binds to an MS4A4A protein, wherein the antibody comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises: HVR-H1 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 94, 108, 116, 146, 147, 308, and 311; HVR-H2 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 96, 97, 98, 99, 110, 111, 118, 119, 120, 121, 122, 149, 150, 151, 152, 153, 309, and 312; and HVR-H3 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: NO: 100, 101, 102, 112, 123, 124, 125, 126, 127, 128, 129, 154, 310 and 313; and the light chain variable region includes: HVR-L1, which includes an amino acid sequence selected from SEQ ID NO: 103, 104, 113, 130, 131, 132, 133, 134, 135, 136, 137, 138, 156, 157, 158, 314 and 317; HVR-L2, which includes an amino acid sequence selected from SEQ ID NO: 105, 106, 114, 139, 140, 141, 142, 143, 159, 160, 161, 315 and 318; and HVR-L3, which includes an amino acid sequence selected from SEQ ID NO: The amino acid sequences of NO:107, 115, 144, 145, 163, 316 and 319.

[0013] In some embodiments that may be combined with any of the embodiments herein, an anti-MS4A4A antibody of the present disclosure is an isolated antibody that binds to an MS4A4A protein, wherein the antibody comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises: HVR-H1 comprising an amino acid sequence of SEQ ID NO: 94; HVR-H2 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 96-99; and HVR-H3 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 100-102; and the light chain variable region comprises: HVR-L1 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 103-104; HVR-L2 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 105-106; and HVR-L3 comprising an amino acid sequence of SEQ ID NO: 107.

[0014] In some embodiments that may be combined with any of the embodiments herein, an anti-MS4A4A antibody of the present disclosure is an isolated antibody that binds to an MS4A4A protein, wherein the antibody comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises: HVR-H1 comprising the amino acid sequence of SEQ ID NO: 308; HVR-H2 comprising the amino acid sequence of SEQ ID NO: 309; and HVR-H3 comprising the amino acid sequence of SEQ ID NO: 310; and the light chain variable region comprises: HVR-L1 comprising the amino acid sequence of SEQ ID NO: 314; HVR-L2 comprising the amino acid sequence of SEQ ID NO: 315; and HVR-L3 comprising the amino acid sequence of SEQ ID NO: 316.

[0015] In some embodiments that may be combined with any of the embodiments herein, an anti-MS4A4A antibody of the present disclosure is an isolated antibody that binds to an MS4A4A protein, wherein the antibody comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises: HVR-H1 comprising the amino acid sequence of SEQ ID NO: 311; HVR-H2 comprising the amino acid sequence of SEQ ID NO: 312; and HVR-H3 comprising the amino acid sequence of SEQ ID NO: 313; and the light chain variable region comprises: HVR-L1 comprising the amino acid sequence of SEQ ID NO: 317; HVR-L2 comprising the amino acid sequence of SEQ ID NO: 318; and HVR-L3 comprising the amino acid sequence of SEQ ID NO: 319.

[0016] In some embodiments that may be combined with any of the embodiments herein, an anti-MS4A4A antibody of the present disclosure is an isolated antibody that binds to an MS4A4A protein, wherein the antibody comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises: HVR-H1 comprising the amino acid sequence of SEQ ID NO: 108; HVR-H2 comprising an amino acid sequence selected from SEQ ID NOs: 110-111; and HVR-H3 comprising the amino acid sequence of SEQ ID NO: 112; and the light chain variable region comprises: HVR-L1 comprising the amino acid sequence of SEQ ID NO: 113; HVR-L2 comprising the amino acid sequence of SEQ ID NO: 114; and HVR-L3 comprising the amino acid sequence of SEQ ID NO: 115.

[0017] In some embodiments that may be combined with any of the embodiments herein, an anti-MS4A4A antibody of the present disclosure is an isolated antibody that binds to an MS4A4A protein, wherein the antibody comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises: HVR-H1 comprising an amino acid sequence of SEQ ID NO: 116; HVR-H2 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 118-122; and HVR-H3 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 123-129; and the light chain variable region comprises: HVR-L1 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 130-138; HVR-L2 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 139-143; and HVR-L3 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 144-145.

[0018] In some embodiments that may be combined with any of the embodiments herein, an anti-MS4A4A antibody of the present disclosure is an isolated antibody that binds to an MS4A4A protein, wherein the antibody comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises: HVR-H1 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 146-147; HVR-H2 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 149-153; and HVR-H3 comprising an amino acid sequence of SEQ ID NO: 154; and the light chain variable region comprises: HVR-L1 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 156-158; HVR-L2 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 159-161; and HVR-L3 comprising an amino acid sequence of SEQ ID NO: 163.

[0019] In some embodiments that may be combined with any of the embodiments herein, the anti-MS4A4A antibody of the present disclosure is an isolated antibody that binds to the MS4A4A protein, wherein the antibody comprises a heavy chain variable region, wherein the heavy chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 5-15, 304, and 306. In some embodiments that may be combined with any of the embodiments herein, the anti-MS4A4A antibody of the present disclosure is an isolated antibody that binds to the MS4A4A protein, wherein the antibody comprises a heavy chain variable region, wherein the heavy chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 24-30. In some embodiments that may be combined with any of the embodiments herein, the anti-MS4A4A antibody of the present disclosure is an isolated antibody that binds to the MS4A4A protein, wherein the antibody comprises a heavy chain variable region, wherein the heavy chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 40-53. In some embodiments that may be combined with any of the embodiments herein, the anti-MS4A4A antibody of the present disclosure is an isolated antibody that binds to an MS4A4A protein, wherein the antibody comprises a heavy chain variable region, wherein the heavy chain variable region comprises an amino acid sequence selected from SEQ ID NOs: 76-84.

[0020] In some embodiments that may be combined with any of the embodiments herein, an anti-MS4A4A antibody of the present disclosure is an isolated antibody that binds to an MS4A4A protein, wherein the antibody comprises a light chain variable region, wherein the light chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 17-22, 305, and 307. In some embodiments that may be combined with any of the embodiments herein, an anti-MS4A4A antibody of the present disclosure is an isolated antibody that binds to an MS4A4A protein, wherein the antibody comprises a light chain variable region, wherein the light chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 32-36. In some embodiments that may be combined with any of the embodiments herein, an anti-MS4A4A antibody of the present disclosure is an isolated antibody that binds to an MS4A4A protein, wherein the antibody comprises a light chain variable region, wherein the light chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 55-74. In some embodiments that may be combined with any of the embodiments herein, the anti-MS4A4A antibody of the present disclosure is an isolated antibody that binds to an MS4A4A protein, wherein the antibody comprises a light chain variable region, wherein the light chain variable region comprises an amino acid sequence selected from SEQ ID NOs: 86-93.

[0021] In some embodiments that may be combined with any of the embodiments herein, an anti-MS4A4A antibody of the present disclosure is an isolated antibody that binds to an MS4A4A protein, wherein the antibody comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 5-15, 304, and 306, and the light chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 17-22, 305, and 307. In some embodiments that may be combined with any of the embodiments herein, an anti-MS4A4A antibody of the present disclosure is an isolated antibody that binds to an MS4A4A protein, wherein the antibody comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 24-30, and the light chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 32-36. In some embodiments that may be combined with any of the embodiments herein, the anti-MS4A4A antibody of the present disclosure is an isolated antibody that binds to the MS4A4A protein, wherein the antibody comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 40-53, and the light chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 55-74. In some embodiments that may be combined with any of the embodiments herein, the anti-MS4A4A antibody of the present disclosure is an isolated antibody that binds to the MS4A4A protein, wherein the antibody comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 76-84, and the light chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 86-93.

[0022] In some embodiments that may be combined with any of the embodiments herein, an anti-MS4A4A antibody of the present disclosure is an isolated antibody that binds to an MS4A4A protein, wherein the antibody comprises a heavy chain and a light chain, wherein the heavy chain comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 320-343. In some embodiments, the heavy chain comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 336-343 (optionally wherein the light chain comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 22, and optionally wherein the light chain further comprises a constant region comprising the amino acid sequence of SEQ ID NO: 344). In some embodiments, the heavy chain comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 320-327 (optionally wherein the light chain comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 304, and optionally wherein the light chain further comprises a constant region comprising the amino acid sequence of SEQ ID NO: 344). In some embodiments, the heavy chain comprises an amino acid sequence selected from SEQ ID NOs: 328-335 (optionally wherein the light chain comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 305, and optionally wherein the light chain further comprises a constant region comprising the amino acid sequence of SEQ ID NO: 344).

[0023] In some embodiments that may be combined with any of the embodiments herein, an anti-MS4A4A antibody of the present disclosure is an isolated antibody that binds to an MS4A4A protein, wherein the antibody comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises HVR-H1 comprising the amino acid sequence of SEQ ID NO: 94, HVR-H2 comprising the amino acid sequence of SEQ ID NO: 96, and HVR-H3 comprising the amino acid sequence of SEQ ID NO: 102; and the light chain variable region comprises HVR-L1 comprising the amino acid sequence of SEQ ID NO: 104, HVR-L2 comprising the amino acid sequence of SEQ ID NO: 105, and HVR-L3 comprising the amino acid sequence of SEQ ID NO: 107. In some embodiments that may be combined with any of the embodiments herein, an anti-MS4A4A antibody of the present disclosure is an isolated antibody that binds to an MS4A4A protein, wherein the antibody comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 14, and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 22. In some embodiments that may be combined with any of the embodiments herein, an anti-MS4A4A antibody of the present disclosure is an isolated antibody that binds to an MS4A4A protein, wherein the antibody comprises a heavy chain and a light chain, wherein the heavy chain comprises the amino acid sequence of SEQ ID NO: 359 or 360, and the light chain comprises the amino acid sequence of SEQ ID NO: 365.

[0024] In some embodiments that may be combined with any of the embodiments herein, an anti-MS4A4A antibody of the present disclosure is an isolated antibody that binds to an MS4A4A protein, wherein the antibody comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises: HVR-H1 comprising the amino acid sequence of SEQ ID NO: 308; HVR-H2 comprising the amino acid sequence of SEQ ID NO: 309; and HVR-H3 including the amino acid sequence of SEQ ID NO: 310; and the light chain variable region comprises: HVR-L1 comprising the amino acid sequence of SEQ ID NO: 314; HVR-L2 comprising the amino acid sequence of SEQ ID NO: 315; and HVR-L3 comprising the amino acid sequence of SEQ ID NO: 316. In some embodiments that may be combined with any of the embodiments herein, an anti-MS4A4A antibody of the present disclosure is an isolated antibody that binds to an MS4A4A protein, wherein the antibody comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 304, and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 305. In some embodiments that may be combined with any of the embodiments herein, an anti-MS4A4A antibody of the present disclosure is an isolated antibody that binds to an MS4A4A protein, wherein the antibody comprises a heavy chain and a light chain, wherein the heavy chain comprises the amino acid sequence of SEQ ID NO: 324 or 325, and the light chain comprises the amino acid sequence of SEQ ID NO: 363.

[0025] In some embodiments that may be combined with any of the embodiments herein, an anti-MS4A4A antibody of the present disclosure is an isolated antibody that binds to an MS4A4A protein, wherein the antibody comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises: HVR-H1 comprising the amino acid sequence of SEQ ID NO: 311; HVR-H2 comprising the amino acid sequence of SEQ ID NO: 312; and HVR-H3 comprising the amino acid sequence of SEQ ID NO: 313; and the light chain variable region comprises: HVR-L1 comprising the amino acid sequence of SEQ ID NO: 317; HVR-L2 comprising the amino acid sequence of SEQ ID NO: 318; and HVR-L3 comprising the amino acid sequence of SEQ ID NO: 319. In some embodiments that may be combined with any of the embodiments herein, an anti-MS4A4A antibody of the present disclosure is an isolated antibody that binds to an MS4A4A protein, wherein the antibody comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 306, and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 307. In some embodiments that may be combined with any of the embodiments herein, an anti-MS4A4A antibody of the present disclosure is an isolated antibody that binds to an MS4A4A protein, wherein the antibody comprises a heavy chain and a light chain, wherein the heavy chain comprises the amino acid sequence of SEQ ID NO: 332 or 333, and the light chain comprises the amino acid sequence of SEQ ID NO: 364.

[0026] In one aspect, the disclosure relates to an isolated antibody that binds to MS4A4A protein, wherein the antibody competitively inhibits binding of one or more of the antibodies described in any of the embodiments herein to MS4A4A.

[0027] In another aspect, the disclosure relates to an isolated antibody that binds to an MS4A4A protein, wherein the antibody binds to substantially the same or overlapping epitope on MS4A4A as the antibody as described in any of the embodiments herein.

[0028] In some embodiments that may be combined with any of the embodiments herein, the anti-MS4A4A antibodies of the present disclosure are isolated antibodies that bind to the MS4A4A protein, wherein the antibody binds to the extracellular domain 1 of human MS4A4A. In some embodiments that may be combined with any of the embodiments herein, the anti-MS4A4A antibodies of the present disclosure are isolated antibodies that bind to the MS4A4A protein, wherein the antibody binds to one or more amino acid residues within the amino acid sequence CMASNTYGSNPIS (SEQ ID NO: 177) of SEQ ID NO: 1. In some embodiments that may be combined with any of the embodiments herein, the anti-MS4A4A antibodies of the present disclosure are isolated antibodies that bind to the MS4A4A protein, wherein the antibody binds to the extracellular domain 2 of human MS4A4A. In some embodiments that may be combined with any of the embodiments herein, the anti-MS4A4A antibodies of the present disclosure are isolated antibodies that bind to the MS4A4A protein, wherein the antibody binds to one or more amino acid residues within the amino acid sequence SFHHPYCNYYGNSNNCHGTMS (SEQ ID NO: 178) of SEQ ID NO: 1. In some embodiments that may be combined with any of the embodiments herein, the anti-MS4A4A antibody of the present disclosure is an isolated antibody that binds to an MS4A4A protein, wherein the antibody binds to one or more amino acid residues within the amino acid sequence SFHHPYCNYYGNSNNCHGTMS (SEQ ID NO: 178) of SEQ ID NO: 1, and further wherein the antibody does not bind to amino acid residue Proline 163 (P163) of SEQ ID NO: 1. In some embodiments that may be combined with any of the embodiments herein, the anti-MS4A4A antibody is a human anti-MS4A4A antibody or a humanized anti-MS4A4A antibody.

[0029] In some embodiments that may be combined with any of the embodiments herein, the anti-MS4A4A antibody of the present disclosure is an isolated antibody that binds to the MS4A4A protein, wherein the antibody reduces cell surface levels of MS4A4A or reduces intracellular levels of MS4A4A.

[0030] In certain embodiments that may be combined with any of the embodiments herein, the MS4A4A protein is a mammalian or human protein. In certain embodiments that may be combined with any of the embodiments herein, the MS4A4A protein is a wild-type protein. In certain embodiments that may be combined with any of the embodiments herein, the MS4A4A protein is a naturally occurring variant.

[0031] In some embodiments that may be combined with any of the embodiments herein, the anti-MS4A4A antibodies of the present disclosure increase soluble TREM2 levels in myeloid cells, increase membrane TREM2 levels, or increase soluble TREM2 and increase membrane TREM2 levels. In some embodiments that may be combined with any of the embodiments herein, the anti-MS4A4A antibodies of the present disclosure increase soluble TREM2 levels. In some embodiments that may be combined with any of the embodiments herein, the anti-MS4A4A antibodies of the present disclosure increase membrane TREM2 levels. In some embodiments that may be combined with any of the embodiments herein, the anti-MS4A4A antibodies of the present disclosure increase soluble TREM2 levels and membrane TREM2 levels. In some embodiments that may be combined with any of the embodiments herein, the anti-MS4A4A antibodies of the present disclosure increase soluble TREM2 levels in vivo. In some embodiments that may be combined with any of the embodiments herein, the anti-MS4A4A antibodies of the present disclosure increase soluble TREM2 levels in serum in vivo. In some embodiments that may be combined with any of the embodiments herein, the anti-MS4A4A antibodies of the present disclosure increase soluble TREM2 levels in cerebrospinal fluid (CSF) in vivo. In some embodiments that may be combined with any of the embodiments herein, the anti-MS4A4A antibodies of the present disclosure increase soluble TREM2 levels in serum and CSF in vivo.

[0032] In some embodiments that may be combined with any of the embodiments herein, the anti-MS4A4A antibodies of the present disclosure reduce the expression of M2 cell surface markers (e.g., receptors) on myeloid cells. In some embodiments that may be combined with any of the embodiments herein, the anti-MS4A4A antibodies of the present disclosure reduce or decrease the level of M2 cell surface markers in myeloid cells (e.g., macrophages).

[0033] In some embodiments that may be combined with any of the embodiments herein, the anti-MS4A4A antibodies of the present disclosure reduce the expression of CD200R, Dectin-1, or CD163 on myeloid cells. In some embodiments that may be combined with any of the embodiments herein, the anti-MS4A4A antibodies of the present disclosure reduce or decrease cell surface levels of CD200R, Dectin-1, and / or CD163 in myeloid cells (e.g., macrophages).

[0034] In some embodiments that may be combined with any of the embodiments herein, the anti-MS4A4A antibodies of the present disclosure increase the mRNA and / or protein expression of gelsolin and / or osteopontin on myeloid cells. In some embodiments that may be combined with any of the embodiments herein, the anti-MS4A4A antibodies of the present disclosure increase the mRNA and / or protein levels of IL1RN on myeloid cells. In some embodiments that may be combined with any of the embodiments herein, the anti-MS4A4A antibodies of the present disclosure increase the mRNA and / or protein levels of CSF1R on myeloid cells. In some embodiments that may be combined with any of the embodiments herein, the anti-MS4A4A antibodies of the present disclosure decrease the mRNA and / or protein levels of the purinergic receptor P2RY12 and / or CX3C chemokine receptor 1 (CX3CR1) on myeloid cells. In some embodiments that may be combined with any of the embodiments herein, the anti-MS4A4A antibodies of the present disclosure increase the mRNA and / or protein levels of 1-phosphatidylinositol-4,5-bisphosphate phosphodiesterase gamma-2 (PLCG2), C-type lectin domain family 7 member A (CLEC7A), inositol 1,4,5-triphosphate receptor 2 (ITPR2), and / or antigen KI-67 (MK167) on myeloid cells. In some embodiments that may be combined with any of the embodiments herein, the anti-MS4A4A antibodies of the present disclosure decrease the mRNA and / or protein levels of transmembrane glycoprotein NMB (GPNMB) on myeloid cells.

[0035] In one aspect, the present disclosure relates to an isolated antibody that binds to the MS4A4A protein, wherein the antibody increases the mRNA and / or protein expression of gelsolin and / or osteopontin on myeloid cells. In some embodiments that may be combined with any of the embodiments herein, the myeloid cells are human macrophages. In certain embodiments that may be combined with any of the embodiments herein, the isolated antibody that binds to the MS4A4A protein is human or humanized. In certain embodiments that may be combined with any of the embodiments herein, the isolated antibody that binds to the MS4A4A protein is a human antibody, a humanized antibody, a bispecific antibody, a monoclonal antibody, a multivalent antibody, a conjugated antibody, or a chimeric antibody. In certain embodiments that may be combined with any of the embodiments herein, the isolated antibody that binds to the MS4A4A protein binds to the same epitope as an antibody selected from 4A-18, 4A-25, 4A-214, 4A-21, and 4A-202. In certain embodiments that may be combined with any of the embodiments herein, the isolated antibody that binds to the MS4A4A protein reduces the expression of an M2 marker on myeloid cells. In certain embodiments that may be combined with any of the embodiments herein, the myeloid cells are human macrophages. In certain embodiments that may be combined with any of the embodiments herein, the M2 marker is CD200R, Dectin-1, and / or CD163. In certain embodiments that may be combined with any of the embodiments herein, the isolated antibody that binds to the MS4A4A protein increases plasma membrane or cell surface TREM2 on human macrophages by at least 50%, 90%, 100%, 150%, 200%, or 250%. In certain embodiments that may be combined with any of the embodiments herein, the isolated antibody that binds to the MS4A4A protein increases soluble TREM2 in the supernatant of human macrophages by at least 15%, 20%, 25%, 30%, 40%, 50%, 60%, or 70%.

[0036] In certain embodiments that may be combined with any of the embodiments herein, the anti-MS4A4A antibody is a bispecific antibody that recognizes a first antigen and a second antigen. In certain embodiments that may be combined with any of the embodiments herein, the first antigen is MS4A4A and the second antigen is an antigen that promotes transport across the blood-brain barrier. In certain embodiments that may be combined with any of the embodiments herein, the second antigen is selected from MS4A4A; transferrin receptor (TR); insulin receptor (HIR); insulin-like growth factor receptor (IGFR); low-density lipoprotein receptor-related proteins 1 and 2 (LPR-1 and LPR-2); diphtheria toxin receptor CRM197; llama single domain antibody TMEM 30 (A); protein transduction domains TAT, Syn-B, penetratin; polyarginine peptides; vascular peptides, basic immunoglobulins, Glut1 and CD98hc; and ANG1005.

[0037] In some embodiments that may be combined with any of the embodiments herein, the anti-MS4A4A antibody of the present disclosure is a monoclonal antibody. In some embodiments that may be combined with any of the embodiments herein, the antibody is a human antibody. In some embodiments that may be combined with any of the embodiments herein, the antibody is a humanized antibody. In some embodiments that may be combined with any of the embodiments herein, the antibody is a bispecific antibody. In some embodiments that may be combined with any of the embodiments herein, the antibody is a multivalent antibody. In some embodiments that may be combined with any of the embodiments herein, the antibody is a chimeric antibody.

[0038] In some embodiments that may be combined with any of the embodiments herein, the anti-MS4A4A antibodies of the present disclosure are of the IgG, IgM, or IgA class. In some embodiments, the antibodies are of the IgG class and have an IgG1, IgG2, or IgG4 isotype. In certain embodiments that may be combined with any of the embodiments herein, the antibodies are antibody fragments. In certain embodiments that may be combined with any of the embodiments herein, the antibodies are antibody fragments that bind to an epitope that includes amino acid residues on human MS4A4A or mammalian MS4A4A proteins. In certain embodiments that may be combined with any of the embodiments herein, the antibody fragments are Fab, Fab', Fab'-SH, F(ab')2, Fv, or scFv fragments.

[0039] In some embodiments that may be combined with any of the embodiments herein, the antibody is a bispecific antibody that recognizes a first antigen and a second antigen. In some embodiments that may be combined with any of the embodiments herein, the first antigen is MS4A4A and the second antigen is:

[0040] (a) Antigens that promote transport across the blood-brain barrier;

[0041] (b) an antigen that promotes transport across the blood-brain barrier selected from the group consisting of transferrin receptor (TR); insulin receptor (HIR); insulin-like growth factor receptor (IGFR); low-density lipoprotein receptor-related proteins 1 and 2 (LPR-1 and LPR-2); diphtheria toxin receptor CRM197; llama single domain antibody TMEM 30 (A); protein transduction domains TAT, Syn-B, penetrant peptides; polyarginine peptides; vascular peptides; and ANG1005;

[0042] (c) a pathogenic agent selected from a pathogenic peptide or protein or a pathogenic nucleic acid, wherein the pathogenic nucleic acid is an antisense GGCCCC (G2C4) repeat expansion RNA, and the pathogenic protein is selected from amyloid beta, oligomeric amyloid beta, amyloid beta plaques, amyloid precursor protein or fragments thereof, Tau, IAPP, α-synuclein, TDP-43, FUS protein, C9orf72 (chromosome 9 open reading frame 72), c9RAN protein, prion protein, PrPSc, huntingtin, calcitonin, superoxide dismutase, ataxia, ataxia 1, ataxia 2, ataxia 3, ataxia 7, ataxia 8, ataxia 10, Lewy bodies, body), atrial natriuretic factor, amylin, insulin, apolipoprotein AI, serum amyloid A, medin, prolactin, transthyretin, lysozyme, β2-microglobulin, gelsolin, corneal epithelial protein, cystatin, immunoglobulin light chain AL, S-IBM protein, repeat-associated non-ATG (RAN) translation products, dipeptide repeat (DPR) peptides, glycine-alanine (GA) repeat peptides, glycine-proline (GP) repeat peptides, glycine-arginine (GR) repeat peptides, proline-alanine (PA) repeat peptides, ubiquitin, and proline-arginine (PR) repeat peptides;

[0043] (d) a ligand and / or protein expressed on an immune cell, wherein the ligand and / or protein is selected from the group consisting of CD40, OX40, ICOS, CD28, CD137 / 4-1BB, CD27, GITR, PD-L1, CTLA-4, PD-L2, PD-1, B7-H3, B7-H4, HVEM, BTLA, KIR, GAL9, TIM3, A2AR, LAG-3, and phosphatidylserine; or

[0044] (e) a protein, lipid, polysaccharide, or glycolipid expressed on one or more tumor cells.

[0045] In another aspect, the present disclosure relates to an isolated nucleic acid comprising a nucleic acid sequence encoding an antibody as described in any one of the preceding embodiments. In some embodiments, the present disclosure relates to a vector comprising a nucleic acid as described in any one of the preceding embodiments. In some embodiments, the present disclosure relates to an isolated host cell comprising a vector as described in any one of the preceding embodiments.

[0046] In another aspect, the present disclosure relates to a method for producing an antibody that binds to human MS4A4A, comprising culturing the host cell of any of the preceding embodiments such that the anti-MS4A4A antibody is produced. In certain embodiments, the method further comprises recovering the anti-MS4A4A antibody produced by the cell.

[0047] In another aspect, the present disclosure relates to a pharmaceutical composition comprising the antibody of any one of the preceding embodiments and a pharmaceutically acceptable carrier.

[0048] In one aspect, the present disclosure relates to a method for preventing, reducing the risk of, or treating an individual suffering from a disease, disorder, or injury selected from Alzheimer's disease, late-onset Alzheimer's disease, and cognitive impairment, the method comprising administering to an individual in need thereof a therapeutically effective amount of an antibody as described in any one of the preceding embodiments. In another aspect, the present disclosure relates to a method for preventing, reducing the risk of, or treating an individual suffering from a disease, disorder, condition, or injury caused by or associated with overexpression or increased activity of MS4A4A, the method comprising administering to an individual in need thereof a therapeutically effective amount of an antibody as described in any one of the preceding embodiments. In some embodiments of the present disclosure, the method further comprises detecting the level of osteopontin, gelsolin, plasma membrane or cell surface TREM2, and / or soluble TREM2 before and / or after administration of the antibody.

[0049] In one aspect, the present disclosure relates to a method of preventing, reducing the risk of, or treating an individual having a CSF1R deficiency disease or condition, the method comprising administering to an individual in need thereof a therapeutically effective amount of an antibody as described in any one of the preceding embodiments. In some embodiments, the CSF1R deficiency disease or condition is adult-onset leukoencephalopathy with axonal balling and pigmented glia (ALSP) or hereditary diffuse leukoencephalopathy (HDLS).

[0050] Other aspects of the present disclosure relate to isolated anti-MS4A4A antibodies produced by the method of any one of the embodiments. Other aspects of the present disclosure relate to pharmaceutical compositions comprising the anti-MS4A4A antibody of any one of the embodiments and a pharmaceutically acceptable carrier.

[0051] In another aspect, the disclosure relates to an antibody that binds to an MS4A4A protein, wherein the antibody comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises HVR-H1, HVR-H2, and HVR-H3 of antibody 4A-301, 4A-302, 4A-303, 4A-304, 4A-305, 4A-306, 4A-307, 4A-308, 4A-309, 4A-310, 4A-311, 4A-312, 4A-313, 4A-314, 4A-419, or 4A-450 (as shown in Tables 1, 5, 30, and 31).

[0052] In another aspect, the disclosure relates to an antibody that binds to an MS4A4A protein, wherein the antibody comprises a heavy chain variable region and a light chain variable region, wherein the light chain variable region comprises HVR-L1, HVR-L2, and HVR-L3 of antibody 4A-301, 4A-302, 4A-303, 4A-304, 4A-305, 4A-306, 4A-307, 4A-308, 4A-309, 4A-310, 4A-311, 4A-312, 4A-313, 4A-314, 4A-419, or 4A-450 (as shown in Tables 1, 5, 30, and 31).

[0053] In another aspect, the disclosure relates to an antibody that binds to an MS4A4A protein, wherein the antibody comprises a heavy chain variable region comprising HVR-H1, HVR-H2, and HVR-H3, and a light chain variable region comprising HVR-L1, HVR-L2, and HVR-L3, wherein the antibody comprises HVR-H1, HVR-H2, HVR-H3, HVR-L1, HVR-L2, and HVR-L3 of antibody 4A-301, 4A-302, 4A-303, 4A-304, 4A-305, 4A-306, 4A-307, 4A-308, 4A-309, 4A-310, 4A-311, 4A-312, 4A-313, 4A-314, 4A-419, or 4A-450 (as shown in Tables 1, 5, 30, and 31).

[0054] In another aspect, the disclosure relates to an antibody that binds to an MS4A4A protein, wherein the antibody comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises HVR-H1, HVR-H2, and HVR-H3 of antibody 4A-315, 4A-316, 4A-317, 4A-318, 4A-319, 4A-320, 4A-321, 4A-322, 4A-323, 4A-324, 4A-325, 4A-326, 4A-327, 4A-328, 4A-329, 4A-330, or 4A-331 (as shown in Tables 2 and 6).

[0055] In another aspect, the disclosure relates to an antibody that binds to an MS4A4A protein, wherein the antibody comprises a heavy chain variable region and a light chain variable region, wherein the light chain variable region comprises HVR-L1, HVR-L2, and HVR-L3 of antibody 4A-315, 4A-316, 4A-317, 4A-318, 4A-319, 4A-320, 4A-321, 4A-322, 4A-323, 4A-324, 4A-325, 4A-326, 4A-327, 4A-328, 4A-329, 4A-330, or 4A-331 (as shown in Tables 2 and 6).

[0056] In another aspect, the disclosure relates to an antibody that binds to an MS4A4A protein, wherein the antibody comprises a heavy chain variable region comprising HVR-H1, HVR-H2, and HVR-H3, and a light chain variable region comprising HVR-L1, HVR-L2, and HVR-L3, wherein the antibody comprises HVR-H1, HVR-H2, HVR-H3, HVR-L1, HVR-L2, and HVR-L3 of antibody 4A-315, 4A-316, 4A-317, 4A-318, 4A-319, 4A-320, 4A-321, 4A-322, 4A-323, 4A-324, 4A-325, 4A-326, 4A-327, 4A-328, 4A-329, 4A-330, or 4A-331 (as shown in Tables 2 and 6).

[0057] In another aspect, the disclosure relates to antibodies that bind to MS4A4A protein, wherein the antibodies comprise a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises antibodies 4A-332, 4A-333, 4A-334, 4A-335, 4A-336, 4A-337, 4A-338, 4A-339, 4A-340, 4A-341, 4A-342, 4A-343, 4A-344, 4A-345, 4A-346, 4A-347, 4A-348, 4A-349, 4A-350, 4A-351, 4A-352, 4A-353, 4A-354, 374, 4A-375, 4A-376, 4A-377, 4A-378, 4A-379, 4A-380, or 4A-381 of HVR-H1, HVR-H2, and HVR-H3 (as shown in Tables 3 and 7).

[0058] In another aspect, the disclosure relates to antibodies that bind to MS4A4A protein, wherein the antibodies comprise a heavy chain variable region and a light chain variable region, wherein the light chain variable region comprises antibodies 4A-332, 4A-333, 4A-334, 4A-335, 4A-336, 4A-337, 4A-338, 4A-339, 4A-340, 4A-341, 4A-342, 4A-343, 4A-344, 4A-345, 4A-346, 4A-347, 4A-348, 4A-349, 4A-350, 4A-351, 4A-352, 4A-353, 4A-354, A-354, 4A-355, 4A-356, 4A-357, 4A-358, 4A-359, 4A-360, 4A-361, 4A-361, 4A-363, 4A-364, 4A-365, 4A-366, 4A-367, 4A-368, 4A- HVR-L1, HVR-L2 and HVR-L3 of 369, 4A-370, 4A-371, 4A-372, 4A-373, 4A-374, 4A-375, 4A-376, 4A-377, 4A-378, 4A-379, 4A-380 or 4A-381 (As shown in Table 3 and Table 7).

[0059] In another aspect, the disclosure relates to antibodies that bind to MS4A4A protein, wherein the antibodies comprise a heavy chain variable region comprising HVR-H1, HVR-H2, and HVR-H3, and a light chain variable region comprising HVR-L1, HVR-L2, and HVR-L3, wherein the antibodies include antibodies 4A-332, 4A-333, 4A-334, 4A-335, 4A-336, 4A-337, 4A-338, 4A-339, 4A-340, 4A-341, 4A-342, 4A-343, 4A-344, 4A-345, 4A-346, 4A-347, 4A-348, 4A-349, 4A-350, 4A-351, 4A-352, 4A-353, 4A-354, 4A-355, 4A-356, 4A-357, 4A-358, 4A-359, 4A-360, 4A-361, 4A-362, 4A-363, 4A-364, 4A-365, 4A-366, 4A-367, 4A-368, 4A-369, 4A-370, 4A-371, 4A-372, 4A-373, 4A-374, 4A-375 352, 4A-353, 4A-354, 4A-355, 4A-356, 4A-357, 4A-358, 4A-359, 4A-360, 4A -361, 4A-361, 4A-363, 4A-364, 4A-365, 4A-366, 4A-367, 4A-368, 4A-369, 4 A-370, 4A-371, 4A-372, 4A-373, 4A-374, 4A-375, 4A-376, 4A-377, 4A-378, HVR-H1, HVR-H2, HVR-H3, HVR-L1, HVR-L2 and HVR-L3 of 4A-379, 4A-380 or 4A-381 (As shown in Table 3 and Table 7).

[0060] In another aspect, the disclosure relates to an antibody that binds to an MS4A4A protein, wherein the antibody comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises HVR-H1, HVR-H2, and HVR-H3 of antibody 4A-382, 4A-383, 4A-384, 4A-385, 4A-386, 4A-387, 4A-388, 4A-389, or 4A-390 (as shown in Tables 4 and 8).

[0061] In another aspect, the disclosure relates to an antibody that binds to an MS4A4A protein, wherein the antibody comprises a heavy chain variable region and a light chain variable region, wherein the light chain variable region comprises HVR-L1, HVR-L2, and HVR-L3 of antibody 4A-382, 4A-383, 4A-384, 4A-385, 4A-386, 4A-387, 4A-388, 4A-389, or 4A-390 (as shown in Tables 4 and 8).

[0062] In another aspect, the disclosure relates to an antibody that binds to an MS4A4A protein, wherein the antibody comprises a heavy chain variable region comprising HVR-H1, HVR-H2, and HVR-H3, and a light chain variable region comprising HVR-L1, HVR-L2, and HVR-L3, wherein the antibody comprises HVR-H1, HVR-H2, HVR-H3, HVR-L1, HVR-L2, and HVR-L3 of antibody 4A-382, 4A-383, 4A-384, 4A-385, 4A-386, 4A-387, 4A-388, 4A-389, or 4A-390 (as shown in Tables 4 and 8).

[0063] In certain embodiments that may be combined with any of the embodiments herein, the present disclosure relates to antibodies that bind to the MS4A4A protein, wherein the antibodies increase mRNA and / or protein expression of gelsolin and / or osteopontin on myeloid cells. In some embodiments, the myeloid cells are macrophages.

[0064] In certain embodiments that may be combined with any of the embodiments herein, the anti-MS4A4A antibody specifically binds to human MS4A4A, mouse MS4A4A, cynomolgus monkey MS4A4A, or a combination thereof.

[0065] It should be understood that one, some or all of the properties of the various embodiments described herein can be combined to form other embodiments of the present disclosure. These and other aspects of the present disclosure will become apparent to those skilled in the art. These and other embodiments of the present disclosure are further described in the following embodiments. BRIEF DESCRIPTION OF THE DRAWINGS

[0066] This patent or application file contains at least one drawing executed in color. Copies of this patent or patent application publication with color drawing(s) will be provided by the Office upon request and payment of the necessary fee.

[0067] Figure 1 The primary amino acid sequence of human MS4A4A is shown. The intracellular domain is italicized, the transmembrane domain is underlined, and the extracellular loops are in bold italics.

[0068] Figure 2A Shown is a schematic representation of the structure of a soluble four-helix bundle scaffold (PDB_ID: 1P68) obtained from the RCSB Protein Data Bank, which has four transmembrane domains shown as helical ribbons in the structure. Figure 2B and Figure 2C The primary amino acid sequences of the soluble four-helix bundle scaffolds PDB-ID: 1P68 and PDB_ID: 1M6T are shown, respectively; the four transmembrane domains are underlined in the amino acid sequences.

[0069] Figure 3A The primary amino acid sequences of the soluble four-helix bundle scaffolds JS1, JS5, and JS6 are shown, with the addition of two extracellular loops of human MS4A4A (shown in bold and underlined). Figure 3B The primary amino acid sequences of the soluble four-helix bundle scaffolds JS4 and JS10, used as negative control variants, are shown.

[0070] Figure 4 Binding of certain anti-MS4A4A antibodies of the present disclosure to recombinant MS4A4A soluble cyclic grafted polypeptides JS1, JS4, JS5, JS6, and JS10 by ELISA is shown.

[0071] Figure 5 Shown are the binding of certain humanized anti-MS4A4A antibody variants of the murine anti-MS4A4A antibody 4A-21 to the recombinant MS4A4A soluble cyclic grafted polypeptide JS1.

[0072] Figure 6 Shown are the binding of certain humanized anti-MS4A4A antibody variants of the murine anti-MS4A4A antibody 4A-202 to the recombinant MS4A4A soluble cyclic grafted polypeptide JS1.

[0073] Figure 7 Shown are the binding of certain humanized and affinity matured anti-MS4A4A antibody variants of the murine anti-MS4A4A antibody 4A-202 to the recombinant MS4A4A soluble cyclic grafted polypeptide JS5.

[0074] Figure 8 Shown are the binding of certain humanized and affinity matured anti-MS4A4A antibody variants of the murine anti-MS4A4A antibody 4A-21 to the recombinant MS4A4A soluble cyclic grafted polypeptide JS5.

[0075] Figure 9 Shown are the binding of certain humanized and affinity matured anti-MS4A4A antibody variants of the murine anti-MS4A4A antibody 4A-21 to the recombinant MS4A4A soluble cyclic grafted polypeptide JS5.

[0076] FIG. 10A to FIG. 10B Shown is the effect of anti-MS4A4A antibody 4A-202 on sTREM2 levels in cynomolgus monkey serum. Figure 10A Provided are the levels of sTREM2 (ng / mL) in cynomolgus monkey serum at the indicated times (hours) following administration of the anti-MS4A4A antibody 4A-202 or an isotype control antibody (huIgG1), both at a dose of 80 mg / kg. Figure 10BProvided are the levels of sTREM2 (percent of baseline) in cynomolgus monkey serum at the indicated times (hours) following administration of the anti-MS4A4A antibody 4A-202 or an isotype control antibody (huIgG1), both at a dose of 80 mg / kg.

[0077] Figures 11A to 11B Shown are the effects of anti-MS4A4A antibody 4A-202 on sTREM2 levels in cynomolgus monkey cerebrospinal fluid (CSF). Figure 11A Provided are the levels of sTREM2 (ng / mL) in cynomolgus monkey CSF at the indicated times (hours) following administration of the anti-MS4A4A antibody 4A-202 or an isotype control antibody (huIgG1), both at a dose of 80 mg / kg. Figure 11B Provided are the levels of sTREM2 (percent of baseline) in cynomolgus monkey CSF at the indicated times (hours) following administration of the anti-MS4A4A antibody 4A-202 or an isotype control antibody (huIgG1), both at a dose of 80 mg / kg.

[0078] Figure 12 Presented are data showing binding curves of anti-MS4A4A antibodies 4A-313 wild-type (WT) huIgG1 and 4A-450 WT huIgG1 to U937 cells expressing recombinant human MS4A4A.

[0079] Figure 13 Presented are data showing membrane TREM2 levels in primary human macrophages treated with anti-MS4A4A antibodies 4A-450 WT huIgG1, 4A-450 NSLF huIgG1, and 4A-450 K322A huIgG1.

[0080] Figure 14 Presented are data showing membrane TREM2 levels in primary human macrophages treated with anti-MS4A4A antibodies 4A-313 WT huIgG1, 4A-313 NSLF huIgG1, and 4A-313 K322A huIgG1.

[0081] Figure 15 Presented are data showing increased levels of soluble TREM2 in supernatants of primary human macrophages treated with anti-MS4A4A antibody 4A-450 huIgG1.

[0082] Figure 16 Data showing increased levels of soluble TREM2 in supernatants of primary human macrophages treated with anti-MS4A4A antibody 4A-313 huIgG1 are presented.

[0083] Figure 17Presented are data showing increased ATP levels in primary human macrophages treated with anti-MS4A4A antibodies 4S-313 NSLF huIgG1, 4A-313 PS huIgG1, 4A-450 NSLF huIgG1, and 4A-450 PS huIgG1.

[0084] Figure 18A 、 Figure 18B and Figure 18C Presented are data showing changes in CD14, CD163, and CD200R levels (respectively) in primary human macrophages treated with anti-MS4A4A antibodies 4A-313 NSLF huIgG1 and 4A-419 WT huIgG1.

[0085] Figure 19 Presented are data showing complement-dependent cytotoxicity (CDC) activity of anti-MS4A4A antibodies 4A-313 NSLFhuIgGl, 4A-313 K322A huIgGl, and 4A-313 WT huIgGl in U937 cells expressing recombinant human MS4A4A as measured by propidium iodide uptake.

[0086] Figure 20 Presented are data showing ADCP activity of anti-MS4A4A antibodies 4A-313 NSLFhuIgGl, 4A-313 K322A huIgGl, and 4A-313 WT huIgGl in primary human macrophages as measured by FcyRIIIa activation.

[0087] Figure 21 Presented are data showing ADCC activity of anti-MS4A4A antibodies 4A-313 NSLFhuIgGl, 4A-313 K322A huIgGl, and 4A-313 WT huIgGl in primary human macrophages as measured by FcyRIIIa activation.

[0088] Figure 22A and Figure 22B Listed are the genes showing that MS4A4A knockout affects membrane TREM2 in primary human macrophages ( Figure 22A ) and soluble TREM2 ( Figure 22B ) data affected by the

[0089] Figure 23A 、 Figure 23B and Figure 23C Listed are data showing the kinetics of changes in soluble TREM2 protein levels, membrane TREM2 protein levels, and TREM2 mRNA levels, respectively, in human macrophages treated with anti-MS4A4A antibodies of the present disclosure.

[0090] Figure 24A and Figure 24B Data showing that siRNA knockdown of MS4A4A increases membrane TREM2 and soluble TREM2 levels in human macrophages are presented.

[0091] Figure 25A 、 Figure 25B and Figure 25C Data showing a dose-dependent increase in membrane TREM2 levels in primary human macrophages from three different donors following addition of an anti-MS4A4A antibody of the present disclosure are presented.

[0092] Figure 26A 、 Figure 26B and Figure 26C Data showing a dose-dependent increase in soluble TREM2 levels in primary human macrophages from three different donors following addition of an anti-MS4A4A antibody of the present disclosure are presented.

[0093] Figure 27A 、 Figure 27B and Figure 27C Presented are data showing a dose-dependent increase in ATP levels in primary human macrophages from three different donors following addition of an anti-MS4A4A antibody of the present disclosure.

[0094] Figure 28A and Figure 28B Data showing that anti-MS4A4A antibodies of the present disclosure effectively rescue cell death induced by CSF1R inhibition are presented.

[0095] Figure 29 Shown is a volcano plot of global proteome effects in cynomolgus monkey CSF following administration of an anti-MS4A4A antibody of the present disclosure.

[0096] Figure 30 Presented are data showing increased sTREM2 levels in cynomolgus monkey serum following administration of an anti-MS4A4A antibody of the present disclosure.

[0097] Figure 31A 、 Figure 31B 、 Figure 31C and Figure 31D Presented are data showing that osteopontin levels are increased in cynomolgus monkey CSF following administration of an anti-MS4A4A antibody of the disclosure.

[0098] Figure 32A and Figure 32B Presented are data showing that osteopontin levels are increased in the frontal cortex and hippocampus, respectively, of cynomolgus monkey brains following administration of an anti-MS4A4A antibody of the present disclosure.

[0099] Figure 33A and Figure 33B Presented are data showing that CSF1R levels are increased in the frontal cortex and hippocampus, respectively, of cynomolgus monkey brains following administration of an anti-MS4A4A antibody of the present disclosure.

[0100] Figure 34A and Figure 34B Data are presented showing that total TREM2 protein levels are increased in the frontal cortex and hippocampus of cynomolgus monkey brains, respectively, following administration of an anti-MS4A4A antibody of the present disclosure.

[0101] Figure 35A 、 Figure 35B and Figure 35C Presented are data showing changes in mRNA levels of markers of microglial activation, migration, and proliferation in microglial cells isolated from the brains of cynomolgus monkeys administered anti-MS4A4A antibodies of the present disclosure.

[0102] Figure 36 Presented are data showing that anti-MS4A4A antibodies of the present disclosure reduce cell surface expression of recombinantly expressed HA-Snorkel-tagged MS4A4A protein in U937 cells.

[0103] Figure 37 Presented are data showing that anti-MS4A4A antibodies of the present disclosure reduce GPNMB cell surface expression in primary human macrophages. DETAILED DESCRIPTION

[0104] The present disclosure relates to anti-MS4A4A antibodies (eg, monoclonal antibodies); methods of making and using such antibodies; pharmaceutical compositions comprising such antibodies; nucleic acids encoding such antibodies; and host cells comprising nucleic acids encoding such antibodies.

[0105] The techniques and procedures described or referred to herein are generally well understood by those skilled in the art and are routinely employed using conventional methods, such as, for example, widely utilized methods such as those described in the following references: Sambrook et al., Molecular Cloning: A Laboratory Manual , 3rd edition (2001) ColdSpring Harbor Laboratory Press, Cold Spring Harbor, NY; Current Protocols in Molecular Biology (F. M. Ausubel et al., eds. (2003); Monoclonal Antibodies: A Practical Approach (Editors P. Shepherd and C. Dean, Oxford University Press, 2000).

[0106] I. Definition

[0107] Unless otherwise specified, the terms "MS4A4A" or "MS4A4A polypeptide" are used interchangeably herein to refer to any native MS4A4A from any vertebrate source, including mammals, such as primates (e.g., humans and cynomolgus monkeys) and rodents (e.g., mice and rats). In some embodiments, the terms encompass both wild-type sequences and naturally occurring variant sequences (e.g., splice variants or allelic variants). In some embodiments, the terms encompass unprocessed "full-length" MS4A4A as well as any form of MS4A4A resulting from processing in cells. In some embodiments, MS4A4A is human MS4A4A. In some embodiments, the amino acid sequence of an exemplary MS4A4A is Uniprot Accession No. Q96JQ5 as of December 1, 2001. In some embodiments, the amino acid sequence of an exemplary human MS4A4A is SEQ ID NO: 1.

[0108] The terms "anti-MS4A4A antibody," "antibody that binds to MS4A4A," and "antibody that specifically binds to MS4A4A" refer to antibodies that bind to MS4A4A with sufficient affinity to allow the antibody to be used as a diagnostic and / or therapeutic agent targeting MS4A4A. In one embodiment, the extent of binding of the anti-MS4A4A antibody to an unrelated, non-MS4A4A polypeptide is less than about 10% of the binding of the antibody to MS4A4A, as measured, for example, by radioimmunoassay (RIA). In certain embodiments, the antibody binds to MS4A4A with an affinity of <1 μM, <100 nM, <10 nM, <1 nM, <0.1 nM, <0.01 nM, or <0.001 nM (e.g., <10 -8 M or smaller, such as 10 -8 M to 10 -13 M, for example 10 -9 M to 10 -13 In certain embodiments, an anti-MS4A4A antibody binds to an epitope of MS4A4A that is conserved among MS4A4As from different species.

[0109] With respect to the binding of an antibody to a target molecule, the term "specific binding" or "specifically binds to" or "specifically for" a particular polypeptide or an epitope on a particular polypeptide target means binding that is measurably different from non-specific interactions. Specific binding can be measured, for example, by determining the binding of a molecule compared to the binding of a control molecule. For example, specific binding can be determined by competition with a control molecule that is similar to the target (e.g., an excess of unlabeled target). In this case, if the binding of the labeled target to the probe is competitively inhibited by an excess of unlabeled target, specific binding is indicated. As used herein, the term "specific binding" or "specifically binds to" or "specifically for" a particular polypeptide or an epitope on a particular polypeptide target can be determined, for example, by determining a KD for the target of about 10 -4 M or lower, 10 -5 M or lower, 10 -6 M or lower, 10 -7 M or lower, 10 -8 M or lower, 10 -9 M or lower, 10 -10 M or lower, 10 -11 M or lower, 10 -12 M or lower or KD in 10 -4 M to 10 -6 M or 10 -6 M to 10 -10 M or 10 -7 M to 10 -9 The term "specific binding" refers to binding of a molecule to a specific polypeptide or epitope on a specific polypeptide without substantially binding to any other polypeptide or polypeptide epitope.

[0110] the term" immunoglobulins ” (Ig) can be used in this article with “ Antibody The term "antibody" as used herein is used in the broadest sense and specifically covers monoclonal antibodies, polyclonal antibodies, multispecific antibodies (e.g., bispecific antibodies) (including those formed from at least two intact antibodies), and antibody fragments, so long as they exhibit the desired biological activity.

[0111] “ natural antibodies"A heterotetrameric glycoprotein, usually about 150,000 daltons, composed of two identical light ("L") chains and two identical heavy ("H") chains. Each light chain is linked to a heavy chain by one covalent disulfide bond, while the number of disulfide linkages varies among the heavy chains of different immunoglobulin isotypes. Each heavy and light chain also has regularly spaced intrachain disulfide bond. Each heavy chain has a variable domain (V H ), followed by multiple constant domains. Each light chain has a variable domain (V L ) and has a constant domain at its other end; the constant domain of the light chain is aligned with the first constant domain of the heavy chain, and the light chain variable domain is aligned with the variable domain of the heavy chain. Specific amino acid residues are thought to form an interface between the light chain and heavy chain variable domains.

[0112] For the structure and properties of different classes of antibodies, see, for example Basic and Clinical Immunology , 8th ed., Daniel P. Stites, Abba I. Terr, and Tristram G. Parslow (eds.), Appleton & Lange, Norwalk, CT, 1994, p. 71 and chap. 6.

[0113] Light chains from any vertebrate species can be assigned to one of two clearly distinct types, called kappa ("κ") and lambda ("λ"), based on the amino acid sequence of the light chain constant domain. Immunoglobulin heavy chain (CH) constant domains can be assigned to different classes or isotypes based on their amino acid sequence. There are five classes of immunoglobulins: IgA, IgD, IgE, IgG, and IgM, whose heavy chains are designated alpha ("α"), delta ("δ"), ipsilon ("ε"), gamma ("γ"), and muon ("μ"), respectively. The gamma and alpha classes are further divided into subclasses (isotypes) based on relatively minor differences in CH sequence and function; for example, humans express the following subclasses: IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2. The subunit structures and three-dimensional configurations of different classes of immunoglobulins are well known and generally described in, for example, Abbas et al., Cellular and Molecular Immunology, 4th ed. (WB Saunders Co., 2000).

[0114] "Antibodies (such as the anti-MS4A4A antibodies of the present disclosure)" variable region "or" variable domain " refers to the amino terminal domain of the heavy or light chain of the antibody. The variable domains of the heavy and light chains may be referred to as "V H ” and “V LThese domains are generally the most variable parts of an antibody (relative to other antibodies of the same class) and contain the antigen-binding site.

[0115] the term" variable " refers to the fact that certain segments of the variable domains differ extensively in sequence between antibodies, such as the anti-MS4A4A antibodies of the present disclosure. The variable domain mediates antigen binding and defines the specificity of a particular antibody for its particular antigen. However, the variability is not evenly distributed across the span of the variable domain. Instead, it is concentrated in three segments called hypervariable regions (HVRs) in both the light and heavy chain variable domains. The more highly conserved portions of the variable domains are called framework regions (FRs). The variable domains of native heavy and light chains each comprise four FR regions connected by three HVRs, which largely adopt a β-sheet configuration. The HVRs form loops connecting, and in some cases forming part of, the β-sheet structure. The HVRs in each chain are held in close proximity by the FR regions and, together with the HVRs from the other chain, contribute to the formation of the antibody's antigen-binding site (see Kabat et al., 2002). Sequences of Immunological Interest , 5th ed., National Institute of Health, Bethesda, MD (1991). The constant domains are not involved directly in binding an antibody to an antigen, but exhibit various effector functions, such as enabling the antibody to participate in antibody-dependent cellular toxicity.

[0116] As used herein, the term " Monoclonal antibodies "Monoclonal" refers to an antibody (such as the monoclonal anti-MS4A4A antibody of the present disclosure) obtained from a population of substantially homogeneous antibodies, i.e., the individual antibodies comprising the population are identical except for possible naturally occurring mutations and / or post-translational modifications (e.g., isomerization, amidation, etc.) that may be present in minor amounts. Monoclonal antibodies are highly specific, being directed against a single antigenic site. In contrast to polyclonal antibody preparations, which typically include different antibodies directed against different determinants (epitopes), each monoclonal antibody is directed against a single determinant on the antigen. In addition to its specificity, monoclonal antibodies are advantageous in that they are synthesized by hybridoma cultures without contamination by other immunoglobulins. The modifier "monoclonal" indicates the characteristic of the antibody as being obtained from a substantially homogeneous population of antibodies and is not to be construed as requiring production of the antibody by any particular method. For example, the monoclonal antibodies used in accordance with the present disclosure can be made by a variety of techniques, including, for example, the hybridoma method, recombinant DNA methods, and techniques for producing human or human-like antibodies in animals that have part or all of the human immunoglobulin loci or genes encoding human immunoglobulin sequences.

[0117] the term" Full-length antibody "," intact antibodies "or" Full antibody" are used interchangeably to refer to an antibody (such as an anti-MS4A4A antibody of the present disclosure) in its substantially intact form, as opposed to an antibody fragment. Specifically, whole antibodies include those having heavy and light chains and including an Fc region. The constant domains may be native sequence constant domains (e.g., human native sequence constant domains) or amino acid sequence variants thereof. In some cases, an intact antibody may have one or more effector functions.

[0118] “ Antibody fragments "Antibody fragments" refer to molecules comprising a portion of an intact antibody, other than an intact antibody, that binds to the antigen to which the intact antibody binds. Examples of antibody fragments include Fab, Fab', F(ab')2, and Fv fragments; diabodies; linear antibodies (see U.S. Pat. No. 5,641,870, Example 2; Zapata et al., Protein Eng. 8(10):1057-1062(1995)); single-chain antibody molecules and multispecific antibodies formed from antibody fragments.

[0119] Papain digestion of antibodies (such as the anti-MS4A4A antibodies of the present disclosure) produces two identical antigen-binding fragments (referred to as " Fab "Fragment) and a remnant" Fc " fragment, whose name reflects the ability to crystallize easily. The Fab fragment consists of the entire light chain and the variable region domain of the heavy chain (V H ) and the first constant domain of a heavy chain (C H 1) Composition. Each Fab fragment is monovalent with respect to antigen binding, i.e., it has a single antigen binding site. Pepsin treatment of an antibody produces a single large F(ab')2 fragment that roughly corresponds to two disulfide-linked Fab fragments with different antigen binding activities and is still able to cross-link antigen. Fab' fragments differ from Fab fragments in that the C H The carboxyl terminus of the constant domain has several additional residues, including one or more cysteines from the antibody hinge region. Herein, Fab'-SH is the designation for Fab' in which one or more cysteine ​​residues of the constant domains bear a free thiol group. F(ab')2 antibody fragments originally were produced as pairs of Fab' fragments with hinge cysteines between them. Other chemical couplings of antibody fragments are also known.

[0120] The Fc fragment comprises the carboxyl-terminal portions of two heavy chains held together by disulfide bonds.The effector functions of an antibody are determined by sequences in the Fc region, which is also recognized by Fc receptors (FcR) found on certain types of cells.

[0121] "Antibodies (such as the anti-MS4A4A antibodies of the present disclosure)" Functional snippets"Antibody fragments" generally comprise a portion of an intact antibody, which portion generally includes the antigen-binding or variable region of the intact antibody or an Fc region of an antibody that retains or has improved FcR binding ability. Examples of antibody fragments include linear antibodies, single-chain antibody molecules, and multispecific antibodies formed from antibody fragments.

[0122] the term" Double Antibody " refers to small antibody fragments prepared by: using V H With V L sFv fragments (see previous paragraph) are constructed using short linkers (about 5-10 residues) between the domains, which allow interchain, rather than intrachain, pairing of the variable domains, thus generating bivalent fragments, i.e., fragments with two antigen-binding sites. Bispecific diabodies are heterodimers of two "crossover" sFv fragments, in which the V domains of the two antibodies are bound to each other. H and V L Domains are present on different polypeptide chains.

[0123] As used herein, “ chimeric antibodies " refers to an antibody (immunoglobulin) in which a portion of the heavy chain and / or light chain is identical or homologous to the corresponding sequence in an antibody derived from a particular species or belonging to a particular antibody class or subclass, and the remainder of the chain is identical or homologous to the corresponding sequence in an antibody derived from another species or belonging to another antibody class or subclass (such as the chimeric anti-MS4A4A antibody of the present disclosure), as well as fragments of such antibodies, as long as they exhibit the desired biological activity. The chimeric antibodies contemplated herein include PRIMATIZED ® Antibodies, wherein the antigen-binding region of the antibody is derived from an antibody produced by, for example, immunizing macaques with an antigen of interest. As used herein, "humanized antibody" is used as a subset of "chimeric antibody."

[0124] Non-human (e.g., mouse) antibodies Humanization "Humanized forms" (such as humanized forms of the anti-MS4A4A antibodies of the present disclosure) are chimeric antibodies comprising amino acid residues from non-human HVRs and amino acid residues from human FRs. In certain embodiments, a humanized antibody will comprise substantially all of at least one, and typically two, variable domains, in which all or substantially all of each HVR (e.g., CDRs) correspond to those of a non-human antibody, and all or substantially all of each FR corresponds to those of a human antibody. A humanized antibody optionally may comprise at least a portion of an antibody constant region derived from a human antibody. A "humanized form" of an antibody (e.g., a non-human antibody) is one that has undergone humanization.

[0125] “ Human antibodies"A human antibody" is an antibody having an amino acid sequence that corresponds to the amino acid sequence of an antibody produced by a human and / or prepared using any of the techniques for making human antibodies as disclosed herein (such as the anti-MS4A4A antibodies of the present disclosure). This definition of a human antibody specifically excludes humanized antibodies comprising non-human antigen-binding residues. Human antibodies can be produced using various techniques known in the art, including phage display libraries and yeast display libraries. Human antibodies can be produced by administering an antigen to transgenic animals (e.g., immunized xenogeneic mice) that have been modified to produce such antibodies in response to antigenic challenge, but whose endogenous loci have been disabled, as well as through human B-cell hybridoma technology.

[0126] the term" Hypervariable region "," HVR "or" HV "HVRs" as used herein refers to the regions of an antibody variable domain (such as the variable domains of the anti-MS4A4A antibodies of the present disclosure) that are hypervariable in sequence and / or form structurally defined loops. Generally, antibodies comprise six HVRs; three in V H (H1, H2, H3), and three in V L (L1, L2, L3). In natural antibodies, H3 and L3 display most of the diversity of the six HVRs, and H3 in particular is believed to play a unique role in conferring fine specificity to antibodies. Natural camelid antibodies composed only of heavy chains are functional and stable in the absence of light chains.

[0127] Various descriptions of HVRs are used and encompassed herein. In some embodiments, HVRs may be Kabat complementarity determining regions (CDRs) based on sequence variability and are most commonly used (Kabat et al., supra). In some embodiments, HVRs may be Chothia CDRs. Chothia refers to the position of the structural loops (Chothia and Lesk, J. Mol. Biol. 196:901-917 (1987). In some embodiments, the HVR may be an AbM HVR. The AbM HVR represents a compromise between the Kabat CDR and the Chothia structural loops and is used in Oxford Molecular's AbM antibody modeling software. In some embodiments, the HVR may be a "contact" HVR. The "contact" HVR is based on analysis of available complex crystal structures. The residues from each of these HVRs are shown below.

[0128]

[0129] The HVRs may comprise "extended HVRs" as follows: 24-36 or 24-34 (L1), 46-56 or 50-56 (L2), and 89-97 or 89-96 (L3) in VL and 26-35 (H1), 50-65 or 49-65 (preferred embodiments) (H2), and 93-102, 94-102, or 95-102 (H3) in VH. For each of these extended HVR definitions, the variable domain residues are numbered according to Kabat et al. (supra).

[0130] “ frame "or" FR "HVR" residues are those variable domain residues other than the HVR residues as herein defined.

[0131] As used herein, “ Recipient Framework " is a V sequence comprising a human immunoglobulin framework or a human consensus framework L or V H The acceptor human framework "derived from" a human immunoglobulin framework or a human consensus framework may comprise the same amino acid sequence thereof, or it may comprise pre-existing amino acid sequence changes. In some embodiments, the number of pre-existing amino acid changes is 10 or less, 9 or less, 8 or less, 7 or less, 6 or less, 5 or less, 4 or less, 3 or less or 2 or less. When pre-existing amino acid changes are present in VH, those changes preferably occur only at three, two or one of positions 71H, 73H and 78H; for example, the amino acid residues at those positions may be 71A, 73T and / or 78A. In one embodiment, the sequence of the acceptor human framework for the VL is identical to that of the VH. L The human immunoglobulin framework sequences or human consensus framework sequences are identical.

[0132] “ Human-shared framework " stands for human immunoglobulin V L or V H The framework sequence is selected from the most common amino acid residues in the framework. L or V H The choice of framework sequence is from a subset of variable domain sequences. Typically, the subset of sequences is as described by Kabat et al. Sequences of Proteins of Immunological Interest , 5th ed., Public Health Service, National Institutes of Health, Bethesda, MD (1991). Examples include, for V L For V, the subgroup may be subgroup κI, κII, κIII or κIV as in Kabat et al. (supra). HFor example, the subgroup may be subgroup I, subgroup II, or subgroup III as in Kabat et al. (supra).

[0133] " Amino acid modification " refers to a substitution or deletion of the specified residue, or an insertion of at least one amino acid residue adjacent to the specified residue. "Adjacent" to the specified residue means insertion within one to two residues thereof. The insertion may be made at the N-terminus or C-terminus of the specified residue. The preferred amino acid modification herein is substitution.

[0134] “ Affinity Maturation "Antibodies, such as the affinity matured anti-MS4A4A antibodies of the present disclosure, are antibodies with one or more alterations in one or more HVRs thereof that result in an improvement in the affinity of the antibody for the antigen, compared to a parent antibody that does not possess those alterations. In one embodiment, the affinity matured antibody has nanomolar or even picomolar affinities for the target antigen. Affinity matured antibodies are produced by procedures known in the art. For example, Marks et al., Bio / Technology 10:779-783 (1992) describes the H and V L Affinity maturation by domain shuffling. Random mutagenesis of HVR and / or framework residues is described, for example, in Barbas et al. Proc Nat. Acad. Sci. USA 91:3809-3813 (1994); Schier et al., Gene 169:147-155 (1995); Yelton et al., J. Immunol. 155:1994-2004 (1995); Jackson et al. J. Immunol. 154(7):3310-9 (1995); and Hawkins et al., J. Mol. Biol. 226:889-896 (1992).

[0135] “ Fv " is the smallest antibody fragment that contains a complete antigen recognition and binding site. This fragment consists of a dimer of one heavy chain variable region domain and one light chain variable region domain in a tight non-covalent association. Six hypervariable loops are generated from the folding of these two domains (3 loops each from the H chain and the L chain), which contribute amino acid residues for antigen binding and confer antigen binding specificity to the antibody. However, even if a single variable domain (or half of an Fv, which contains only three HVRs specific for an antigen) has the ability to recognize and bind to an antigen, its affinity is lower than that of the entire binding site.

[0136] “ single-chain Fv ” is also abbreviated as “ FWf "or" scFv ", which is an antibody fragment comprising the VH and VL antibody domains linked into a single polypeptide chain. Preferably, the sFv polypeptide also comprises a V H With V L A polypeptide linker between the domains that enables the sFv to form the desired antigen-binding structure.

[0137] Antibody" Effector function ” refers to those biological activities attributable to the Fc region (a native sequence Fc region or amino acid sequence variant Fc region) of an antibody, and varies with the antibody isotype.

[0138] the term" Fc region " is used herein to define the C-terminal region of an immunoglobulin heavy chain, which includes native sequence Fc regions and variant Fc regions. Although the boundaries of the immunoglobulin heavy chain Fc region can vary, the human IgG heavy chain Fc region is generally defined as stretching from an amino acid residue at position Cys226, or from Pro230, to its carboxyl terminus. The C-terminal lysine of the Fc region (residue 447 according to the EU numbering system) can be removed, for example, during production or purification of the antibody, or by recombinant engineering of the nucleic acid encoding the antibody heavy chain. Thus, a repertoire of intact antibodies can include antibody populations with all K447 residues removed, antibody populations with the K447 residue not removed, and antibody populations having a mixture of antibodies with and without the K447 residue. Suitable native sequence Fc regions for use in the antibodies of the present disclosure include human IgG1, IgG2, IgG3, and IgG4.

[0139] “ native sequence Fc region " comprises an amino acid sequence identical to the amino acid sequence of an Fc region found in nature. Native sequence human Fc regions include native sequence human IgG1 Fc region (non-A and A allotypes); native sequence human IgG2 Fc region; native sequence human IgG3 Fc region; and native sequence human IgG4 Fc region, as well as naturally occurring variants thereof.

[0140] “ Variant Fc region " comprises an amino acid sequence that differs from a native sequence Fc region by virtue of at least one amino acid modification, preferably one or more amino acid substitutions. Preferably, the variant Fc region has at least one amino acid substitution compared to a native sequence Fc region or compared to the Fc region of a parent polypeptide, e.g., from about one to about ten amino acid substitutions in a native sequence Fc region or the Fc region of a parent polypeptide, and preferably from about one to about five amino acid substitutions. The variant Fc region herein will preferably have at least about 80% homology to a native sequence Fc region and / or to the Fc region of a parent polypeptide, and most preferably at least about 90% homology thereto, and more preferably at least about 95% homology thereto.

[0141] “ Fc receptors "or" FqV " describes a receptor that binds to the Fc region of an antibody. The preferred FcR is a native sequence human FcR. Furthermore, the preferred FcR is an FcR that binds to an IgG antibody (gamma receptor) and includes receptors of the FcγRI, FcγRII, and FcγRIII subclasses, including allelic variants and alternatively spliced ​​forms of these receptors, the FcγRII receptors including FcγRIIA ("activating receptor") and FcγRIIB ("inhibiting receptor"), both of which have similar amino acid sequences that differ primarily in their cytoplasmic domains. The activating receptor FcγRIIA contains an immunoreceptor tyrosine-based activation motif ("ITAM") in its cytoplasmic domain. The inhibitory receptor FcγRIIB contains an immunoreceptor tyrosine-based inhibition motif ("ITIM") in its cytoplasmic domain. The term "FcR" herein encompasses other FcRs, including those identified in the future. FcRs may also extend the serum half-life of antibodies.

[0142] As used herein, "a peptide, polypeptide, or antibody sequence" Amino acid sequence identity percentage (%) "and" Homologous sex " refers to the percentage of amino acid residues in a candidate sequence that are identical with the amino acid residues in a specified peptide or polypeptide sequence, after aligning the sequences and introducing gaps (if necessary) to achieve the maximum percent sequence identity, and not considering any conservative substitutions as part of the sequence identity. For purposes of determining percent amino acid sequence identity, alignment can be achieved in various ways that are within the skill in the art, for instance, using publicly available computer software such as BLAST, BLAST-2, ALIGN, or MEGALIGN TM (DNASTAR) software. Those skilled in the art can determine appropriate parameters for measuring alignment, including any algorithms known in the art needed to achieve maximal alignment over the full length of the sequences being compared.

[0143] the term" compete"When used in the context of antibodies that compete for the same epitope (e.g., neutralizing antibodies), refers to competition between antibodies as determined by an assay in which the test antibody prevents or inhibits (e.g., reduces) specific binding of a reference molecule (e.g., a ligand or reference antibody) to a common antigen (e.g., MS4A4A or a fragment thereof). Various types of competitive binding assays can be used to determine whether antibodies compete with each other, for example: solid phase direct or indirect radioimmunoassays (RIAs), solid phase direct or indirect enzyme immunoassays (EIAs), sandwich competition assays (see, e.g., Stahli et al., 1983, Methods in Enzymology 9:242-253); solid phase direct biotin-avidin EIA (see, e.g., Kirkland et al., 1986, J. Immunol. 137:3614-3619); solid phase direct labeled assays, solid phase direct labeled sandwich assays (see, e.g., Harlow and Lane, 1988, Antibodies, A Laboratory Manual, Cold Spring Harbor Laboratory Press, 1992 ... Press); solid phase direct labeling RIA using I-125 labeling (see, e.g., Morel et al., 1988, Molec. Immunol. 25:7-15); solid phase direct biotin-avidin EIA (see, e.g., Cheung et al., 1990, Virology 176:546-552); and direct labeling RIA (Moldenhauer et al., 1990, Scand. J. Immunol. 32:77-82). Typically, such analysis involves the use of purified antigens bound to a solid surface or cell carrying any of these unlabeled test antibodies and labeled reference antibodies. Competitive inhibition is measured by determining the amount of labeling bound to the solid surface or cell in the presence of the test antibody. Typically, the test antibody is present in excess. Antibodies identified by competitive assays (competing antibodies) include antibodies that bind to the same epitope as the reference antibody and antibodies that bind to adjacent epitopes that are sufficiently close to the epitope bound by the reference antibody due to steric hindrance. Additional details about methods for determining competitive binding are provided in Examples herein. Typically, when the competitive antibody is present in excess, it will inhibit (e.g., reduce) the specific binding of the reference antibody to the common antigen by at least 20%, 30%, 40%, 50%, 60%, 70%, 80%, 85%, 90%, 95%, 97.5% and / or approach 100%.

[0144] As used herein, a "relationship" between an MS4A4A polypeptide and a second polypeptide interaction" encompasses, but is not limited to, protein-protein interactions, physical interactions, chemical interactions, binding, covalent binding, and ionic binding. As used herein, an antibody "inhibits an interaction between two polypeptides" when it disrupts, reduces, or completely eliminates the interaction between the two polypeptides. An antibody of the present disclosure "inhibits an interaction between two polypeptides" when it binds to one of the two polypeptides. In some embodiments, the interaction can be inhibited by at least about any of 20%, 30%, 40%, 50%, 60%, 70%, 80%, 85%, 90%, 95%, 97.5%, and / or close to 100%.

[0145] the term" Epitope " includes any determinant capable of being bound by an antibody. An epitope is a region of an antigen that is bound by an antibody that targets the antigen, and when the antigen is a polypeptide, it includes specific amino acids that directly contact the antibody. In most cases, epitopes are located on polypeptides, but in some cases they can be located on other types of molecules (such as nucleic acids). Epitope determinants may include chemically active surface groups of molecules (such as amino acids, sugar side chains, phosphoryl or sulfonyl groups) and may have specific three-dimensional structural characteristics and / or specific charge characteristics. Generally, an antibody specific for a particular target antigen will preferentially recognize an epitope on the target antigen in a complex mixture of polypeptides and / or macromolecules.

[0146] “ agonists "Antibodies or" Activation An "antibody" is an antibody that induces (e.g., increases) one or more activities or functions of an antigen upon binding of the antibody to the antigen.

[0147] “ antagonists "Antibodies or" Blocking "Antibodies or" Inhibitory An antibody is an antibody that reduces, inhibits, and / or eliminates (e.g., reduces) the binding of an antigen to one or more ligands and / or reduces, inhibits, and / or eliminates (e.g., reduces) one or more activities or functions of an antigen after the antibody binds to the antigen. In some embodiments, an antagonist antibody, a blocking antibody, or an inhibitory antibody substantially or completely inhibits the binding of an antigen to one or more ligands and / or one or more activities or functions of an antigen.

[0148] “ SeparationAn antibody, such as an isolated anti-MS4A4A antibody of the present disclosure, is one that has been identified, separated, and / or recovered from a component of its production environment (e.g., natural or recombinant). Preferably, the isolated antibody is free from all other contaminating components from its production environment. Contaminating components from the environment in which the isolated antibody is produced (such as those produced by recombinant transfected cells) are materials that would generally interfere with research, diagnostic, or therapeutic uses of the antibody and may include enzymes, hormones, and other proteinaceous or nonproteinaceous solutes. In preferred embodiments, the antibody is purified to the following degrees: (1) greater than 95% by weight, and in some embodiments greater than 99% by weight, as determined by, for example, the Lowry method; (2) to a degree sufficient to obtain at least 15 N-terminal or internal amino acid sequence residues by use of a spinning cup sequenator, or (3) by SDS-PAGE under non-reducing or reducing conditions using Coomassie Blue. The homogeneity of the isolated polypeptide or antibody is preferably blue) or preferably silver stain. Isolation of antibodies includes the in situ antibodies in the recombinant T cells, because at least one component in the antibody natural environment will not exist. However, isolation of polypeptide or antibody will usually be prepared by at least one purification step.

[0149] "a" encoding an antibody (such as an anti-MS4A4A antibody of the present disclosure) Separation A nucleic acid molecule is one that has been identified and separated from at least one contaminating nucleic acid molecule with which it is normally associated in its production environment. Preferably, an isolated nucleic acid is free from association with all components associated with the production environment. An isolated nucleic acid molecule encoding the polypeptides and antibodies herein is in a form other than that in which it is found in nature. Thus, an isolated nucleic acid molecule is distinguished from nucleic acids encoding the polypeptides and antibodies herein that are naturally present in cells.

[0150] As used herein, the term " carrier " is intended to refer to a nucleic acid molecule capable of transporting another nucleic acid to which it is linked. One type of vector is a "plasmid," which refers to a circular double-stranded DNA into which additional DNA segments can be ligated. Another type of vector is a bacteriophage vector. Another type of vector is a viral vector, where additional DNA segments can be ligated into the viral genome. Certain vectors are capable of autonomous replication in a host cell into which they have been introduced (e.g., bacterial vectors having a bacterial origin of replication and episomal mammalian vectors). Other vectors (e.g., non-episomal mammalian vectors) can be integrated into the genome of a host cell upon introduction into the host cell and are thereby replicated along with the host genome. Furthermore, certain vectors are capable of directing the expression of genes to which they are operably linked. Such vectors are referred to herein as "recombinant expression vectors," or simply "expression vectors." In general, expression vectors useful in recombinant DNA techniques are often in the form of plasmids. In this specification, "plasmid" and "vector" are used interchangeably, as plasmids are the most commonly used form of vectors.

[0151] As used interchangeably herein, “ polynucleotides "or" Nucleic Acids " refers to polymers of nucleotides of any length and includes DNA and RNA. The nucleotides can be deoxyribonucleotides, ribonucleotides, modified nucleotides or bases and / or their analogs or any substrate that can be incorporated into a polymer by DNA or RNA polymerase or by a synthetic reaction.

[0152] “ host cells "Host cells" include any individual cell or cell culture that can become or has become a recipient of a vector for incorporating a polynucleotide insert. Host cells include progeny of a single host cell, and the progeny may not necessarily be completely identical (in morphology or in genomic DNA complement) to the original parent cell due to natural, accidental, or deliberate mutation. Host cells include cells transfected in vivo with a polynucleotide of the disclosure.

[0153] As used herein, the terms "membrane TREM2," "plasma membrane TREM2," and "cell surface TREM2" are used interchangeably.

[0154] As used herein, “ carrier " includes pharmaceutically acceptable carriers, excipients, or stabilizers that are non-toxic to the cells or mammals exposed thereto at the dosages and concentrations employed.

[0155] As used herein, the term “ prevention " includes providing prevention of the occurrence or recurrence of a particular disease, disorder or condition in an individual. An individual may be predisposed to, susceptible to, or at risk of developing a particular disease, disorder or condition but has not yet been diagnosed with the disease, disorder or condition.

[0156] As used herein, a person is at risk of developing a particular disease, condition, or disorder. Under risk An individual who is "at risk" may or may not have detectable disease or disease symptoms and may or may not have exhibited detectable disease or disease symptoms prior to the treatment methods described herein. As known in the art, "at risk" means that the individual has one or more risk factors, which are measurable parameters that correlate with the development of a particular disease, disorder, or condition. An individual who has one or more of these risk factors has a higher probability of developing the particular disease, disorder, or condition than an individual who does not have one or more of these risk factors.

[0157] As used herein, the term “ treat"Treatment" refers to a clinical intervention designed to alter the natural course of the clinical pathological course of the treated individual. Desirable therapeutic effects include reducing the rate of progression of the specific disease, disorder or condition, ameliorating or lessening its pathological state, and alleviating or improving its prognosis. For example, a subject is successfully "treated" if one or more symptoms associated with a specific disease, disorder or condition are alleviated or eliminated.

[0158] “ effective dose "" refers to an amount effective to achieve the desired therapeutic or preventive result, at least at the dosage and for the period of time required. An effective amount can be provided in one or more administrations. An effective amount herein may vary according to factors such as the disease state, the age, sex, and weight of the individual, and the ability of the treatment to elicit a desired response in the individual. An effective amount is also one in which any toxic or detrimental effects of the treatment are outweighed by the beneficial effects of the treatment. For prophylactic use, beneficial or desired results include results such as eliminating or reducing the risk of, lessening the severity of, or delaying the onset of the disease, including its biochemical, histological, and / or behavioral symptoms, its complications, and intermediate pathological phenotypes that manifest during the course of the disease. For therapeutic use, beneficial or desired results include results such as Such as the following clinical results: such as reducing one or more symptoms caused by the disease, improving the quality of life of the patient, reducing the dosage of other drugs required for treating the disease, and enhancing the effect of another drug via targeting, delaying the progression of the disease and / or prolonging survival. The effective amount of a drug, compound, or pharmaceutical composition is an amount sufficient to directly or indirectly achieve a preventive or therapeutic treatment. As understood in clinical situations, the effective amount of a drug, compound, or pharmaceutical composition may be achieved in conjunction with another drug, compound, or pharmaceutical composition or may not be achieved in conjunction with it. Therefore, it can be considered that an "effective amount" is performed in the context of administering one or more therapeutic agents, and if the desired result can be achieved or achieved in conjunction with one or more other agents, it can be considered that a single agent is given in an effective amount.

[0159] For treatment, prevention or risk reduction purposes individual " refers to any animal classified as a mammal, including humans, domestic and farm animals, and zoo animals, sport animals, or pets, such as dogs, horses, rabbits, cows, pigs, hamsters, gerbils, mice, ferrets, rats, cats, and the like. In some embodiments, the subject is a human.

[0160] As used herein, "combination with another compound or composition" joint "Administration" includes simultaneous administration and / or administration at different times. Co-administration also encompasses administration as a co-formulation or as separate compositions, including administration at different dosing frequencies or intervals and using the same route of administration or different routes of administration. In some embodiments, co-administration is administration as part of the same treatment regimen.

[0161] As used herein, the term " about ” refers to the usual error range of the corresponding value that is easily known to those skilled in the art. about "A value or parameter includes (and describes) embodiments involving the value or parameter itself.

[0162] As used herein and in the appended claims, the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise. For example, a reference to an "antibody" is a reference to one to a number of antibodies, such as molar amounts, and includes equivalents thereof known to those skilled in the art, etc.

[0163] It is to be understood that aspects and embodiments of the present disclosure described herein include "comprising," "consisting of," and "consisting essentially of" aspects and embodiments.

[0164] II. Anti-MS4A4A Antibodies

[0165] Anti-MS4A4A antibodies are provided herein. The antibodies provided herein can be used, for example, to diagnose or treat MS4A4A-associated disorders.

[0166] In one aspect, the present disclosure provides isolated (e.g., monoclonal) antibodies that bind to an epitope within the MS4A4A proteins of the present disclosure. The MS4A4A proteins of the present disclosure include, but are not limited to, mammalian MS4A4A proteins, human MS4A4A proteins, mouse MS4A4A proteins, and cynomolgus monkey MS4A4A proteins. The MS4A4A proteins of the present disclosure include naturally occurring variants of MS4A4A.

[0167] Human MS4A4A is a 239-amino acid protein encoding a membrane glycoprotein. The amino acid sequence of human MS4A4A is shown in SEQ ID NO: 1:

[0168] MHQTYSRHCRPEESTFSAAMTTMQGMEQAMPGAGPGVPQLGNMAVIHSHLWKGLQEKFLKGEPKVLGVVQILTALMSLSMGITMMCMASNTYGSNPISVYIGYTIWGSVMFIISGSLSI AAGIRTTKGLVRGSLGMNITSSVLAASGILINTFSLAFYSFHHPYCNYYGNSNNCHGTMSILMGLDGMVLLLSVLEFCIAVSLSAFGCKVLCCTPGGVVLILPSHSHMAETASPTPLNEV

[0169] In addition, the amino acid sequence of mouse MS4A4A is shown in SEQ ID NO: 2:

[0170] MLVIQGTEQSALEAGYGAQQNGQPLYVNSHSWKRMTEKFLKGEPKILGIVQIVIAIMNLSIGIMMIATVSTGEIPPSSVYIGYPIWGSLMFIISGSFSIVAGRRTTKGLVRSSLGLN ITSSVFAFSGIVISSLSPGIYSFHVYYCTYRGSSEGCHMTLSILMGLDIVVVVLSVLEFCIGVSLSAFGCRVMCCNPGGVMIIMPSNPTKAETANPVTLQSGLMPPEHQERNVPENMH

[0171] In addition, the amino acid sequence of cynomolgus (cynomolgus, cyno) MS4A4A is shown in SEQ ID NO: 3:

[0172] HQTYRRHCRPEESTFSAAMTTMQGMEQATPGAGPGVPQLGNMAVVHSHLWKGLQEKFLKGEPKVLGVVQILIALMSLSMGITMMMCVAFSAYGHYPISVYIGYTIWGSVMFIISGSLSIA AGIRTTKGLVRGSLGMNITSSVLAVSAILINTISLTIYSFYHRYCNYYGNPNNCHGTVSILMGMDGMVLLLSVLEFCIAVSLSAFGCKAICCTPGGVVLIIPSNSHMAEAAPLTPLNEV

[0173] In some embodiments, MS4A4A is expressed in cells. In some embodiments, MS4A4A is expressed in myeloid cells. In some embodiments, MS4A4A is expressed in brain cells. In some embodiments, MS4A4A is expressed in astrocytes, including but not limited to mature astrocytes. In some embodiments, MS4A4A is expressed in oligodendrocytes. In some embodiments, MS4A4A is expressed in microglia. In some embodiments, MS4A4A is expressed in immune cells, including but not limited to macrophages, eosinophils, mast cells, dendritic cells, natural killer cells, neutrophils, and T cells. In some embodiments, MS4A4A is expressed in olfactory cells. In some embodiments, MS4A4A is expressed on the surface of cells.

[0174] The MS4A4A proteins of the present disclosure include several domains, including, but not limited to, a cytoplasmic domain (amino acid residues 1-64 of human MS4A4A; see SEQ ID NO: 1); a transmembrane domain (amino acid residues 65-85 of human MS4A4A); an extracellular domain (extracellular domain 1; ECL1), which corresponds to amino acid residues 86-98 of human MS4A4A; a transmembrane domain (amino acid residues 99-119 of human MS4A4A); a cytoplasmic domain (amino acid residues 120-137 of human MS4A4A); a transmembrane domain (amino acid residues 138-158 of human MS4A4A); an extracellular domain (extracellular domain 2; ECL2), which corresponds to amino acid residues 159-179; a transmembrane domain (amino acid residues 180-200 of human MS4A4A); and a cytoplasmic domain (amino acid residues 201-239 of human MS4A4A). In addition, the MS4A4A proteins of the present disclosure are expressed in a variety of tissues and cells, including but not limited to the brain, neurons, glial cells, endothelial cells, pericytes, pericytes, and the like.

[0175] Role of MS4A4A in macrophage and microglial cell function

[0176] The macrophages and myeloid cells (such as microglial cells) of the central nervous system (CNS) have inherent plasticity in their phenotype and function. In vitro macrophages can be divided into M1 macrophages and M2 macrophages, which have different phagocytic and inflammatory potentials, phenotypes and activities. For example, in peripheral organs, it is believed that macrophages with an M1 phenotype have proinflammatory and antimicrobial phenotypes and functions, while it is believed that macrophages with an M2 phenotype are in a more stable state, with an anti-inflammatory phenotype and function.

[0177] Compared to M1 markers (e.g., CD16, MHC class II, CD86), microglia associated with healthy steady-state conditions express more M2 markers (e.g., CD200R, CD163, and CD115) on their cell surface (Ginhoux and Prinz, 2015, Cold Spring Harb Perspect Biol, 7: a020537). However, in both Alzheimer's disease mouse models and human Alzheimer's disease, disease-associated microglia (DAMs) are in a proinflammatory or activated state. Disease-associated microglia in a proinflammatory or activated state are considered beneficial because they play an active role in reducing the pathology associated with Alzheimer's disease and other neurodegenerative disorders.

[0178] MS4A4A expression is elevated in M2 macrophages in vitro, and it has been shown that MS4A4A is a novel cell surface marker for M2 macrophages. In addition, MS4A4A has also been shown to regulate the cell surface trafficking of cKit on mast cells, suggesting a role for MS4A4A in regulating mast cell degranulation and survival (Cruse et al., 2015, Molecular Biol Cell, 26: 1711-1727). Taken together, these reported findings suggest that targeting MS4A4A can affect the recycling, expression, and / or degradation of various macrophage surface receptors associated with the M1 and M2 macrophage phenotypes, thereby affecting their function and activity.

[0179] The anti-MS4A4A antibodies of the present disclosure affect the expression of M2 macrophage surface markers. Specifically, the anti-MS4A4A antibodies of the present disclosure reduce the cell surface expression of CD200R, Dectin-1, and CD163 in macrophages, suggesting that the anti-MS4A4A antibodies modulate macrophage polarization, function, and / or activity by reducing the expression of M2 macrophage surface receptors. The anti-MS4A4A antibodies of the present disclosure reduce M2 macrophage surface receptors, suggesting that the anti-MS4A4A antibodies of the present disclosure effectively alter the physiological state of microglia to a more protective phenotype, such as to a more proinflammatory or more activated state (e.g., to a more M1-biased phenotype). Therefore, the anti-MS4A4A antibodies of the present disclosure may be used to treat Alzheimer's disease and other neurodegenerative disorders, in part by altering the phenotype of macrophages and microglia to a proinflammatory and activated state.

[0180] The anti-MS4A4A antibodies disclosed herein affect the activation state of microglia. The anti-MS4A4A antibodies disclosed herein increase mRNA levels of proteins associated with microglial activation (e.g., C-type lectin domain family 7 member A (CLEC7A)), microglial migration (e.g., inositol 1,4,5-triphosphate receptor 2 (ITPR2)), and microglial proliferation (e.g., antigen KI-67 (MKI67)) in vivo. In addition, the anti-MS4A4A antibodies disclosed herein increase mRNA levels of microglial activation markers IL1RN, SPP1, and PLCG2. The anti-MS4A4A antibodies disclosed herein also reduce mRNA levels of steady-state microglial markers purinergic receptor P2RY12 and CX3C chemokine receptor 1 (CX3CR1). Thus, the anti-MS4A4A antibodies of the present disclosure effectively activate microglia in vivo, as demonstrated by increased levels of various mRNAs for proteins associated with microglial activation, migration, and proliferation, and decreased levels of various mRNAs for proteins associated with microglial homeostasis.

[0181] MS4A4A and TREM2 expression

[0182] Neurodegenerative diseases are characterized, in part, by immune deficiencies in the central nervous system (CNS). For example, it is believed that a reduction in the viability and function of the CNS myeloid cell compartment (including but not limited to microglial cells) contributes to susceptibility to neurodegenerative conditions such as Alzheimer's disease. Pharmacological interventions that increase the viability and / or function of myeloid cells would provide effective treatments to improve the onset, severity, or progression of such neurodegenerative diseases and conditions.

[0183] Triggering receptor expressed on myeloid cells -2 (TREM2) is an immunoglobulin-like receptor expressed mainly on myeloid cells, such as macrophages, dendritic cells, monocytes, Langerhans cells of the skin (Langerhans cell), Kupffer cells (Kupffer cell), osteoclasts and microglia. During experimental autoimmune encephalomyelitis and Alzheimer's disease, TREM2 is highly expressed on microglia and infiltrating macrophages in CNS (Piccio et al., 2007, Eur J Immunol, 37: 1290-1301; Wang, 2015, Cell, 160: 1061-1071). TREM2 pathway is considered a key regulator of CNS myeloid cell viability and function.

[0184] Data from human genetic studies have shown a strong genetic association between MS4A4A and TREM2 and with susceptibility to Alzheimer's disease (Piccio et al., 2016, Acta Neuropathol, 131:925-9330). Specifically, the MS4A4A allele, which is protective against Alzheimer's disease, is associated with increased levels of sTREM2 in the cerebrospinal fluid of patients.

[0185] The anti-MS4A4A antibodies of the present invention increase cellular ATP levels in macrophages, indicating that the anti-MS4A4A antibodies effectively increase, maintain, or enhance cell (e.g., macrophage, myeloid cell, microglia) viability and function. In addition, compared to the previously reported commercially available anti-MS4A4A antibody 5C12, which reduced sTREM2 levels in cultured human macrophage supernatants, the anti-MS4A4A antibodies of the present invention increased sTREM2 and mTREM2 levels in macrophages (Deming et al., 2018, bioRxiv, doi: dx doi org / 10.1101 / 352179). Since the MS4A4A protective allele for Alzheimer's disease is associated with increased sTREM2 levels, the results provided herein indicate that the anti-MS4A4A antibodies of the present invention mimic or replicate a protective phenotype in neurodegenerative diseases and disorders such as Alzheimer's disease by increasing sTREM2 and mTREM2 levels. In some embodiments, the anti-MS4A4A antibodies of the present disclosure increase cell surface expression of TREM2 in myeloid cells (e.g., macrophages, human macrophages, microglia) by at least 10%, at least 20%, at least 25%, at least 50%, at least 75%, at least 90%, at least 100%, at least 125%, at least 150%, at least 200%, or at least 250%. In some embodiments, the anti-MS4A4A antibodies of the present disclosure increase soluble TREM2 levels in myeloid cells (e.g., macrophages, human macrophages, microglia) by at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 100%.

[0186] In some embodiments, the anti-MS4A4A antibodies of the present disclosure increase mTREM protein levels in myeloid cells (e.g., macrophages, human macrophages, microglia) with an EC50 of about 0.028 μg / ml to about 0.039 μg / ml. In some embodiments, the anti-MS4A4A antibodies of the present disclosure increase sTREM protein levels in myeloid cells (e.g., macrophages, human macrophages, microglia) with an EC50 of about 0.025 μg / ml to about 0.069 μg / ml. In other embodiments, the anti-MS4A4A antibodies of the present disclosure increase ATP levels in myeloid cells (e.g., macrophages, human macrophages, microglia) with an EC50 of about 0.010 μg / ml to about 0.021 μg / ml. In some embodiments, an anti-MS4A4A antibody of the present disclosure increases ATP levels in myeloid cells (e.g., macrophages, human macrophages, microglia) to about 1.2-fold, about 1.4-fold, or about 1.7-fold the ATP level in such cells in the absence of the anti-MS4A4A antibody.

[0187] In some embodiments, the anti-MS4A4A antibodies of the present disclosure increase soluble TREM2 levels in vivo (e.g., in non-human primates or humans). In some embodiments, the anti-MS4A4A antibodies of the present disclosure increase soluble TREM2 levels in serum in vivo by at least 10%, at least 20%, at least 25%, at least 50%, at least 75%, at least 90%, at least 100%, at least 125%, or at least 150% from baseline soluble TREM2 levels in serum in vivo prior to administration of the anti-MS4A4A antibodies of the present disclosure. In some embodiments, the anti-MS4A4A antibodies of the present disclosure increase soluble TREM2 levels in serum in vivo by about 50% from baseline soluble TREM2 levels in serum in vivo prior to administration of the anti-MS4A4A antibodies of the present disclosure.

[0188] In some embodiments, the anti-MS4A4A antibodies of the present disclosure increase the level of soluble TREM2 in serum in vivo by at least 1.1-fold, at least 1.2-fold, at least 1.3-fold, at least 1.4-fold, at least 1.5-fold, at least 1.6-fold, at least 1.7-fold, at least 1.8-fold, at least 1.9-fold, or at least 2-fold the level of soluble TREM2 in serum in vivo prior to administration of the anti-MS4A4A antibodies of the present disclosure. In some embodiments, the anti-MS4A4A antibodies of the present disclosure increase the level of soluble TREM2 in serum in vivo by about 1.5-fold the level of soluble TREM2 in serum in vivo prior to administration of the anti-MS4A4A antibodies of the present disclosure.

[0189] In some embodiments, the anti-MS4A4A antibodies of the present disclosure increase soluble TREM2 levels in serum in vivo for at least 1 day, at least 2 days, at least 3 days, at least 4 days, at least 5 days, at least 6 days, at least 7 days, at least 8 days, at least 9 days, at least 10 days, at least 11 days, at least 12 days, at least 13 days, at least 14 days, at least 15 days, at least 16 days, at least 17 days, at least 18 days, at least 19 days, or at least 20 days, compared to the level of soluble TREM2 in serum in vivo before administration of the anti-MS4A4A antibodies of the present disclosure. In some embodiments, the anti-MS4A4A antibodies of the present disclosure increase soluble TREM2 levels in serum in vivo for at least 20 days, compared to the level of soluble TREM2 in serum in vivo before administration of the anti-MS4A4A antibodies of the present disclosure. In some embodiments, the anti-MS4A4A antibodies of the present disclosure increase soluble TREM2 levels in serum in vivo for at least 24 hours, at least 48 hours, at least 72 hours, at least 96 hours, at least 120 hours, at least 144 hours, at least 168 hours, at least 192 hours, at least 216 hours, at least 240 hours, at least 264 hours, at least 288 hours, at least 312 hours, at least 336 hours, at least 360 hours, at least 384 hours, at least 408 hours, at least 432 hours, at least 456 hours, or at least 480 hours, compared to the level of soluble TREM2 in serum in vivo before administration of the anti-MS4A4A antibodies of the present disclosure. In some embodiments, the anti-MS4A4A antibodies of the present disclosure increase soluble TREM2 levels in serum in vivo for at least 480 hours, compared to the level of soluble TREM2 in serum in vivo before administration of the anti-MS4A4A antibodies of the present disclosure.

[0190] In some embodiments, the anti-MS4A4A antibodies of the present disclosure increase soluble TREM2 levels in CSF in vivo by at least 10%, at least 20%, at least 25%, at least 50%, at least 75%, at least 90%, at least 100%, at least 125%, at least 150%, at least 200%, at least 225%, at least 250%, at least 275%, at least 300%, at least 325%, at least 350%, at least 375%, or at least 400% from baseline soluble TREM2 levels in CSF in vivo prior to administration of the anti-MS4A4A antibodies of the present disclosure. In some embodiments, the anti-MS4A4A antibodies of the present disclosure increase soluble TREM2 levels in CSF in vivo by about 300% from baseline soluble TREM2 levels in CSF in vivo prior to administration of the anti-MS4A4A antibodies of the present disclosure.

[0191] In some embodiments, the anti-MS4A4A antibodies of the present disclosure increase the level of soluble TREM2 in CSF in vivo by at least 1.4-fold, at least 1.6-fold, at least 1.8-fold, at least 2.0-fold, at least 2.2-fold, at least 2.4-fold, at least 2.6-fold, at least 2.8-fold, at least 3.0-fold, at least 3.2-fold, at least 3.4-fold, at least 3.6-fold, at least 3.8-fold, or at least 4.0-fold the level of soluble TREM2 in CSF in vivo prior to administration of the anti-MS4A4A antibodies of the present disclosure. In some embodiments, the anti-MS4A4A antibodies of the present disclosure increase the level of soluble TREM2 in CSF in vivo by about 2-fold to about 4-fold the level of soluble TREM2 in CSF in vivo prior to administration of the anti-MS4A4A antibodies of the present disclosure.

[0192] In some embodiments, the anti-MS4A4A antibodies of the present disclosure increase soluble TREM2 levels in CSF in vivo for at least 1 day, at least 2 days, at least 3 days, or at least 4 days, compared to the level of soluble TREM2 in CSF in vivo before administration of the anti-MS4A4A antibodies of the present disclosure. In some embodiments, the anti-MS4A4A antibodies of the present disclosure increase soluble TREM2 levels in CSF in vivo for at least 4 days, compared to the level of soluble TREM2 in CSF in vivo before administration of the anti-MS4A4A antibodies of the present disclosure. In some embodiments, the anti-MS4A4A antibodies of the present disclosure increase soluble TREM2 levels in CSF in vivo for at least 24 hours, at least 48 hours, at least 72 hours, or at least 96 hours, compared to the level of soluble TREM2 in CSF in vivo before administration of the anti-MS4A4A antibodies of the present disclosure. In some embodiments, the anti-MS4A4A antibodies of the present disclosure increase soluble TREM2 levels in CSF in vivo for at least 96 hours, compared to soluble TREM2 levels in CSF in vivo before administration of the anti-MS4A4A antibodies of the present disclosure.

[0193] The evidence provided herein demonstrates that the anti-MS4A4A antibodies of the present disclosure affect TREM2, but does not exclude antibodies that act via other pathways.

[0194] Antagonist antibodies

[0195] In some embodiments, antibodies that bind to the MS4A4A protein may include antagonist antibodies that bind to MS4A4A and inhibit one or more MS4A4A activities by preventing interaction between MS4A4A and its ligand or by preventing transduction of MS4A4A signals into the cytoplasm in the presence of a ligand. In some embodiments, the antagonist antibodies of the present disclosure may have the epitope specificity of the agonist antibodies of the present disclosure, but have an Fc domain that is incapable of binding to an Fcg receptor and, therefore, is unable to aggregate the MS4A4A receptor, for example.

[0196] In some embodiments, the antibodies of the present disclosure are antagonist antibodies. In some embodiments, the antagonist antibodies inhibit one or more MS4A4A activities. In some embodiments, the antagonist antibodies reduce the activity of one or more MS4A4A-dependent genes. In some embodiments, the antagonist antibodies inhibit the interaction between MS4A4A and one or more MS4A4A ligands. In some embodiments, the antagonist antibodies inhibit MS4A4A signaling. In some embodiments, the antagonist antibodies inhibit the interaction between MS4A4A and one or more MS4A4A ligands and inhibit MS4A4A signaling.

[0197] In some embodiments, downregulation of MS4A4A protein levels or reduction of MS4A4A activity is achieved by downregulating or reducing MS4A4A protein levels in cells using anti-MS4A4A antibodies. In some embodiments, downregulation of MS4A4A protein levels or reduction of MS4A4A activity is achieved by downregulating MS4A4A nucleic acid expression or levels (e.g., by using antisense approaches, gene therapy, etc.) using methods known and available to those skilled in the art. Thus, in some embodiments, reduction of MS4A4A protein levels or activity is achieved using anti-MS4A4A antibodies of the present disclosure or by reducing MS4A4A nucleic acid (e.g., mRNA) expression or levels.

[0198] In some embodiments, agonist antibody function requires antibody cross-linking. Antibody cross-linking can occur by binding to a secondary antibody in vitro or by binding to an Fc receptor in vivo. For example, antagonist antibodies can be converted into agonist antibodies in vitro via biotin / streptavidin cross-linking or secondary antibody binding (see, for example, Gravestein et al., 1996, J. Exp. Med. 184: 675-685; Gravestein et al., 1994, International Immunol, 7: 551-557). Agonist antibodies can exert their activity by simulating the biological activity of receptor ligands or by enhancing receptor aggregation, thereby activating receptor signaling. In some embodiments, antagonist activity requires the absence of antibody cross-linking. Antagonist antibodies can exert their activity by blocking receptor-ligand interactions.

[0199] Phenotypic simulation of gelsolin, osteopontin, and Alzheimer's disease protective alleles

[0200] Three SNPs in the MS4A gene cluster are associated with an increased risk of late-onset Alzheimer's disease. These SNPs include rs4938933 in MS4A4A, rs670139 in MS4A4E, and rs610932 in MS4A6A (Hollingworth et al., 2011, Nat Genetics, 43:429-435; Naj et al., 2011, Nature Genetics, 43:436-441; Antunez et al., 2011, Genome Medicine, 3, Article 33). In addition, the MS4A4A locus SNPs (rs2304933 and rs2304935) are associated with higher levels of MS4A4A and increased risk of Alzheimer's disease, including late-onset Alzheimer's disease (LOAD) (Allen et al., 2012, Neurology, 79:221-228).

[0201] Alzheimer's disease-associated genetic variants (SNPs) associated with the MS4A gene cluster have been identified. One of those variant alleles is rs1582763, which is associated with increased CSF sTREM2 levels and reduced risk of Alzheimer's disease and delayed age of onset, and is therefore considered a protective allele. (Deming et al., 2018, bioRxiv, doi: dxdoi org / 10.1101 / 352179). The rs1582763 protective allele is associated with reduced levels of MS4A4A mRNA in the blood. The findings further suggest that the rs1582763 allele plays a protective role by reducing MS4A4A levels and reducing the risk or severity of Alzheimer's disease. Protective rs1582763 is also associated with increased expression levels of osteopontin and increased levels of gelsolin. The anti-MS4A4A antibodies of the present invention are effective in phenotypically mimicking the protective alleles, at least in terms of reduced MS4A4A expression, increased osteopontin expression, increased gelsolin expression, and / or increased sTREM levels. In some aspects of the present disclosure, anti-MS4A4A antibodies are provided, wherein the antibodies phenotypically mimic one or more alleles of MS4A that are associated with a reduced risk of Alzheimer's disease and / or a delayed age of onset of Alzheimer's disease.

[0202] The anti-MS4A4A antibodies of the present disclosure increase the mRNA level of osteopontin (SPP1) and increase the mRNA level of gelsolin (GSN) in human peripheral blood monocyte-derived macrophages. In some embodiments, the anti-MS4A4A antibodies of the present disclosure increase the mRNA and / or protein level of osteopontin. In some embodiments, the anti-MS4A4A antibodies of the present disclosure increase the mRNA and / or protein level of gelsolin. In some embodiments, the anti-MS4A4A antibodies of the present disclosure increase the mRNA and / or protein level of osteopontin in myeloid cells (e.g., macrophages, human macrophages, microglia) by at least 10%, at least 20%, at least 25%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 75%, at least 80%, at least 90%, at least 100%, at least 125%, at least 150%, at least 200%, or at least 250%. In some embodiments, the anti-MS4A4A antibodies of the present disclosure increase gelsolin mRNA and / or protein levels in myeloid cells (e.g., macrophages, human macrophages, microglia) by at least 10%, at least 20%, at least 25%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 75%, at least 80%, at least 90%, at least 100%, at least 125%, at least 150%, at least 200%, or at least 250%.

[0203] The anti-MS4A4A antibodies of the present disclosure increase osteopontin (SSP1) levels in non-human primates. In some embodiments, the anti-MS4A4A antibodies of the present disclosure increase osteopontin mRNA and / or protein levels in vivo (e.g., in non-human primates or humans). In some embodiments, the anti-MS4A4A antibodies of the present disclosure increase osteopontin mRNA and / or protein levels in serum and / or CSF. In some embodiments, the anti-MS4A4A antibodies of the present disclosure increase osteopontin mRNA and / or protein levels in serum and / or CSF by at least 10%, at least 20%, at least 25%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 75%, at least 80%, at least 90%, at least 100%, at least 125%, at least 150%, at least 200%, or at least 250%. In some embodiments, the anti-MS4A4A antibodies of the present disclosure increase osteopontin protein levels in the brain. In some embodiments, an anti-MS4A4A antibody of the present disclosure increases osteopontin protein levels in the frontal cortex and / or in the hippocampus.

[0204] In some embodiments, the anti-MS4A4A antibodies of the present disclosure phenotypically mimic the protective rs1582763 allele in terms of increasing expression of osteopontin and gelsolin. In other aspects, the anti-MS4A4A antibodies of the present disclosure effectively increase expression of osteopontin, gelsolin and / or sTREM2 and have biological activity in reducing the risk and / or severity of Alzheimer's disease, similar to the protective rs1582763 allele. The data indicate that SPP1 and GSN are pharmacodynamic markers of protective biological activity associated with the rs158273 allele.

[0205] CSFR1 and IL1RN expression

[0206] The anti-MS4A4A antibodies of the present disclosure increase CSF1R levels in non-human primates. In some embodiments, the anti-MS4A4A antibodies of the present disclosure increase CSF1R mRNA and / or protein levels in vivo (e.g., in non-human primates or humans). In some embodiments, the anti-MS4A4A antibodies of the present disclosure increase CSF1R mRNA and / or protein levels in serum and / or CSF. In some embodiments, the anti-MS4A4A antibodies of the present disclosure increase CSF1R mRNA and / or protein levels in serum and / or CSF by at least 10%, at least 20%, at least 25%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 75%, at least 80%, at least 90%, at least 100%, at least 125%, at least 150%, at least 200%, or at least 250%. In some embodiments, the anti-MS4A4A antibodies of the present disclosure increase CSF1R protein levels in the brain. In some embodiments, an anti-MS4A4A antibody of the present disclosure increases protein levels of CSF1R in the frontal cortex and / or in the hippocampus.

[0207] CSF1R deficiency negatively affects the development of microglia in the brain (Swerdlow et al. (2000) Neurology, 111:300-311; Baba et al. (2006) Acta Neuropath, 111:300-311), and recent studies have linked mutations in the CSF1R gene to various conditions, including adult-onset leukoencephalopathy with axonal spheroids and pigmented glial cells (ALSP) and hereditary diffuse leukoencephalopathy with spheroids (HDLS) (Oosterhof et al. (2019) Am J Hum Genet, 104:936-947; Rademaker et al. (2011) Nat Genet, 44:200-205; Nicholson et al. (2013) Neurology, 80:1033-1040). The anti-MS4A4A antibodies of the present disclosure reduce cell death and maintain the survival of human macrophages after CSF1R inhibition. Thus, in some embodiments, the anti-MS4A4A antibodies of the present disclosure can be used to treat an individual suffering from a CSF1R deficiency disease or disorder, such as ALSP or HDLS.

[0208] The anti-MS4A4A antibodies of the present disclosure increase IL1RN levels in human peripheral blood monocyte-derived macrophages. In some embodiments, the anti-MS4A4A antibodies of the present disclosure increase IL1RN mRNA and / or protein levels. In some embodiments, the anti-MS4A4A antibodies of the present disclosure increase IL1RN mRNA and / or protein levels. In some embodiments, the anti-MS4A4A antibodies of the present disclosure increase IL1RN mRNA and / or protein levels in myeloid cells (e.g., macrophages, human macrophages, microglia) by at least 10%, at least 20%, at least 25%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 75%, at least 80%, at least 90%, at least 100%, at least 125%, at least 150%, at least 200%, or at least 250%.

[0209] GPNMB expression

[0210] GPNMB (glycoprotein non-metastatic melanoma protein B) is a surface glycoprotein expressed in a variety of cell types (including tissue macrophages and microglial cells). Several genetic variants are associated with Parkinson's disease (Parkinson's disease, PD) risk. GPNMB protein levels in the substantia nigra of PD patients are elevated, and GPNMB levels increase after lysosomal stress (Moloney et al., 2018, Neurobio Dis. 120: 1-11). In addition, increased expression of GPNMB is associated with SNP rs199347, and this risk SNP is located within the GPNMB gene (Murthy et al., Neurogenetics, 2017, 18: 121-133).

[0211] The anti-MS4A4A antibodies disclosed herein reduce GPNMB cell surface protein levels in primary human macrophages. In some embodiments, the anti-MS4A4A antibodies disclosed herein reduce GPNMB cell surface protein levels in myeloid cells (e.g., macrophages, human macrophages, microglia) by at least 10%, at least 20%, at least 30%, at least 40%, or at least 50%. Because increased GPNMB levels are associated with risk alleles for PD, a reduction in GPNMB following the addition of anti-MS4A4A antibodies may provide a therapeutic approach for PD.

[0212] A. Exemplary Antibodies and Certain Other Antibody Embodiments

[0213] In some embodiments, provided herein are anti-MS4A4A antibodies comprising at least one, two, three, four, five, or six HVRs selected from the group consisting of: (a) an HVR-H1 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 94, 108, 116, 146, 147, 308, and 311, or an amino acid having at least about 95% homology to an amino acid selected from the group consisting of SEQ ID NOs: 94, 108, 116, 146, 147, 308, and 311; (b) an HVR-H2 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 96, 97, 98, 99, 110, 111, 118, 119, 120, 121, 122, 149, 150, 151, 152, 153, 309, and 312, or an amino acid having at least about 95% homology to an amino acid selected from the group consisting of SEQ ID NOs: (c) an HVR-H3 comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 100, 101, 102, 112, 123, 124, 125, 126, 127, 128, 129, 154, 310, and 313, or a sequence homologous to an amino acid selected from the group consisting of SEQ ID NO: 100, 101, 102, 112, 123, 124, 125, 126, 127, 128, 129, 154, 310, and 313; (d) an HVR-L1 comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 103, 104, 113, 130, 131, 132, 133, 134, 135, 136, 137, 138, 156, 157, 158, 314, and 317 or a sequence identical to an amino acid selected from the group consisting of SEQ ID NO: 103, 104, 113, 130, 131, 132, 133, 134, 135, 136, 137, 138, 156, 157, 158, 314, and 317; (e) an HVR-L2 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 105, 106, 114, 139, 140, 141, 142, 143, 160, 161, 315, and 318, or an amino acid with at least about 95% homology to an amino acid selected from the group consisting of SEQ ID NOs: 105, 106, 114, 139, 140, 141, 142, 143, 160, 161, 315, and 318;and (f) HVR-L3 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 107, 115, 144, 145, 163, 316, and 319, or an amino acid having at least about 95% homology to an amino acid selected from the group consisting of SEQ ID NOs: 107, 115, 144, 145, 163, 316, and 319;

[0214] In some embodiments, provided herein are anti-MS4A4A antibodies comprising at least one, two, three, four, five, or six HVRs selected from the group consisting of: (a) an HVR-H1 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 94 and 308, or an amino acid having at least about 95% homology to an amino acid sequence selected from the group consisting of SEQ ID NOs: 94 and 308; (b) an HVR-H2 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 96, 97, 98, 99, and 309, or an amino acid having at least about 95% homology to an amino acid selected from the group consisting of SEQ ID NOs: 96, 97, 98, 99, and 309; (c) an HVR-H3 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 100, 101, 102, and 310, or an amino acid having at least about 95% homology to an amino acid selected from the group consisting of SEQ ID NOs: 100, 101, 102, and 310; (d) an HVR-H4 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 100, 101, 102, and 310, or an amino acid having at least about 95% homology to an amino acid selected from the group consisting of SEQ ID NOs: 100, 101, 102, and 310; (e) HVR-L2 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 105, 106 and 315, or an amino acid with at least about 95% homology to an amino acid selected from the group consisting of SEQ ID NOs: 105, 106 and 315; and (f) HVR-L3 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 107 and 316, or an amino acid sequence with at least about 95% homology to an amino acid selected from the group consisting of SEQ ID NOs: 107 and 316.

[0215] In some embodiments, provided herein are anti-MS4A4A antibodies comprising at least one, two, three, four, five, or six HVRs selected from the group consisting of: (a) an HVR-H1 comprising the amino acid sequence of SEQ ID NO: 108, or an amino acid having at least about 95% homology to an amino acid of SEQ ID NO: 108; (b) an HVR-H2 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 110 and 111, or an amino acid having at least about 95% homology to an amino acid selected from the group consisting of SEQ ID NOs: 110 and 111; (c) an HVR-H3 comprising the amino acid sequence of SEQ ID NO: 112, or an amino acid having at least about 95% homology to an amino acid of SEQ ID NO: 112; (d) an HVR-L1 comprising the amino acid sequence of SEQ ID NO: 113, or an amino acid having at least about 95% homology to an amino acid of SEQ ID NO: 113; (e) an HVR-L2 comprising the amino acid sequence of SEQ ID NO: 114, or an amino acid having at least about 95% homology to an amino acid of SEQ ID NO: NO: 114; and (f) HVR-L3 comprising the amino acid sequence of SEQ ID NO: 115 or an amino acid having at least about 95% homology to the amino acid of SEQ ID NO: 115.

[0216] In some embodiments, provided herein are anti-MS4A4A antibodies comprising at least one, two, three, four, five, or six HVRs selected from the group consisting of: (a) an HVR-H1 comprising an amino acid sequence of SEQ ID NO: 116, or an amino acid having at least about 95% homology to an amino acid of SEQ ID NO: 116; (b) an HVR-H2 comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 118, 119, 120, 121, and 122, or an amino acid having at least about 95% homology to an amino acid selected from the group consisting of SEQ ID NO: 118, 119, 120, 121, and 122; (c) an HVR-H3 comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 123, 124, 125, 126, 127, 128, and 129, or an amino acid having at least about 95% homology to an amino acid selected from the group consisting of SEQ ID NO: 129; (d) an HVR-L1 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 130, 131, 132, 133, 134, 135, 136, 137, and 138, or an amino acid with at least about 95% homology to an amino acid selected from the group consisting of SEQ ID NOs: 130, 131, 132, 133, 134, 135, 136, 137, and 138; (e) an HVR-L2 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 139, 140, 141, 142, and 143, or an amino acid with at least about 95% homology to an amino acid selected from the group consisting of SEQ ID NOs: 139, 140, 141, 142, and 143; and (f) HVR-L3 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 144 and 145, or an amino acid having at least about 95% homology to an amino acid selected from the group consisting of SEQ ID NOs: 144 and 145.

[0217] In some embodiments, provided herein are anti-MS4A4A antibodies comprising at least one, two, three, four, five, or six HVRs selected from the group consisting of: (a) an HVR-H1 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 146 and 147, or an amino acid having at least about 95% homology to an amino acid selected from the group consisting of SEQ ID NOs: 146 and 147; (b) an HVR-H2 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 149, 150, 151, 152, and 153, or an amino acid having at least about 95% homology to an amino acid selected from the group consisting of SEQ ID NOs: 149, 150, 151, 152, and 153; (c) an HVR-H3 comprising an amino acid sequence of SEQ ID NO: 154, or an amino acid having at least about 95% homology to an amino acid selected from SEQ ID NO: 154; (d) an HVR-L1 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: NO: 156, 157 and 158, or an amino acid with at least about 95% homology to an amino acid selected from the group consisting of SEQ ID NO: 156, 157 and 158; (e) HVR-L2 comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 159, 160 and 161, or an amino acid with at least about 95% homology to an amino acid selected from the group consisting of SEQ ID NO: 159, 160 and 161; and (f) HVR-L3 comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 163, or an amino acid with at least about 95% homology to an amino acid selected from the group consisting of SEQ ID NO: 163.

[0218] In some embodiments, provided herein are anti-MS4A4A antibodies comprising: (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 94; (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO: 96; (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO: 100; (d) HVR-L1 comprising the amino acid sequence of SEQ ID NO: 103; (e) HVR-L2 comprising the amino acid sequence of SEQ ID NO: 105; and (f) HVR-L3 comprising the amino acid sequence of SEQ ID NO: 107. ID NO:105; and (f) HVR-L3 comprising the amino acid sequence of SEQ ID NO:107; (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO:94; (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO:98; (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO:100; (d) HVR-L1 comprising the amino acid sequence of SEQ ID NO:103; (e) HVR-L2 comprising the amino acid sequence of SEQ ID NO:105; and (f) HVR-L3 comprising the amino acid sequence of SEQ ID NO:107; (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO:94; (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO:96; (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO:101; (d) HVR-L1 comprising the amino acid sequence of SEQ ID NO: NO:103; (e) HVR-L2 comprising the amino acid sequence of SEQ ID NO:106; and (f) HVR-L3 comprising the amino acid sequence of SEQ ID NO:107; (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO:94; (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO:96; (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO:102; (d) HVR-L1 comprising the amino acid sequence of SEQ ID NO:104;(e) HVR-L2 comprising the amino acid sequence of SEQ ID NO: 105; and (f) HVR-L3 comprising the amino acid sequence of SEQ ID NO: 107; or (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 94; (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO: 99; (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO: 100; (d) HVR-L1 comprising the amino acid sequence of SEQ ID NO: 104; (e) HVR-L2 comprising the amino acid sequence of SEQ ID NO: 105; and (f) HVR-L3 comprising the amino acid sequence of SEQ ID NO: 107.

[0219] In some embodiments, provided herein are anti-MS4A4A antibodies comprising: (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 308; (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO: 309; (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO: 310; (d) HVR-L1 comprising the amino acid sequence of SEQ ID NO: 314; (e) HVR-L2 comprising the amino acid sequence of SEQ ID NO: 315; and (f) HVR-L3 comprising the amino acid sequence of SEQ ID NO: 316; or (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 311; (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO: 312; (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO: 313; (d) HVR-L1 comprising the amino acid sequence of SEQ ID NO: 317; (e) (f) HVR-L2 comprising the amino acid sequence of SEQ ID NO: 318; and (f) HVR-L3 comprising the amino acid sequence of SEQ ID NO: 319.

[0220] In some embodiments, provided herein are anti-MS4A4A antibodies comprising: (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 108; (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO: 110; (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO: 112; (d) HVR-L1 comprising the amino acid sequence of SEQ ID NO: 113; (e) HVR-L2 comprising the amino acid sequence of SEQ ID NO: 114; and (f) HVR-L3 comprising the amino acid sequence of SEQ ID NO: 115; or (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 108; (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO: 111; (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO: 112; (d) HVR-L1 comprising the amino acid sequence of SEQ ID NO: 113; (e) (f) HVR-L2 comprising the amino acid sequence of SEQ ID NO: 114; and (f) HVR-L3 comprising the amino acid sequence of SEQ ID NO: 115.

[0221] In some embodiments, provided herein are anti-MS4A4A antibodies comprising: (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 116; (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO: 118; (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO: 123; (d) HVR-L1 comprising the amino acid sequence of SEQ ID NO: 130; (e) HVR-L2 comprising the amino acid sequence of SEQ ID NO: 139; and (f) HVR-L3 comprising the amino acid sequence of SEQ ID NO: 144. HVR-L2 comprising the amino acid sequence of SEQ ID NO: 139; and (f) HVR-L3 comprising the amino acid sequence of SEQ ID NO: 144; (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 116; (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO: 120; (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO: 123; (d) HVR-L1 comprising the amino acid sequence of SEQ ID NO: 130; (e) HVR-L2 comprising the amino acid sequence of SEQ ID NO: 139; and (f) HVR-L3 comprising the amino acid sequence of SEQ ID NO: 144; (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 116; (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO: 121; (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO: 123; (d) HVR-L1 comprising the amino acid sequence of SEQ ID NO: 130; (e) HVR-L2 comprising the amino acid sequence of SEQ ID NO: 139; and (f) HVR-L3 comprising the amino acid sequence of SEQ ID NO: 144; (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 116; (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO: 118; (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO: 124; (d) HVR-L1 comprising the amino acid sequence of SEQ ID NO: 130;(e) HVR-L2 comprising the amino acid sequence of SEQ ID NO: 139; and (f) HVR-L3 comprising the amino acid sequence of SEQ ID NO: 144; (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 116; (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO: 118; (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO: 124; (d) HVR-L1 comprising the amino acid sequence of SEQ ID NO: 131; (e) HVR-L2 comprising the amino acid sequence of SEQ ID NO: 139; and (f) HVR-L3 comprising the amino acid sequence of SEQ ID NO: 144; (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 116; (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO: 118; (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO: NO:124; (d) HVR-L1 comprising the amino acid sequence of SEQ ID NO:132; (e) HVR-L2 comprising the amino acid sequence of SEQ ID NO:139; and (f) HVR-L3 comprising the amino acid sequence of SEQ ID NO:144; (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO:116; (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO:118; (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO:124; (d) HVR-L1 comprising the amino acid sequence of SEQ ID NO:133; (e) HVR-L2 comprising the amino acid sequence of SEQ ID NO:139; and (f) HVR-L3 comprising the amino acid sequence of SEQ ID NO:144; (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO:116; (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO:118 NO:118; (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO:124; (d) HVR-L1 comprising the amino acid sequence of SEQ ID NO:134; (e) HVR-L2 comprising the amino acid sequence of SEQ ID NO:139; and (f) HVR-L3 comprising the amino acid sequence of SEQ ID NO:144; (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO:116; (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO:118; (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO:124;(d) HVR-L1 comprising the amino acid sequence of SEQ ID NO: 135; (e) HVR-L2 comprising the amino acid sequence of SEQ ID NO: 139; and (f) HVR-L3 comprising the amino acid sequence of SEQ ID NO: 144; (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 116; (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO: 118; (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO: 124; (d) HVR-L1 comprising the amino acid sequence of SEQ ID NO: 136; (e) HVR-L2 comprising the amino acid sequence of SEQ ID NO: 139; and (f) HVR-L3 comprising the amino acid sequence of SEQ ID NO: 144; NO:118; (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO:124; (d) HVR-L1 comprising the amino acid sequence of SEQ ID NO:130; (e) HVR-L2 comprising the amino acid sequence of SEQ ID NO:140; and (f) HVR-L3 comprising the amino acid sequence of SEQ ID NO:144; (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO:116; (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO:118; (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO:124; (d) HVR-L1 comprising the amino acid sequence of SEQ ID NO:130; (e) HVR-L2 comprising the amino acid sequence of SEQ ID NO:139; and (f) HVR-L3 comprising the amino acid sequence of SEQ ID NO:145; NO:116; (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO:118; (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO:124; (d) HVR-L1 comprising the amino acid sequence of SEQ ID NO:137; (e) HVR-L2 comprising the amino acid sequence of SEQ ID NO:139; and (f) HVR-L3 comprising the amino acid sequence of SEQ ID NO:144; (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO:116; (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO:118;(c) HVR-H3 comprising the amino acid sequence of SEQ ID NO: 124; (d) HVR-L1 comprising the amino acid sequence of SEQ ID NO: 138; (e) HVR-L2 comprising the amino acid sequence of SEQ ID NO: 141; and (f) HVR-L3 comprising the amino acid sequence of SEQ ID NO: 144; (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 116; (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO: 118; (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO: 124; (d) HVR-L1 comprising the amino acid sequence of SEQ ID NO: 130; (e) HVR-L2 comprising the amino acid sequence of SEQ ID NO: 142; and (f) HVR-L3 comprising the amino acid sequence of SEQ ID NO: 144; NO:116; (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO:118; (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO:124; (d) HVR-L1 comprising the amino acid sequence of SEQ ID NO:130; (e) HVR-L2 comprising the amino acid sequence of SEQ ID NO:141; and (f) HVR-L3 comprising the amino acid sequence of SEQ ID NO:144; (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO:116; (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO:118; (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO:124; (d) HVR-L1 comprising the amino acid sequence of SEQ ID NO:130; (e) HVR-L2 comprising the amino acid sequence of SEQ ID NO:143; and (f) HVR-L3 comprising the amino acid sequence of SEQ ID NO: NO:144; (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO:116; (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO:118; (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO:125; (d) HVR-L1 comprising the amino acid sequence of SEQ ID NO:137; (e) HVR-L2 comprising the amino acid sequence of SEQ ID NO:139; and (f) HVR-L3 comprising the amino acid sequence of SEQ ID NO:144; (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO:116;(b) HVR-H2 comprising the amino acid sequence of SEQ ID NO: 118; (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO: 125; (d) HVR-L1 comprising the amino acid sequence of SEQ ID NO: 130; (e) HVR-L2 comprising the amino acid sequence of SEQ ID NO: 143; and (f) HVR-L3 comprising the amino acid sequence of SEQ ID NO: 144; (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 116; (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO: 118; (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO: 125; (d) HVR-L1 comprising the amino acid sequence of SEQ ID NO: 131; (e) HVR-L2 comprising the amino acid sequence of SEQ ID NO: 139; and (f) HVR-L3 comprising the amino acid sequence of SEQ ID NO: NO:144; (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO:116; (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO:118; (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO:125; (d) HVR-L1 comprising the amino acid sequence of SEQ ID NO:130; (e) HVR-L2 comprising the amino acid sequence of SEQ ID NO:139; and (f) HVR-L3 comprising the amino acid sequence of SEQ ID NO:144; (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO:116; (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO:118; (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO:125; (d) HVR-L1 comprising the amino acid sequence of SEQ ID NO:130; (e) HVR-L2 comprising the amino acid sequence of SEQ ID NO: NO:142; and (f) HVR-L3 comprising the amino acid sequence of SEQ ID NO:144; (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO:116; (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO:118; (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO:125; (d) HVR-L1 comprising the amino acid sequence of SEQ ID NO:130; (e) HVR-L2 comprising the amino acid sequence of SEQ ID NO:139; and (f) HVR-L3 comprising the amino acid sequence of SEQ ID NO:145;(a) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 116; (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO: 118; (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO: 126; (d) HVR-L1 comprising the amino acid sequence of SEQ ID NO: 138; (e) HVR-L2 comprising the amino acid sequence of SEQ ID NO: 141; and (f) HVR-L3 comprising the amino acid sequence of SEQ ID NO: 144; (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 116; (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO: 118; (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO: 126; (d) HVR-L1 comprising the amino acid sequence of SEQ ID NO: 130; (e) HVR-L2 comprising the amino acid sequence of SEQ ID NO: NO:139; and (f) HVR-L3 comprising the amino acid sequence of SEQ ID NO:144; (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO:116; (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO:118; (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO:126; (d) HVR-L1 comprising the amino acid sequence of SEQ ID NO:130; (e) HVR-L2 comprising the amino acid sequence of SEQ ID NO:141; and (f) HVR-L3 comprising the amino acid sequence of SEQ ID NO:144; (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO:116; (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO:118; (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO:127; (d) HVR-L1 comprising the amino acid sequence of SEQ ID NO:130 NO:131; (e) HVR-L2 comprising the amino acid sequence of SEQ ID NO:139; and (f) HVR-L3 comprising the amino acid sequence of SEQ ID NO:144; (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO:116; (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO:122; (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO:127; (d) HVR-L1 comprising the amino acid sequence of SEQ ID NO:131; (e) HVR-L2 comprising the amino acid sequence of SEQ ID NO:139;and (f) HVR-L3 comprising the amino acid sequence of SEQ ID NO: 144; (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 116; (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO: 118; (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO: 128; (d) HVR-L1 comprising the amino acid sequence of SEQ ID NO: 131; (e) HVR-L2 comprising the amino acid sequence of SEQ ID NO: 139; and (f) HVR-L3 comprising the amino acid sequence of SEQ ID NO: 144; (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 116; (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO: 118; (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO: 129; (d) HVR-L1 comprising the amino acid sequence of SEQ ID NO: 131; NO:131; (e) HVR-L2 comprising the amino acid sequence of SEQ ID NO:139; and (f) HVR-L3 comprising the amino acid sequence of SEQ ID NO:144; (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO:116; (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO:118; (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO:129; (d) HVR-L1 comprising the amino acid sequence of SEQ ID NO:132; (e) HVR-L2 comprising the amino acid sequence of SEQ ID NO:139; and (f) HVR-L3 comprising the amino acid sequence of SEQ ID NO:144; (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO:116; (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO:118; (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO: NO:129; (d) HVR-L1 comprising the amino acid sequence of SEQ ID NO:133; (e) HVR-L2 comprising the amino acid sequence of SEQ ID NO:139; and (f) HVR-L3 comprising the amino acid sequence of SEQ ID NO:144; (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO:116; (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO:118; (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO:129; (d) HVR-L1 comprising the amino acid sequence of SEQ ID NO:134;(e) HVR-L2 comprising the amino acid sequence of SEQ ID NO: 139; and (f) HVR-L3 comprising the amino acid sequence of SEQ ID NO: 144; (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 116; (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO: 118; (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO: 129; (d) HVR-L1 comprising the amino acid sequence of SEQ ID NO: 135; (e) HVR-L2 comprising the amino acid sequence of SEQ ID NO: 139; and (f) HVR-L3 comprising the amino acid sequence of SEQ ID NO: 144; (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 116; (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO: 118; (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO: NO:129; (d) HVR-L1 comprising the amino acid sequence of SEQ ID NO:136; (e) HVR-L2 comprising the amino acid sequence of SEQ ID NO:139; and (f) HVR-L3 comprising the amino acid sequence of SEQ ID NO:144.

[0222] In some embodiments, provided herein are anti-MS4A4A antibodies comprising: (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 147; (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO: 149; (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO: 154; (d) HVR-L1 comprising the amino acid sequence of SEQ ID NO: 156; (e) HVR-L2 comprising the amino acid sequence of SEQ ID NO: 160; and (f) HVR-L3 comprising the amino acid sequence of SEQ ID NO: 163; (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 146; (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO: 150; (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO: 154; (d) HVR-L1 comprising the amino acid sequence of SEQ ID NO: 156; (e) HVR-L2 comprising the amino acid sequence of SEQ ID NO: 160; and (f) HVR-L3 comprising the amino acid sequence of SEQ ID NO: 163; (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 147; (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO: 149; (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO: 154; (d) HVR-L1 comprising the amino acid sequence of SEQ ID NO: 157; (e) HVR-L2 comprising the amino acid sequence of SEQ ID NO: 159; and (f) HVR-L3 comprising the amino acid sequence of SEQ ID NO: 163; (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 147; (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO: 151; (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO: 154; (d) HVR-L1 comprising the amino acid sequence of SEQ ID NO: 156; (e) HVR-L2 comprising the amino acid sequence of SEQ ID NO: 160; and (f) HVR-L3 comprising the amino acid sequence of SEQ ID NO: 163; (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 147; (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO: 151; (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO: 154; (d) HVR-L1 comprising the amino acid sequence of SEQ ID NO: 158;(e) HVR-L2 comprising the amino acid sequence of SEQ ID NO: 159; and (f) HVR-L3 comprising the amino acid sequence of SEQ ID NO: 163; (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 146; (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO: 152; (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO: 154; (d) HVR-L1 comprising the amino acid sequence of SEQ ID NO: 156; (e) HVR-L2 comprising the amino acid sequence of SEQ ID NO: 161; and (f) HVR-L3 comprising the amino acid sequence of SEQ ID NO: 163; (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 146; (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO: 153; (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO: NO:154; (d) HVR-L1 comprising the amino acid sequence of SEQ ID NO:158; (e) HVR-L2 comprising the amino acid sequence of SEQ ID NO:159; and (f) HVR-L3 comprising the amino acid sequence of SEQ ID NO:163.

[0223] In some embodiments, provided herein are anti-MS4A4A antibodies comprising at least one, at least two, or all three V selected from the group consisting of: H HVR sequences: (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO:94 or an amino acid sequence that is at least about 95% homologous to the amino acid sequence of SEQ ID NO:94; (b) HVR-H2 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs:96, 97, 98 and 99 or an amino acid sequence that is at least about 95% homologous to an amino acid sequence selected from the group consisting of SEQ ID NOs:96, 97, 98 and 99; and (c) HVR-H3 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs:100, 101 and 102 or an amino acid sequence that is at least about 95% homologous to an amino acid sequence selected from the group consisting of SEQ ID NOs:100, 101 and 102.

[0224] In some embodiments, provided herein are anti-MS4A4A antibodies comprising at least one, at least two, or all three V selected from the group consisting of: HHVR sequences: (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 308 or an amino acid sequence that is at least about 95% homologous to the amino acid sequence of SEQ ID NO: 308; (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO: 309 or an amino acid sequence that is at least about 95% homologous to the amino acid sequence of SEQ ID NO: 309; and (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO: 310 or an amino acid sequence that is at least about 95% homologous to the amino acid sequence of SEQ ID NO: 310.

[0225] In some embodiments, provided herein are anti-MS4A4A antibodies comprising at least one, at least two, or all three V selected from the group consisting of: H HVR sequences: (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 311 or an amino acid sequence that is at least about 95% homologous to the amino acid sequence of SEQ ID NO: 311; (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO: 312 or an amino acid sequence that is at least about 95% homologous to the amino acid sequence of SEQ ID NO: 312; and (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO: 313 or an amino acid sequence that is at least about 95% homologous to the amino acid sequence of SEQ ID NO: 313.

[0226] In some embodiments, provided herein are anti-MS4A4A antibodies comprising at least one, at least two, or all three V selected from the group consisting of: L HVR sequences: (a) HVR-L1 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 103 and 104, or an amino acid sequence having at least about 95% homology to an amino acid sequence selected from the group consisting of SEQ ID NOs: 103 and 104; (b) HVR-L2 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 105 and 106, or an amino acid sequence having at least about 95% homology to an amino acid sequence selected from the group consisting of SEQ ID NOs: 105 and 106; and (c) HVR-L3 comprising an amino acid sequence of SEQ ID NO: 107, or an amino acid sequence having at least about 95% homology to an amino acid sequence of SEQ ID NO: 107.

[0227] In some embodiments, provided herein are anti-MS4A4A antibodies comprising at least one, at least two, or all three V selected from the group consisting of: LHVR sequences: (a) HVR-L1 comprising an amino acid sequence selected from SEQ ID NO: 314 or an amino acid sequence that is at least about 95% homologous to an amino acid sequence selected from SEQ ID NO: 314; (b) HVR-L2 comprising an amino acid sequence of SEQ ID NO: 315 or an amino acid sequence that is at least about 95% homologous to an amino acid sequence of SEQ ID NO: 315; and (c) HVR-L3 comprising an amino acid sequence of SEQ ID NO: 316 or an amino acid sequence that is at least about 95% homologous to an amino acid sequence of SEQ ID NO: 316.

[0228] In some embodiments, provided herein are anti-MS4A4A antibodies comprising at least one, at least two, or all three V selected from the group consisting of: L HVR sequences: (a) HVR-L1 comprising an amino acid sequence selected from SEQ ID NO: 317 or an amino acid sequence that is at least about 95% homologous to an amino acid sequence selected from SEQ ID NO: 317; (b) HVR-L2 comprising an amino acid sequence of SEQ ID NO: 318 or an amino acid sequence that is at least about 95% homologous to an amino acid sequence of SEQ ID NO: 318; and (c) HVR-L3 comprising an amino acid sequence of SEQ ID NO: 319 or an amino acid sequence that is at least about 95% homologous to an amino acid sequence of SEQ ID NO: 319.

[0229] In some embodiments, provided herein are anti-MS4A4A antibodies comprising (a) V H A domain comprising at least one, at least two or all three V H HVR sequences: (i) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 94 or an amino acid sequence at least about 95% homologous to the amino acid sequence of SEQ ID NO: 94, (ii) HVR-H2 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 96, 97, 98 and 99 or an amino acid sequence at least about 95% homologous to an amino acid sequence selected from the group consisting of SEQ ID NOs: 96, 97, 98 and 99, and (iii) HVR-H3 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 100, 101 and 102 or an amino acid sequence at least about 95% homologous to an amino acid sequence selected from the group consisting of SEQ ID NOs: 100, 101 and 102; and (b) V L A domain comprising at least one, at least two or all three V LHVR sequences: (i) HVR-L1 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 103 and 104, or an amino acid sequence having at least about 95% homology to an amino acid sequence selected from the group consisting of SEQ ID NOs: 103 and 104, (ii) HVR-L2 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 105 and 106, or an amino acid sequence having at least about 95% homology to an amino acid sequence selected from the group consisting of SEQ ID NOs: 105 and 106, and (iii) HVR-L3 comprising an amino acid sequence of SEQ ID NO: 107, or an amino acid sequence having at least about 95% homology to an amino acid sequence of SEQ ID NO: 107.

[0230] In some embodiments, provided herein are anti-MS4A4A antibodies comprising (a) V H A domain comprising at least one, at least two or all three V H HVR sequences: (i) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 308 or an amino acid sequence at least about 95% homologous to the amino acid sequence of SEQ ID NO: 308, (ii) HVR-H2 comprising the amino acid sequence of SEQ ID NO: 309 or an amino acid sequence at least about 95% homologous to the amino acid sequence of SEQ ID NO: 309, and (iii) HVR-H3 comprising the amino acid sequence of SEQ ID NO: 310 or an amino acid sequence at least about 95% homologous to the amino acid sequence of SEQ ID NO: 310; and (b) HVR-H4 comprising the amino acid sequence of SEQ ID NO: 311 or an amino acid sequence at least about 95% homologous to the amino acid sequence of SEQ ID NO: 312; L A domain comprising at least one, at least two or all three V L HVR sequences: (i) HVR-L1 comprising the amino acid sequence of SEQ ID NO: 314 or an amino acid sequence that is at least about 95% homologous to the amino acid sequence of SEQ ID NO: 314, (ii) HVR-L2 comprising the amino acid sequence of SEQ ID NO: 315 or an amino acid sequence that is at least about 95% homologous to an amino acid sequence selected from SEQ ID NO: 315, and (iii) HVR-L3 comprising the amino acid sequence of SEQ ID NO: 316 or an amino acid sequence that is at least about 95% homologous to the amino acid sequence of SEQ ID NO: 316.

[0231] In some embodiments, provided herein are anti-MS4A4A antibodies comprising (a) V H A domain comprising at least one, at least two or all three VH HVR sequences: (i) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 311 or an amino acid sequence at least about 95% homologous to the amino acid sequence of SEQ ID NO: 311, (ii) HVR-H2 comprising the amino acid sequence of SEQ ID NO: 312 or an amino acid sequence at least about 95% homologous to the amino acid sequence of SEQ ID NO: 312, and (iii) HVR-H3 comprising the amino acid sequence of SEQ ID NO: 313 or an amino acid sequence at least about 95% homologous to the amino acid sequence of SEQ ID NO: 313; and (b) HVR-H4 comprising the amino acid sequence of SEQ ID NO: 314 or an amino acid sequence at least about 95% homologous to the amino acid sequence of SEQ ID NO: 314; L A domain comprising at least one, at least two or all three V L HVR sequences: (i) HVR-L1 comprising the amino acid sequence of SEQ ID NO: 317 or an amino acid sequence that is at least about 95% homologous to the amino acid sequence of SEQ ID NO: 317, (ii) HVR-L2 comprising the amino acid sequence of SEQ ID NO: 318 or an amino acid sequence that is at least about 95% homologous to an amino acid sequence selected from SEQ ID NO: 318, and (iii) HVR-L3 comprising the amino acid sequence of SEQ ID NO: 316 or an amino acid sequence that is at least about 95% homologous to the amino acid sequence of SEQ ID NO: 316.

[0232] In some embodiments, provided herein are anti-MS4A4A antibodies comprising at least one, at least two, or all three V selected from the group consisting of: H HVR sequences: (a) HVR-H1 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 24, 25, 26, 27, 28, 29 and 30, or an amino acid sequence having at least about 95% homology to an amino acid sequence selected from the group consisting of SEQ ID NOs: 24, 25, 26, 27, 28, 29 and 30; (b) HVR-H2 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 110 and 111, or an amino acid sequence having at least about 95% homology to an amino acid sequence selected from the group consisting of SEQ ID NOs: 110 and 111; and (c) HVR-H3 comprising an amino acid sequence of SEQ ID NO: 112, or an amino acid sequence having at least about 95% homology to an amino acid sequence of SEQ ID NO: 112.

[0233] In some embodiments, provided herein are anti-MS4A4A antibodies comprising at least one, at least two, or all three V selected from the group consisting of: LHVR sequences: (a) HVR-L1 comprising the amino acid sequence of SEQ ID NO: 113 or an amino acid sequence that is at least about 95% homologous to the amino acid sequence of SEQ ID NO: 113; (b) HVR-L2 comprising the amino acid sequence of SEQ ID NO: 114 or an amino acid sequence that is at least about 95% homologous to the amino acid sequence of SEQ ID NO: 114; and (c) HVR-L3 comprising the amino acid sequence of SEQ ID NO: 115 or an amino acid sequence that is at least about 95% homologous to the amino acid sequence of SEQ ID NO: 115.

[0234] In some embodiments, provided herein are anti-MS4A4A antibodies comprising (a) V H A domain comprising at least one, at least two or all three V H HVR sequences: (i) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 108 or an amino acid sequence at least about 95% homologous to the amino acid sequence of SEQ ID NO: 108, (ii) HVR-H2 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 110 and 111 or an amino acid sequence at least about 95% homologous to the amino acid sequence of SEQ ID NOs: 110 and 111, and (iii) HVR-H3 comprising the amino acid sequence of SEQ ID NO: 112 or an amino acid sequence at least about 95% homologous to the amino acid sequence of SEQ ID NO: 112; and (b) HVR-H4 comprising an amino acid sequence of SEQ ID NO: 113 or an amino acid sequence at least about 95% homologous to the amino acid sequence of SEQ ID NO: 113; L A domain comprising at least one, at least two or all three V L HVR sequences: (i) HVR-L1 comprising the amino acid sequence of SEQ ID NO: 113 or an amino acid sequence that is at least about 95% homologous to the amino acid sequence of SEQ ID NO: 113, (ii) HVR-L2 comprising the amino acid sequence of SEQ ID NO: 114 or an amino acid sequence that is at least about 95% homologous to the amino acid sequence of SEQ ID NO: 114, and (iii) HVR-L3 comprising the amino acid sequence of SEQ ID NO: 115 or an amino acid sequence that is at least about 95% homologous to the amino acid sequence of SEQ ID NO: 115.

[0235] In some embodiments, provided herein are anti-MS4A4A antibodies comprising at least one, at least two, or all three V selected from the group consisting of: HHVR sequences: (a) HVR-H1 comprising an amino acid sequence of SEQ ID NO: 116, or an amino acid sequence that is at least about 95% homologous to an amino acid sequence of SEQ ID NO: 116; (b) HVR-H2 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 118, 119, 120, 121, and 122, or an amino acid sequence that is at least about 95% homologous to an amino acid sequence selected from the group consisting of SEQ ID NOs: 118, 119, 120, 121, and 122; and (c) HVR-H3 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 123, 124, 125, 126, 127, 128, and 129, or an amino acid sequence that is at least about 95% homologous to an amino acid sequence selected from the group consisting of SEQ ID NOs: 123, 124, 125, 126, 127, 128, and 129.

[0236] In some embodiments, provided herein are anti-MS4A4A antibodies comprising at least one, at least two, or all three V selected from the group consisting of: L HVR sequences: (a) HVR-L1 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 130, 131, 132, 133, 134, 135, 136, 137, and 138, or an amino acid sequence having at least about 95% homology to an amino acid sequence selected from the group consisting of SEQ ID NOs: 130, 131, 132, 133, 134, 135, 136, 137, and 138; (b) HVR-L2 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 139, 140, 141, 142, and 143, or an amino acid sequence having at least about 95% homology to an amino acid sequence selected from the group consisting of SEQ ID NOs: 139, 140, 141, 142, and 143; and (c) HVR-L3 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 144 and 145, or an amino acid sequence having at least about 95% homology to an amino acid sequence selected from the group consisting of SEQ ID NOs: The amino acid sequences of the group consisting of NO: 144 and 145 have amino acid sequences with at least about 95% homology.

[0237] In some embodiments, provided herein are anti-MS4A4A antibodies comprising (a) V H A domain comprising at least one, at least two or all three V HHVR sequences: (i) HVR-H1 comprising an amino acid sequence of SEQ ID NO: 116, or an amino acid sequence at least about 95% homologous to an amino acid sequence of SEQ ID NO: 116, (ii) HVR-H2 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 118, 119, 120, 121, and 122, or an amino acid sequence at least about 95% homologous to an amino acid sequence selected from the group consisting of SEQ ID NOs: 118, 119, 120, 121, and 122, and (iii) HVR-H3 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 123, 124, 125, 126, 127, 128, and 129, or an amino acid sequence at least about 95% homologous to an amino acid sequence selected from the group consisting of SEQ ID NOs: 123, 124, 125, 126, 127, 128, and 129; and (b) HVR-H4 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 124, 125, 126, 127, 128, and 129; L A domain comprising at least one, at least two or all three V L HVR sequences: (i) HVR-L1 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 130, 131, 132, 133, 134, 135, 136, 137, and 138, or an amino acid sequence having at least about 95% homology to an amino acid sequence selected from the group consisting of SEQ ID NOs: 130, 131, 132, 133, 134, 135, 136, 137, and 138, (ii) HVR-L2 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 139, 140, 141, 142, and 143, or an amino acid sequence having at least about 95% homology to an amino acid sequence selected from the group consisting of SEQ ID NOs: 139, 140, 141, 142, and 143, and (iii) HVR-L3 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 144 and 145, or an amino acid sequence having at least about 95% homology to an amino acid sequence selected from the group consisting of SEQ ID NOs: The amino acid sequences of the group consisting of 144 and 145 have amino acid sequences with at least about 95% homology.

[0238] In some embodiments, provided herein are anti-MS4A4A antibodies comprising at least one, at least two, or all three V selected from the group consisting of: HHVR sequences: (a) HVR-H1 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 146 and 147, or an amino acid sequence having at least about 95% homology to an amino acid sequence selected from the group consisting of SEQ ID NOs: 146 and 147; (b) HVR-H2 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 149, 150, 151, 152, and 153, or an amino acid sequence having at least about 95% homology to an amino acid sequence selected from the group consisting of SEQ ID NOs: 149, 150, 151, 152, and 153; and (c) HVR-H3 comprising an amino acid sequence of SEQ ID NO: 154, or an amino acid sequence having at least about 95% homology to an amino acid sequence of SEQ ID NO: 154.

[0239] In some embodiments, provided herein are anti-MS4A4A antibodies comprising at least one, at least two, or all three V selected from the group consisting of: L HVR sequences: (a) HVR-L1 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 156, 157, and 158, or an amino acid sequence having at least about 95% homology to an amino acid sequence selected from the group consisting of SEQ ID NOs: 156, 157, and 158; (b) HVR-L2 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 160 and 161, or an amino acid sequence having at least about 95% homology to an amino acid sequence selected from the group consisting of SEQ ID NOs: 160 and 161; and (c) HVR-L3 comprising an amino acid sequence of SEQ ID NO: 163, or an amino acid sequence having at least about 95% homology to an amino acid sequence of SEQ ID NO: 163.

[0240] In some embodiments, provided herein are anti-MS4A4A antibodies comprising (a) V H A domain comprising at least one, at least two or all three V HHVR sequences: (i) HVR-H1 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 146 and 147, or an amino acid sequence having at least about 95% homology to an amino acid sequence selected from the group consisting of SEQ ID NOs: 146 and 147, (ii) HVR-H2 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 149, 150, 151, 152, and 153, or an amino acid sequence having at least about 95% homology to an amino acid sequence selected from the group consisting of SEQ ID NOs: 149, 150, 151, 152, and 153, and (iii) HVR-H3 comprising an amino acid sequence of SEQ ID NO: 154, or an amino acid sequence having at least about 95% homology to an amino acid sequence of SEQ ID NO: 154; and (b) HVR-H4 comprising an amino acid sequence of SEQ ID NO: 155, or an amino acid sequence having at least about 95% homology to an amino acid sequence of SEQ ID NO: 155; L A domain comprising at least one, at least two or all three V L HVR sequences: (i) HVR-L1 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 156, 157, and 158, or an amino acid sequence having at least about 95% homology to an amino acid sequence selected from the group consisting of SEQ ID NOs: 156, 157, and 158, (ii) HVR-L2 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 160 and 161, or an amino acid sequence having at least about 95% homology to an amino acid sequence selected from the group consisting of SEQ ID NOs: 160 and 161, and (iii) HVR-L3 comprising an amino acid sequence of SEQ ID NO: 163, or an amino acid sequence having at least about 95% homology to an amino acid sequence of SEQ ID NO: 163.

[0241] In another aspect, the anti-MS4A4A antibody comprises a heavy chain variable domain (V ) having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to an amino acid sequence selected from the group consisting of SEQ ID NO: 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 24, 25, 26, 27, 28, 29, 30, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 76, 77, 78, 79, 80, 81, 82, 83, 84, 304, and 306. HIn certain embodiments, V is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to an amino acid sequence selected from the group consisting of SEQ ID NO: 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 24, 25, 26, 27, 28, 29, 30, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 76, 77, 78, 79, 80, 81, 82, 83, 84, 304, and 306. H The sequences contain substitutions (e.g., conservative substitutions), insertions, or deletions relative to the reference sequence, but anti-MS4A4A antibodies comprising the sequences retain the ability to bind to MS4A4A. In certain embodiments, a total of 1 to 10 amino acids have been substituted, inserted, and / or deleted in SEQ ID NO: 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 24, 25, 26, 27, 28, 29, 30, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 76, 77, 78, 79, 80, 81, 82, 83, 84, 304, or 306. In certain embodiments, a total of 1 to 5 amino acids have been substituted, inserted, and / or deleted in SEQ ID NO: 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 24, 25, 26, 27, 28, 29, 30, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 76, 77, 78, 79, 80, 81, 82, 83, 84, 304, or 306. In certain embodiments, substitutions, insertions, or deletions occur in regions outside of the HVRs (i.e., in the FRs). Optionally, the anti-MS4A4A antibody comprises a V sequence of SEQ ID NO: 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 24, 25, 26, 27, 28, 29, 30, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 76, 77, 78, 79, 80, 81, 82, 83, 84, 304, or 306. H Sequence, including post-translational modifications of said sequence. In a specific embodiment, V Hcomprising one, two or three HVRs selected from the following: (a) HVR-H1 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 94, 108, 116, 146, 147 and 308, and 311, (b) HVR-H2 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 96, 97, 98, 99, 110, 111, 118, 119, 120, 121, 122, 149, 150, 151, 152, 153, 309 and 312, and (c) HVR-H3 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 100, 101, 102, 112, 123, 124, 125, 126, 127, 128, 129, 154, 310 and 313.

[0242] In another aspect, an anti-MS4A4A antibody is provided, wherein the antibody comprises a light chain variable domain (V LLD) having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to an amino acid sequence selected from the group consisting of SEQ ID NO: 17, 18, 19, 20, 21, 22, 32, 33, 34, 35, 36, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 86, 87, 88, 89, 90, 91, 92, 93, 305 and 307. L In certain embodiments, V is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to an amino acid sequence selected from the group consisting of SEQ ID NO: 17, 18, 19, 20, 21, 22, 32, 33, 34, 35, 36, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 86, 87, 88, 89, 90, 91, 92, 93, 305, and 307. LThe sequences contain substitutions (e.g., conservative substitutions), insertions, or deletions relative to the reference sequence, but anti-MS4A4A antibodies comprising the sequences retain the ability to bind to MS4A4A. In some embodiments, a total of 1 to 10 amino acids have been substituted, inserted, and / or deleted in SEQ ID NO: 17, 18, 19, 20, 21, 22, 32, 33, 34, 35, 36, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 86, 87, 88, 89, 90, 91, 92, 93, 305, or 307. In certain embodiments, a total of 1 to 5 amino acids have been substituted, inserted, and / or deleted in SEQ ID NO: 17, 18, 19, 20, 21, 22, 32, 33, 34, 35, 36, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 86, 87, 88, 89, 90, 91, 92, 93, 305, or 307. In certain embodiments, substitutions, insertions, or deletions occur in regions outside of the HVRs (i.e., in the FRs). Optionally, the anti-MS4A4A antibody comprises a V sequence of SEQ ID NO: 17, 18, 19, 20, 21, 22, 32, 33, 34, 35, 36, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 86, 87, 88, 89, 90, 91, 92, 93, 305, or 307. L Sequence, including post-translational modifications of said sequence. In a specific embodiment, V L comprising one, two or three HVRs selected from the group consisting of: (a) HVR-L1 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 103, 104, 113, 130, 131, 132, 133, 134, 135, 136, 137, 138, 156, 157, 158, 314 and 317, (b) HVR-L2 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 105, 106, 114, 139, 140, 141, 142, 143, 160, 161, 315 and 318, and (c) HVR-L3 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 107, 115, 144, 145, 163, 316 and 319.

[0243] In some embodiments, an anti-MS4A4A antibody is provided, wherein the antibody comprises V as in any of the embodiments provided above. Hand V in any one of the embodiments provided above L In some embodiments, provided herein are anti-MS4A4A antibodies, wherein the antibody comprises V as in any of the embodiments provided above. H and V in any one of the embodiments provided above L In one embodiment, the antibody comprises V sequences in SEQ ID NOs: 5-15 and 17-22, respectively. H and V L Sequences, including post-translational modifications of those sequences. In one embodiment, the antibody comprises V sequences in SEQ ID NOs: 24-30 and 32-36, respectively. H and V L Sequences, including post-translational modifications of those sequences. In one embodiment, the antibody comprises V sequences in SEQ ID NOs: 40-53 and 55-74, respectively. H and V L Sequences, including post-translational modifications of those sequences. In one embodiment, the antibody comprises V sequences in SEQ ID NOs: 76-84 and 86-93, respectively. H and V L Sequences, including post-translational modifications of those sequences. In one embodiment, the antibody comprises V sequences in SEQ ID NO: 304 and SEQ ID NO: 305, respectively. H and V L Sequences, including post-translational modifications of those sequences. In one embodiment, the antibody comprises V sequences in SEQ ID NO: 306 and SEQ ID NO: 307, respectively. H and V L sequences, including post-translational modifications of those sequences.

[0244] In some embodiments, provided herein are anti-MS4A4A antibodies comprising a heavy chain variable domain (V H ) and light chain variable domain (V L ), wherein the V H and the V L Selected from the group consisting of: V comprising the amino acid sequence of SEQ ID NO: 5 H and V comprising the amino acid sequence of SEQ ID NO: 17 L ; V comprising the amino acid sequence of SEQ ID NO: 5 H and V comprising the amino acid sequence of SEQ ID NO: 18 L ; V comprising the amino acid sequence of SEQ ID NO: 6H and V comprising the amino acid sequence of SEQ ID NO: 17 L ; V comprising the amino acid sequence of SEQ ID NO: 5 H and V comprising the amino acid sequence of SEQ ID NO: 19 L ; V comprising the amino acid sequence of SEQ ID NO: 7 H and V comprising the amino acid sequence of SEQ ID NO: 17 L ; V comprising the amino acid sequence of SEQ ID NO: 5 H and V comprising the amino acid sequence of SEQ ID NO: 20 L ; V comprising the amino acid sequence of SEQ ID NO: 8 H and V comprising the amino acid sequence of SEQ ID NO: 17 L ; V comprising the amino acid sequence of SEQ ID NO: 9 H and V comprising the amino acid sequence of SEQ ID NO: 17 L ; V comprising the amino acid sequence of SEQ ID NO: 10 H and V comprising the amino acid sequence of SEQ ID NO: 18 L ; V comprising the amino acid sequence of SEQ ID NO: 11 H and V comprising the amino acid sequence of SEQ ID NO: 17 L ; V comprising the amino acid sequence of SEQ ID NO: 12 H and V comprising the amino acid sequence of SEQ ID NO: 17 L ; V comprising the amino acid sequence of SEQ ID NO: 13 H and V comprising the amino acid sequence of SEQ ID NO: 21 L ; V comprising the amino acid sequence of SEQ ID NO: 14 H and V comprising the amino acid sequence of SEQ ID NO: 22 L ; V comprising the amino acid sequence of SEQ ID NO: 15 H and V comprising the amino acid sequence of SEQ ID NO: 122 L ; V comprising the amino acid sequence of SEQ ID NO: 304 H and V comprising the amino acid sequence of SEQ ID NO: 305 L ; and V comprising the amino acid sequence of SEQ ID NO: 306 H and V comprising the amino acid sequence of SEQ ID NO: 307 L .

[0245] In some embodiments, provided herein are anti-MS4A4A antibodies comprising a heavy chain variable domain (V H ) and light chain variable domain (V L ), wherein the V H and the V L Selected from the group consisting of: V comprising the amino acid sequence of SEQ ID NO: 24 H and V comprising the amino acid sequence of SEQ ID NO: 32 L ; V comprising the amino acid sequence of SEQ ID NO: 25 H and V comprising the amino acid sequence of SEQ ID NO: 33 L ; V comprising the amino acid sequence of SEQ ID NO: 25 H and V comprising the amino acid sequence of SEQ ID NO: 32 L ; V comprising the amino acid sequence of SEQ ID NO: 26 H and V comprising the amino acid sequence of SEQ ID NO: 33 L ; V comprising the amino acid sequence of SEQ ID NO: 26 H and V comprising the amino acid sequence of SEQ ID NO: 32 L ; V comprising the amino acid sequence of SEQ ID NO: 27 H and V comprising the amino acid sequence of SEQ ID NO: 32 L ; V comprising the amino acid sequence of SEQ ID NO: 27 H and V comprising the amino acid sequence of SEQ ID NO: 34 L ; V comprising the amino acid sequence of SEQ ID NO: 28 H and V comprising the amino acid sequence of SEQ ID NO: 33 L ; V comprising the amino acid sequence of SEQ ID NO: 28 H and V comprising the amino acid sequence of SEQ ID NO: 32 L ; V comprising the amino acid sequence of SEQ ID NO: 28 H and V comprising the amino acid sequence of SEQ ID NO: 35 L ; V comprising the amino acid sequence of SEQ ID NO: 28 H and V comprising the amino acid sequence of SEQ ID NO: 34 L ; V comprising the amino acid sequence of SEQ ID NO: 29 H and V comprising the amino acid sequence of SEQ ID NO: 32 L; V comprising the amino acid sequence of SEQ ID NO: 29 H and V comprising the amino acid sequence of SEQ ID NO: 36 L ; V comprising the amino acid sequence of SEQ ID NO: 30 H and V comprising the amino acid sequence of SEQ ID NO: 37 L ; V comprising the amino acid sequence of SEQ ID NO: 30 H and V comprising the amino acid sequence of SEQ ID NO: 32 L ; V comprising the amino acid sequence of SEQ ID NO: 30 H and V comprising the amino acid sequence of SEQ ID NO: 38 L ; and V comprising the amino acid sequence of SEQ ID NO: 30 H and V comprising the amino acid sequence of SEQ ID NO: 36 L .

[0246] In some embodiments, provided herein are anti-MS4A4A antibodies comprising a heavy chain variable domain (V H ) and light chain variable domain (V L ), wherein the V H and the V L Selected from the group consisting of: V comprising the amino acid sequence of SEQ ID NO: 40 H and V comprising the amino acid sequence of SEQ ID NO: 55 L ; V comprising the amino acid sequence of SEQ ID NO: 41 H and V comprising the amino acid sequence of SEQ ID NO: 55 L ; V comprising the amino acid sequence of SEQ ID NO: 40 H and V comprising the amino acid sequence of SEQ ID NO: 56 L ; V comprising the amino acid sequence of SEQ ID NO: 40 H and V comprising the amino acid sequence of SEQ ID NO: 57 L ; V comprising the amino acid sequence of SEQ ID NO: 40 H and V comprising the amino acid sequence of SEQ ID NO: 58 L ; V comprising the amino acid sequence of SEQ ID NO: 41 H and V comprising the amino acid sequence of SEQ ID NO: 56 L ; V comprising the amino acid sequence of SEQ ID NO: 41 H and V comprising the amino acid sequence of SEQ ID NO: 57L ; V comprising the amino acid sequence of SEQ ID NO: 41 H and V comprising the amino acid sequence of SEQ ID NO: 58 L ; V comprising the amino acid sequence of SEQ ID NO: 42 H and V comprising the amino acid sequence of SEQ ID NO: 59 L ; V comprising the amino acid sequence of SEQ ID NO: 43 H and V comprising the amino acid sequence of SEQ ID NO: 59 L ; V comprising the amino acid sequence of SEQ ID NO: 44 H and V comprising the amino acid sequence of SEQ ID NO: 60 L ; V comprising the amino acid sequence of SEQ ID NO: 45 H and V comprising the amino acid sequence of SEQ ID NO: 60 L ; V comprising the amino acid sequence of SEQ ID NO: 46 H and V comprising the amino acid sequence of SEQ ID NO: 60 L ; V comprising the amino acid sequence of SEQ ID NO: 43 H and V comprising the amino acid sequence of SEQ ID NO: 61 L ; V comprising the amino acid sequence of SEQ ID NO: 46 H and V comprising the amino acid sequence of SEQ ID NO: 61 L ; V comprising the amino acid sequence of SEQ ID NO: 42 H and V comprising the amino acid sequence of SEQ ID NO: 61 L ; V comprising the amino acid sequence of SEQ ID NO: 44 H and V comprising the amino acid sequence of SEQ ID NO: 61 L ; V comprising the amino acid sequence of SEQ ID NO: 45 H and V comprising the amino acid sequence of SEQ ID NO: 61 L ; V comprising the amino acid sequence of SEQ ID NO: 47 H and V comprising the amino acid sequence of SEQ ID NO: 55 L ; V comprising the amino acid sequence of SEQ ID NO: 47 H and V comprising the amino acid sequence of SEQ ID NO: 62 L ; V comprising the amino acid sequence of SEQ ID NO: 47 Hand V comprising the amino acid sequence of SEQ ID NO: 63 L ; V comprising the amino acid sequence of SEQ ID NO: 47 H and V comprising the amino acid sequence of SEQ ID NO: 64 L ; V comprising the amino acid sequence of SEQ ID NO: 47 H and V comprising the amino acid sequence of SEQ ID NO: 65 L ; V comprising the amino acid sequence of SEQ ID NO: 47 H and V comprising the amino acid sequence of SEQ ID NO: 66 L ; V comprising the amino acid sequence of SEQ ID NO: 47 H and V comprising the amino acid sequence of SEQ ID NO: 67 L ; V comprising the amino acid sequence of SEQ ID NO: 47 H and V comprising the amino acid sequence of SEQ ID NO: 68 L ; V comprising the amino acid sequence of SEQ ID NO: 47 H and V comprising the amino acid sequence of SEQ ID NO: 69 L ; V comprising the amino acid sequence of SEQ ID NO: 47 H and V comprising the amino acid sequence of SEQ ID NO: 70 L ; V comprising the amino acid sequence of SEQ ID NO: 47 H and V comprising the amino acid sequence of SEQ ID NO: 71 L ; V comprising the amino acid sequence of SEQ ID NO: 47 H and V comprising the amino acid sequence of SEQ ID NO: 72 L ; V comprising the amino acid sequence of SEQ ID NO: 47 H and V comprising the amino acid sequence of SEQ ID NO: 73 L ; V comprising the amino acid sequence of SEQ ID NO: 47 H and V comprising the amino acid sequence of SEQ ID NO: 74 L ; V comprising the amino acid sequence of SEQ ID NO: 48 H and V comprising the amino acid sequence of SEQ ID NO: 70 L ; V comprising the amino acid sequence of SEQ ID NO: 48 H and V comprising the amino acid sequence of SEQ ID NO: 74 L ; V comprising the amino acid sequence of SEQ ID NO: 48H and V comprising the amino acid sequence of SEQ ID NO: 62 L ; V comprising the amino acid sequence of SEQ ID NO: 48 H and V comprising the amino acid sequence of SEQ ID NO: 55 L ; V comprising the amino acid sequence of SEQ ID NO: 48 H and V comprising the amino acid sequence of SEQ ID NO: 72 L ; V comprising the amino acid sequence of SEQ ID NO: 48 H and V comprising the amino acid sequence of SEQ ID NO: 69 L ; V comprising the amino acid sequence of SEQ ID NO: 49 H and V comprising the amino acid sequence of SEQ ID NO: 71 L ; V comprising the amino acid sequence of SEQ ID NO: 49 H and V comprising the amino acid sequence of SEQ ID NO: 55 L ; V comprising the amino acid sequence of SEQ ID NO: 49 H and V comprising the amino acid sequence of SEQ ID NO: 73 L ; V comprising the amino acid sequence of SEQ ID NO: 50 H and V comprising the amino acid sequence of SEQ ID NO: 62 L ; V comprising the amino acid sequence of SEQ ID NO: 51 H and V comprising the amino acid sequence of SEQ ID NO: 62 L ; V comprising the amino acid sequence of SEQ ID NO: 52 H and V comprising the amino acid sequence of SEQ ID NO: 62 L ; V comprising the amino acid sequence of SEQ ID NO: 53 H and V comprising the amino acid sequence of SEQ ID NO: 62 L ; V comprising the amino acid sequence of SEQ ID NO: 53 H and V comprising the amino acid sequence of SEQ ID NO: 63 L ; V comprising the amino acid sequence of SEQ ID NO: 53 H and V comprising the amino acid sequence of SEQ ID NO: 64 L ; V comprising the amino acid sequence of SEQ ID NO: 53 H and V comprising the amino acid sequence of SEQ ID NO: 65 L; V comprising the amino acid sequence of SEQ ID NO: 53 H and V comprising the amino acid sequence of SEQ ID NO: 66 L ; and V comprising the amino acid sequence of SEQ ID NO: 53 H and V comprising the amino acid sequence of SEQ ID NO: 67 L .

[0247] In some embodiments, provided herein are anti-MS4A4A antibodies comprising a heavy chain variable domain (V H ) and light chain variable domain (V L ), wherein the V H and the V L Selected from the group consisting of: V comprising the amino acid sequence of SEQ ID NO: 76 H and V comprising the amino acid sequence of SEQ ID NO: 86 L ; V comprising the amino acid sequence of SEQ ID NO: 77 H and V comprising the amino acid sequence of SEQ ID NO: 87 L ; V comprising the amino acid sequence of SEQ ID NO: 78 H and V comprising the amino acid sequence of SEQ ID NO: 88 L ; V comprising the amino acid sequence of SEQ ID NO: 79 H and V comprising the amino acid sequence of SEQ ID NO: 89 L ; V comprising the amino acid sequence of SEQ ID NO: 80 H and V comprising the amino acid sequence of SEQ ID NO: 90 L ; V comprising the amino acid sequence of SEQ ID NO: 81 H and V comprising the amino acid sequence of SEQ ID NO: 91 L ; V comprising the amino acid sequence of SEQ ID NO: 82 H and V comprising the amino acid sequence of SEQ ID NO: 91 L ; V comprising the amino acid sequence of SEQ ID NO: 83 H and V comprising the amino acid sequence of SEQ ID NO: 92 L ; and V comprising the amino acid sequence of SEQ ID NO: 84 H and V comprising the amino acid sequence of SEQ ID NO: 93 L .

[0248] In another aspect, an anti-MS4A4A antibody comprises a heavy chain variable domain (V ) having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 304 and 306. H In certain embodiments, V is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to an amino acid sequence selected from the group consisting of SEQ ID NOs: 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 304 and 306. H The sequence contains substitutions (e.g., conservative substitutions), insertions, or deletions relative to the reference sequence, but the anti-MS4A4A antibody comprising the sequence retains the ability to bind to MS4A4A. In certain embodiments, a total of 1 to 10 amino acids have been substituted, inserted, and / or deleted in SEQ ID NO: 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 304, or 306. In certain embodiments, a total of 1 to 5 amino acids have been substituted, inserted, and / or deleted in SEQ ID NO: 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 304, or 306. In certain embodiments, the substitutions, insertions, or deletions occur in regions outside of the HVRs (i.e., in the FRs). Optionally, the anti-MS4A4A antibody comprises the V region of SEQ ID NO: 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 304, or 306. H Sequence, including post-translational modifications of said sequence. In a specific embodiment, V H comprising one, two or three HVRs selected from the group consisting of: (a) HVR-H1 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 94 and 308, (b) HVR-H2 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 96, 97, 98, 99 and 309, and (c) HVR-H3 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 100, 101, 102 and 310.

[0249] In another aspect, an anti-MS4A4A antibody is provided, wherein the antibody comprises a light chain variable domain (V LLD) having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 17, 18, 19, 20, 21, 22, 305 and 307.L In certain embodiments, V is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to an amino acid sequence selected from the group consisting of SEQ ID NOs: 17, 18, 19, 20, 21, 22, 305 and 307. L The sequence contains substitutions (e.g., conservative substitutions), insertions, or deletions relative to the reference sequence, but the anti-MS4A4A antibody comprising the sequence retains the ability to bind to MS4A4A. In some embodiments, a total of 1 to 10 amino acids have been substituted, inserted, and / or deleted in SEQ ID NO: 17, 18, 19, 20, 21, 22, 305, or 307. In certain embodiments, a total of 1 to 5 amino acids have been substituted, inserted, and / or deleted in SEQ ID NO: 17, 18, 19, 20, 21, 22, 305, or 307. In certain embodiments, the substitutions, insertions, or deletions occur in regions outside the HVRs (i.e., in the FRs). Optionally, the anti-MS4A4A antibody comprises the V region of SEQ ID NO: 17, 18, 19, 20, 21, 22, 305, or 307. L Sequence, including post-translational modifications of said sequence. In a specific embodiment, V L comprising one, two or three HVRs selected from the group consisting of: (a) HVR-L1 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 103, 104 and 314, (b) HVR-L2 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 105, 106 and 315, and (c) HVR-L3 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 107 and 316.

[0250] In another aspect, an anti-MS4A4A antibody comprises a heavy chain variable domain (V ) having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 24, 25, 26, 27, 28, 29 and 30. H In certain embodiments, V is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to an amino acid sequence selected from the group consisting of SEQ ID NOs: 24, 25, 26, 27, 28, 29 and 30. HThe sequence contains substitutions (e.g., conservative substitutions), insertions, or deletions relative to the reference sequence, but the anti-MS4A4A antibody comprising the sequence retains the ability to bind to MS4A4A. In certain embodiments, in SEQ ID NO: 24, 25, 26, 27, 28, 29, or 30, a total of 1 to 10 amino acids have been substituted, inserted, and / or deleted. In certain embodiments, in SEQ ID NO: 24, 25, 26, 27, 28, 29, or 30, a total of 1 to 5 amino acids have been substituted, inserted, and / or deleted. In certain embodiments, the substitutions, insertions, or deletions occur in regions outside the HVRs (i.e., in the FRs). Optionally, the anti-MS4A4A antibody comprises the Vs of SEQ ID NO: 24, 25, 26, 27, 28, 29, or 30. H Sequence, including post-translational modifications of said sequence. In a specific embodiment, V H comprising one, two or three HVRs selected from the following: (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 108, (b) HVR-H2 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 110 and 111, and (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO: 112.

[0251] In another aspect, an anti-MS4A4A antibody is provided, wherein the antibody comprises a light chain variable domain (V LLD) having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 32, 33, 34, 35 and 36. L In certain embodiments, V is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to an amino acid sequence selected from the group consisting of SEQ ID NOs: 32, 33, 34, 35 and 36. LThe sequence contains substitutions (e.g., conservative substitutions), insertions, or deletions relative to the reference sequence, but the anti-MS4A4A antibody comprising the sequence retains the ability to bind to MS4A4A. In some embodiments, in SEQ ID NO: 32, 33, 34, 35, or 36, a total of 1 to 10 amino acids have been substituted, inserted, and / or deleted. In certain embodiments, in SEQ ID NO: 32, 33, 34, 35, or 36, a total of 1 to 5 amino acids have been substituted, inserted, and / or deleted. In certain embodiments, the substitutions, insertions, or deletions occur in regions outside the HVRs (i.e., in the FRs). Optionally, the anti-MS4A4A antibody comprises the Vs of SEQ ID NO: 32, 33, 34, 35, or 36. L Sequence, including post-translational modifications of said sequence. In a specific embodiment, V L comprising one, two or three HVRs selected from the group consisting of: (a) HVR-L1 comprising the amino acid sequence of SEQ ID NO: 113, (b) HVR-L2 comprising the amino acid sequence of SEQ ID NO: 114, and (c) HVR-L3 comprising the amino acid sequence of SEQ ID NO: 115.

[0252] In another aspect, an anti-MS4A4A antibody comprises a heavy chain variable domain (V ) having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, and 53. H In certain embodiments, V is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to an amino acid sequence selected from the group consisting of SEQ ID NOs: 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, and 53. HThe sequences contain substitutions (e.g., conservative substitutions), insertions, or deletions relative to the reference sequence, but anti-MS4A4A antibodies comprising the sequences retain the ability to bind to MS4A4A. In certain embodiments, a total of 1 to 10 amino acids have been substituted, inserted, and / or deleted in SEQ ID NO: 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, or 53. In certain embodiments, a total of 1 to 5 amino acids have been substituted, inserted, and / or deleted in SEQ ID NO: 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, or 53. In certain embodiments, the substitutions, insertions, or deletions occur in regions outside of the HVRs (i.e., in the FRs). Optionally, the anti-MS4A4A antibody comprises a V sequence of SEQ ID NO: 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, or 53. H Sequence, including post-translational modifications of said sequence. In a specific embodiment, V H comprising one, two or three HVRs selected from the following: (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 116, (b) HVR-H2 comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 118, 119, 120, 121 and 122, and (c) HVR-H3 comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 123, 124, 125, 126, 127, 128 and 129.

[0253] In another aspect, an anti-MS4A4A antibody is provided, wherein the antibody comprises a light chain variable domain (V LLD) having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to an amino acid sequence selected from the group consisting of SEQ ID NO: 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73 and 74. L In certain embodiments, V is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to an amino acid sequence selected from the group consisting of SEQ ID NOs: 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, and 74. LThe sequences contain substitutions (e.g., conservative substitutions), insertions, or deletions relative to the reference sequence, but anti-MS4A4A antibodies comprising the sequences retain the ability to bind to MS4A4A. In some embodiments, a total of 1 to 10 amino acids have been substituted, inserted, and / or deleted in SEQ ID NO: 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, or 74. In certain embodiments, a total of 1 to 5 amino acids have been substituted, inserted, and / or deleted in SEQ ID NO: 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, or 74. In certain embodiments, the substitutions, insertions, or deletions occur in regions outside the HVRs (i.e., in the FRs). Optionally, the anti-MS4A4A antibody comprises a V sequence of SEQ ID NO: 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73 or 74. L Sequence, including post-translational modifications of said sequence. In a specific embodiment, V L comprising one, two or three HVRs selected from the group consisting of: (a) HVR-L1 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 130, 131, 132, 133, 134, 135, 136, 137 and 138, (b) HVR-L2 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 139, 140, 141, 142 and 143, and (c) HVR-L3 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 144 and 145.

[0254] In another aspect, an anti-MS4A4A antibody comprises a heavy chain variable domain (V ) having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 76, 77, 78, 79, 80, 81, 82, 83, and 84. H In certain embodiments, V is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to an amino acid sequence selected from the group consisting of SEQ ID NOs: 76, 77, 78, 79, 80, 81, 82, 83 and 84. HThe sequence contains substitutions (e.g., conservative substitutions), insertions, or deletions relative to the reference sequence, but the anti-MS4A4A antibody comprising the sequence retains the ability to bind to MS4A4A. In certain embodiments, a total of 1 to 10 amino acids have been substituted, inserted, and / or deleted in SEQ ID NO: 76, 77, 78, 79, 80, 81, 82, 83, or 84. In certain embodiments, a total of 1 to 5 amino acids have been substituted, inserted, and / or deleted in SEQ ID NO: 76, 77, 78, 79, 80, 81, 82, 83, or 84. In certain embodiments, the substitutions, insertions, or deletions occur in regions outside the HVRs (i.e., in the FRs). Optionally, the anti-MS4A4A antibody comprises the V sequences of SEQ ID NO: 76, 77, 78, 79, 80, 81, 82, 83, or 84. H Sequence, including post-translational modifications of said sequence. In a specific embodiment, V H comprising one, two or three HVRs selected from the following: (a) HVR-H1 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 146 and 147, (b) HVR-H2 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 149, 150, 151, 152 and 153, and (c) HVR-H3 comprising an amino acid sequence of SEQ ID NO: 154.

[0255] In another aspect, an anti-MS4A4A antibody is provided, wherein the antibody comprises a light chain variable domain (V LLD) having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 86, 87, 88, 89, 90, 91, 92, and 93. L In certain embodiments, V is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to an amino acid sequence selected from the group consisting of SEQ ID NOs: 86, 87, 88, 89, 90, 91, 92, and 93. LThe sequence contains substitutions (e.g., conservative substitutions), insertions, or deletions relative to the reference sequence, but the anti-MS4A4A antibody comprising the sequence retains the ability to bind to MS4A4A. In some embodiments, in SEQ ID NO: 86, 87, 88, 89, 90, 91, 92, or 93, a total of 1 to 10 amino acids have been substituted, inserted, and / or deleted. In certain embodiments, in SEQ ID NO: 86, 87, 88, 89, 90, 91, 92, or 93, a total of 1 to 5 amino acids have been substituted, inserted, and / or deleted. In certain embodiments, the substitutions, insertions, or deletions occur in regions outside the HVRs (i.e., in the FRs). Optionally, the anti-MS4A4A antibody comprises the V sequences of SEQ ID NO: 86, 87, 88, 89, 90, 91, 92, or 93. L Sequence, including post-translational modifications of said sequence. In a specific embodiment, V L comprises one, two or three HVRs selected from the following: (a) HVR-L1 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 156, 157 and 158, (b) HVR-L2 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 160 and 161, and (c) HVR-L3 comprising an amino acid sequence of SEQ ID NO: 163.

[0256] In another aspect, the anti-MS4A4A antibody comprises a full-length heavy chain amino acid sequence that has at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 320-335 and 355-362. In certain embodiments, the full-length heavy chain amino acid sequence that has at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 320-335 and 355-362 contains substitutions (e.g., conservative substitutions), insertions, or deletions relative to the reference sequence, but the anti-MS4A4A antibody comprising such a sequence retains the ability to bind to MS4A4A. In certain embodiments, a total of 1 to 10 amino acids have been substituted, inserted, and / or deleted in SEQ ID NOs: 320-335 and 355-362. In certain embodiments, in SEQ ID NO: 320-335 and 355-362, a total of 1 to 5 amino acids have been substituted, inserted and / or deleted. In certain embodiments, the substitution, insertion or deletion occurs in a region outside the HVR (i.e., in the FR).

[0257] In another aspect, the anti-MS4A4A antibody comprises a full-length light chain amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to an amino acid sequence selected from the group consisting of SEQ ID NOs: 363-365. In certain embodiments, the full-length heavy chain amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to an amino acid sequence selected from the group consisting of SEQ ID NOs: 363-365 contains substitutions (e.g., conservative substitutions), insertions, or deletions relative to the reference sequence, but the anti-MS4A4A antibody comprising such a sequence retains the ability to bind to MS4A4A. In certain embodiments, a total of 1 to 10 amino acids have been substituted, inserted, and / or deleted in SEQ ID NOs: 363-365. In certain embodiments, a total of 1 to 5 amino acids have been substituted, inserted, and / or deleted in SEQ ID NOs: 363-365. In certain embodiments, the substitutions, insertions, or deletions occur in regions outside of the LVRs (i.e., in the FRs).

[0258] In some embodiments, the anti-MS4A4A antibody comprises the full-length heavy chain amino acid sequence of SEQ ID NOs: 355-362 and the full-length light chain amino acid sequence of SEQ ID NO: 365. In some embodiments, the anti-MS4A4A antibody comprises the full-length heavy chain amino acid sequence of SEQ ID NOs: 320-327 and the full-length light chain amino acid sequence of SEQ ID NO: 363. In some embodiments, the anti-MS4A4A antibody comprises the full-length heavy chain amino acid sequence of SEQ ID NOs: 328-335 and the full-length light chain amino acid sequence of SEQ ID NO: 364.

[0259] In some embodiments, an anti-MS4A4A antibody of the present disclosure competitively inhibits binding of at least one reference antibody selected from: 4A-202, 4A-301, 4A-302, 4A-303, 4A-304, 4A-305, 4A-306, 4A-307, 4A-308, 4A-309, 4A-310, 4A-311, 4A-312, 4A-313, 4A-314, 4A-419, and 4A-450. In some embodiments, an anti-MS4A4A antibody of the present disclosure competitively inhibits binding of at least one reference antibody selected from: 4A-18, 4A-315, 4A-316, 4A-317, 4A-318, 4A-319, 4A-320, 4A-321, 4A-322, 4A-323, 4A-324, 4A-325, 4A-326, 4A-327, 4A-328, 4A-329, 4A-330, and 4A-331. In some embodiments, an anti-MS4A4A antibody of the present disclosure competitively inhibits binding of at least one reference antibody selected from: 4A-21, 4A-332, 4A-333, 4A-334, 4A-335, 4A-336, 4A-337, 4A-338, 4A-339, 4A-340, 4A-341, 4A-342, 4A-343, 4A-344, 4A-345, 4A-346, 4A-347, 4A-348, 4A-349, 4A-350, 4A-351, 4A-352, 4A-353, 4A-354, 4A-355, 4A-356, 4A-357, 4A-358, 4A-359, 4A-360, 4A-361, 4A-362, 4A-363, 4A-364, 4A-365 -353, 4A-354, 4A-355, 4A-356, 4A-357, 4A-358, 4A-359, 4A-360, 4A-361, 4A-361, 4A-363, 4A-364, 4A-365, 4A-366, 4A-3 67, 4A-368, 4A-369, 4A-370, 4A-371, 4A-372, 4A-373, 4A-374, 4A-375, 4A-376, 4A-377, 4A-378, 4A-379, 4A-380 and 4A-381. In some embodiments, an anti-MS4A4A antibody of the present disclosure competitively inhibits binding of at least one reference antibody selected from: 4A-25, 4A-382, 4A-383, 4A-384, 4A-385, 4A-386, 4A-387, 4A-388, 4A-389, 4A-390.

[0260] In some embodiments, an anti-MS4A4A antibody of the present disclosure binds to an epitope of human MS4A4A that is identical to or overlaps with an epitope of MS4A4A bound by at least one reference antibody selected from the group consisting of: 4A-202, 4A-301, 4A-302, 4A-303, 4A-304, 4A-305, 4A-306, 4A-307, 4A-308, 4A-309, 4A-310, 4A-311, 4A-312, 4A-313, 4A-314, 4A-18, 4A-19. -315, 4A-316, 4A-317, 4A-318, 4A-319, 4A-320, 4A-321, 4A-322, 4A-323, 4A-324, 4A-325, 4A-326, 4A-327, 4A- 328, 4A-329, 4A-330, 4A-331, 4A-21, 4A-332, 4A-333, 4A-334, 4A-335, 4A-336, 4A-337, 4A-338, 4A-339, 4A-340 , 4A-341, 4A-342, 4A-343, 4A-344, 4A-345, 4A-346, 4A-347, 4A-348, 4A-349, 4A-350, 4A-351, 4A-352, 4A-353, 4A-354, 4A-355, 4A-356, 4A-357, 4A-358, 4A-359, 4A-360, 4A-361, 4A-361, 4A-363, 4A-364, 4A-365, 4A-366, 4A Detailed exemplary methods for locating epitopes to which antibodies bind are provided in Morris (1996) "Epitope Mapping Protocols," Methods in Molecular Biology , vol. 66 (Humana Press, Totowa, NJ).

[0261] In some embodiments, an anti-MS4A4A antibody of the present disclosure competitively inhibits binding of at least one reference antibody selected from 4A-202, 4A-301, 4A-302, 4A-303, 4A-304, 4A-305, 4A-306, 4A-307, 4A-308, 4A-309, 4A-310, 4A-311, 4A-312, 4A-313, 4A-314, 4A-419, and 4A-450, and any combination thereof, with respect to binding to MS4A4A. In some embodiments, an anti-MS4A4A antibody of the present disclosure competitively inhibits binding of at least one reference antibody selected from 4A-18, 4A-315, 4A-316, 4A-317, 4A-318, 4A-319, 4A-320, 4A-321, 4A-322, 4A-323, 4A-324, 4A-325, 4A-326, 4A-327, 4A-328, 4A-329, 4A-330, and 4A-331, and any combination thereof, for binding to MS4A4A. In some embodiments, an anti-MS4A4A antibody of the present disclosure competitively inhibits binding of at least one reference antibody selected from the group consisting of: 4A-21, 4A-332, 4A-333, 4A-334, 4A-335, 4A-336, 4A-337, 4A-338, 4A-339, 4A-340, 4A-341, 4A-342, 4A-343, 4A-344, 4A-345, 4A-346, 4A-347, 4A-348, 4A-349, 4A-350, 4A-351, 4A-352, 4A-353, 4A-354, 4A-355, 4A-356, 4A-357, 4A-358, 4A-359, 4A-360, 4A-361, 4A-361, 4A-363, 4A-364, 4A-365, 4A-366, 4A-367, 4 and any combination thereof. In some embodiments, an anti-MS4A4A antibody of the present disclosure competitively inhibits binding of at least one reference antibody selected from 4A-25, 4A-382, 4A-383, 4A-384, 4A-385, 4A-386, 4A-387, 4A-388, 4A-389, or 4A-390, and any combination thereof, with respect to binding to MS4A4A.

[0262] In some embodiments, for binding to MS4A4A, the anti-MS4A4A antibodies of the present disclosure have the same or overlapping epitope on MS4A4A as at least one reference antibody selected from the group consisting of: 4A-202, 4A-301, 4A-302, 4A-303, 4A-304, 4A-305, 4A-306, 4A-307, 4A-308, 4A-309, 4A-310, 4A-311, 4A-312, 4A-313, 4A-314, 4A-419, and 4A-450, and any combination thereof. In some embodiments, the anti-MS4A4A antibodies of the present disclosure have, for binding to MS4A4A, an identical or overlapping epitope on MS4A4A as at least one reference antibody selected from the group consisting of: 4A-18, 4A-315, 4A-316, 4A-317, 4A-318, 4A-319, 4A-320, 4A-321, 4A-322, 4A-323, 4A-324, 4A-325, 4A-326, 4A-327, 4A-328, 4A-329, 4A-330, and 4A-331, and any combination thereof. In some embodiments, for binding to MS4A4A, an anti-MS4A4A antibody of the present disclosure has an epitope on MS4A4A that is identical or overlapping with at least one reference antibody selected from the group consisting of: 4A-21, 4A-332, 4A-333, 4A-334, 4A-335, 4A-336, 4A-337, 4A-338, 4A-339, 4A-340, 4A-341, 4A-342, 4A-343, 4A-344, 4A-345, 4A-346, 4A-347, 4A-348, 4A-349, 4A-350, 4A-351, 4A-352, 4A-353, 4A-354, 4A-355, 4A-356, 4A-357, 4A-358, 4A-359, 4A-360, 4A-361, 4A-362, 4A-363, 4A-364, 4A-365, 4A-366, 4A-367, 4A-368, 4A-369, 4A-370, 4A-371, 4A-372, 4A-373, 4A-374, 4A-375 352, 4A-353, 4A-354, 4A-355, 4A-356, 4A-357, 4A-358, 4A-359, 4A-360, 4A-361, 4A-361, 4A-363, 4A-364, 4A-365, 4A-366, 4A-367 , 4A-368, 4A-369, 4A-370, 4A-371, 4A-372, 4A-373, 4A-374, 4A-375, 4A-376, 4A-377, 4A-378, 4A-379, 4A-380 and 4A-381 and any combination thereof. In some embodiments, the anti-MS4A4A antibodies of the present disclosure have, for binding to MS4A4A, an identical or overlapping epitope on MS4A4A as at least one reference antibody selected from the group consisting of: 4A-25, 4A-382, 4A-383, 4A-384, 4A-385, 4A-386, 4A-387, 4A-388, 4A-389, and 4A-390, and any combination thereof.

[0263] In some embodiments, the anti-MS4A4A antibodies of the present disclosure bind to the extracellular domain 1 (ECL1) of MS4A4A. In some embodiments, the anti-MS4A4A antibodies of the present disclosure bind to one or more amino acids within the amino acid sequence CMASNTYGSNPIS (SEQ ID NO: 177) of SEQ ID NO: 1. In some embodiments, the anti-MS4A4A antibodies of the present disclosure bind to the extracellular domain 2 (ECL2) of MS4A4A. In some embodiments, the anti-MS4A4A antibodies of the present disclosure bind to one or more amino acids within the amino acid sequence SFHHPYCNYYGNSNNCHGTMS (SEQ ID NO: 178) of SEQ ID NO: 1.

[0264] Any suitable competition assay or MS4A4A binding assay known in the art (such as BIAcore analysis, ELISA analysis, or flow cytometry) can be used to determine whether an anti-MS4A4A antibody competes for binding to MS4A4A with (or competitively inhibits binding to) one or more reference antibodies selected from the group consisting of: 4A-202, 4A-301, 4A-302, 4A-303, 4A-304, 4A-305, 4A-306, 4A-307, 4A-308, 4A-309, 4A-310, 4A-311, 4A-312, 4A-313, 4A-314, 4A-315, 4A-316, 4A-317, 4A-318, 4A-319, 4A-320, 4A-321, 4A-322, 4A-323, 4A-324, 4A-325, 4A-326, 4A-327, 4A-328, 4A-329, 4A-330 312, 4A-313, 4A-314, 4A-18, 4A-315, 4A-316, 4A-317, 4A-318, 4A-319, 4A-320, 4A-321, 4A-322, 4A-323, 4A-324, 4A-3 25. 4A-326, 4A-327, 4A-328, 4A-329, 4A-330, 4A-331, 4A-21, 4A-332, 4A-333, 4A-334, 4A-335, 4A-336, 4A-337, 4A-338 , 4A-339, 4A-340, 4A-341, 4A-342, 4A-343, 4A-344, 4A-345, 4A-346, 4A-347, 4A-348, 4A-349, 4A-350, 4A-351, 4A-352 , 4A-353, 4A-354, 4A-355, 4A-356, 4A-357, 4A-358, 4A-359, 4A-360, 4A-361, 4A-361, 4A-363, 4A-364, 4A-365, 4A-366, 4A-367, 4A-368, 4A-369, 4A-370, 4A-371, 4A-372, 4A-373, 4A-374, 4A-375, 4A-376, 4A-377, 4A-378, 4A-379, 4A-380, 4A-381, 4A-25, 4A-382, 4A-383, 4A-384, 4A-385, 4A-386, 4A-387, 4A-388, 4A-389, 4A-390, 4A-419 and 4A-450 and any combination thereof. In an exemplary competition assay, immobilized MS4A4A or cells expressing MS4A4A on the cell surface are incubated in a solution containing a first labeled antibody that binds to MS4A4A (e.g., human or non-human primate) and a second unlabeled antibody that is being tested for its ability to compete with the first antibody for binding to MS4A4A. The second antibody can be present in the hybridoma supernatant. As a control, immobilized MS4A4A or cells expressing MS4A4A are incubated in a solution containing the first labeled antibody but not the second unlabeled antibody.After incubation under conditions permissive for binding of the primary antibody to MS4A4A, excess unbound antibody is removed, and the amount of label associated with immobilized MS4A4A or cells expressing MS4A4A is measured. If the amount of label associated with immobilized MS4A4A or cells expressing MS4A4A is substantially reduced in the test sample relative to the control sample, this indicates that the secondary antibody is competing with the primary antibody for binding to MS4A4A. See Harlow and Lane (1988). Antibodies: A Laboratory Manual Chapter 14 (Cold Spring Harbor Laboratory, Cold Spring Harbor, NY).

[0265] Further provided herein are anti-MS4A4A antibodies that competitively inhibit binding of and / or compete with binding of an anti-MS4A4A antibody, comprising (a) V H A domain comprising (i) HVR-H1 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 94 and 308, (ii) HVR-H2 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 96, 97, 98, 99 and 309, and (iii) HVR-H3 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 100, 101, 102 and 310; and (b) V L A domain comprising (i) HVR-L1 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 103, 104, and 314, (ii) HVR-L2 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 105, 106, and 315, and (iii) HVR-L3 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 107 and 316. In some embodiments, the antibody comprises the V sequences in SEQ ID NOs: 5-15 and 304 and SEQ ID NOs: 17-22 and 305, respectively. H and V L sequence.

[0266] Provided herein are anti-MS4A4A antibodies that bind to an epitope of human MS4A4A that is identical to or overlaps with an epitope bound by an anti-MS4A4A antibody, comprising (a) V HA domain comprising (i) HVR-H1 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 94 and 308, (ii) HVR-H2 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 96, 97, 98, 99 and 309, and (iii) HVR-H3 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 100, 101, 102 and 310; and (b) V L A domain comprising (i) HVR-L1 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 103, 104, and 314, (ii) HVR-L2 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 105, 106, and 315, and (iii) HVR-L3 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 107 and 316. In some embodiments, the antibody comprises the V sequences in SEQ ID NOs: 5-15 and 304 and SEQ ID NOs: 17-22 and 305, respectively. H and V L In some embodiments, the epitope of human MS4A4A is the same as the epitope bound by the anti-MS4A4A antibody.

[0267] Further provided herein are anti-MS4A4A antibodies that competitively inhibit binding of and / or compete with binding of an anti-MS4A4A antibody, comprising (a) V H A domain comprising (i) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 108, (ii) HVR-H2 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 110 and 111, and (iii) HVR-H3 comprising the amino acid sequence of SEQ ID NO: 112; and (b) V L A domain comprising (i) HVR-L1 comprising the amino acid sequence of SEQ ID NO: 113, (ii) HVR-L2 comprising the amino acid sequence of SEQ ID NO: 114, and (iii) HVR-L3 comprising the amino acid sequence of SEQ ID NO: 115. In some embodiments, the antibody comprises the V sequences in SEQ ID NOs: 24-30 and 32-36, respectively. H and V L sequence.

[0268] Provided herein are anti-MS4A4A antibodies that bind to an epitope of human MS4A4A that is identical to or overlaps with an epitope bound by an anti-MS4A4A antibody, comprising (a) V H a domain comprising (i) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 108, (ii) HVR-H2 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 110 and 111, and (iii) HVR-H3 comprising the amino acid sequence of SEQ ID NO: 112; and (b) V L A domain comprising (i) HVR-L1 comprising the amino acid sequence of SEQ ID NO: 113, (ii) HVR-L2 comprising the amino acid sequence of SEQ ID NO: 114, and (iii) HVR-L3 comprising the amino acid sequence of SEQ ID NO: 115. In some embodiments, the antibody comprises the V domains in SEQ ID NOs: 24-30 and 32-36, respectively. H and V L In some embodiments, the epitope of human MS4A4A is the same as the epitope bound by the anti-MS4A4A antibody.

[0269] Further provided herein are anti-MS4A4A antibodies that competitively inhibit binding of and / or compete with binding of an anti-MS4A4A antibody, comprising (a) V H A domain comprising (i) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 116, (ii) HVR-H2 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 118, 119, 120, 121, and 122, and (iii) HVR-H3 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 123, 124, 125, 126, 127, 128, and 129; and (b) V L A domain comprising (i) HVR-L1 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 130, 131, 132, 133, 134, 135, 136, 137, and 138, (ii) HVR-L2 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 139, 140, 141, 142, and 143, and (iii) HVR-L3 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 144 and 145. In some embodiments, the antibody comprises V sequences in SEQ ID NOs: 40-53 and 55-74, respectively.H and V L sequence.

[0270] Provided herein are anti-MS4A4A antibodies that bind to an epitope of human MS4A4A that is identical to or overlaps with an epitope bound by an anti-MS4A4A antibody, comprising (a) V H A domain comprising (i) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 116, (ii) HVR-H2 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 118, 119, 120, 121 and 122, and (iii) HVR-H3 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 123, 124, 125, 126, 127, 128 and 129; and (b) V L A domain comprising (i) HVR-L1 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 130, 131, 132, 133, 134, 135, 136, 137, and 138, (ii) HVR-L2 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 139, 140, 141, 142, and 143, and (iii) HVR-L3 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 144 and 145. In some embodiments, the antibody comprises V sequences in SEQ ID NOs: 40-53 and 55-74, respectively. H and V L In some embodiments, the epitope of human MS4A4A is the same as the epitope bound by the anti-MS4A4A antibody.

[0271] Further provided herein are anti-MS4A4A antibodies that competitively inhibit binding of and / or compete with binding of an anti-MS4A4A antibody, comprising (a) V H A domain comprising (i) HVR-H1 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 146 and 147, (ii) HVR-H2 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 149, 150, 151, 152, and 153, and (iii) HVR-H3 comprising an amino acid sequence of SEQ ID NO: 154; and (b) V LA domain comprising (i) HVR-L1 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 156, 157, and 158, (ii) HVR-L2 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 160 and 161, and (iii) HVR-L3 comprising an amino acid sequence of SEQ ID NO: 163. In some embodiments, the antibody comprises V sequences of SEQ ID NOs: 76-84 and 86-93, respectively. H and V L sequence.

[0272] Provided herein are anti-MS4A4A antibodies that bind to an epitope of human MS4A4A that is identical to or overlaps with an epitope bound by an anti-MS4A4A antibody, comprising (a) V H A domain comprising (i) HVR-H1 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 146 and 147, (ii) HVR-H2 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 149, 150, 151, 152, and 153, and (iii) HVR-H3 comprising an amino acid sequence of SEQ ID NO: 154; and (b) V L A domain comprising (i) HVR-L1 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 156, 157, and 158, (ii) HVR-L2 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 160 and 161, and (iii) HVR-L3 comprising an amino acid sequence of SEQ ID NO: 163. In some embodiments, the antibody comprises the V sequences in SEQ ID NOs: 76-84 and 86-93, respectively. H and V L In some embodiments, the epitope of human MS4A4A is the same as the epitope bound by the anti-MS4A4A antibody.

[0273] In some embodiments, the anti-MS4A4A antibody according to any of the above embodiments is a monoclonal antibody, including humanized and / or human antibodies. In some embodiments, the anti-MS4A4A antibody is an antibody fragment, such as a Fv, Fab, Fab', scFv, diabody, or F(ab')2 fragment. In some embodiments, the anti-MS4A4A antibody is a substantially full-length antibody, such as an IgG1 antibody, an IgG2a antibody, or other antibody class or isotype as defined herein.

[0274] In some embodiments, the anti-MS4A4A antibody according to any one of the above embodiments may incorporate any one of the features described in items 1 to 7 below, singly or in combination:

[0275] (1) Anti-MS4A4A antibody binding affinity

[0276] In some embodiments of any of the antibodies provided herein, the antibody has a dissociation constant (Kd) of <1 μM, <100 nM, <10 nM, <1 nM, <0.1 nM, <0.01 nM, or <0.001 nM (e.g., 10 -8 M or smaller, such as 10 -8 M to 10 -13 M, for example 10 -9 M to 10 -13 M). The dissociation constant can be determined by any analytical technique, including any biochemical or biophysical technique, such as ELISA, surface plasmon resonance (SPR), biolayer interferometry (see, e.g., Octet system (ForteBio)), isothermal titration calorimetry (ITC), differential scanning calorimetry (DSC), circular dichroism (CD), stopped-flow analysis, and colorimetric or fluorescent protein melting analysis. In one embodiment, Kd is measured by a radiolabeled antigen binding assay (RIA). In some embodiments, an RIA is performed using a Fab form of the antibody of interest and its antigen, e.g., as described in Chen et al., J. Mol. Biol. 293:865-881 (1999). In some embodiments, Kd is measured using a BIACORE surface plasmon resonance assay, e.g., an assay using a BIACORE-2000 or BIACORE-3000 (BIAcore, Inc., Piscataway, NJ) at 25°C with immobilized antigen CM5 chips at approximately 10 response units (RU). In some embodiments, Kd is measured using a monovalent antibody (e.g., Fab) or a full-length antibody. D In some embodiments, K is determined using a full-length antibody in a monovalent format. D .

[0277] (2) Antibody fragments

[0278] In some embodiments of any of the antibodies provided herein, the antibody is an antibody fragment. Antibody fragments include, but are not limited to, Fab, Fab', Fab'-SH, F(ab')2, Fv, and scFv fragments, as well as other fragments described below. For a review of certain antibody fragments, see Hudson et al., Nat. Med.9:129-134 (2003). For a review of scFv fragments, see, for example, WO 93 / 16185; and U.S. Patent Nos. 5,571,894 and 5,587,458. For a discussion of Fab and F(ab')2 fragments comprising salvage receptor binding epitope residues and having extended in vivo half-lives, see U.S. Patent No. 5,869,046.

[0279] Diabodies are antibody fragments that have two antigen binding sites and can be bivalent or bispecific. See, for example, EP404097; WO 1993 / 01161; Hudson et al., Nat. Med. 9:129-134 (2003). Hudson et al. Nat. Med Trivalent antibodies and tetravalent antibodies are also described in . 9:129-134 (2003). Single-domain antibodies are antibody fragments that contain all or a portion of the heavy chain variable domain or all or a portion of the light chain variable domain of an antibody. In certain embodiments, the single-domain antibody is a human single-domain antibody (see, e.g., U.S. Patent No. 6,248,516).

[0280] As described herein, antibody fragments can be made by a variety of techniques including, but not limited to, proteolytic digestion of intact antibodies and production of fragments by recombinant host cells, such as E. coli ( E. coli ) or phage) production.

[0281] (3) Chimeric and humanized antibodies

[0282] In some embodiments of any of the antibodies provided herein, the antibody is a chimeric antibody. Certain chimeric antibodies are described, for example, in U.S. Patent No. 4,816,567. In one example, a chimeric antibody comprises a non-human variable region (e.g., a variable region derived from a mouse, rat, hamster, rabbit, or non-human primate (such as a monkey)) and a human constant region. In another example, a chimeric antibody is a "class-switched" antibody in which the class or subclass has changed from that of the parent antibody. Chimeric antibodies include antigen-binding fragments thereof.

[0283] In some embodiments of any of the antibodies provided herein, the antibody is a humanized antibody. Typically, a non-human antibody is humanized to reduce immunogenicity to humans while retaining the specificity and affinity of the parent non-human antibody. In certain embodiments, a humanized antibody is substantially non-immunogenic in humans. In certain embodiments, a humanized antibody has substantially the same affinity for a target as an antibody from another species from which the humanized antibody is derived. See, for example, U.S. Patent Nos. 5,530,101; 5,693,761; 5,693,762; and 5,585,089. In certain embodiments, amino acids in the antibody variable domains are identified that can be modified without reducing the natural affinity of the antigen-binding domain while reducing its immunogenicity. See, for example, U.S. Patent Nos. 5,766,886 and 5,869,619. Typically, a humanized antibody comprises one or more variable domains, wherein HVRs (or portions thereof) are derived from non-human antibodies, and FRs (or portions thereof) are derived from human antibody sequences. The humanized antibody optionally also comprises at least a portion of a human constant region. In some embodiments, some FR residues in a humanized antibody are substituted by corresponding residues from a non-human antibody (eg, the antibody from which the HVR residues are derived), for example, to restore or improve antibody specificity or affinity.

[0284] Humanized antibodies and methods for their preparation are reviewed in, for example, Almagro et al. Front. Biosci. 13:1619-1633 (2008), and further described in, for example, U.S. Patent Nos. 5821337, 7527791, 6982321, and 7087409. Human framework regions useful for humanization include, but are not limited to, framework regions selected using the "best fit" method (see, for example, Sims et al., J. Immunol. 151:2296 (1993)); framework regions derived from the consensus sequence of human antibodies of a particular subgroup of light or heavy chain variable regions (see, e.g., Carter et al., Proc. Natl. Acad. Sci. USA 89:4285 (1992); and Presta et al., J. Immunol . 151: 2623 (1993)); human mature (somatically mutated) framework regions or human germline framework regions (see, e.g., Almagro and Fransson, Front. Biosci. 13:1619-1633 (2008)); and framework regions derived from screening FR libraries (see, e.g., Baca et al., J. Biol. Chem. 272:10678-10684 (1997) and Rosok et al. J. Biol. Chem . 271:22611-22618 (1996)).

[0285] (4) Human antibodies

[0286] In some embodiments of any of the antibodies provided herein, the antibody is a human antibody. Human antibodies can be produced using various techniques known in the art. Human antibodies are generally described in van Dijk et al., Curr. Opin. Pharmacol .5:368-74 (2001) and Lonberg Curr. Opin. Immunol . 20:450-459 (2008).

[0287] Human antibodies can be prepared by administering immunogens to transgenic animals, which have been improved to produce complete human antibodies or complete antibodies with human variable regions in response to antigenic attacks. Larger fragments of human Ig loci can be used to engineer defective mouse strains for mouse antibody production, and it is expected that such mice will produce human antibodies in the absence of mouse antibodies. Larger human Ig fragments can maintain larger variable gene diversity and the appropriate regulation of antibody production and expression. By utilizing the mouse machinery (machinery) for antibody diversification and selection and the lack of immune tolerance to human proteins, the human antibody repertoire replicated in these mouse strains can produce high-affinity fully human antibodies for any antigen (including human antigens) of interest. Using hybridoma technology, antigen-specific human MAbs with desired specificity can be produced and selected. Some exemplary methods are described in U.S. Patent number 5545807, EP 546073 and EP 546073. See also, for example, U.S. Patent Nos. 6,075,181 and 6,150,584, which describe XENOMOUSE™ technology; 5,770,429, which describes HUMAB® technology; 7,041,870, which describes KM MOUSE® technology; and U.S. Patent Application Publication No. US 2007 / 0061900, which describes VELOCIMOUSE® technology. Human variable regions from intact antibodies generated by such animals can be further modified, for example, by combining with different human constant regions.

[0288] Human antibodies can also be made by hybridoma-based methods. Human myeloma and mouse-human heteromyeloma cell lines for producing human monoclonal antibodies have been described. (See, for example, Kozbor J. Immunol. 133:3001 (1984) and Boerner et al., J. Immunol. 147:86 (1991)). Human antibodies produced via human B cell hybridoma technology are also described in Li et al., Proc. Natl. Acad. Sci. USA, 1 03:3557-3562 (2006). Other methods include, for example, those described in U.S. Patent No. 7,189,826 (describing the production of monoclonal human IgM antibodies from hybridoma cell lines). Human hybridoma technology (Trioma technology) is also described in Vollmers et al., Histology and Histopathology 20 (3): 927-937 (2005) and Vollmers et al., Methods and Findings in Experimental and Clinical Pharmacology 27 (3): 185-91 (2005). Human antibodies can also be produced by isolating Fv clone variable domain sequences selected from human phage display libraries. Such variable domain sequences can then be combined with desired human constant domains. Techniques for selecting human antibodies from antibody libraries are described below.

[0289] In some embodiments of any of the antibodies provided herein, the antibodies are human antibodies isolated by in vitro methods and / or screening for antibodies with one or more desired activities in combinatorial libraries. Suitable examples include, but are not limited to, phage display (CAT, Morphosys, Dyax, Biosite / Medarex, Xoma, Symphogen, Alexion (formerly Proliferon), Affimed), ribosome display (CAT), yeast display (Adimab), and the like. In certain phage display methods, repertoires of VH and VL genes are individually cloned by polymerase chain reaction (PCR) and randomly recombined in phage libraries, which can then be displayed as in Winter et al., Ann. Rev. Immunol . 12: 433-455 (1994) as described in screening antigen-binding phage. For example, various methods are known in the art for generating phage display libraries and screening such libraries for antibodies with desired binding characteristics. See also Sidhu et al., J. Mol. Biol. 338(2): 299-310,2004;Lee et al. J. Mol. Biol.340(5): 1073-1093, 2004;Fellouse Proc. Natl. Acad. Sci. USA 101(34):12467-12472 (2004); and Lee et al., J. Immunol. Methods 284(-2):1 19-132 (2004). Phages typically display antibody fragments in the form of single-chain Fv (scFv) fragments or Fab fragments. Libraries from immune sources can provide high-affinity antibodies to the immunogen without the need to construct hybridomas. Alternatively, as in Griffiths et al., EMBO J . 12: 725-734 (1993), the natural repertoire can be cloned (e.g., from humans) to provide a single source of antibodies against a wide variety of non-self antigens as well as self antigens without any immunization. Finally, it is also possible, as described by Hoogenboom et al., J. Mol. Biol As described in U.S. Pat. No. 5,750,373 and U.S. Patent Publication Nos. 2007 / 0292936 and 2009 / 0002360, natural libraries are prepared synthetically by cloning unrearranged V-gene segments from stem cells and using PCR primers containing random sequences encoding the hypervariable HVR3 regions to achieve rearrangement in vitro. Patent publications describing human antibody phage libraries include, for example, U.S. Pat. No. 5,750,373 and U.S. Patent Publication Nos. 2007 / 0292936 and 2009 / 0002360. Antibodies isolated from human antibody libraries are referred to herein as human antibodies or human antibody fragments.

[0290] (5) Constant region including Fc region

[0291] In some embodiments of any of the antibodies provided herein, the antibody comprises an Fc. In some embodiments, the Fc is a human IgG1, IgG2, IgG3, and / or IgG4 isotype. In some embodiments, the antibody is an IgG class, an IgM class, or an IgA class.

[0292] In certain embodiments of any of the antibodies provided herein, the antibody has an IgG2 isotype. In some embodiments, the antibody contains a human IgG2 constant region. In some embodiments, the human IgG2 constant region includes an Fc region. In some embodiments, the antibody induces one or more MS4A4A activities or independently binds to an Fc receptor. In some embodiments, the antibody binds to an inhibitory Fc receptor. In certain embodiments, the inhibitory Fc receptor is inhibitory Fc-gamma receptor IIB (FcγIIB).

[0293] In certain embodiments of any of the antibodies provided herein, the antibody has an IgG1 isotype. In some embodiments, the antibody contains a mouse IgG1 constant region. In some embodiments, the antibody contains a human IgG1 constant region. In some embodiments, the human IgG1 constant region includes an Fc region. In some embodiments, the human IgG1 light chain constant region comprises the amino acid sequence of SEQ ID NO: 344. In some embodiments, the antibody binds to an inhibitory Fc receptor. In certain embodiments, the inhibitory Fc receptor is inhibitory Fc-gamma receptor IIB (FcγIIB).

[0294] In certain embodiments of any of the antibodies provided herein, the antibody has an IgG4 isotype. In some embodiments, the antibody contains a human IgG4 constant region. In some embodiments, the human IgG4 constant region includes an Fc region. In some embodiments, the antibody binds to an inhibitory Fc receptor. In certain embodiments, the inhibitory Fc receptor is an inhibitory Fc-gamma receptor IIB (FcγIIB).

[0295] In certain embodiments of any one of the antibodies provided herein, the antibody has a hybrid IgG2 / 4 isotype. In some embodiments, the antibody includes an amino acid sequence comprising EU numbering amino acids 118 to 260 according to human IgG2 and EU numbering amino acids 261 to 447 according to human IgG4 (WO 1997 / 11971; WO 2007 / 106585).

[0296] In some embodiments, the Fc region increases aggregation without complement activation compared to a corresponding antibody comprising an Fc region that does not comprise the amino acid substitution. In some embodiments, the antibody induces one or more activities of the target to which the antibody specifically binds. In some embodiments, the antibody binds to MS4A4A.

[0297] It may also be desirable to modify the anti-MS4A4A antibodies of the present disclosure to improve effector function and / or extend the serum half-life of the antibodies. For example, the Fc receptor binding sites on the constant region may be modified or mutated to remove or reduce binding affinity to certain Fc receptors (such as FcγRI, FcγRII and / or FcγRIII) to reduce antibody-dependent cell-mediated cytotoxicity. In some embodiments, effector function is impaired by removing N-glycosylation from the Fc region of the antibody (e.g., in the CH2 domain of IgG). In some embodiments, as described in WO 99 / 58572 and Armour et al., Molecular Immunology 40:585-593 (2003); Reddy et al. J. ImmunologyEffector function is impaired by modifying regions such as 233-236, 297, and / or 327-331 of human IgG, as described in SEQ ID NO: 164:1925-1933 (2000). In other embodiments, it may also be desirable to modify the anti-MS4A4A antibodies of the present disclosure to improve effector function, thereby increasing selectivity for ITIM-containing FcgRIIb (CD32b) to increase the accumulation of MS4A4A antibodies on neighboring cells without activating humoral responses (including antibody-dependent cell-mediated cytotoxicity and antibody-dependent cellular phagocytosis).

[0298] To extend the serum half-life of antibodies, salvage receptor binding epitopes can be incorporated into antibodies (particularly antibody fragments) as described, for example, in U.S. Pat. No. 5,739,277. As used herein, the term " salvage receptor binding epitope " refers to an epitope of the Fc region of an IgG molecule (eg, IgG1, IgG2, IgG3, or IgG4) that is responsible for extending the serum half-life of the IgG molecule in vivo. Other amino acid sequence modifications.

[0299] (6) Multispecific antibodies

[0300] Multispecific antibodies are antibodies that have binding specificity for at least two different epitopes, including those on the same or another polypeptide (e.g., one or more MS4A4A polypeptides disclosed herein). In some embodiments, multispecific antibodies may be bispecific antibodies. In some embodiments, multispecific antibodies may be trispecific antibodies. In some embodiments, multispecific antibodies may be tetraspecific antibodies. Such antibodies may be derived from full-length antibodies or antibody fragments (e.g., F(ab')2 bispecific antibodies). In some embodiments, the multispecific antibody comprises a first antigen-binding region that binds to a first site on MS4A4A and a second antigen-binding region that binds to a second site on MS4A4A. In some embodiments, the multispecific antibody comprises a first antigen-binding region that binds to MS4A4A and a second antigen-binding region that binds to a second polypeptide.

[0301] Provided herein are multispecific antibodies comprising a first antigen-binding region that binds to MS4A4A, wherein the first antigen-binding region comprises six HVRs of an antibody described herein; and a second antigen-binding region that binds to a second polypeptide. In some embodiments, the first antigen-binding region comprises VVRs of an antibody described herein. H or V L .

[0302] In some embodiments of any of the multispecific antibodies, the second polypeptide is a) an antigen that promotes transport across the blood-brain barrier; (b) an antigen that promotes transport across the blood-brain barrier selected from the group consisting of transferrin receptor (TR); insulin receptor (HIR); insulin-like growth factor receptor (IGFR); low-density lipoprotein receptor-related proteins 1 and 2 (LPR-1 and LPR-2); diphtheria toxin receptor CRM197; llama single domain antibody TMEM 30 (A); protein transduction domain TAT, Syn-B, transmembrane peptide; polyarginine peptide; vascular peptide and ANG1005; (c) a pathogenic protein selected from the group consisting of amyloid beta, oligomeric amyloid beta, amyloid beta plaques, amyloid precursor protein or fragments thereof, Tau, IAPP, alpha-synuclein, TDP-43, FUS protein, C9orf72 (chromosome 9 open reading frame 72), c9RAN protein, prion protein, PrPSc, huntingtin protein, calcitonin, superoxide dismutase, ataxia, ataxia 1, ataxia 2, ataxia 3, ataxia 7, ataxia 8, ataxia 10, Lewy bodies, atrial natriuretic factor, islet amylin, insulin, apolipoprotein AI, serum amyloid A, medin, prolactin, transthyretin, lysozyme, β 2 microglobulin, gelsolin, corneal epithelial protein, cystatin, immunoglobulin light chain AL, S-IBM protein, repeat sequence associated non-ATG (RAN) translation product, dipeptide repeat sequence (DPR) peptide, glycine-alanine (GA) repeat sequence peptide, glycine-proline (GP) repeat sequence peptide, glycine-arginine (GR) repeat sequence peptide, proline-alanine (PA) repeat sequence peptide, ubiquitin and proline-arginine (PR) repeat sequence peptide; (d) ligands and / or proteins expressed on immune cells, wherein the ligands and / or proteins are selected from CD40, OX40, ICOS, CD28, CD137 / 4-1BB, CD27 , GITR, PD-L1, CTLA-4, PD-L2, PD-1, B7-H3, B7-H4, HVEM, BTLA, KIR, GAL9, TIM3, A2AR, LAG-3 and phosphatidylserine; and / or (e) proteins, lipids, polysaccharides or glycolipids expressed on one or more tumor cells and any combination thereof.

[0303] A variety of antigens that facilitate transport across the blood-brain barrier are known in the art (see, e.g., Gabathuler R. Neurobiol. Dis.37:48-57 (2010). Such second antigens include, but are not limited to, transferrin receptor (TR), insulin receptor (HIR), insulin-like growth factor receptor (IGFR), low-density lipoprotein receptor-related proteins 1 and 2 (LPR-1 and LPR-2), diphtheria toxin receptor (including CRM197 (a non-toxic mutant of diphtheria toxin)), llama single domain antibodies (such as TMEM 30(A) (Flippase)), protein transduction domains (such as TAT, Syn-B or transmembrane peptides), polyarginine or generally positively charged peptides, vascular peptides (such as ANG1005) (see, e.g., Gabathuler, 2010), and other cell surface proteins enriched on blood-brain barrier endothelial cells (see, e.g., Daneman et al., 2010). PLoS One 5(10):e13741 (2010)).

[0304] The multivalent antibody may recognize the MS4A4A antigen and other antigens, including but not limited to, Aβ peptide, antigen, α-synuclein antigen, Tau protein antigen, TDP-43 protein antigen, prion protein antigen, huntingtin protein antigen, RAN, translation product antigens, including dipeptide repeats (DPR peptides) consisting of glycine-alanine (GA), glycine-proline (GP), glycine-arginine (GR), proline-alanine (PA), or proline-arginine (PR), insulin receptor, insulin-like growth factor receptor. Transferrin receptor or any other antigen that promotes antibody transfer across the blood-brain barrier. In some embodiments, the second polypeptide is transferrin. In some embodiments, the second polypeptide is Tau. In some embodiments, the second polypeptide is Aβ. In some embodiments, the second polypeptide is TREM2. In some embodiments, the second polypeptide is α-synuclein.

[0305] A multivalent antibody comprises at least one polypeptide chain (and preferably two polypeptide chains), wherein the one or more polypeptide chains comprise two or more variable domains. For example, the one or more polypeptide chains may comprise VD1-(X1) n -VD2-(X2) n -Fc, wherein VD1 is the first variable domain, VD2 is the second variable domain, Fc is a polypeptide chain of the Fc region, X1 and X2 represent amino acids or polypeptides, and n is 0 or 1. Similarly, the one or more polypeptide chains may comprise V H -C H 1-Flexible joint-V H -C H 1-Fc region chain; or V H -C H 1-V H -C H1-Fc region chain. The multivalent antibody herein preferably further comprises at least two (and preferably four) light chain variable domain polypeptides. The multivalent antibody herein may, for example, comprise from about two to about eight light chain variable domain polypeptides. The light chain variable domain polypeptides contemplated herein comprise a light chain variable domain and, optionally, further comprise a CL domain.

[0306] Techniques for preparing multispecific antibodies include, but are not limited to, recombinant co-expression of two immunoglobulin heavy chain-light chain pairs with different specificities (see Milstein and Cuello, 1996). Nature 305:537 (1983)), WO 93 / 08829 and Traunecker et al., EMBO J. 10:3655 (1991)) and "knob-in-hole" engineering (see, e.g., U.S. Pat. No. 5,731,168). See also WO 2013 / 026833 (CrossMab). Multispecific antibodies can also be prepared by engineering electrostatic steering effects for preparing antibody Fc-heterodimer molecules (WO 2009 / 089004A1); cross-linking two or more antibodies (see, e.g., U.S. Pat. No. 4,676,980); using leucine; using "diabody" technology for preparing bispecific antibody fragments (see, e.g., Hollinger et al., Proc. Natl. Acad. Sci. USA 90:6444-6448 (1993)); and using single-chain Fv (scFv) dimers (see, e.g., Gruber et al., J. Immunol. 152:5368 (1994); and as, for example, Tutt et al., J. Immunol. Trispecific antibodies were prepared as described in 147: 60 (1991).

[0307] Also included herein are engineered antibodies with three or more functional antigen-binding sites, including "octopus antibodies" (see, e.g., US 2006 / 0025576). Antibodies herein also include "dual-acting FAbs" or "DAFs," which contain antigen-binding sites that bind to multiple MS4A4A (see, e.g., US 2008 / 0069820).

[0308] (7) Antibody variants

[0309] In some embodiments of any of the antibodies provided herein, amino acid sequence variants of the antibodies are contemplated. For example, it may be desirable to improve the binding affinity and / or other biological properties of the antibodies.

[0310] (i) Substitution, insertion and deletion variants

[0311] In some embodiments of any one of the antibodies provided herein, antibody variants with one or more amino acid substitutions are provided. The amino acid sequence variants of the antibody can be prepared by introducing suitable modifications into the nucleotide sequence encoding the antibody or by peptide synthesis. Such modifications include, for example, deletions and / or insertions and / or substitutions of residues in the amino acid sequence of the antibody.

[0312] Table A: Amino Acid Substitutions

[0313]

[0314] Substantial modification of the biological properties of antibodies is achieved by selecting substitutions that differ significantly in their effect on preserving (a) the structure of the polypeptide backbone in the area of ​​the substitution, such as a sheet or helical conformation, (b) the charge or hydrophobicity of the molecule at the target site, or (c) the bulk of the side chain. Natural residues are divided into the following groups based on common side chain properties:

[0315] (1) Hydrophobicity: norleucine, Met, Ala, Val, Leu, Ile;

[0316] (2) Neutral hydrophilicity: Cys, Ser, Thr, Asn, Gln;

[0317] (3) Acidic: Asp, Glu;

[0318] (4) Basic: His, Lys, Arg;

[0319] (5) Residues that affect chain orientation: Gly, Pro; and

[0320] (6) Aromatic: Trp, Tyr, Phe.

[0321] For example, non-conservative substitutions may involve exchanging a member of one of these classes for a member of another class.Such substitution residues may be introduced into regions of a human antibody, for example, that are homologous to a non-human antibody, or into non-homologous regions of the molecule.

[0322] According to certain embodiments, when making changes to the polypeptides or antibodies described herein, the hydropathic index of the amino acids may be considered. Each amino acid has been assigned a hydropathic index based on its hydrophobicity and charge characteristics. The hydropathicity indices are: isoleucine (+4.5); valine (+4.2); leucine (+3.8); phenylalanine (+2.8); cysteine / cystine (+2.5); methionine (+1.9); alanine (+1.8); glycine (−0.4); threonine (−0.7); serine (−0.8); tryptophan (−0.9); tyrosine (−1.3); proline (−1.6); histidine (−3.2); glutamate (−3.5); glutamine (−3.5); aspartate (−3.5); asparagine (−3.5); lysine (−3.9); and arginine (−4.5).

[0323] The importance of the hydropathic amino acid index in conferring interactive biological function on proteins is well understood in the art. Kyte et al., J. Mol. Biol ., 157:105-131 (1982). It is known that certain amino acids can be substituted for other amino acids with similar hydropathicity indices or scores and still retain similar biological activity. When making changes based on the hydropathicity index, in certain embodiments, substitutions of amino acids with hydropathicity indices within ±2 are included. In certain embodiments, substitutions of amino acids within ±1 are included, and in certain embodiments, substitutions of amino acids within ±0.5 are included.

[0324] It is also understood in the art that substitution of similar amino acids can be effectively made based on hydrophilicity, particularly where the resulting biologically functional protein or peptide is intended for use in immunological embodiments, as is the case with the present invention. In certain embodiments, the greatest local average hydrophilicity of a protein (as governed by the hydrophilicity of its neighboring amino acids) correlates with its immunogenicity and antigenicity (i.e., the biological properties of the protein).

[0325] The following hydrophilicity values ​​have been assigned to the amino acid residues: arginine (+3.0); lysine (+3.0 ± 1); aspartate (+3.0 ± 1); glutamate (+3.0 ± 1); serine (+0.3); asparagine (+0.2); glutamine (+0.2); glycine (0); threonine (−0.4); proline (−0.5 ± 1); alanine (−0.5); histidine (−0.5); cysteine ​​(−1.0); methionine (−1.3); valine (−1.5); leucine (−1.8); isoleucine (−1.8); tyrosine (−2.3); phenylalanine (−2.5) and tryptophan (−3.4). When making changes based on similar hydrophilicity values, in certain embodiments, substitution of amino acids with hydrophilicity values ​​within ±2 is included, in certain embodiments, substitution of those amino acids within ±1 is included, and in certain embodiments, substitution of those amino acids within ±0.5 is included. Epitopes can also be identified from the primary amino acid sequence based on hydrophilicity. These regions are also referred to as "epitope core regions."

[0326] In certain embodiments, substitutions, insertions or deletions may occur within one or more HVRs, as long as such changes do not substantially reduce the ability of the antibody to bind to antigen. For example, conservative changes that do not substantially reduce binding affinity may be made to HVRs (e.g., conservative substitutions as provided herein). Such changes may, for example, be outside the antigen contact residues in HVRs. In certain embodiments of the variant VH and VL sequences provided above, each HVR is unchanged or contains no more than one, two or three amino acid substitutions.

[0327] Amino acid sequence insertions include amino-terminal and / or carboxyl-terminal fusions (ranging in length from one residue to polypeptides containing hundreds or more residues) as well as intrasequence insertions of single or multiple amino acid residues. Examples of terminal insertions include antibodies with an N-terminal methionyl residue. Other insertion variants of the antibody molecule include fusions of the N-terminus or C-terminus of the antibody with an enzyme (e.g., an enzyme for ADEPT) or a polypeptide that extends the serum half-life of the antibody.

[0328] Any cysteine ​​residue that is not involved in maintaining the proper conformation of the antibody may also be substituted with serine, to improve the oxidative stability of the molecule and prevent abnormal cross-linking. Conversely, one or more cysteine ​​bonds may be added to the antibody to improve its stability (especially when the antibody is an antibody fragment (such as an Fv fragment)).

[0329] (ii) Glycosylation variants

[0330] In some embodiments of any of the antibodies provided herein, the antibody is altered to increase or decrease the degree of glycosylation of the antibody. The addition or deletion of glycosylation sites in an antibody can be conveniently achieved by altering the amino acid sequence to create or remove one or more glycosylation sites.

[0331] Glycosylation of antibodies is typically either N-linked or O-linked. N-linked refers to the attachment of a carbohydrate moiety to the side chain of an asparagine residue. The tripeptide sequences asparagine-X-serine and asparagine-X-threonine (where X is any amino acid except proline) are the recognition sequences for enzymatic attachment of the carbohydrate moiety to the asparagine side chain. Therefore, the presence of either of these tripeptide sequences in a polypeptide creates a potential glycosylation site. O-linked glycosylation refers to the attachment of one of the sugars N-acetylgalactosamine, galactose, or xylose to a hydroxyamino acid (most commonly serine or threonine, but 5-hydroxyproline or 5-hydroxylysine may also be used).

[0332] Addition of glycosylation sites to the antibody can be conveniently accomplished by altering the amino acid sequence such that it contains one or more of the above-described tripeptide sequences (for N-linked glycosylation sites). The alteration can also be accomplished by adding or substituting one or more serine or threonine residues to the sequence of the original antibody (for O-linked glycosylation sites).

[0333] When the antibody comprises the Fc district, the carbohydrate connected thereto can be changed. The natural antibody produced by mammalian cells generally comprises the biantennary oligosaccharide of side chain, which is generally connected to the Asn297 (according to Kabat numbering) of the CH2 domain in the Fc district by N linkage. The oligosaccharide may include various carbohydrates, such as mannose, N-acetylglucosamine (GlcNAc), galactose and sialic acid and the fucose of the GlcNAc in the " stem " of the biantennary oligosaccharide structure. In some embodiments, the oligosaccharide in the disclosed antibody may be modified to produce antibody variants with some improved properties.

[0334] In one embodiment, antibody variants are provided that have a carbohydrate structure that lacks fucose attached (directly or indirectly) to the Fc region. See, for example, U.S. Patent Publication Nos. 2003 / 0157108 and 2004 / 0093621. Examples of publications related to "defucosylated" or "fucose-deficient" antibody variants include: US 2003 / 0157108; US 2003 / 0115614; US 2002 / 0164328; US 2004 / 0093621; US ​​2004 / 0132140; US 2004 / 0110704; US 2004 / 0110282; US 2004 / 0109865; Okazaki et al., J. Mol. Biol. 336:1239-1249 (2004); Yamane-Ohnuki et al., Biotech. Bioeng . 87:614 (2004). Examples of cell lines capable of producing defucosylated antibodies include Led 3 CHO cells that lack protein fucosylation (Ripka et al., Arch. Biochem. Biophys . 249:533-545 (1986); US 2003 / 0157108), and knockout cell lines (such as α-1,6-fucosyltransferase gene FUT8 knockout CHO cells) (see, for example, Yamane-Ohnuki et al., Biotech. Bioeng . 87:614 (2004) and Kanda et al., Biotechnol. Bioeng . 94(4):680-688 (2006)).

[0335] (iii) Modified constant region

[0336] In some embodiments of any of the antibodies provided herein, the antibody Fc is an antibody Fc isotype and / or modified form. In some embodiments, the antibody Fc isotype and / or modified form is capable of binding to an Fc gamma receptor.

[0337] In some embodiments of any of the antibodies provided herein, the modified antibody Fc is an IgG1 modified Fc. In some embodiments, the IgG1 modified Fc comprises one or more modifications. For example, in some embodiments, the IgG1 modified Fc comprises one or more amino acid substitutions (e.g., relative to a wild-type Fc region of the same isotype). In some embodiments, the one or more amino acid substitutions are selected from N297A (Bolt S et al., (1993) Eur J Immunol 23:403-411), D265A (Shields et al., (2001) RJ Biol. Chem.276, 6591-6604), L234A, L235A (Hutchins et al., (1995) Proc Natl Acad Sci USA, 92:11980-11984; Alegre et al., (1994) Transplantation 57:1537-1543. 31; Xu et al., (2000) Cell Immunol , 200:16-26), G237A (Alegre et al., (1994) Transplantation 57:1537-1543. 31; Xu et al., (2000) Cell Immunol , 200:16-26), C226S, C229S, E233P, L234V, L234F, L235E (McEarchern et al., (2007) Blood , 109:1185-1192), P331S (Sazinsky et al., (2008) Proc Natl Acad Sci USA 2008, 105:20167-20172), S267E, L328F, A330L, M252Y, S254T and / or T256E, wherein amino acid positions are according to the EU numbering convention.

[0338] In some embodiments of any of the IgG1 modified Fcs, the Fc comprises an N297A mutation according to EU numbering. In some embodiments of any of the IgG1 modified Fcs, the Fc comprises a D265A and N297A mutation according to EU numbering. In some embodiments of any of the IgG1 modified Fcs, the Fc comprises an N297A mutation according to EU numbering. In some embodiments of any of the IgG1 modified Fcs, the Fc comprises a K322A mutation according to EU numbering. In some embodiments of any of the IgG1 modified Fcs, the Fc comprises an N297A mutation according to EU numbering. In some embodiments of any of the IgG1 modified Fcs, the Fc comprises a P331S mutation according to EU numbering. In some embodiments of any of the IgG1 modified Fcs, the Fc comprises a D270A mutation according to EU numbering. In some embodiments, the IgG1 modified Fc comprises an L234A and L235A mutation according to EU numbering. In some embodiments of any one of the Fc modified by IgG1, Fc comprises L234A and G237A mutations according to EU numbering. In some embodiments of any one of the Fc modified by IgG1, Fc comprises L234A, L235A and G237A mutations according to EU numbering. In some embodiments of any one of the Fc modified by IgG1, Fc comprises one or more (including all) in P238D, L328E, E233, G237D, H268D, P271G and A330R mutations according to EU numbering. In some embodiments of any one of the Fc modified by IgG1, Fc comprises one or more in S267E / L328F mutations according to EU numbering. In some embodiments of any one of the Fc modified by IgG1, Fc comprises N325S and L328F mutations (N325S / L328F) according to EU numbering. In some embodiments of any one of the IgG1 modified Fc, the Fc comprises P238D, L328E, E233D, G237D, H268D, P271G, and A330R mutations according to EU numbering. In some embodiments of any one of the IgG1 modified Fc, the Fc comprises P238D, L328E, G237D, H268D, P271G, and A330R mutations according to EU numbering. In some embodiments of any one of the IgG1 modified Fc, the Fc comprises P238D, S267E, L328E, E233D, G237D, H268D, P271G, and A330R mutations according to EU numbering. In some embodiments of any of the IgG1 modified Fcs, the Fc comprises P238D, S267E, L328E, G237D, H268D, P271G, and A330R mutations according to EU numbering.In some embodiments of any one of the Fc modified by IgG1, Fc comprises C226S, C229S, E233P, L234V and L235A mutations according to EU numbering. In some embodiments of any one of the Fc modified by IgG1, Fc comprises L234F, L235E and P331S mutations according to EU numbering. In some embodiments of any one of the Fc modified by IgG1, Fc comprises S267E and L328F mutations according to EU numbering. In some embodiments of any one of the Fc modified by IgG1, Fc comprises S267E mutations according to EU numbering. In some embodiments of any one of the Fc modified by IgG1, Fc comprises CH1 and hinge region (according to EU numbering, amino acid 118-230 of IgG2) replacing IgG1 with constant heavy chain 1 (CH1) and hinge region of IgG2 with κ light chain.

[0339] In some embodiments of any one of the Fc modified by IgG1, the Fc includes two or more amino acid replacements that increase antibody aggregation without complement activation compared to the corresponding antibody with the Fc region that does not include two or more amino acid replacements. Therefore, in some embodiments of any one of the Fc modified by IgG1, the Fc modified by IgG1 is an antibody comprising an Fc region, wherein the antibody comprises an amino acid replacement at position E430G according to EU numbering and one or more amino acid replacements at residue positions in the Fc region selected from the group consisting of L234F, L235A, L235E, S267E, K322A, L328F, A330S, P331S, and any combination thereof. In some embodiments, the Fc modified by IgG1 comprises an amino acid replacement at position E430G, L234A, L235A, and P331S according to EU numbering. In some embodiments, the Fc modified by IgG1 comprises an amino acid replacement at position L234A, L235A, and P331S according to EU numbering. In some embodiments, the IgG1 modified Fc comprises amino acid substitutions at positions E430G and P331S according to EU numbering. In some embodiments, the IgG1 modified Fc comprises amino acid substitutions at positions E430G and K322A according to EU numbering. In some embodiments, the IgG1 modified Fc comprises amino acid substitutions at positions E430G, A330S, and P331S according to EU numbering. In some embodiments, the IgG1 modified Fc comprises amino acid substitutions at positions E430G, K322A, A330S, and P331S according to EU numbering. In some embodiments, the IgG1 modified Fc comprises amino acid substitutions at positions E430G, K322A, and A330S according to EU numbering. In some embodiments, the IgG1 modified Fc comprises amino acid substitutions at positions E430G, K322A, and P331S according to EU numbering.

[0340] In some embodiments of any of the IgG1 modified Fcs, the IgG1 modified Fc may further comprise an A330L mutation according to the EU numbering convention (Lazar et al., Proc Natl Acad Sci USA , 103:4005-4010 (2006)), or one or more of the L234F, L235E and / or P331S mutations (Sazinsky et al., Proc Natl Acad Sci USA, 105:20167-20172 (2008)), to eliminate complement activation. In some embodiments of any one of the IgG1 modified Fc, the IgG1 modified Fc may also include one or more of A330L, A330S, L234F, L235E and / or P331S according to EU numbering. In some embodiments of any one of the IgG1 modified Fc, the IgG1 modified Fc may also include one or more mutations to extend the antibody half-life in human serum (e.g., according to EU numbering convention, one or more (including all) of M252Y, S254T and T256E mutations). In some embodiments of any one of the IgG1 modified Fc, the IgG1 modified Fc may also include one or more of E430G, E430S, E430F, E430T, E345K, E345Q, E345R, E345Y, S440Y and / or S440W according to EU numbering.

[0341] Other aspects of the present disclosure relate to antibodies with modified constant regions (i.e., Fc regions). If an antibody that relies on binding to FcgR receptors for activation by the targeted receptor is engineered to eliminate FcgR binding, its agonist activity may be lost (see, e.g., Wilson et al., CancerCell 19:101-113 (2011); Armour et al. Immunology 40:585-593 (2003); and White et al., Cancer Cell 27:138-148 (2015). Therefore, when the anti-MS4A4A antibodies of the present disclosure with the correct epitope specificity have an Fc domain (CH1 and hinge region) from the human IgG2 isotype or another type of Fc domain or variant thereof that is capable of preferentially binding to the inhibitory FcgRIIB r receptor, it is believed that the antibodies can activate the target antigen with minimal adverse effects.

[0342] In some embodiments of any of the antibodies provided herein, the modified antibody Fc is an IgG2 modified Fc. In some embodiments, the IgG2 modified Fc comprises one or more modifications. For example, in some embodiments, the IgG2 modified Fc comprises one or more amino acid substitutions (e.g., relative to a wild-type Fc region of the same isotype). In some embodiments of any of the IgG2 modified Fc, the one or more amino acid substitutions are selected from V234A according to the EU numbering convention (Alegre et al., Transplantation 57:1537-1543 (1994); Xu et al., Cell Immunol , 200:16-26 (2000)); G237A (Cole et al., Transplantation, 68:563-571 (1999)); H268Q, V309L, A330S, P331S (US 2007 / 0148167; Armour et al., Eur J Immunol 29: 2613-2624 (1999); Armour et al. The Haematology Journal 1 (Suppl 1):27 (2000); Armour et al., The Haematology Journal 1 (Suppl 1):27 (2000)), C219S and / or C220S (White et al., Cancer Cell 27, 138-148 (2015)); S267E, L328F (Chu et al., Mol Immunol , 45:3926-3933 (2008)); and M252Y, S254T and / or T256E. In some embodiments of any of the IgG2 modified Fcs, the Fc comprises amino acid substitutions at positions V234A and G237A according to EU numbering. In some embodiments of any of the IgG2 modified Fcs, the Fc comprises amino acid substitutions at positions C219S or C220S according to EU numbering. In some embodiments of any of the IgG2 modified Fcs, the Fc comprises amino acid substitutions at positions A330S and P331S according to EU numbering. In some embodiments of any of the IgG2 modified Fcs, the Fc comprises amino acid substitutions at positions S267E and L328F according to EU numbering.

[0343] In some embodiments of any of the IgG2 modified Fcs, the Fc comprises a C127S amino acid substitution according to the EU numbering convention (White et al., (2015) Cancer Cell 27, 138-148; Lightle et al. Protein Sci. 19:753-762 (2010); and WO 2008 / 079246). In some embodiments of any of the IgG2 modified Fc, the antibody has an IgG2 isotype in which the kappa light chain constant domain comprises a C214S amino acid substitution according to the EU numbering convention (White et al., Cancer Cell 27:138-148 (2015); Lightle et al. Protein Sci. 19:753-762(2010); and WO 2008 / 079246).

[0344] In some embodiments of any of the IgG2 modified Fcs, the Fc comprises a C220S amino acid substitution according to the EU numbering convention. In some embodiments of any of the IgG2 modified Fcs, the antibody has an IgG2 isotype with the kappa light chain constant domain comprising a C214S amino acid substitution according to the EU numbering convention.

[0345] In some embodiments of any of the IgG2 modified Fc, the Fc comprises a C219S amino acid substitution according to the EU numbering convention. In some embodiments of any of the IgG2 modified Fc, the antibody has an IgG2 isotype with the kappa light chain constant domain comprising a C214S amino acid substitution according to the EU numbering convention.

[0346] In some embodiments of any of the IgG2 modified Fcs, the Fc comprises an IgG2 isotype heavy chain constant domain 1 (CH1) and a hinge region (White et al., Cancer Cell 27:138-148 (2015). In certain embodiments of any of the IgG2 modified Fcs, the IgG2 isotype CH1 and hinge regions comprise an amino acid sequence according to EU numbering 118-230. In some embodiments of any of the IgG2 modified Fcs, the antibody Fc region comprises an S267E amino acid substitution, an L328F amino acid substitution, or both, and / or an N297A or N297Q amino acid substitution according to the EU numbering convention.

[0347] In some embodiments of any of the IgG2 modified Fcs, the Fc further comprises one or more amino acid substitutions at the following positions according to EU numbering: E430G, E430S, E430F, E430T, E345K, E345Q, E345R, E345Y, S440Y, and S440W. In some embodiments of any of the IgG2 modified Fcs, the Fc further comprises one or more mutations to extend the antibody half-life in human serum (e.g., one or more (including all) of the M252Y, S254T, and T256E mutations according to the EU numbering convention). In some embodiments of any of the IgG2 modified Fcs, the Fc further comprises A330S and P331S.

[0348] In some embodiments of any of the IgG2 modified Fcs, the Fc is an IgG2 / 4 hybrid Fc. In some embodiments, the IgG2 / 4 hybrid Fc comprises IgG2 aa 118 to 260 and IgG4 aa 261 to 447. In some embodiments of any of the IgG2 modified Fcs, the Fc comprises one or more amino acid substitutions at positions H268Q, V309L, A330S, and P331S according to EU numbering.

[0349] In some embodiments of any of the IgG1 and / or IgG2 modified Fc, the Fc comprises one or more additional amino acid substitutions selected from A330L, L234F, L235E, or P331S according to EU numbering, and any combination thereof.

[0350] In certain embodiments of any one of the IgG1 and / or IgG2 modified Fcs, the Fc comprises one or more amino acid substitutions according to EU numbering at residue positions selected from the group consisting of C127S, L234A, L234F, L235A, L235E, S267E, K322A, L328F, A330S, P331S, E345R, E430G, S440Y, and any combination thereof. In some embodiments of any one of the IgG1 and / or IgG2 modified Fcs, the Fc comprises amino acid substitutions at positions E430G, L243A, L235A, and P331S according to EU numbering. In some embodiments of any one of the IgG1 and / or IgG2 modified Fcs, the Fc comprises amino acid substitutions at positions E430G and P331S according to EU numbering. In some embodiments of any one of the IgG1 and / or IgG2 modified Fcs, the Fc comprises amino acid substitutions at positions E430G and K322A according to EU numbering. In some embodiments of any one of the IgG1 and / or IgG2 modified Fcs, the Fc comprises amino acid substitutions at positions E430G, A330S, and P331S according to EU numbering. In some embodiments of any one of the IgG1 and / or IgG2 modified Fcs, the Fc comprises amino acid substitutions at positions E430G, K322A, A330S, and P331S according to EU numbering. In some embodiments of any one of the IgG1 and / or IgG2 modified Fcs, the Fc comprises amino acid substitutions at positions E430G, K322A, A330S, and P331S according to EU numbering. In some embodiments of any one of the IgG1 and / or IgG2 modified Fcs, the Fc comprises amino acid substitutions at positions E430G, K322A, and A330S according to EU numbering. In some embodiments of any one of the IgG1 and / or IgG2 modified Fcs, the Fc comprises amino acid substitutions at positions E430G, K322A, and P331S according to EU numbering. In some embodiments of any of the IgG1 and / or IgG2 modified Fc, the Fc comprises amino acid substitutions at positions S267E and L328F according to EU numbering. In some embodiments of any of the IgG1 and / or IgG2 modified Fc, the Fc comprises amino acid substitutions at positions C127S according to EU numbering. In some embodiments of any of the IgG1 and / or IgG2 modified Fc, the Fc comprises amino acid substitutions at positions E345R, E430G, and S440Y according to EU numbering.

[0351] In some embodiments of any of the antibodies provided herein, the modified antibody Fc is an IgG4 modified Fc. In some embodiments, the IgG4 modified Fc comprises one or more modifications. For example, in some embodiments, the IgG4 modified Fc comprises one or more amino acid substitutions (e.g., relative to a wild-type Fc region of the same isotype). In some embodiments of any of the IgG4 modified Fc, the one or more amino acid substitutions are selected from L235A, G237A, S229P, L236E according to the EU numbering convention (Reddy et al., J Immunol 164:1925-1933 (2000)), S267E, E318A, L328F, M252Y, S254T and / or T256E. In some embodiments of any of the IgG4 modified Fcs, the Fc may further comprise L235A, G237A and E318A according to the EU numbering convention. In some embodiments of any of the IgG4 modified Fcs, the Fc may further comprise S228P and L235E according to the EU numbering convention. In some embodiments of any of the IgG4 modified Fcs, the IgG4 modified Fc may further comprise S267E and L328F according to the EU numbering convention.

[0352] In some embodiments of any of the IgG4 modified Fcs, the IgG4 modified Fc can be combined with an S228P mutation according to the EU numbering convention (Angal et al., Mol Immunol. 30:105-108 (1993)) and / or with (Peters et al., J Biol Chem. 287(29):24525-33 (2012)) to enhance antibody stability.

[0353] In some embodiments of any of the IgG4 modified Fcs, the IgG4 modified Fc may further comprise one or more mutations to extend the half-life of the antibody in human serum (e.g., one or more (including all) of the M252Y, S254T, and T256E mutations according to the EU numbering convention).

[0354] In some embodiments of any one of the Fc modified by IgG4, Fc comprises L235E according to EU numbering. In some embodiments of any one of the Fc modified by IgG4, Fc comprises S228P mutation according to EU numbering. In some embodiments of any one of the Fc modified by IgG4, Fc comprises S267E and L328F mutation according to EU numbering. In certain embodiments of any one of the Fc modified by IgG4, Fc comprises one or more amino acid substitutions selected from the following residue positions according to EU numbering: C127S, F234A, L235A, L235E, S267E, K322A, L328F, E345R, E430G, S440Y and any combination thereof. In some embodiments of any one of the Fc modified by IgG4, Fc comprises amino acid substitutions at positions E430G, L243A, L235A and P331S according to EU numbering. In some embodiments of any of the IgG4 modified Fcs, the Fc comprises amino acid substitutions at positions E430G and P331S according to EU numbering. In some embodiments of any of the IgG4 modified Fcs, the Fc comprises amino acid substitutions at positions E430G and K322A according to EU numbering. In some embodiments of any of the IgG4 modified Fcs, the Fc comprises amino acid substitutions at positions E430 according to EU numbering. In some embodiments of any of the IgG4 modified Fcs, the Fc region comprises amino acid substitutions at positions E430G and K322A according to EU numbering. In some embodiments of any of the IgG4 modified Fcs, the Fc comprises amino acid substitutions at positions S267E and L328F according to EU numbering. In some embodiments of any of the IgG4 modified Fcs, the Fc comprises amino acid substitutions at positions C127S according to EU numbering. In some embodiments of any of the IgG4 modified Fcs, the Fc comprises amino acid substitutions at positions E345R, E430G, and S440Y according to EU numbering.

[0355] In some embodiments, the antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 336. In some embodiments, the antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 328. In some embodiments, the antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 320.

[0356] In some embodiments, the antibody has a human IgG1 heavy chain that does not contain a C-terminal lysine. In some embodiments, the antibody has a human IgG1 heavy chain that contains a P331S mutation. In some embodiments, the antibody has a human IgG1 heavy chain that contains a P331S mutation and does not contain a C-terminal lysine. In some embodiments, the antibody has a human IgG1 heavy chain that contains a N325S and L328F mutation (N325S / L328F). In some embodiments, the antibody has a human IgG1 heavy chain that contains a N325S / L328F mutation and does not contain a C-terminal lysine. In some embodiments, the antibody has a human IgG1 heavy chain that contains a K322A mutation. In some embodiments, the antibody has a human IgG1 heavy chain that contains a K322A mutation and does not contain a C-terminal lysine. In the aforementioned embodiments, mutations are represented according to EU numbering.

[0357] In some embodiments, the antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 337. In some embodiments, the antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 338. In some embodiments, the antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 339. In some embodiments, the antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 340. In some embodiments, the antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 341. In some embodiments, the antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 342. In some embodiments, the antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 343.

[0358] In some embodiments, the antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 329. In some embodiments, the antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 330. In some embodiments, the antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 331. In some embodiments, the antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 332. In some embodiments, the antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 333. In some embodiments, the antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 334. In some embodiments, the antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 335.

[0359] In some embodiments, the antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 321. In some embodiments, the antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 322. In some embodiments, the antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 323. In some embodiments, the antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 324. In some embodiments, the antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 325. In some embodiments, the antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 326. In some embodiments, the antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 327.

[0360] (8) Other antibody modifications

[0361] In some embodiments of any one of the antibodies, the antibody is a derivative. The term "derivative" refers to a molecule that is chemically modified except for the insertion, deletion or substitution of amino acids (or nucleic acids). In certain embodiments, the derivative comprises a covalent modification, including but not limited to chemical bonding to a polymer, lipid or other organic or inorganic moiety. In certain embodiments, the chemically modified antigen-binding protein may have a longer circulation half-life than an antigen-binding protein that is not chemically modified. In certain embodiments, the chemically modified antigen-binding protein may have improved targeting capabilities to desired cells, tissues and / or organs. In some embodiments, the derivative antigen-binding protein is covalently modified to include one or more water-soluble polymer linkers, including but not limited to polyethylene glycol, polyoxyethylene glycol or polypropylene glycol. See, for example, U.S. Patent Nos. 4,640,835, 4,496,689, 4,301,144, 4,670,417, 4,791,192 and 4,179,337. In certain embodiments, the derivative antigen binding protein comprises one or more polymers including, but not limited to, monomethoxy-polyethylene glycol, dextran, cellulose, ethylene glycol / propylene glycol copolymers, carboxymethyl cellulose, polyvinyl pyrrolidone, poly-1,3-dioxolane, poly-1,3,6-trioxane, ethylene / maleic anhydride copolymers, polyamino acids (homopolymers or random copolymers), poly-(N-vinyl pyrrolidone)-polyethylene glycol, propylene glycol homopolymers, polypropylene oxide / ethylene oxide copolymers, polyoxyethylated polyols (e.g., glycerol), and polyvinyl alcohol, as well as mixtures of such polymers.

[0362] In certain embodiments, derivatives are covalently modified with polyethylene glycol (PEG) subunits. In certain embodiments, one or more water-soluble polymers are bonded at one or more specific positions of the derivative (e.g., at the amino terminal end). In certain embodiments, one or more water-soluble polymers are randomly attached to one or more side chains of the derivative. In certain embodiments, PEG is used to improve the therapeutic ability of antigen-binding proteins. In certain embodiments, PEG is used to improve the therapeutic ability of humanized antibodies. Some such methods are discussed in, for example, U.S. Patent No. 6,133,426, which is incorporated herein by reference for any purpose.

[0363] Peptide analogs are commonly used in the pharmaceutical industry as non-peptide drugs whose properties are similar to those of the template peptide. These types of non-peptide compounds are called "peptide mimetics (peptide mimetic or peptidomimetic)". Fauchere, J. Adv. Drug Res ., 15:29 (1986); and Evans et al., J. Med. Chem ., 30:1229 (1987), which is incorporated herein by reference for any purpose. Such compounds are generally developed with the help of computerized molecular modeling. Peptide mimetics that are structurally similar to therapeutically useful peptides can be used to produce similar therapeutic or preventive effects. Typically, peptide mimetics are structurally similar to paradigm polypeptides (i.e., polypeptides having biochemical properties or pharmacological activity), such as human antibodies, but one or more peptide linkages thereof are optionally replaced by linkages selected from the following by methods well known in the art: -CH2NH-, -CH2S-, -CH2-CH2-, -CH═CH- (cis and trans), -COCH2-, -CH(OH)CH2-, and -CH2SO-. In certain embodiments, systematic substitution of one or more amino acids of a consensus sequence with D-amino acids of the same type (e.g., D-lysine instead of L-lysine) can be used to produce more stable peptides. In addition, restricted peptides comprising a consensus sequence or substantially identical consensus sequence variations can be produced by methods known in the art (Rizo and Gierasch, Ann.Rev. Biochem ., 61:387 (1992), which is incorporated herein by reference for any purpose); for example, by adding internal cysteine ​​residues capable of forming intramolecular disulfide bonds that cyclize the peptide.

[0364] Drug conjugation involves coupling a bioactive cytotoxic (anticancer) payload or drug to an antibody that specifically targets a tumor marker (e.g., a polypeptide ideally found only in or on tumor cells). The antibodies track these proteins in the body and attach themselves to the surface of cancer cells. The biochemical reaction between the antibody and the target protein (antigen) triggers a signal in the tumor cell, which then absorbs or internalizes the antibody along with the cytotoxin. After the ADC is internalized, the cytotoxic drug is released and the cancer is killed. Due to this targeting, the drug ideally has lower side effects and gives a wider therapeutic window than other chemotherapeutic agents. The technology for conjugating the disclosed antibodies is known in the art (see, for example, Jane de Lartigue, OncLive July 5, 2012; ADC Review on antibody-drug conjugates; and Ducry et al., Bioconjugate Chemistry 21 (1):5-13 (2010).

[0365] III. Nucleic Acids, Vectors, and Host Cells

[0366] The anti-MS4A4A antibodies of the present disclosure can be produced using, for example, recombinant methods and compositions as described in U.S. Patent No. 4,816,567. In some embodiments, isolated nucleic acids are provided having a nucleotide sequence encoding any of the anti-MS4A4A antibodies of the present disclosure. Such nucleic acids may encode V sequences comprising the anti-MS4A4A antibodies. L The amino acid sequence and / or V sequences constituting the anti-MS4A4A antibody H In some embodiments, one or more vectors (e.g., expression vectors) comprising such nucleic acids are provided. In some embodiments, a host cell comprising such nucleic acids is also provided. In some embodiments, the host cell comprises the following vectors (e.g., has been transduced with the following vectors): (1) a vector comprising the following nucleic acid: encoding the V L The amino acid sequence and V H or (2) a first vector comprising a V encoding an antibody constituting the L A nucleic acid containing an amino acid sequence of H In some embodiments, the host cell is a eukaryotic cell, such as a Chinese hamster ovary (CHO) cell or a lymphoid cell (e.g., a Y0, NS0, Sp20 cell). The host cells of the present disclosure also include (but are not limited to) isolated cells, in vitro cultured cells, and ex vivo cultured cells.

[0367] Methods for preparing the anti-MS4A4A antibodies of the present disclosure are provided. In some embodiments, the methods comprise culturing a host cell of the present disclosure comprising a nucleic acid encoding the antibody under conditions suitable for expression of the anti-MS4A4A antibody. In some embodiments, the antibody is subsequently recovered from the host cell (or host cell culture medium).

[0368] To recombinantly produce the anti-MS4A4A antibodies of the present disclosure, nucleic acid encoding the anti-MS4A4A antibodies is isolated and inserted into one or more vectors for further cloning and / or expression in host cells. This nucleic acid can be readily isolated and sequenced using conventional procedures (e.g., by using oligonucleotide probes that are capable of binding specifically to genes encoding the heavy and light chains of the antibody).

[0369] Suitable vectors containing nucleic acid sequences encoding any of the anti-MS4A4A antibodies disclosed herein or cell surface expression fragments or polypeptides described herein (including antibodies) include, but are not limited to, cloning vectors and expression vectors. Suitable cloning vectors can be constructed according to standard techniques or can be selected from a large number of cloning vectors available in the art. Although the cloning vector of choice can vary depending on the host cell to be used, useful cloning vectors generally have the ability to self-replicate, may have a single target for a specific restriction endonuclease, and / or may carry a gene for a marker that can be used to select clones containing the vector. Suitable examples include plasmids and bacterial viruses, such as pUC18, pUC19, Bluescript (e.g., pBS SK+) and its derivatives, mpl8, mpl9, pBR322, pMB9, ColE1, pCR1, RP4, phage DNA, and shuttle vectors (such as pSA3 and pAT28). These and many other cloning vectors are available from commercial suppliers such as BioRad, Strategene, and Invitrogen.

[0370] Suitable host cells for cloning or expressing antibody-encoding vectors include prokaryotic or eukaryotic cells. For example, the anti-MS4A4A antibodies of the present disclosure can be produced in bacteria, especially when glycosylation and Fc effector functions are not desired. Expression of antibody fragments and polypeptides in bacteria is described (e.g., U.S. Patent Nos. 5,648,237, 5,789,199, and 5,840,523). Following expression, the antibody can be isolated from the bacterial cell mass as a soluble fraction and further purified.

[0371] In addition to prokaryotes, eukaryotic microorganisms (such as filamentous fungi or yeast) are also suitable cloning or expression hosts for antibody-encoding vectors, including fungal and yeast strains whose glycosylation pathways have been "humanized" to produce antibodies with partially or fully human glycosylation patterns (e.g., Gerngross, Nat. Biotech. 22:1409-1414 (2004); and Li et al., Nat. Biotech. 24:210-215 (2006)).

[0372] Suitable host cells for expressing glycosylated antibodies can also be derived from multicellular organisms (invertebrates and vertebrates). Examples of invertebrate cells include plant and insect cells. Spodoptera frugiperda Plant cell cultures can also be used as hosts (e.g., U.S. Patent Nos. 5,959,177, 6,040,498, 6,420,548, 7,125,978, and 6,417,429, which describe the PLANTIBODIES™ technology for producing antibodies in transgenic plants).

[0373] Vertebrate cells can also be used as hosts. For example, mammalian cell lines adapted for growth in suspension c...

Claims

1. An isolated antibody that binds to MS4A4A protein, wherein the antibody comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the HVR-H1 amino acid sequence of SEQ ID NO:94, the HVR-H2 amino acid sequence of SEQ ID NO:96, and the HVR-H3 amino acid sequence of SEQ ID NO:102; and the light chain variable region comprises the HVR-L1 amino acid sequence of SEQ ID NO:104, the HVR-L2 amino acid sequence of SEQ ID NO:105, and the HVR-L3 amino acid sequence of SEQ ID NO:

107.

2. The antibody of claim 1, wherein the antibody comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 14 and the light chain variable region comprises the amino acid sequence of SEQ ID NO:

22.

3. The antibody of claim 1, wherein the antibody comprises a heavy chain and a light chain, wherein the heavy chain comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 355-362 and the light chain comprises the amino acid sequence of SEQ ID NO:

365.

4. The antibody of claim 3, wherein the heavy chain comprises the amino acid sequence of SEQ ID NO: 360 and the light chain comprises the amino acid sequence of SEQ ID NO:

365.

5. The antibody of claim 3, wherein the heavy chain comprises the amino acid sequence of SEQ ID NO: 362 and the light chain comprises the amino acid sequence of SEQ ID NO:

365.

6. The antibody of claim 3, wherein the heavy chain comprises the amino acid sequence of SEQ ID NO: 358 and the light chain comprises the amino acid sequence of SEQ ID NO:

365.

7. The antibody of claim 1, wherein the antibody is humanized.

8. The antibody of any one of claims 1-7, wherein the antibody is a monoclonal antibody.

9. The antibody of any one of claims 1, 2 and 7, wherein the antibody is of the IgG class and has an IgG1, IgG2 or IgG4 isotype.

10. The antibody of claim 9, wherein the antibody is of IgG1 isotype having N325S and L328F mutations according to EU numbering.

11. The antibody of any one of claims 1, 2 and 7, wherein the antibody is an antibody fragment.

12. The antibody of claim 11, wherein the antibody fragment is a Fab, Fab', Fab'-SH, F(ab')2, Fv or scFv fragment.

13. A pharmaceutical composition comprising the antibody according to any one of claims 1 to 12 and a pharmaceutically acceptable carrier.

14. An isolated nucleic acid comprising a nucleic acid sequence encoding the antibody of any one of claims 1-12.

15. An isolated host cell comprising the nucleic acid of claim 14.

16. A method for producing an antibody that binds to human MS4A4A, comprising culturing the cell according to claim 15 so that the antibody is produced.

17. Use of the antibody of any one of claims 1 to 12 or the pharmaceutical composition of claim 13 in the preparation of a medicament for treating an individual suffering from a disease, disorder or injury selected from the group consisting of Alzheimer's disease and cognitive impairment.

18. Use of the antibody of any one of claims 1 to 12 or the pharmaceutical composition of claim 13 in the preparation of a medicament for treating an individual suffering from late-onset Alzheimer's disease.

19. Use of the antibody of any one of claims 1 to 12 or the pharmaceutical composition of claim 13 in the preparation of a medicament for treating an individual suffering from a CSF1R deficiency disease or condition, wherein the CSF1R deficiency disease or condition is adult-onset leukoencephalopathy with axonal spheroids and pigmented glial cells (ALSP) or hereditary diffuse leukoencephalopathy (HDLS).

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