Anti-ms4a4a antibodies and methods of use thereof

Abstract

To provide therapies that are capable of modulating (e.g., inhibiting or reducing; activating or enhancing) the activity of MS4A4A, in order to treat various diseases, disorders, and conditions associated with MS4A4A activity.SOLUTION: The present disclosure is directed to compositions that include antibodies, e.g., monoclonal antibodies, humanized antibodies and antibody fragments, that specifically bind a MS4A4A polypeptide, e.g., a mammalian MS4A4A or human MS4A4A, and use of such compositions in preventing, reducing risk, or treating an individual in need thereof.SELECTED DRAWING: None

Description

[Technical Field]

[0001] Submitting a sequence listing as an ASCII text file The entire contents of the following submission in an ASCII text file are incorporated herein by reference: Computer Readable (CRF) Sequence Listing (File Name: 4503_008PC06_Seqlisting_ST25.TXT, Created: July 28, 2020, Size: 289,448 bytes).

[0002] Technical Field of the Disclosure The present disclosure relates to anti-MS4A4A antibodies and therapeutic uses of such antibodies. [Background technology]

[0003] Background of the Disclosure The four-transmembrane domain subfamily A (MS4A) gene cluster is located on chromosome 11q12 and contains 18 genes. This MS4A gene family generally encodes membrane proteins with a tetraspanin 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). These transmembrane domains are interconnected by one intracellular loop and two extracellular loops, and both the N- and C-termini are cytoplasmic. Most MS4A proteins share amino acid sequence homology (20-30%) with MS4A1 (CD20), with the highest sequence identity observed in the first three transmembrane domains. Highly conserved motifs within these transmembrane regions across different MS4A proteins suggest that the transmembrane domains play an important general role in the function of MS4A proteins. The greatest areas of variability among MS4A proteins are found in the N- and C-terminal cytoplasmic domains and the second putative extracellular loop (Ishibashi et al., 2001, Gene, 265:87-93), suggesting that these regions confer unique functional properties.

[0004] Despite this diversity, MS4A domains share several common elements. For example, one notable feature conserved among MS4A proteins (except for MS4A8B and MS4A12) is the conservation of two cysteine ​​residues in the second putative extracellular loop, which may form disulfide bridges. The N- and C-terminal domains of MS4A proteins are also rich in proline residues, although the functional significance of this remains to be elucidated (Hulett et al., 2001, Genomics, 72:119-127). However, proline-rich regions are commonly involved in various cellular processes, such as cytoskeletal reorganization, transcription initiation, signal transduction cascades, and associating with SH3 domains as part of adaptor systems that facilitate protein-protein interactions (Kay et al., 2000, FASEB J, 14:231-241).

[0005] The MS4A protein family has some notable exceptions: MS4A1 (CD20) is expressed exclusively in B lymphocytes and its function remains poorly characterized, except for its B cell antigen receptor and calcium influx signaling functions. CD20 is the target of immunotherapeutic antibodies used to deplete pathogenic B cells in chronic lymphocytic leukemia, lymphoma, autoimmune diseases, and solid organ transplantation. MS4A2 (FcεRβ) is a signaling subunit of the high-affinity IgE receptor (FcεRI) and low-affinity IgG receptor (FcεRIII) on mast cells and plays an important role in hypersensitivity and allergic reactions. MS4A2 is an ITAM domain protein that amplifies signals via 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 for cell cycle regulation.

[0006] Although most members of the MS4A family remain uncharacterized, reports suggest that MS4A proteins function as chemosensors and chemoreceptors for various exogenous and endogenous ligands, such as fatty acids, peptides, and sulfated steroids, and may be involved in mediating calcium influx, regulating endocytosis, trafficking, and functioning as adaptors in signaling 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).

[0007] Specific MS4A genes have been genetically associated with various disorders and diseases, particularly neurodegenerative disorders. For example, genome-wide significance association studies have identified the MS4A gene cluster on chromosome 11q12 as one of the most important Alzheimer's disease loci. One particularly interesting identified gene 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).

[0008] Thus, there is a need for therapies that target MS4A4A, such as antibodies that specifically bind to MS4A4A, and / or that can modulate (e.g., inhibit or suppress, activate or enhance) the activity of MS4A4A, e.g., by suppressing or increasing MS4A4A protein levels or activity, to treat various diseases, disorders, and conditions associated with MS4A4A activity. All documents cited herein, including patent applications and publications, are hereby incorporated by reference in their entirety.

Prior Technology Literature

Non-licensed literature

[0009] [Non-licensed document 1] Ishibashi et al,2001,Gene,265:87-93 [Non-licensed document 2] Liang and Tedder,2001,Genomics,72:119-127 [Non-licensed document 3] Efthymiou and Goate,2017,Molecular Neurodegeneration,12:43

Non-licensed Document 4

Non-licensed Document 5

Non-licensed Document 6

Non-licensed Document 7

Non-licensed literature 9

Non-licensed literature 10

Non-licensed Document 11

Non-licensed Document 12

[0010] Summary of the Disclosure The present disclosure generally relates to anti-MS4A4A antibodies and methods of using such antibodies. The methods provided herein find use in preventing, reducing the risk of, or treating individuals with neurodegenerative diseases, disorders, or conditions. In some embodiments, the present disclosure provides methods of preventing, reducing the risk of, or treating individuals with a neurodegenerative disease, disorder, or condition selected from Alzheimer's disease, late-onset Alzheimer's disease, dementia, and cognitive impairment, comprising administering to an individual in need thereof a therapeutically effective amount of an anti-MS4A4A antibody. In some embodiments, the present disclosure provides methods of preventing, reducing the risk of, or treating individuals with a disease, disorder, or condition associated with elevated MS4A4A expression or activity, comprising administering to an individual in need thereof a therapeutically effective amount of an anti-MS4A4A antibody.

[0011] In one aspect, the disclosure relates to an isolated antibody that binds to an MS4A4A protein, the antibody comprising 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 SEQ ID NOs: 94, 108, 116, 146, 147, 308, and 311; HVR-H2 comprising an amino acid sequence selected from 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 SEQ ID NOs: 100, 101, 102, 112, 123, 124, 125, 126, 127, 128, 129, 154, 310. and 313; and the light chain variable region comprises: HVR-L1 comprising an amino acid sequence selected from SEQ ID NOs: 103, 104, 113, 130, 131, 132, 133, 134, 135, 136, 137, 138, 156, 157, 158, 314, and 317; HVR-L2 comprising an amino acid sequence selected from SEQ ID NOs: 105, 106, 114, 139, 140, 141, 142, 143, 159, 160, 161, 315, and 318; and HVR-L3 comprising an amino acid sequence selected from SEQ ID NOs: 107, 115, 144, 145, 163, 316, and 319.

[0012] In some embodiments that may be combined with any embodiment herein, the anti-MS4A4A antibody of the present disclosure is an isolated antibody that binds to MS4A4A protein, the antibody comprising 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 an amino acid sequence selected from SEQ ID NOs: 96 to 99; and HVR-H3 comprising an amino acid sequence selected from SEQ ID NOs: 100 to 102; and the light chain variable region comprises: HVR-L1 comprising an amino acid sequence selected from SEQ ID NOs: 103 to 104; HVR-L2 comprising an amino acid sequence selected from SEQ ID NOs: 105 to 106; and HVR-L3 comprising the amino acid sequence of SEQ ID NO: 107.

[0013] 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 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.

[0014] 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 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.

[0015] In some embodiments that may be combined with any embodiment herein, the anti-MS4A4A antibody of the present disclosure is 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: 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.

[0016] In some embodiments that may be combined with any embodiment herein, the anti-MS4A4A antibody of the present disclosure is an isolated antibody that binds to MS4A4A protein, the antibody comprising 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: 116; HVR-H2 comprising an amino acid sequence selected from SEQ ID NOs: 118 to 122; and HVR-H3 comprising an amino acid sequence selected from SEQ ID NOs: 123 to 129; and the light chain variable region comprises: HVR-L1 comprising an amino acid sequence selected from SEQ ID NOs: 130 to 138; HVR-L2 comprising an amino acid sequence selected from SEQ ID NOs: 139 to 143; and HVR-L3 comprising an amino acid sequence selected from SEQ ID NOs: 144 to 145.

[0017] In some embodiments that may be combined with any embodiment herein, the anti-MS4A4A antibody of the present disclosure is an isolated antibody that binds to MS4A4A protein, the antibody comprising 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 SEQ ID NOs: 146 to 147; HVR-H2 comprising an amino acid sequence selected from SEQ ID NOs: 149 to 153; and HVR-H3 comprising the amino acid sequence of SEQ ID NO: 154; and the light chain variable region comprises: HVR-L1 comprising an amino acid sequence selected from SEQ ID NOs: 156 to 158; HVR-L2 comprising an amino acid sequence selected from SEQ ID NOs: 159 to 161; and HVR-L3 comprising the amino acid sequence of SEQ ID NO: 163.

[0018] 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, the antibody comprising a heavy chain variable region, wherein the heavy chain variable region comprises an amino acid sequence selected from SEQ ID NOs: 5 to 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, the antibody comprising a heavy chain variable region, wherein the heavy chain variable region comprises an amino acid sequence selected from SEQ ID NOs: 24 to 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, the antibody comprising a heavy chain variable region, wherein the heavy chain variable region comprises an amino acid sequence selected from SEQ ID NOs: 40 to 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 MS4A4A protein, wherein the antibody comprises a heavy chain variable region, and the heavy chain variable region comprises an amino acid sequence selected from SEQ ID NOs: 76 to 84.

[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, the antibody comprising a light chain variable region, wherein the light chain variable region comprises an amino acid sequence selected from SEQ ID NOs: 17-22, 305, and 307. 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, the antibody comprising a light chain variable region, wherein the light chain variable region comprises an amino acid sequence selected from 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, the antibody comprising a light chain variable region, wherein the light chain variable region comprises an amino acid sequence selected from 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 MS4A4A protein, wherein the antibody comprises a light chain variable region, and the light chain variable region comprises an amino acid sequence selected from SEQ ID NOs: 86 to 93.

[0020] 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 MS4A4A protein, the antibody comprising a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises an amino acid sequence selected from SEQ ID NOs: 5-15, 304, and 306, and the light chain variable region comprises an amino acid sequence selected from SEQ ID NOs: 17-22, 305, and 307. 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 MS4A4A protein, the antibody comprising a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises an amino acid sequence selected from SEQ ID NOs: 24-30, and the light chain variable region comprises an amino acid sequence selected from SEQ ID NOs: 32-36. In some embodiments that can be combined with any of the embodiments herein, the anti-MS4A4A antibody of the present disclosure is an isolated antibody that binds to MS4A4A protein, the antibody comprising a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises an amino acid sequence selected from SEQ ID NOs: 40 to 53, and the light chain variable region comprises an amino acid sequence selected from SEQ ID NOs: 55 to 74. In some embodiments that can be combined with any of the embodiments, the anti-MS4A4A antibody of the present disclosure is an isolated antibody that binds to MS4A4A protein, the antibody comprising a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises an amino acid sequence selected from SEQ ID NOs: 76 to 84, and the light chain variable region comprises an amino acid sequence selected from SEQ ID NOs: 86 to 93.

[0021] In some embodiments that may be combined with any embodiment herein, an anti-MS4A4A antibody of the present disclosure is an isolated antibody that binds to MS4A4A protein, the antibody comprising a heavy chain and a light chain, wherein the heavy chain comprises an amino acid sequence selected from SEQ ID NOs: 320-343. In some embodiments, the heavy chain comprises an amino acid sequence selected from SEQ ID NOs: 336-343 (optionally, the light chain comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 22, and optionally, the light chain 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: 320-327 (optionally, the light chain comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 304, and optionally, the light chain 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, the light chain comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 305, and further optionally, the light chain comprises a constant region comprising the amino acid sequence of SEQ ID NO: 344).

[0022] 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 MS4A4A protein, the antibody comprising 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, the anti-MS4A4A antibody of the present disclosure is an isolated antibody that binds to MS4A4A protein, the antibody comprising 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, the anti-MS4A4A antibody of the present disclosure is an isolated antibody that binds to 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.

[0023] 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 MS4A4A protein, the antibody comprising 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. 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 MS4A4A protein, the antibody comprising 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, the anti-MS4A4A antibody of the present disclosure is an isolated antibody that binds to 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.

[0024] 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 MS4A4A protein, the antibody comprising 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, the anti-MS4A4A antibody of the present disclosure is an isolated antibody that binds to MS4A4A protein, the antibody comprising 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, the anti-MS4A4A antibody of the present disclosure is an isolated antibody that binds to 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.

[0025] In one aspect, the disclosure relates to an isolated antibody that binds to an MS4A4A protein, wherein the antibody competitively inhibits the binding of one or more antibodies of any of the embodiments herein with respect to binding to MS4A4A.

[0026] In another aspect, the present disclosure relates to an isolated antibody that binds to MS4A4A protein, wherein the antibody binds to an epitope of MS4A4A that is essentially the same as or overlaps with an antibody of any embodiment herein.

[0027] 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 binds to extracellular domain 1 of human MS4A4A. 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 binds to one or more amino acid residues in 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 antibody of the present disclosure is an isolated antibody that binds to the MS4A4A protein, wherein the antibody binds to extracellular domain 2 of human MS4A4A. 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 binds to one or more amino acid residues in 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 MS4A4A protein, wherein the antibody binds to one or more amino acid residues in 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.

[0028] 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 MS4A4A protein, and the antibody reduces cell surface levels of MS4A4A or reduces intracellular levels of MS4A4A.

[0029] In certain embodiments that may be combined with any of the embodiments herein, the MS4A4A protein is a mammalian protein or a 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.

[0030] 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, increase membrane TREM2 levels, or increase soluble TREM2 and increase membrane TREM2 levels in bone marrow cells. 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.

[0031] In some embodiments that may be combined with any of the embodiments herein, the anti-MS4A4A antibodies of the present disclosure inhibit 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 inhibit the levels of M2 cell surface markers on myeloid cells, e.g., macrophages.

[0032] In some embodiments that may be combined with any of the embodiments herein, the anti-MS4A4A antibodies of the present disclosure inhibit 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 inhibit the cell surface levels of CD200R, Dectin-1, and / or CD163 on myeloid cells, e.g., macrophages.

[0033] In some embodiments that may be combined with any of the embodiments herein, an anti-MS4A4A antibody of the present disclosure increases the mRNA and / or protein expression of gelsolin and / or osteopontin in bone marrow cells. In some embodiments that may be combined with any of the embodiments herein, an anti-MS4A4A antibody of the present disclosure increases the mRNA and / or protein level of IL1RN in bone marrow cells. In some embodiments that may be combined with any of the embodiments herein, an anti-MS4A4A antibody of the present disclosure increases the mRNA and / or protein level of CSF1R in bone marrow cells. In some embodiments that may be combined with any of the embodiments herein, an anti-MS4A4A antibody of the present disclosure decreases the mRNA and / or protein level of the purinergic receptor P2RY12 and / or CX3C chemokine receptor 1 (CX3CR1) in bone marrow cells. In some embodiments that may be combined with any of the embodiments herein, an anti-MS4A4A antibody of the present disclosure increases the mRNA and / or protein levels of 1-phosphotidylinositol-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) in myeloid cells. In some embodiments that may be combined with any of the embodiments herein, an anti-MS4A4A antibody of the present disclosure decreases the mRNA and / or protein levels of transmembrane glycoprotein NMB (GPNMB) in myeloid cells.

[0034] In one aspect, the present disclosure relates to an isolated antibody that binds to the MS4A4A protein, wherein the antibody increases the expression of gelsolin and / or osteopontin mRNA and / or protein in bone marrow cells. In some embodiments that may be combined with any of the embodiments herein, the bone marrow 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 inhibits expression of the M2 marker in 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 in 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 human macrophage supernatants by at least 15%, 20%, 25%, 30%, 40%, 50%, 60%, or 70%.

[0035] 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 2), diphtheria toxin receptor, CRM197, llama single-domain antibody, TMEM30(A), protein transduction domain, TAT, Syn-B, penetratin, poly-arginine peptide, angiopep peptide, basigin, Glut1, CD98hc, and ANG1005.

[0036] 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.

[0037] In some embodiments that may be combined with any of the embodiments herein, the anti-MS4A4A antibody of the present disclosure belongs to the IgG class, the IgM class, or the IgA class. In some embodiments, the antibody belongs to the IgG class and has an IgG1, IgG2, or IgG4 isotype. In certain embodiments that may be combined with any of the embodiments herein, the antibody is an antibody fragment. In certain embodiments that may be combined with any of the embodiments herein, the antibody is an antibody fragment that binds to an epitope comprising amino acid residues of human MS4A4A or a mammalian MS4A4A protein. In certain embodiments that may be combined with any of the embodiments herein, the antibody fragment is a Fab, Fab', Fab'-SH, F(ab')2, Fv, or scFv fragment.

[0038] 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: (a) antigens that facilitate transport across the blood-brain barrier; (b) an antigen that promotes transport across the blood-brain barrier, selected from transferrin receptor (TR), insulin receptor (HIR), insulin-like growth factor receptor (IGFR), low-density lipoprotein receptor-related proteins 1 and 2 (LPR-1 and 2), diphtheria toxin receptor, CRM197, llama single domain antibody, TMEM30(A), protein transduction domain, TAT, Syn-B, penetratin, polyarginine peptide, angiopeptide, and ANG1005; (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 amyloid beta, oligomeric amyloid beta, amyloid beta plaque, amyloid precursor protein or a fragment thereof, Tau, IAPP, alpha-synuclein, TDP-43, FUS protein, C9orf72 (chromosome 9 open reading frame 72), c9RAN protein, prion protein, PrPSc, or Hunchback. the pathogenic agent is selected from insulin, calcitonin, superoxide dismutase, ataxin, ataxin 1, ataxin 2, ataxin 3, ataxin 7, ataxin 8, ataxin 10, Lewy bodies, atrial natriuretic factor, islet amyloid polypeptide, insulin, apolipoprotein AI, serum amyloid A, medin, prolactin, transthyretin, lysozyme, beta 2 microglobulin, gelsolin, keratoepithelin, 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; (d) a ligand and / or protein expressed on an immune cell, the ligand and / or protein being 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; or (e) a protein, lipid, polysaccharide, or glycolipid expressed by one or more tumor cells.

[0039] In another aspect, the disclosure relates to an isolated nucleic acid comprising a nucleic acid sequence encoding the antibody of any of the preceding embodiments. In some embodiments, the disclosure relates to a vector comprising the nucleic acid of any of the preceding embodiments. In some embodiments, the disclosure relates to an isolated host cell comprising the vector of any of the preceding embodiments.

[0040] In another aspect, the disclosure relates to a method of producing an antibody that binds to a human MS4A4A antibody, comprising culturing the host cell of any of the preceding embodiments to produce the anti-MS4A4A antibody. In certain embodiments, the method further comprises recovering the anti-MS4A4A antibody produced by the cell.

[0041] 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.

[0042] In one aspect, the present disclosure relates to a method of preventing, reducing the risk of, or treating an individual having a disease, disorder, or injury selected from Alzheimer's disease, late-onset Alzheimer's disease, and cognitive impairment, comprising administering to an individual in need thereof a therapeutically effective amount of the antibody of any one of the preceding embodiments. In another aspect, the present disclosure relates to a method of preventing, reducing the risk of, or treating an individual having a disease, disorder, condition, or injury caused by or associated with increased overexpression or activity of MS4A4A, comprising administering to an individual in need thereof a therapeutically effective amount of the antibody of any one of the preceding embodiments. In some embodiments of the present disclosure, the method further comprises detecting levels of osteopontin, gelsolin, plasma membrane or cell surface TREM2, and / or soluble TREM2 before and / or after administration of the antibody.

[0043] In certain aspects, the disclosure relates to a method of preventing, reducing the risk of, or treating an individual having a CSF1R-deficient disease or disorder, comprising administering to an individual in need thereof a therapeutically effective amount of the antibody of any one of the preceding embodiments, hi some embodiments, the CSF1R-deficient disease or disorder is adult-onset leukoencephalopathy with axonal spheroids and pigmented glia (ALSP) or hereditary diffuse leukoencephalopathy (HDLS).

[0044] Other aspects of the disclosure relate to an isolated anti-MS4A4A antibody produced by the method of any of the preceding embodiments. Other aspects of the disclosure relate to a pharmaceutical composition comprising the anti-MS4A4A antibody of any of the preceding embodiments and a pharmaceutically acceptable carrier.

[0045] In another aspect, the disclosure relates to an antibody that binds to MS4A4A protein, the antibody comprising 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 (shown in Table 1, Table 5, Table 30, and Table 31).

[0046] In another aspect, the disclosure relates to an antibody that binds to MS4A4A protein, the antibody comprising 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 (shown in Table 1, Table 5, Table 30, and Table 31).

[0047] In another aspect, the disclosure relates to an antibody that binds to MS4A4A protein, the antibody comprising 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, the antibody being identified as 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 HVR-H1, HVR-H2, HVR-H3, HVR-L1, HVR-L2, and HVR-L3.

[0048] In another aspect, the disclosure relates to an antibody that binds to MS4A4A protein, the antibody comprising 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 (shown in Table 2 and Table 6).

[0049] In another aspect, the disclosure relates to an antibody that binds to MS4A4A protein, the antibody comprising 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 (shown in Table 2 and Table 6).

[0050] In another aspect, the disclosure relates to an antibody that binds to MS4A4A protein, the antibody comprising 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, the antibody comprising 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 (shown in Table 2 and Table 6).

[0051] In another aspect, the disclosure relates to an antibody that binds to MS4A4A protein, the antibody comprising a heavy chain variable region and a light chain variable region, the heavy chain variable region being selected from the group consisting of 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, 4A-376, 4A-377, 4A-378, 4A-379, 4A-380, 4A-381, 4A-382, 4A-383, 4A-384, 4A-385, 4A-386, 4A-387, 4A-388, 4A-38 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-3 69, 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 including HVR-H1, HVR-H2, and HVR-H3.

[0052] In another aspect, the disclosure relates to an antibody that binds to MS4A4A protein, the antibody comprising a heavy chain variable region and a light chain variable region, the light chain variable region being selected from the group consisting of 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, 4A-376, 4A-377, 4A-378, 4A-379, 4A-380, 4A-381, 4A-382, 4A-383, 4A-384, 4A-385, 4A-386, 4A-387, 4A-388, 4A-38 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-3 69, 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 HVR-L1, HVR-L2, and HVR-L3.

[0053] In another aspect, the disclosure relates to an antibody that binds to MS4A4A protein, the antibody comprising 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, the antibody being selected from the group consisting of 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, 4A-376, 4A-377, 4A-378, 4A-379, 4A-380, 4A-381, 4A-382, 4A-383, 4A-384, 4 A-351, 4A-352, 4A-353, 4A-354, 4A-355, 4A-356, 4A-357, 4A-358, 4A-359, 4A-3 60, 4A-361, 4A-361, 4A-363, 4A-364, 4A-365, 4A-366, 4A-367, 4A-368, 4A-369, 4 HVR-H1, HVR-H2, HVR-H3, HVR-L1, HVR-L2, and HVR-L3 of 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.

[0054] In another aspect, the disclosure relates to an antibody that binds to MS4A4A protein, the antibody comprising 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 (shown in Tables 4 and 8).

[0055] In another aspect, the disclosure relates to an antibody that binds to MS4A4A protein, the antibody comprising 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 (shown in Tables 4 and 8).

[0056] In another aspect, the disclosure relates to an antibody that binds to 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 (shown in Table 4 and Table 8).

[0057] In certain embodiments that may be combined with any of the embodiments herein, the present disclosure relates to an antibody that binds to the MS4A4A protein, wherein the antibody increases expression of gelsolin and / or osteopontin mRNA and / or protein in bone marrow cells. In some embodiments, the bone marrow cells are macrophages.

[0058] 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.

[0059] [The present invention 1001] An isolated antibody that binds to MS4A4A protein, the antibody comprising 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 the group consisting of SEQ ID NOs: 102, 100, 101, 102, 112, 123, 124, 125, 126, 127, 128, 129, 154, 310, and 313. and an HVR-H3 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 104, 103, 113, 130, 131, 132, 133, 134, 135, 136, 137, 138, 156, 157, 158, 314, and 317; an HVR-L1 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 105, 106, 114, 139, 140, 141, 142, 143, 159, 160, 161, 315, and 318; and an HVR-L3 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 107, 115, 144, 145, 163, 316, and 319. [The present invention 1002] The antibody of the present invention 1001, 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 an amino acid sequence selected from the group consisting of SEQ ID NOs: 96 to 99; and HVR-H3 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 102, 100, and 101, and the light chain variable region comprises: HVR-L1 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 104 and 103; HVR-L2 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 105 to 106; and HVR-L3 comprising the amino acid sequence of SEQ ID NO: 107. [The present invention 1003] The antibody of the present invention 1001, 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 the group consisting of SEQ ID NOs: 110 to 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. [The present invention 1004] The antibody of the present invention 1001, 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: 116; HVR-H2 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 118 to 122; and HVR-H3 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 123 to 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 to 138; HVR-L2 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 139 to 143; and HVR-L3 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 144 to 145. [The present invention 1005] The antibody of the present invention 1001, 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 to 147; HVR-H2 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 149 to 153; and HVR-H3 comprising the 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 to 158; HVR-L2 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 159 to 161; and HVR-L3 comprising the amino acid sequence of SEQ ID NO: 163. [The present invention 1006] The antibody of the present invention 1001, 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. [The present invention 1007] The antibody of the present invention 1001, 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. [The present invention 1008] The antibody of the present invention 1001, 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. [The present invention 1009] An antibody that binds to MS4A4A protein, the antibody comprising 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. [The present invention 1010] An antibody that binds to MS4A4A protein, the antibody comprising a heavy chain variable region and a light chain variable region, and the light chain variable region comprising 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. [The present invention 1011] An antibody that binds to MS4A4A protein, the antibody comprising 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, the antibody comprising 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. [The present invention 1012] An antibody that binds to MS4A4A protein, the antibody comprising 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. [The present invention 1013] An antibody that binds to MS4A4A protein, the antibody comprising 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. [The present invention 1014] An antibody that binds to MS4A4A protein, the antibody comprising 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. [The present invention 1015] An antibody that binds to MS4A4A protein, the antibody comprising a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region is selected from the group consisting of 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, The antibody, comprising HVR-H1, HVR-H2, and HVR-H3 of 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, 4A-376, 4A-377, 4A-378, 4A-379, 4A-380, or 4A-381. [The present invention 1016] An antibody that binds to MS4A4A protein, the antibody comprising a heavy chain variable region and a light chain variable region, wherein the light chain variable region is selected from the group consisting of 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, The antibody, comprising HVR-L1, HVR-L2, and HVR-L3 of 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, 4A-376, 4A-377, 4A-378, 4A-379, 4A-380, or 4A-381. [The present invention 1017] An antibody that binds to MS4A4A protein, the antibody comprising 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, the antibody being selected from the group consisting of 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, 4A-376, 4A-377, 4A-378, 4A-379, 4A-380, 4A-381, 4A-382, 4A-383, 4A-384, 4A-385, 4 A-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, 4A-376, 4A-377, 4A-378, 4A-379, 4A-3 80, or 4A-381, HVR-H1, HVR-H2, HVR-H3, HVR-L1, HVR-L2, and HVR-L3. [The present invention 1018] An antibody that binds to MS4A4A protein, the antibody comprising 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. [The present invention 1019] An antibody that binds to MS4A4A protein, wherein the antibody comprises a heavy chain variable region and a light chain variable region, and 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. [The present invention 1020] An antibody that binds to MS4A4A protein, the antibody comprising 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, the antibody comprising 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. [The present invention 1021] The antibody of the present invention 1001 or 1009, wherein the antibody comprises a heavy chain variable region, and the heavy chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 5 to 15, 304, and 306. [The present invention 1022] The antibody of the present invention 1001 or 1012, wherein the antibody comprises a heavy chain variable region, and the heavy chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 24 to 30. [The present invention 1023] The antibody of the present invention 1001 or 1015, wherein the antibody comprises a heavy chain variable region, and the heavy chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 40 to 53. [The present invention 1024] The antibody of the present invention 1001 or 1018, wherein the antibody comprises a heavy chain variable region, and the heavy chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 76 to 84. [The present invention 1025] The antibody of the present invention 1001 or 1010, wherein the antibody comprises a light chain variable region, and the light chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 17 to 22, 305, and 307. [The present invention 1026] The antibody of the present invention 1001 or 1013, wherein the antibody comprises a light chain variable region, and the light chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 32 to 36. [The present invention 1027] The antibody of the present invention 1001 or 1016, wherein the antibody comprises a light chain variable region, and the light chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 55 to 74. [The present invention 1028] The antibody of the present invention 1001 or 1019, wherein the antibody comprises a light chain variable region, and the light chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 86 to 93. [The present invention 1029] The antibody of the present invention 1001 or 1011, wherein the antibody comprises a heavy chain variable region and a light chain variable region, the heavy chain variable region comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 5 to 15, 304, and 306, and the light chain variable region comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 17 to 22, 305, and 307. [The present invention 1030] The antibody of the present invention 1001 or 1014, wherein the antibody comprises a heavy chain variable region and a light chain variable region, the heavy chain variable region comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 24 to 30, and the light chain variable region comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 32 to 36. [The present invention 1031] The antibody of the present invention 1001 or 1017, wherein the antibody comprises a heavy chain variable region and a light chain variable region, the heavy chain variable region comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 40 to 53, and the light chain variable region comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 55 to 74. [The present invention 1032] The antibody of the present invention 1001 or 1020, wherein the antibody comprises a heavy chain variable region and a light chain variable region, the heavy chain variable region comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 76 to 84, and the light chain variable region comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 86 to 93. [The present invention 1033] The antibody of any one of the antibodies of the present invention 1001, 1002, 1004, 1006 to 1011, 1021, 1025, and 1029, wherein the antibody comprises a heavy chain and a light chain, and the heavy chain comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 320 to 343. [The present invention 1034] Any one of the antibodies of the present invention 1001, 1002, 1006, 1009 to 1011, 1021, 1025, and 1029, wherein the antibody comprises a heavy chain and a light chain, the heavy chain comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 336 to 343, and the light chain comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO: 22, and optionally the light chain further comprising a constant region comprising the amino acid sequence of SEQ ID NO: 344. [This invention 1035] Any of the antibodies of the present invention 1001, 1007, 1009 to 1011, 1021, 1025, and 1029, wherein the antibody comprises a heavy chain and a light chain, the heavy chain comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 320 to 327, the light chain comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO: 305, and optionally the light chain further comprising a constant region comprising the amino acid sequence of SEQ ID NO: 344. [The present invention 1036] Any of the antibodies of the present invention 1001, 1008 to 1011, 1021, 1025, and 1029, wherein the antibody comprises a heavy chain and a light chain, the heavy chain comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 328 to 335, the light chain comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO: 307, and optionally the light chain further comprising a constant region comprising the amino acid sequence of SEQ ID NO: 344. [This invention 1037] Any of the antibodies of the present invention 1001, 1002, 1006, 1009 to 1011, 1021, 1025, and 1029, wherein the antibody comprises a heavy chain and a light chain, the heavy chain comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 355 to 362, and the light chain comprising the amino acid sequence of SEQ ID NO: 365. [The present invention 1038] Any of the antibodies of the present invention 1001, 1007, 1009 to 1011, 1021, 1025, and 1029, wherein the antibody comprises a heavy chain and a light chain, the heavy chain comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 320 to 327, and the light chain comprising the amino acid sequence of SEQ ID NO: 363. [This invention 1039] Any of the antibodies of the present invention 1001, 1008 to 1011, 1021, 1025, and 1029, wherein the antibody comprises a heavy chain and a light chain, the heavy chain comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 328 to 335, and the light chain comprising the amino acid sequence of SEQ ID NO: 364. [The present invention 1040] the antibody: a. heavy and light chain variable regions, 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; b. 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; or c. An antibody of the present invention, comprising 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. [This invention 1041] the antibody: a. 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; or b. The antibody of the present invention, comprising 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. [The present invention 1042] the antibody: a. 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; or b. The antibody of the present invention, comprising 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. [This invention 1043] An isolated antibody that binds to MS4A4A protein, wherein the antibody competitively inhibits the binding to MS4A4A of one or more of the antibodies of any one of claims 1001 to 1042 of the present invention. [This invention 1044] An isolated antibody that binds to MS4A4A protein, said antibody binding to an epitope of MS4A4A that is essentially the same as or overlaps with any of antibodies 1001 to 1042 of the present invention. [This invention 1045] 1044. The antibody of the present invention, wherein the antibody binds to one or more amino acid residues of the amino acid sequence SFHHPYCNYYGNSNNCHGTMS (SEQ ID NO: 178) of SEQ ID NO: 1, and the antibody does not bind to amino acid residue proline 163 of SEQ ID NO: 1. [The present invention 1046] The antibody of any one of claims 1001 to 1042, which binds to the extracellular domain 1 of human MS4A4A. [This invention 1047] The antibody of any one of claims 1001 to 1042, wherein the antibody binds to one or more amino acid residues of the amino acid sequence CMASNTYGSNPIS of SEQ ID NO: 1 (SEQ ID NO: 177). [This invention 1048] The antibody of any one of claims 1001 to 1042, which binds to the extracellular domain 2 of human MS4A4A. [This invention 1049] The antibody of any one of claims 1001 to 1042, wherein the antibody binds to one or more amino acid residues of the amino acid sequence SFHHPYCNYYGNSNNCHGTMS of SEQ ID NO: 1 (SEQ ID NO: 178). [The present invention 1050] The antibody of any one of claims 1001 to 1049, wherein the antibody is human or humanized. [This invention 1051] The antibody of any one of claims 1001 to 1050, wherein the antibody reduces the cell surface level of MS4A4A or reduces the intracellular level of MS4A4A. [This invention 1052] The antibody of any one of claims 1001 to 1051, wherein the antibody increases soluble TREM2 levels, increases membrane TREM2 levels, or increases soluble TREM2 and increases membrane TREM2 levels in bone marrow cells. [This invention 1053] The antibody of any one of claims 1001 to 1052, wherein the antibody suppresses the expression of an M2 cell surface marker in bone marrow cells. [This invention 1054] The antibody of the present invention, wherein the antibody inhibits the expression of CD200R, Dectin-1, or CD163 in bone marrow cells. [This invention 1055] The antibody of any one of claims 1001 to 1050, wherein the antibody enhances the expression of gelsolin and / or osteopontin mRNA and / or protein in bone marrow cells. [This invention 1056] 1. An isolated antibody that binds to MS4A4A protein, wherein said antibody increases expression of gelsolin and / or osteopontin mRNA and / or protein in bone marrow cells. [This invention 1057] The antibody of the present invention 1056, wherein said bone marrow cells are human macrophages. [This invention 1058] The antibody of the invention 1056 or 1057, wherein said antibody is human or humanized. [This invention 1059] The antibody according to any one of claims 1056 to 1058, which binds to the same epitope as an antibody selected from the group consisting of 4A-18, 4A-25, 4A-214, 4A-21, and 4A-202. [The present invention 1060] The antibody of any one of 1056 to 1059 of the present invention, which suppresses the expression of an M2 marker in bone marrow cells. [This invention 1061] The antibody of the present invention, wherein the bone marrow cells are human macrophages. [This invention 1062] The antibody of the present invention 1061, wherein the M2 marker is CD200R, Dectin-1, and / or CD163. [This invention 1063] The antibody of any of claims 1056 to 1062, wherein the antibody increases membrane TREM2 in human macrophages by at least 50%, 90%, 100%, 150%, 200%, or 250%. [This invention 1064] The antibody of any of claims 1056 to 1063, wherein the antibody increases soluble TREM2 in human macrophage supernatant by at least 15%, 20%, 25%, 30%, 40%, 50%, 60%, or 70%. [This invention 1065] The antibody of any one of claims 1001 to 1064, wherein the antibody is a monoclonal antibody. [The present invention 1066] The antibody of any one of claims 1001 to 1065 of the present invention, which binds to human MS4A4A of SEQ ID NO:1. [This invention 1067] 1066. The antibody of the present invention, wherein said antibody further binds to cynomolgus monkey MS4A4A of SEQ ID NO:3. [The present invention 1068] The antibody of any one of claims 1001 to 1067, wherein the antibody belongs to the IgG class and has an IgG1, IgG2, or IgG4 isotype. [The present invention 1069] The antibody of the present invention, wherein said antibody has an IgG1 isotype. [The present invention 1070] 1069. The antibody of the present invention, wherein the antibody comprises a modified Fc comprising N325S and L328F mutations as defined by the EU numbering system. [This invention 1071] The antibody of any one of claims 1001 to 1070, wherein the antibody is an antibody fragment. [This invention 1072] The antibody of the present invention 1071, wherein the antibody fragment is a Fab, Fab', Fab'-SH, F(ab')2, Fv, or scFv fragment. [This invention 1073] The antibody of any of claims 1001 to 1072, wherein the antibody is a bispecific antibody that recognizes a first antigen and a second antigen. [This invention 1074] The first antigen is MS4A4A and the second antigen is: (a) antigens that facilitate 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 2), diphtheria toxin receptor, CRM197, llama single domain antibody, TMEM30(A), protein transduction domain, TAT, Syn-B, penetratin, polyarginine peptide, angiopeptide, and ANG1005; (c) a pathogenic agent selected from the group consisting of 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 amyloid beta, oligomeric amyloid beta, amyloid beta plaque, amyloid precursor protein or a fragment thereof, Tau, IAPP, alpha-synuclein, TDP-43, FUS protein, C9orf72 (chromosome 9 open reading frame 72), c9RAN protein, prion protein, PrPSc, Ha the pathogenic agent is selected from the group consisting of insulin, calcitonin, superoxide dismutase, ataxin, ataxin 1, ataxin 2, ataxin 3, ataxin 7, ataxin 8, ataxin 10, Lewy bodies, atrial natriuretic factor, islet amyloid polypeptide, insulin, apolipoprotein AI, serum amyloid A, medin, prolactin, transthyretin, lysozyme, beta 2 microglobulin, gelsolin, keratoepithelin, 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; (d) a ligand and / or protein expressed on an immune cell, the ligand and / or protein being 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 (e) The antibody of the present invention 1073, which is a protein, lipid, polysaccharide, or glycolipid expressed in one or more tumor cells. [This invention 1075] An isolated nucleic acid comprising a nucleic acid sequence encoding any of the preceding antibodies of the present invention. [This invention 1076] A vector comprising the nucleic acid of the present invention. [This invention 1077] An isolated host cell comprising a nucleic acid of the invention 1075 or a vector of the invention 1076. [This invention 1078] A method for producing an antibody that binds to human MS4A4A, comprising culturing the cell of the present invention 1077 to produce the antibody. [This invention 1079] The method of claim 1078, further comprising recovering the antibody produced by the cells. [The present invention 1080] A pharmaceutical composition comprising any one of the antibodies of the present invention 1001 to 1074 and a pharmaceutically acceptable carrier. [This invention 1081] A method for preventing, reducing the risk of, or treating an individual having a disease, disorder, or injury selected from the group consisting of Alzheimer's disease, late-onset Alzheimer's disease, and cognitive impairment, the method comprising administering a therapeutically effective amount of any of the antibodies 1001 to 1074 of the present invention to an individual in need thereof. [This invention 1082] A method for preventing, reducing the risk of, or treating an individual with a disease, disorder, pathology, or injury caused by or associated with overexpression or increased activity of MS4A4A, comprising administering a therapeutically effective amount of any of the antibodies of the present inventions 1001 to 1074 to an individual in need thereof. [This invention 1083] A method for preventing, suppressing the risk of, or treating an individual with a CSF1R-deficient disease or disorder, comprising administering a therapeutically effective amount of any of the antibodies of the present inventions 1001 to 1074 to an individual in need thereof. [This invention 1084] The method of claim 1083, wherein the CSF1R-deficient disease or disorder is adult-onset leukoencephalopathy with axonal spheroids and pigmented glia (ALSP) or hereditary diffuse leukoencephalopathy (HDLS). [This invention 1085] The method of any of claims 1081 to 1084, further comprising detecting levels of osteopontin, gelsolin, membrane TREM2, and / or soluble TREM2 before and / or after administration of said antibody. It should be understood that one, some, or all of the properties of the various embodiments described herein may be combined to form other embodiments of the present disclosure. These and other aspects of the present disclosure will be apparent to those skilled in the art. These and other embodiments of the present disclosure are described in further detail below.

[0060] A patent or application file shall contain at least one drawing printed in color. Copies of the patent or patent application publication containing such drawing(s) in color will be provided by the Office upon request and payment of the required fee. [Brief explanation of the drawings]

[0061] [Figure 1] The primary amino acid sequence of human MS4A4A is shown, with the intracellular domain in italics, the transmembrane domain underlined, and the extracellular loops in bold italics. [Figure 2] A shows a structural schematic of a soluble four-helix bundle scaffold (PDB_ID: 1P68) obtained from the RCSB Protein Data Bank, with four transmembrane domains visible as helical ribbons within the structure. B and C show the primary amino acid sequences of the soluble four-helix bundle scaffolds PDB-ID: 1P68 and PDB_ID: 1M6T, respectively; the four transmembrane domains are underlined in the amino acid sequence. [Figure 3A]A shows the primary amino acid sequences of the soluble four-helix bundle scaffolds JS1, JS5, and JS6 plus the two extracellular loops (bold underlined) of human MS4A4A. [Figure 3B] B shows the primary amino acid sequences of the soluble four-helix bundle scaffolds JS4 and JS10 used as negative control variants. [Figure 4] 1 shows that specific anti-MS4A4A antibodies of the disclosure bind to recombinant MS4A4A soluble loop-grafted polypeptides JS1, JS4, JS5, JS6, and JS10 by ELISA. [Figure 5] 1 shows specific humanized anti-MS4A4A antibody variants of the murine anti-MS4A4A antibody 4A-21 binding to the recombinant MS4A4A soluble loop-grafted polypeptide JS1. [Figure 6] 1 shows specific humanized anti-MS4A4A antibody variants of the murine anti-MS4A4A antibody 4A-202 binding to the recombinant MS4A4A soluble loop-grafted polypeptide JS1. [Figure 7] 1 shows specific humanized and affinity matured anti-MS4A4A antibody variants of the murine anti-MS4A4A antibody 4A-202 binding to the recombinant MS4A4A soluble loop-grafted polypeptide JS5. [Figure 8] 1 shows specific humanized and affinity matured anti-MS4A4A antibody variants of the murine anti-MS4A4A antibody 4A-21 binding to the recombinant MS4A4A soluble loop-grafted polypeptide JS5. [Figure 9] 1 shows specific humanized and affinity matured anti-MS4A4A antibody variants of the murine anti-MS4A4A antibody 4A-21 binding to the recombinant MS4A4A soluble loop-grafted polypeptide JS5. [Figure 10A]1 shows the effect of anti-MS4A4A antibody 4A-202 on sTREM2 levels in cynomolgus monkey serum. Anti-MS4A4A antibody 4A-202 or an isotype control antibody (huIgG1), both administered at a dose of 80 mg / kg, shows sTREM2 levels (ng / mL) in cynomolgus monkey serum at the indicated time points (hours). [Figure 10B] 1 shows the effect of anti-MS4A4A antibody 4A-202 on sTREM2 levels in serum of cynomolgus monkeys. Anti-MS4A4A antibody 4A-202 or an isotype control antibody (huIgG1), both administered at a dose of 80 mg / kg, shows the levels of sTREM2 (percent of baseline) in serum of cynomolgus monkeys at the indicated time points (hours). [Figure 11A] 1 shows the effect of anti-MS4A4A antibody 4A-202 on sTREM2 levels in the cerebrospinal fluid (CSF) of cynomolgus monkeys. Anti-MS4A4A antibody 4A-202 or an isotype control antibody (huIgG1), both administered at a dose of 80 mg / kg, shows the levels of sTREM2 (ng / mL) in the CSF of cynomolgus monkeys at the indicated time points (hours). [Figure 11B] 1 shows the effect of anti-MS4A4A antibody 4A-202 on sTREM2 levels in the cerebrospinal fluid (CSF) of cynomolgus monkeys. Anti-MS4A4A antibody 4A-202 or an isotype control antibody (huIgG1), both administered at a dose of 80 mg / kg, shows the levels of sTREM2 (percent of baseline) in the CSF of cynomolgus monkeys at the indicated time points (hours). [Figure 12] 1 shows data depicting the binding curves of anti-MS4A4A antibodies 4A-313 wild-type (WT) huIgG1 and 4A-450 WT huIgG1 to U937 cells expressing recombinant human MS4A4A. [Figure 13] 1 shows data depicting 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. [Figure 14] 1 shows data depicting 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. [Figure 15] Figure 1 shows data demonstrating increased soluble TREM2 levels in the supernatants of primary human macrophages treated with anti-MS4A4A antibody 4A-450 huIgG1. [Figure 16] Figure 1 shows data demonstrating increased levels of soluble TREM2 in the supernatants of primary human macrophages treated with the anti-MS4A4A antibody 4A-313 huIgG1. [Figure 17] 1 shows data demonstrating 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. [Figure 18] AC show data depicting changes in CD14, CD163, and CD200R levels in primary human macrophages treated with anti-MS4A4A antibodies 4A-313 NSLF huIgG1 and 4A-419 WT huIgG1 (respectively). [Figure 19] 1 shows data demonstrating the complement-dependent cytotoxicity (CDC) activity of anti-MS4A4A antibodies 4A-313 NSLF huIgG1, 4A-313 K322A huIgG1, and 4A-313 WT huIgG1 in U937 cells expressing recombinant human MS4A4A, as measured by propidium iodide incorporation. [Figure 20] 1 shows data demonstrating the ADCP activity of anti-MS4A4A antibodies 4A-313 NSLF huIgG1, 4A-313 K322A huIgG1, and 4A-313 WT huIgG1 in primary human macrophages, as measured by FcyRIIIa activation. [Figure 21]1 shows data demonstrating the ADCC activity of anti-MS4A4A antibodies 4A-313 NSLF huIgG1, 4A-313 K322A huIgG1, and 4A-313 WT huIgG1 in primary human macrophages, as measured by FcyRIIIa activation. [Figure 22] AB show data depicting the effect of MS4A4A knockout on membrane TREM2 (A) and soluble TREM2 (B) in primary human macrophages. [Figure 23A] Panels A to C show 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 the anti-MS4A4A antibody of the present disclosure. [Figure 23B] Panels A to C show 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 the anti-MS4A4A antibody of the present disclosure. [Figure 23C] Panels A to C show 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 the anti-MS4A4A antibody of the present disclosure. [Figure 24] AB show data demonstrating that siRNA knockdown of MS4A4A increased membrane and soluble TREM2 levels in human macrophages. [Figure 25A] AC show data demonstrating a dose-dependent increase in membrane TREM2 levels in primary human macrophages from three different donors after addition of an anti-MS4A4A antibody of the present disclosure. [Figure 25B] AC show data demonstrating a dose-dependent increase in membrane TREM2 levels in primary human macrophages from three different donors after addition of an anti-MS4A4A antibody of the present disclosure. [Figure 25C]AC show data demonstrating a dose-dependent increase in membrane TREM2 levels in primary human macrophages from three different donors after addition of an anti-MS4A4A antibody of the present disclosure. [Figure 26A] AC show data demonstrating a dose-dependent increase in soluble TREM2 levels in primary human macrophages from three different donors after addition of an anti-MS4A4A antibody of the present disclosure. [Figure 26B] AC show data demonstrating a dose-dependent increase in soluble TREM2 levels in primary human macrophages from three different donors after addition of an anti-MS4A4A antibody of the present disclosure. [Figure 26C] AC show data demonstrating a dose-dependent increase in soluble TREM2 levels in primary human macrophages from three different donors after addition of an anti-MS4A4A antibody of the present disclosure. [Figure 27A] AC show data demonstrating a dose-dependent increase in ATP levels in primary human macrophages from three different donors after addition of an anti-MS4A4A antibody of the present disclosure. [Figure 27B] AC show data demonstrating a dose-dependent increase in ATP levels in primary human macrophages from three different donors after addition of an anti-MS4A4A antibody of the present disclosure. [Figure 27C] AC show data demonstrating a dose-dependent increase in ATP levels in primary human macrophages from three different donors after addition of an anti-MS4A4A antibody of the present disclosure. [Figure 28] Panels A-B show data demonstrating that anti-MS4A4A antibodies of the present disclosure are effective in rescuing cell death induced by CSF1R inhibition. [Figure 29] 1 shows a volcano plot of the proteome-wide effect in cynomolgus monkey CSF after administration of an anti-MS4A4A antibody of the present disclosure. [Figure 30] 1 shows data demonstrating increased sTREM2 levels in cynomolgus monkey serum after administration of an anti-MS4A4A antibody of the present disclosure. [Figure 31A]5A-5D show data demonstrating increased osteopontin levels in cynomolgus monkey CSF after administration of an anti-MS4A4A antibody of the present disclosure. [Figure 31B] 5A-5D show data demonstrating increased osteopontin levels in cynomolgus monkey CSF after administration of an anti-MS4A4A antibody of the present disclosure. [Figure 31C] 5A-5D show data demonstrating increased osteopontin levels in cynomolgus monkey CSF after administration of an anti-MS4A4A antibody of the present disclosure. [Figure 31D] 5A-5D show data demonstrating increased osteopontin levels in cynomolgus monkey CSF after administration of an anti-MS4A4A antibody of the present disclosure. [Figure 32A] AB show data showing an increase in osteopontin levels in the frontal lobe and hippocampus, respectively, of the brain of cynomolgus monkeys after administration of an anti-MS4A4A antibody of the present disclosure. [Figure 32B] AB show data showing an increase in osteopontin levels in the frontal lobe and hippocampus, respectively, of the brain of cynomolgus monkeys after administration of an anti-MS4A4A antibody of the present disclosure. [Figure 33A] AB show data showing an increase in CSF1R levels in the frontal lobe and hippocampus, respectively, of the brain of cynomolgus monkeys after administration of an anti-MS4A4A antibody of the present disclosure. [Figure 33B] AB show data showing an increase in CSF1R levels in the frontal lobe and hippocampus, respectively, of the brain of cynomolgus monkeys after administration of an anti-MS4A4A antibody of the present disclosure. [Figure 34] AB show data showing increased levels of total TREM2 protein in the frontal lobe and hippocampus, respectively, of the cynomolgus monkey brain after administration of an anti-MS4A4A antibody of the present disclosure. [Figure 35A] 1A-C show data depicting changes in mRNA levels of markers of microglial activation, migration, and proliferation in microglia isolated from the brains of cynomolgus monkeys administered an anti-MS4A4A antibody of the present disclosure. [Figure 35B]1A-C show data depicting changes in mRNA levels of markers of microglial activation, migration, and proliferation in microglia isolated from the brains of cynomolgus monkeys administered an anti-MS4A4A antibody of the present disclosure. [Figure 35C] 1A-C show data depicting changes in mRNA levels of markers of microglial activation, migration, and proliferation in microglia isolated from the brains of cynomolgus monkeys administered an anti-MS4A4A antibody of the present disclosure. [Figure 36] 1 shows data demonstrating that anti-MS4A4A antibodies of the present disclosure inhibit cell surface expression of recombinantly expressed HA-snorkel-tagged MS4A4A protein in U937 cells. [Figure 37] 1 presents data demonstrating that anti-MS4A4A antibodies of the present disclosure suppressed GPNMB cell surface expression in primary human macrophages. DETAILED DESCRIPTION OF THE INVENTION

[0062] The present disclosure relates to anti-MS4A4A antibodies (e.g., monoclonal antibodies); methods of making and using these antibodies; pharmaceutical compositions containing these antibodies; nucleic acids encoding these antibodies; and host cells containing nucleic acids encoding these antibodies.

[0063] The techniques and procedures described or referenced herein are generally well understood, and those skilled in the art are familiar with conventional methodologies that are widely used methodologies, such as those described in, for example, Sambrook et al. Molecular Cloning: A Laboratory Manual 3rd edition (2001) Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY; Current Protocols in Molecular Biology (F.M. Ausubel, et al. eds., (2003); Monoclonal Antibodies: A Practical Approach (P. Shepherd and C. Dean, eds., Oxford University Press, 2000).

[0064] I. Definition The terms "MS4A4A" or "MS4A4A polypeptide" are used interchangeably herein and refer to any naturally occurring MS4A4A of any vertebrate origin, including mammals such as primates (e.g., humans and cynomolgus monkeys) and rodents (e.g., mice and rats), unless otherwise specified. In some embodiments, the term includes both wild-type sequences and naturally occurring mutant sequences, e.g., splice variants or allelic variants. In some embodiments, the term includes "full-length," unprocessed MS4A4A, as well as all forms of MS4A4A that involve intracellular processing. In some embodiments, the 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.

[0065] The terms "anti-MS4A4A antibody," "antibody that binds to MS4A4A," and "antibody that specifically binds to MS4A4A" refer to an antibody that is useful as a diagnostic and / or therapeutic agent targeting MS4A4A and is capable of binding to MS4A4A with sufficient affinity. In certain embodiments, the extent of binding of an anti-MS4A4A antibody to an unrelated, non-MS4A4A polypeptide is less than about 10% of the binding of the antibody to MS4A4A, as determined, for example, by radioimmunoassay (RIA). In certain embodiments, an antibody that binds to MS4A4A has 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 less, e.g., 10 -8 M~10 -13 M, e.g., 10 -9 M~10 -13 In certain embodiments, the anti-MS4A4A antibody binds to an epitope of MS4A4A that is conserved among MS4A4A from different species.

[0066] With respect to antibody binding to a target molecule, the terms "specific binding" or "specifically binds," or "specific" for a particular polypeptide or epitope on a particular polypeptide target, refer to measurable binding that is distinct from non-specific interactions. Specific binding can be measured, for example, by determining binding of a molecule compared to 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, specific binding is indicated if excess unlabeled target competitively inhibits binding of the labeled target to the probe. As used herein, the terms "specific binding" or "specifically binds," or "specific" for a particular polypeptide or epitope on a particular polypeptide target, refer to binding that is distinct from non-specific interactions, for example, approximately 10 -4 M or less, 10 -5 M or less, 10 -6 M or less, 10 -7 M or less, 10 -8 M or less, 10-9 M or less, 10 -10 M or less, 10 -11 M or less, 10 -12 KD for any target less than or equal to M, or 10 -4 M~10 -6 M or 10 -6 M~10 -10 M or 10 -7 M~10 -9 A molecule can be expressed as having a KD in the range of M. Those skilled in the art will understand that affinity and KD values ​​are inversely proportional. A high affinity for an antigen is reflected in a low KD value. In one embodiment, the term "specific binding" refers to binding of a molecule to a particular polypeptide or epitope on a particular polypeptide without substantially binding to any other polypeptides or polypeptide epitopes.

[0067] As used herein, the term "immunoglobulin" (Ig) is used interchangeably with "antibody." As used herein, the term "antibody" is used in the broadest sense to specifically include monoclonal antibodies, polyclonal antibodies, multispecific antibodies (e.g., bispecific antibodies) formed from at least two intact antibodies, and antibody fragments so long as they exhibit the desired biological activity.

[0068] "Native antibodies" are usually heterotetrameric glycoproteins of 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 chain and each light chain also have regularly spaced intrachain disulfide bridges. Each heavy chain contains at one end a variable domain (V H ), followed by several constant domains. Each light chain has at one end a variable domain (V L) at one end and a constant domain at the other end; the light-chain constant domain 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. Particular amino acid residues are believed to form an interface between the light-chain variable domain and the heavy-chain variable domain.

[0069] For the structure and properties of different antibody classes, see, e.g., Basic and Clinical Immunology, 8th Ed., Daniel P. Stites, Abba I. Terr and Tristram G. Parslow (eds.), Appleton & Lange, Norwalk, CT, 1994, page 71 and Chapter 6.

[0070] 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 their constant domain. Immunoglobulins can be assigned to different classes or isotypes depending on the amino acid sequence of the constant domain (CH) of their heavy chains. There are five classes of immunoglobulins: IgA, IgD, IgE, IgG, and IgM, with heavy chains designated alpha ("α"), delta ("δ"), epsilon ("ε"), gamma ("γ"), and mu ("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; see, for example, Abbas et al., Cellular and Molecular Immunology, 4 th This is outlined in Saunders Co., ed. (WBSaunders Co., 2000).

[0071] The "variable region" or "variable domain" of an antibody, such as the anti-MS4A4A antibody of the present disclosure, refers to the amino-terminal domains of the heavy or light chain of the antibody. The variable domains of the heavy and light chains are referred to as "V H " and "V L These domains are generally the most variable parts of an antibody (compared to other antibodies of the same class) and contain the antigen-binding sites.

[0072] The term "variable" refers to the fact that certain segments of the variable domains vary significantly in sequence among antibodies, such as the anti-MS4A4A antibodies of the present disclosure. The variable domains mediate antigen binding and define the specificity of a particular antibody for its particular antigen. However, the variability is not uniformly distributed throughout the variable domains. Rather, it is concentrated in three segments called hypervariable regions (HVRs) in both the light-chain and heavy-chain variable domains. The highly conserved portions of the variable domains are referred to as framework regions (FRs). Naturally occurring heavy-chain and light-chain variable domains each contain four FR regions primarily in a beta-sheet configuration, connected by three HVRs, which form loops that connect to, and in some cases form part of, the beta-sheet structure. The HVRs in each chain are held together in close proximity by the FR regions and, together with the HVRs from the other chain, contribute to the formation of the antigen-binding site of the antibody (see Kabat et al., Sequences of Immunological Interest, Fifth Edition, National Institutes of Health, Bethesda, MD (1991)). The constant domains are not directly involved in binding the antibody to an antigen but exhibit various effector functions, such as participation of the antibody in antibody-dependent cellular cytotoxicity.

[0073] As used herein, the term "monoclonal antibody" refers to an antibody obtained from a substantially homogeneous antibody population, such as the monoclonal anti-MS4A4A antibody of the present disclosure; i.e., the individual antibodies comprising the population are identical except for minor naturally occurring mutations and / or post-translational modifications (e.g., isomerization, amidation, etc.). Monoclonal antibodies are highly specific, being directed against one or more antigenic sites. 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 their specificity, monoclonal antibodies are advantageous in that they are synthesized in a hybridoma culture, uncontaminated by other immunoglobulins. The modifier "monoclonal" indicates the character of the antibody as being obtained from a substantially homogeneous antibody population, and should not be construed as requiring production of the antibody by any particular method. For example, monoclonal antibodies for use in accordance with the present disclosure may be produced by a variety of techniques, including, for example, hybridoma methods, recombinant DNA methods, and techniques for producing human or human-like antibodies in animals that contain some or all of the human immunoglobulin loci or genes encoding human immunoglobulin sequences.

[0074] The terms "full length antibody," "intact antibody," or "whole 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, a whole antibody includes one having heavy and light chains, 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, the intact antibody may have one or more effector functions.

[0075] "Antibody fragment" refers to a molecule other than an intact antibody that contains a portion of the 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.

[0076] Papain digestion of antibodies, such as the anti-MS4A4A antibody of the present disclosure, produces two identical antigen-binding fragments called "Fab" fragments and a residual "Fc" fragment, a name reflecting its ability to crystallize readily. The Fab fragment contains the entire light chain as well as the variable region domain of the heavy chain (V H ) and the first constant domain of one heavy chain (C H 1). Each Fab fragment is monovalent with respect to antigen binding; i.e., it has a single antigen-binding site. Pepsin treatment of an antibody yields a single large F(ab')2 fragment, which roughly corresponds to two disulfide-linked Fab fragments with different antigen-binding activities and are still capable of cross-linking antigen. Fab' fragments are C H F(ab')2 antibody fragments differ from Fab fragments by having several additional residues at the carboxy terminus of one domain, including one or more cysteines from the antibody hinge region. Fab'-SH is the designation used herein for Fab' in which the cysteine ​​residue(s) of the constant domains bear a free thiol group. F(ab')2 antibody fragments were originally produced as pairs of Fab' fragments that have hinge cysteines between them. Other chemical couplings of antibody fragments are also known.

[0077] The Fc fragment contains the carboxy-terminal portions of both heavy chains held together by disulfides. The effector functions of an antibody are determined by sequences in the Fc region, which is also recognized by Fc receptors (FcRs) found on certain cell types.

[0078] A "functional fragment" of an antibody, such as the anti-MS4A4A antibody of the present disclosure, comprises a portion of the intact antibody, generally comprising the antigen-binding or variable region of the intact antibody, or the Fc region of the antibody that retains or has altered FcR binding ability. Examples of antibody fragments include linear antibodies, single-chain antibody molecules, and multispecific antibodies formed from antibody fragments.

[0079] The term "diabody" refers to a group of V domains that are fragmented such that inter-chain, but not intra-chain, pairing of the variable domains is achieved, thereby resulting in a bivalent fragment, i.e., a fragment with two antigen-binding sites. H Domains and V L This refers to small antibody fragments prepared by constructing sFv fragments (see previous paragraph) with a short linker (approximately 5-10 residues) between the domains. Bispecific diabodies are small antibody fragments prepared by constructing sFv fragments (see previous paragraph) with a short linker (approximately 5-10 residues) between the domains. H Domain and V L It is a heterodimer of two "crossover" sFv fragments in which the domains are present on different polypeptide chains.

[0080] As used herein, "chimeric antibody" refers to antibodies (immunoglobulins), such as the chimeric anti-MS4A4A antibodies of the present disclosure, in which a portion of the heavy and / or light chain is identical to or homologous to corresponding sequences in antibodies from a particular species or belonging to a particular antibody class or subclass, and the remainder of the chain(s) is / are identical to or homologous to corresponding sequences in antibodies from another species or belonging to another antibody class or subclass, as well as fragments of such antibodies, so long as they exhibit the desired biological activity. Chimeric antibodies of interest herein include PRIMATIZED® antibodies, whose antigen-binding regions are derived from, for example, antibodies produced by immunizing macaque monkeys with the antigen of interest. As used herein, "humanized antibody" is used as a subset of "chimeric antibody."

[0081] "Humanized" forms of non-human (e.g., murine) antibodies, such as humanized anti-MS4A4A antibodies of the present disclosure, are chimeric antibodies comprising amino acid residues derived from non-human HVRs and human FRs. In certain embodiments, a humanized antibody comprises substantially all of at least one, and typically two, variable domains, with all or substantially all of each HVR (e.g., CDRs) corresponding to HVRs of a non-human antibody and all or substantially all of each FR corresponding to FRs of a human antibody. A humanized antibody may optionally 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, refers to an antibody that has undergone humanization.

[0082] A "human antibody" is an antibody, such as the anti-MS4A4A antibody of the present disclosure, that has an amino acid sequence corresponding to that of an antibody produced in a human and / or that has been produced using any of the techniques disclosed herein for generating human antibodies. This definition of a human antibody specifically excludes humanized antibodies that contain non-human antigen-binding residues. Human antibodies can be produced using a variety of techniques known in the art, including phage display libraries. Human antibodies have been modified to produce such antibodies in response to antigen challenge, and can be prepared, for example, by administering the antigen to immunized xeno-mouse and transgenic animals that have been generated via human B-cell hybridomas and have their endogenous loci disabled.

[0083] As used herein, the term "hypervariable region," "HVR," or "HV" refers to a region of an antibody variable domain, such as the region of an antibody variable domain of an anti-MS4A4A antibody of the present disclosure, that is hypervariable in sequence and / or forms structurally defined loops. Generally, antibodies have six HVRs; three are V H (H1, H2, H3), and three are in V L In native antibodies, H3 and L3 are the most diverse of the six HVRs, and H3 in particular is thought to play a unique role in conferring superior specificity to antibodies. Natural camelid antibodies, consisting only of heavy chains, are functional and stable even without light chains.

[0084] A number of HVR descriptions are used and included herein. In some embodiments, these HVRs may be Kabat complementarity determining regions (CDRs) based on sequence variability and are the most commonly used (Kabat et al. al, supra). In some embodiments, these HVRs may be Chothia CDRs. Chothia refers more to the location of structural loops (Chothia and Lesk J. Mol. Biol. 196:901-917 (1987)). In some embodiments, these HVRs may be AbM HVRs. These AbM HVRs represent a compromise between Kabat CDRs and Chothia structural loops and are used in Oxford Molecular's AbM antibody modeling software. In some embodiments, these HVRs may be "contact" HVRs. These "contact" HVRs are based on analysis of available complex crystal structures. The respective residues of these HVRs are shown below. [Table 1]

[0085] HVRs may include the following "extended HVRs": 24-36 or 24-34 (L1), 46-56 or 50-56 (L2), and 89-97 or 89-96 (L3) in the VL, and 26-35 (H1), 50-65 or 49-65 (preferred embodiment) (H2), and 93-102, 94-102, or 95-102 (H3) in the VH. The variable domain residues are numbered according to Kabat et al., supra, for each of these extended HVR definitions.

[0086] "Framework" or "FR" residues are those variable domain residues other than the HVR residues as herein defined.

[0087] As used herein, an "acceptor human framework" refers to a V-framework derived from a human immunoglobulin framework or a human consensus framework. L or V HA framework comprising the amino acid sequence of a framework. An acceptor human framework "derived" from a human immunoglobulin framework or a human consensus framework may contain the same amino acid sequence as the framework, or may contain 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, preferred amino acid changes occur at only three, two, or one of positions 71H, 73H, and 78H; for example, the amino acid residues at these positions may be 71A, 73T, and / or 78A. In some embodiments, a VL acceptor human framework comprises, in sequence, V L It is identical to a human immunoglobulin framework sequence or a human consensus framework sequence.

[0088] The "human consensus framework" is a V-type of human immunoglobulin L or V H A framework representing the most commonly occurring amino acid residues in a framework sequence selection. L or V H The framework sequences are selected from a subgroup of variable domain sequences. Typically, the subgroup of sequences is a subgroup such as that described in Kabat et al., Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, MD (1991). For example, the V L For V, the subgroup can be subgroup kappa I, kappa II, kappa III, or kappa IV, such as Kabat et al., supra. H With respect to, the subgroup can be subgroup I, subgroup II, or subgroup III, such as in Kabat et al., supra.

[0089] For example, an "amino acid modification" at a specified position, such as an amino acid modification of an anti-MS4A4A antibody of the present disclosure, 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" a specified residue means an insertion of up to one to two residues therefrom. The insertion may be made on the N-terminal or C-terminal side of the specified residue. A preferred amino acid modification herein is a substitution.

[0090] An "affinity matured" antibody, such as, for example, an affinity matured anti-MS4A4A antibody of the present disclosure, is an antibody in which one or more HVRs of the antibody contain one or more modifications such that the affinity of the antibody for antigen is improved compared to the parent antibody lacking that modification(s). In certain embodiments, the affinity matured antibody has nanomolar or even picomolar affinity for the target antigen. Affinity matured antibodies are produced by techniques known in the art. For example, Marks et al., Bio / Technology 10:779-783 (1992) describe V H - and V L Affinity maturation using domain shuffling has been described. 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).

[0091] An "Fv" is the minimum antibody fragment containing a complete antigen-recognition and antigen-binding site. This fragment consists of a dimer of one heavy-chain variable region domain and one light-chain variable region domain in tight, non-covalent association. The folding of these two domains results in six hypervariable loops (three loops each from the H and L chains) that provide the amino acid residues for antigen binding and confer antigen-binding specificity to the antibody. However, even a single variable domain (or half an Fv containing only three antigen-specific HVRs) has the ability to recognize and bind antigen, albeit with lower affinity than the entire binding site.

[0092] "Single-chain Fv," also abbreviated as "sFv" or "scFv," is a V-like molecule connected in the form of a single polypeptide chain. H and V L Preferably, the sFv polypeptide is an antibody fragment containing the V H and V L It further comprises a polypeptide linker between the domains which enables the sFv to form the desired structure for antigen binding.

[0093] Antibody "effector functions" refer to those biological activities involving the Fc region of an antibody (a native sequence Fc region or amino acid sequence variant Fc region), and vary depending on the antibody isotype.

[0094] As used herein, the term "Fc region" is used to define the C-terminal region of an immunoglobulin heavy chain, including native-sequence Fc regions and variant Fc regions. Although the boundaries of the Fc region of an immunoglobulin heavy chain vary, the human IgG heavy chain Fc region is usually defined as extending from amino acid residue Cys226 or Pro230 to the carboxyl terminus. The C-terminal lysine (residue 447 in the EU numbering system) of the Fc region can be removed, for example, during antibody production or purification, or by recombinantly engineering the nucleic acid encoding the antibody heavy chain. Thus, intact antibody compositions can include antibody populations in which all K447 residues have been removed, antibody populations in which K447 residues have not been removed, and antibody populations having a mixture of antibodies with and without the K447 residue. Native-sequence Fc regions suitable for use in the antibodies of the present disclosure include human IgG1, IgG2, IgG3, and IgG4.

[0095] A "native-sequence Fc region" comprises an amino acid sequence identical to that of an Fc region found in nature. Native-sequence human Fc regions include native-sequence human IgG1 Fc regions (non-A and A allotypes), native-sequence human IgG2 Fc regions, native-sequence human IgG3 Fc regions, and native-sequence human IgG4 Fc regions, as well as naturally occurring variants thereof.

[0096] A "variant Fc region" comprises an amino acid sequence that differs from that of a native-sequence Fc region by at least one amino acid modification, preferably one or more amino acid substitution(s). Preferably, the variant Fc region comprises at least one amino acid substitution compared to a native-sequence Fc region or the Fc region of a parent polypeptide, e.g., about one to about ten amino acid substitutions, preferably about one to about five amino acid substitutions, in the native-sequence Fc region or the Fc region of a parent polypeptide. A variant Fc region herein preferably has at least about 80% homology to a native-sequence Fc region and / or 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.

[0097] "Fc receptor" or "FcR" refers to a receptor that binds to the Fc region of an antibody. A preferred FcR is a native-sequence human FcR. Furthermore, a preferred FcR is an antibody that binds IgG antibodies (gamma receptors), and includes receptors of the FcγRI, FcγRII, and FcγRIII subclasses, including allelic variants and alternatively spliced ​​forms of these receptors. FcγRII receptors include FcγRIIA (an "activating receptor") and FcγRIIB (an "inhibiting receptor"), which have a similar amino acid sequence but differ primarily in their cytoplasmic domains. Activating receptor FcγRIIA contains an immunoreceptor tyrosine-based activation motif ("ITAM") in its cytoplasmic domain. Inhibiting receptor FcγRIIB contains an immunoreceptor tyrosine-based inhibitory motif ("ITIM") in its cytoplasmic domain. Other FcRs, including those yet to be identified, are encompassed by the term "FcR" herein. FcRs can also increase the serum half-life of antibodies.

[0098] As used herein, "percent (%) amino acid sequence identity" and "homology" with respect to peptide, polypeptide, or antibody sequences refer to the percentage of amino acid residues in a candidate sequence that are identical to those in a specified peptide or polypeptide sequence, after aligning the sequences and introducing gaps, if necessary, to achieve the maximum percent sequence identity, without considering any conservative substitutions as part of the sequence identity. Alignment to determine percent amino acid sequence identity can be accomplished using a variety of methods within the skill of the art, for example, publicly available computer software such as BLAST, BLAST-2, ALIGN, or MEGALIGN™ (DNASTAR) software. Those skilled in the art can determine appropriate parameters for assessing alignment, including any algorithms known in the art that are needed to achieve maximal alignment over the entire length of the sequences being compared.

[0099] The term "compete," as used in the context of competing antibodies (e.g., neutralizing antibodies) for the same epitope, refers to competition between antibodies as determined by an assay in which the antibody being tested prevents or inhibits (e.g., suppresses) specific binding of a reference molecule (e.g., a ligand or reference antibody) to a common antigen (e.g., MS4A4A or a fragment thereof). Numerous types of competitive binding assays can be used to determine whether an antibody competes with another antibody, such as; solid-phase direct or indirect radioimmunoassay (RIA), solid-phase direct or indirect enzyme immunoassay (EIA), sandwich competition assay (see, e.g., Stahl 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 label assay, solid-phase direct label sandwich assay (see, e.g., Harlow and Lane, 1988, Antibodies, A Laboratory Manual, Cold Spring Harbor Press); solid-phase direct label RIA using 1-125 label (see, e.g., Morel et al., 1988, Antibodies, A Laboratory Manual, Cold Spring Harbor Press); 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 assays involve the use of purified antigen bound to a solid surface or cells bearing either an unlabeled test antibody and a labeled reference antibody. Competitive inhibition is measured by determining the amount of label bound to the solid surface or cells in the presence of the test antibody. The test antibody is usually present in excess. Antibodies identified in competitive assays (competing antibodies) include those that bind to the same epitope as the reference antibody and those that bind to an adjacent epitope sufficiently close to the epitope bound by the reference antibody to cause steric hindrance.Further details regarding methods for determining competitive binding are provided in the Examples herein. Typically, when present in excess, a competing antibody inhibits (e.g., suppresses) specific binding of a reference antibody to a common antigen by at least 20%, 30%, 40%, 50%, 60%, 70%, 80%, 85%, 90%, 95%, 97.5%, and / or nearly 100%.

[0100] As used herein, an "interaction" between an MS4A4A polypeptide and a second polypeptide includes, but is not limited to, a protein-protein interaction, a physical interaction, a chemical interaction, a bond, a covalent bond, and an ionic bond. As used herein, an antibody "inhibits an interaction" between two polypeptides if the antibody disrupts, prevents, or completely eliminates the interaction between the two polypeptides. An antibody of the present disclosure "inhibits an interaction" between two polypeptides if the antibody binds to one of the two polypeptides. In some embodiments, the interaction can be inhibited by at least about 20%, 30%, 40%, 50%, 60%, 70%, 80%, 85%, 90%, 95%, 97.5%, and / or approximately 100%.

[0101] The term "epitope" refers to any determinant capable of binding by an antibody. An epitope is the region of an antigen to which an antibody directed against that antigen binds, and, if the antigen is a polypeptide, includes specific amino acids that make direct contact with the antibody. Most often, epitopes reside on polypeptides, but in some cases, they can reside on other types of molecules, such as nucleic acids. Epitope determinants include chemically active surface moieties on molecules, such as amino acids, sugar side chains, phosphoryl or sulfonyl groups, and can have specific three-dimensional structural characteristics and / or specific charge characteristics. Generally, antibodies specific for a particular target antigen preferentially recognize epitopes associated with the target antigen in complex mixtures of polypeptides and / or macromolecules.

[0102] An "agonist" or "activating" antibody is an antibody that induces (eg, enhances) one or more activities or functions of an antigen after the antibody binds to the antigen.

[0103] An "antagonist" or "blocking" or "inhibitory" antibody is an antibody that inhibits, inhibits, and / or eliminates (e.g., attenuates) antigen binding to one or more ligands and / or inhibits, inhibits, and / or eliminates (e.g., attenuates) one or more activities or functions of the antigen after the antibody binds to the antigen. In some embodiments, an antagonist, blocking, or inhibitory antibody substantially or completely inhibits antigen binding to one or more ligands and / or one or more activities or functions of the antigen.

[0104] An "isolated" antibody, such as an isolated anti-MS4A4A antibody of the present disclosure, is one that has been identified, separated, and / or recovered (e.g., naturally or recombinantly) from components of its production environment. Preferably, the isolated antibody is free from association with all other contaminating components from its production environment, such as those obtained from recombinantly transfected cells, which generally interfere with research, diagnostic, or therapeutic uses of the antibody and may include enzymes, hormones, and other proteinaceous or non-proteinaceous solutes. In preferred embodiments, the antibody is purified: (1) to greater than 95% by weight, and in some embodiments, greater than 99% by weight, of the antibody, e.g., as determined by the Lowry assay; (2) to a degree sufficient to obtain at least 15 residues of N-terminal or internal amino acid sequence using a spinning cup sequencer; or (3) to homogeneity by SDS-PAGE under non-reducing or reducing conditions using Coomassie blue or, preferably, silver staining. Isolated antibody includes the antibody in its original situ within recombinant T cells, since at least one component of the antibody's natural environment will not be present. Ordinarily, however, isolated polypeptide or antibody will be prepared by at least one purification step.

[0105] An "isolated" nucleic acid molecule encoding an antibody, such as the anti-MS4A4A antibody of the present disclosure, is a nucleic acid molecule that has been identified and separated from at least one contaminant nucleic acid molecule with which it is ordinarily associated in the environment in which it is produced. Preferably, the isolated nucleic acid is free from association with all components associated with its production environment. Isolated nucleic acid molecules encoding the polypeptides and antibodies of the present disclosure are present in a form that is different from the form or setting in which it is found in nature. Thus, isolated nucleic acid molecules are distinguished from nucleic acids encoding the polypeptides and antibodies of the present disclosure that exist naturally in cells.

[0106] As used herein, the term "vector" refers to a nucleic acid molecule capable of transporting another nucleic acid to which it is linked. A "plasmid" is a type of vector and refers to a circular double-stranded DNA into which additional DNA segments can be ligated. A phage vector is another type of vector. Another type of vector is a viral vector, into which additional DNA segments can be ligated. Certain vectors (e.g., bacterial vectors having a bacterial origin of replication and episomal mammalian vectors) are capable of autonomous replication in a host cell into which they are introduced. Other vectors (e.g., non-episomal mammalian vectors) are integrated into the genome of a host cell upon introduction into the host cell, and thereby are replicated along with the host genome. Moreover, 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." Generally, expression vectors utilized in recombinant DNA techniques are often in the form of plasmids. As used herein, "plasmid" and "vector" may be used interchangeably as the plasmid is the most commonly used form of vector.

[0107] The terms "polynucleotide" and "nucleic acid," as used interchangeably herein, refer to a polymer of nucleotides of any length, and include 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.

[0108] A "host cell" includes an individual cell or cell culture that can be or has been the recipient of a vector(s) for incorporating a polynucleotide insert. A host cell includes the progeny of a single host cell, and the progeny may not necessarily be completely identical (morphologically or in complementary strands of genomic DNA) to the original parent cell due to natural, accidental, or deliberate mutation. A host cell includes cells transfected in vivo with a polynucleotide(s) of the present disclosure.

[0109] As used herein, the terms "membrane TREM2," "plasma membrane TREM2," and "cell surface TREM2" are used interchangeably.

[0110] As used herein, a "carrier" includes a pharmaceutically acceptable carrier, excipient, or stabilizer that is non-toxic to cells or mammals exposed thereto at the dosages and concentrations employed.

[0111] As used herein, the term "preventing" includes providing protection against the occurrence or recurrence of a particular disease, disorder, or condition in an individual who is predisposed to, susceptible to, or at risk of developing a particular disease, disorder, or condition, but who has not yet been diagnosed with the disease, disorder, or condition.

[0112] As used herein, an individual "at risk" of developing a particular disease, disorder, or condition may or may not have detectable disease or disease symptoms, and may or may not exhibit detectable disease or disease symptoms prior to administering the treatments described herein. "At risk" indicates that an individual has one or more risk factors known in the art, which are measurable parameters that correlate with the development of a particular disease, disorder, or condition. Individuals who have one or more of these risk factors have a higher likelihood of developing a particular disease, disorder, or condition than individuals who do not have one or more of these risk factors.

[0113] As used herein, the term "treatment" refers to a clinical intervention designed to alter the natural course of a clinical pathology in the individual being treated. Desirable effects of treatment include slowing the rate of progression, improving or alleviating the pathological condition, and achieving remission or improving the prognosis of a particular disease, disorder, or condition. For example, an individual is successfully "treated" if one or more symptoms associated with a particular disease, disorder, or condition are alleviated or eliminated.

[0114] An "effective amount" refers to at least an amount effective, at dosages and for periods of time necessary, to achieve the desired therapeutic or prophylactic result. An effective amount can be provided in one or more administrations. The effective amount described herein can vary depending on factors such as the individual's condition, age, sex, and weight, as well as the ability of the treatment to elicit a desired response in the individual. An effective amount is also an amount in which any toxic or detrimental effects of the treatment are outweighed by the therapeutically beneficial effects. For prophylactic use, beneficial or desired results include elimination or reduction of the risk of, reduction in the severity of, or delay in the onset of, the biochemical, histological, and / or behavioral symptoms of the disease, its complications, and intermediate pathological phenotypes during the course of the disease. For therapeutic use, beneficial or desired results include suppression of one or more symptoms associated with the disease, improvement in the quality of life of those afflicted with the disease, reduction in the dosage of other drugs required to treat the disease, improvement in the effectiveness of other drugs, such as through targeting, delay in disease progression, and / or prolongation of survival. An effective amount of a drug, compound, or pharmaceutical composition is an amount sufficient to achieve prophylactic or therapeutic treatment either directly or indirectly. As understood in a clinical context, an effective amount of a drug, compound, or pharmaceutical composition may or may not be achieved in combination with another drug, compound, or pharmaceutical composition. Thus, an "effective amount" may be considered in the context of administering one or more therapeutic agents, and a single agent may be considered to be given in an effective amount if it can, or does, achieve the desired result in combination with one or more other agents.

[0115] An "individual" for purposes of treatment, prevention, or risk reduction refers to any animal classified as a mammal, including humans, domestic and farm animals, as well as zoo animals, sport or pet animals, such as dogs, horses, rabbits, cows, pigs, hamsters, gerbils, mice, ferrets, rats, cats, etc. In some embodiments, the individual is a human.

[0116] As used herein, administration "in combination with" another compound or composition includes simultaneous administration and / or administration at different times. Co-administration also includes administration as a combined preparation or as separate compositions, at different dosing frequencies or intervals, and using the same or different routes of administration. In some embodiments, co-administration is administration as part of the same therapeutic regimen.

[0117] As used herein, the term "about" refers to a normal error range for each numerical value, which would be readily understood by one of ordinary skill in the art. Reference herein to a value or parameter with "about" includes (and describes) embodiments directed to that numerical value or parameter itself.

[0118] As used in this specification and the appended claims, the singular forms "a," "an," and "the" include the plural unless the context clearly indicates otherwise. For example, reference to an "antibody" can refer to one antibody to molar quantities of many antibodies, and includes equivalents thereof known to those skilled in the art, and so forth.

[0119] It should be understood that the aspects and embodiments of the present disclosure described herein include "comprising," "consisting of," and "consisting essentially of" aspects and embodiments.

[0120] II. Anti-MS4A4A antibody Provided herein are anti-MS4A4A antibodies, which are useful, for example, for the diagnosis or treatment of MS4A4A-associated disorders.

[0121] In one aspect, the present disclosure provides an isolated (e.g., monoclonal) antibody that binds to an epitope of an MS4A4A protein of the present disclosure. The MS4A4A protein of the present disclosure includes, but is not limited to, a mammalian MS4A4A protein, a human MS4A4A protein, a mouse MS4A4A protein, and a cynomolgus monkey MS4A4A protein. The MS4A4A protein of the present disclosure includes naturally occurring variants of MS4A4A.

[0122] Human MS4A4A is a 239 amino acid protein that encodes a membrane glycoprotein. The amino acid sequence of human MS4A4A is shown in SEQ ID NO: 1: MHQTYSRHCRPEESTFSAAMTTMQGMEQAMPGAGPGVPQLGNMAVIHSHLWKGLQEKFLKGEPKVLGVVQILTALMSLSMGITMMCMASNTYGSNPISVYIGYTIWGSVMFIISGSSLSI AAGIRTTKGLVRGSLGMNITSSVLAASGILINTFSLAFYSFHHPYCNYYGNSNNCHGTMSILMGLDGMVLLLSVLEFCIAVSLSAFGCKVLCCTPGGVVLILPSHSHMAETASPTPLNEV

[0123] Additionally, the amino acid sequence of mouse MS4A4A is shown in SEQ ID NO:2: MLVIQGTEQSALEAGYGAQQNGQPLYVNSHSWKRMTEKFLKGEPKILGIVQIVIAIMNLSIGIMMIIATVSTGEIPPSSVYIGYPIWGSLMFIISGSFSIVAGRRTTKGLVRSSLGLN ITSSVFAFSGIVISSLSPGIYSFHVYYCTYRGSSEGCHMTLSILMGLDIVVVVLSVLEFCIGVSLSAFGCRVMCCNPGGVMIIMPSNPTKAETANPVTLQSGLMPPEHQERNVPENMH

[0124] Additionally, the amino acid sequence of cynomolgus monkey (cyno) MS4A4A is shown in SEQ ID NO:3: HQTYRRHCRPEESTFSAAMTTMQGMEQATPGAGPGVPQLGNMAVVHSHLWKGLQEKFLKGEPKVLGVVQILIALMSLSMGITMMCVAFSAYGHYPISVYIGYTIWGSVMFIISGSLSIA AGIRTTKGLVRGSLGMNITSSVLAVSAILINTISLTIYSFYHRYCNYYGNPNNCHGTVSILMGMDGMVLLLSVLEFCIAVSLSAFGCKAICCTPGGVVLIIPSNSHMAEAAPLTPLNEV

[0125] In some embodiments, MS4A4A is expressed in cells. In some embodiments, MS4A4A is expressed in bone marrow 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 microglial cells. 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 cell surface.

[0126] The MS4A4A protein of the present disclosure has several domains, including, but not limited to, a cytoplasmic domain (amino acid residues 1 to 64 of human MS4A4A; see SEQ ID NO: 1); a transmembrane domain (amino acid residues 65 to 85 of human MS4A4A); an extracellular domain (extracellular domain 1; ECL1) corresponding to amino acid residues 86 to 98 of human MS4A4A; a transmembrane domain (amino acid residues 99 to 119 of human MS4A4A); a cytoplasmic domain (amino acid residues 120 to 137 of human MS4A4A); a transmembrane domain (amino acid residues 138 to 158 of human MS4A4A); an extracellular domain (extracellular domain 2; ECL2) corresponding to amino acid residues 159 to 179; a transmembrane domain (amino acid residues 180 to 200 of human MS4A4A); and a cytoplasmic domain (amino acid residues 201 to 239 of human MS4A4A). Additionally, the MS4A4A protein of the present disclosure is expressed in numerous tissues and cells, including, but not limited to, the brain, neurons, glial cells, endothelial cells, perivascular cells, and pericytes.

[0127] MS4A4A in macrophage and microglial cell function Macrophages and myeloid cells of the central nervous system (CNS), such as microglia, possess inherently diverse phenotypes and functions. Macrophages can be divided into M1 and M2 macrophages in vitro, which differ in their phagocytic and potentially inflammatory phenotypes and activities. For example, in peripheral organs, macrophages with the M1 phenotype are thought to have a pro-inflammatory and antibacterial phenotype and function, while macrophages with the M2 phenotype have an anti-inflammatory phenotype and function and are thought to be in a homeostatic state.

[0128] Healthy, homeostatic microglia express more M2 markers (e.g., CD200R, CD163, and CD115) on their cell surface compared with M1 markers (e.g., CD16, MHC class II, and CD86) (Ginhoux and Prinz, 2015, Cold Spring Harb Perspect Biol, 7:a020537). However, disease-associated microglia (DAMs) in both mouse models of Alzheimer's disease and in humans with Alzheimer's disease are in an inflammatory or activated state. Inflammatory or activated disease-associated microglia are thought to play a positive role in suppressing pathology associated with Alzheimer's disease and other neurodegenerative disorders and are therefore beneficial.

[0129] Expression of MS4A4A is elevated in M2 macrophages in vitro, suggesting that MS4A4A is a novel cell surface marker for M2 macrophages. Additionally, MS4A4A has been shown to regulate the cell surface trafficking of cKit in mast cells, suggesting a role for MS4A4A in regulating mast cell degranulation and survival (Cruse et al., 2015, Molecular Biol Cell, 26:1711-1727). Collectively, these findings suggest that targeting MS4A4A may affect the recycling, expression, and / or degradation of various macrophage cell surface receptors associated with M1 and M2 macrophage phenotypes, thereby potentially affecting their function and activity.

[0130] The anti-MS4A4A antibodies of the present disclosure affect the expression of M2 macrophage cell surface markers. In particular, the anti-MS4A4A antibodies of the present disclosure inhibit the cell surface expression of CD200R, Dectin-1, and CD163 on macrophages, suggesting that the anti-MS4A4A antibodies regulate macrophage polarization, function, and / or activity by inhibiting the expression of M2 macrophage cell surface receptors. The anti-MS4A4A antibodies of the present disclosure inhibit M2 macrophage cell surface receptors, suggesting that the anti-MS4A4A antibodies of the present disclosure are effective in altering the physiological state of microglial cells, for example, toward a more protective phenotype (e.g., toward an increased M1 phenotype) with increased inflammatory or activation. Thus, the anti-MS4A4A antibodies of the present disclosure are useful in the treatment of Alzheimer's disease and other neurodegenerative disorders, in part by altering the phenotype of macrophages and microglia to a pro-inflammatory and activated state.

[0131] The anti-MS4A4A antibodies of the present disclosure affect the activation state of microglia. In vivo, the anti-MS4A4A antibodies increased the 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-trisphosphate receptor 2 (ITPR2)), and microglial proliferation (e.g., antigen KI-67 (MKI67)). Additionally, the anti-MS4A4A antibodies of the present disclosure increased the mRNA levels of microglial activation markers IL1RN, SPP1, and PLCG2. Furthermore, the anti-MS4A4A antibodies of the present disclosure decreased the mRNA levels of the homeostatic microglial markers purinergic receptor P2RY12 and CX3C chemokine receptor 1 (CX3CR1). Thus, the anti-MS4A4A antibodies of the present disclosure are effective in activating microglia in vivo, as evidenced by increasing various mRNA levels of proteins associated with microglial activation, migration, and proliferation; and decreasing various mRNA levels of proteins associated with microglial homeostasis.

[0132] Expression of MS4A4A and TREM2 Neurodegenerative diseases are characterized, in part, by defective immune function in the central nervous system (CNS). For example, reduced viability and function of some CNS myeloid cells, including but not limited to microglia, is thought to contribute to susceptibility to neurodegenerative diseases such as Alzheimer's disease. Pharmacological interventions that enhance myeloid cell viability and / or function may provide effective treatments for ameliorating the onset, severity, or progression of such neurodegenerative diseases and disorders.

[0133] Triggering receptor expressed on myeloid cells 2 (TREM2) is an immunoglobulin-like receptor (IGR) predominantly expressed in myeloid cells, including macrophages, dendritic cells, monocytes, dermal Langerhans cells, Kupffer cells, osteoclasts, and microglia. TREM2 is highly expressed in CNS microglia and infiltrating macrophages during experimental autoimmune encephalomyelitis and Alzheimer's disease (Piccio et al., 2007, Eur J Immunol, 37:1290-1301; Wang, 2015, Cell, 160:1061-1071). The TREM2 pathway is thought to be a key regulator of CNS myeloid cell viability and function.

[0134] Data from human genetic studies suggest a strong genetic association between MS4A4A and TREM2 and susceptibility to Alzheimer's disease (Piccio et al., 2016, Acta Neuropathol, 131:925-9330). Notably, the protective MS4A4A allele for Alzheimer's disease is associated with increased levels of sTREM2 in the cerebrospinal fluid of patients.

[0135] The anti-MS4A4A antibodies of the present invention increased cellular ATP levels in macrophages, indicating that they are effective in increasing, maintaining, or enhancing the viability and function of cells (e.g., macrophages, myeloid cells, and microglia). In addition, the anti-MS4A4A antibodies of the present invention increased sTREM2 and mTREM2 levels in macrophages, in contrast to a previous report that reported that the commercially available anti-MS4A4A antibody 5C12 suppressed sTREM2 levels in the supernatant of cultured human macrophages (Deming et al., 2018, bioRxiv, doi:dxdoiorg / 10.1101 / 352179). Because the MS4A4A protective allele for Alzheimer's disease is associated with increased levels of sTREM2, the results provided herein demonstrate that the anti-MS4A4A antibodies of the present disclosure mimic or replicate the protective phenotype in neurodegenerative diseases and disorders, such as Alzheimer's disease, following increased levels of sTREM2 and mTREM2. 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%.

[0136] In some embodiments, 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, 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 yet other embodiments, 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, the anti-MS4A4A antibodies of the present disclosure increase ATP levels in myeloid cells (e.g., macrophages, human macrophages, microglia) by about 1.2-fold, about 1.4-fold, or about 1.7-fold compared to the levels of ATP in those cells in the absence of the anti-MS4A4A antibody.

[0137] In some embodiments, anti-MS4A4A antibodies of the present disclosure increase soluble TREM2 levels in vivo (e.g., in non-human primates or humans). In some embodiments, 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% compared to baseline soluble TREM2 levels in serum in vivo before administration of an anti-MS4A4A antibody of the present disclosure. In some embodiments, anti-MS4A4A antibodies of the present disclosure increase soluble TREM2 levels in serum in vivo by about 50% compared to baseline soluble TREM2 levels in serum in vivo before administration of an anti-MS4A4A antibody of the present disclosure.

[0138] In some embodiments, an anti-MS4A4A antibody of the present disclosure increases in vivo serum soluble TREM2 levels 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 compared to the soluble TREM2 levels in vivo in serum before administration of an anti-MS4A4A antibody of the present disclosure. In some embodiments, an anti-MS4A4A antibody of the present disclosure increases in vivo serum soluble TREM2 levels by about 1.5-fold compared to the soluble TREM2 levels in vivo in serum before administration of an anti-MS4A4A antibody of the present disclosure.

[0139] In some embodiments, an anti-MS4A4A antibody of the present disclosure increases 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 soluble TREM2 levels in serum in vivo before administration of an anti-MS4A4A antibody of the present disclosure. In some embodiments, an anti-MS4A4A antibody of the present disclosure increases soluble TREM2 levels in serum in vivo for at least 20 days compared to the soluble TREM2 levels in serum in vivo before administration of an anti-MS4A4A antibody of the present disclosure. In some embodiments, an anti-MS4A4A antibody of the present disclosure elevates 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, 432 hours, at least 456 hours, or at least 480 hours compared to soluble TREM2 levels in serum in vivo before administration of an anti-MS4A4A antibody of the present disclosure. In some embodiments, an anti-MS4A4A antibody of the present disclosure elevates soluble TREM2 levels in serum in vivo for at least 480 hours compared to soluble TREM2 levels in serum in vivo before administration of an anti-MS4A4A antibody of the present disclosure.

[0140] In some embodiments, an anti-MS4A4A antibody of the present disclosure increases soluble TREM2 levels in the 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 the CSF in vivo before administering an anti-MS4A4A antibody of the present disclosure. In some embodiments, an anti-MS4A4A antibody of the present disclosure increases soluble TREM2 levels in the CSF in vivo by about 300% from baseline soluble TREM2 levels in the CSF in vivo before administering an anti-MS4A4A antibody of the present disclosure.

[0141] In some embodiments, an anti-MS4A4A antibody of the present disclosure increases soluble TREM2 levels in the 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 compared to the soluble TREM2 levels in the CSF in vivo before administration of an anti-MS4A4A antibody of the present disclosure. In some embodiments, an anti-MS4A4A antibody of the present disclosure increases soluble TREM2 levels in the CSF in vivo by about 2-fold to about 4-fold compared to the soluble TREM2 levels in the CSF in vivo before administration of an anti-MS4A4A antibody of the present disclosure.

[0142] In some embodiments, an anti-MS4A4A antibody of the present disclosure elevates soluble TREM2 levels in the CSF in vivo for at least 1 day, at least 2 days, at least 3 days, or at least 4 days compared to the soluble TREM2 levels in the CSF in vivo before administering an anti-MS4A4A antibody of the present disclosure. In some embodiments, an anti-MS4A4A antibody of the present disclosure elevates soluble TREM2 levels in the CSF in vivo for at least 4 days compared to the soluble TREM2 levels in the CSF in vivo before administering an anti-MS4A4A antibody of the present disclosure. In some embodiments, an anti-MS4A4A antibody of the present disclosure elevates soluble TREM2 levels in the CSF in vivo for at least 24 hours, at least 48 hours, at least 72 hours, or at least 96 hours compared to the soluble TREM2 levels in the CSF in vivo before administering an anti-MS4A4A antibody of the present disclosure. In some embodiments, an anti-MS4A4A antibody of the present disclosure increases soluble TREM2 levels in the CSF in vivo for at least 96 hours compared to soluble TREM2 levels in the CSF in vivo before administration of an anti-MS4A4A antibody of the present disclosure.

[0143] 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.

[0144] Antagonist antibodies In some embodiments, antibodies that bind to MS4A4A protein may be antagonistic antibodies that bind to MS4A4A and inhibit one or more MS4A4A activities by preventing the interaction between MS4A4A and its ligand(s) or by preventing MS4A4A from transmitting signals to the cytoplasm in the presence of a ligand. In some embodiments, antagonistic antibodies of the present disclosure may have the epitope specificity of agonistic antibodies of the present disclosure, but have an Fc domain that is incapable of binding to Fcg receptors and therefore, for example, incapable of clustering MS4A4A receptors.

[0145] In some embodiments, the antibodies of the present disclosure are antagonistic antibodies. In some embodiments, the antagonistic antibodies inhibit one or more MS4A4A activities. In some embodiments, the antagonistic antibodies suppress the activity of one or more MS4A4A-dependent genes. In some embodiments, the antagonistic antibodies inhibit the interaction between MS4A4A and one or more MS4A4A ligands. In some embodiments, the antagonistic antibodies inhibit MS4A4A signaling. In some embodiments, the antagonistic antibodies inhibit the interaction between MS4A4A and one or more MS4A4A ligands and inhibit MS4A4A signaling.

[0146] In some embodiments, downregulation of MS4A4A protein levels or inhibition of MS4A4A activity is achieved with an anti-MS4A4A antibody that downregulates or inhibits MS4A4A protein levels in cells. In some embodiments, downregulation of MS4A4A protein levels or inhibition of MS4A4A activity is achieved by downregulating MS4A4A nucleic acid expression or levels using methods known and available to those of skill in the art, e.g., using antisense methodology, gene therapy, etc. Thus, in some embodiments, inhibition of MS4A4A protein levels or activity is achieved using an anti-MS4A4A antibody of the present disclosure or by reducing the expression or level of MS4A4A nucleic acid (e.g., mRNA).

[0147] In some embodiments, antibody cross-linking is required for agonist antibody function. Antibody cross-linking can be achieved through binding to a secondary antibody in vitro or through binding to an Fc receptor in vivo. For example, antagonist antibodies can be converted into agonist antibodies through in vitro biotin / streptavidin cross-linking or secondary antibody binding (see, e.g., Gravestein et al., 1996, J. Exp. Med. 184:675-685; Gravestein et al., 1994, International Immunol. 7:551-557). Agonist antibodies may exert their activity by mimicking the biological activity of receptor ligands or enhancing receptor aggregation, thereby activating receptor signaling. In some embodiments, the absence of antibody cross-linking is required for antagonist activity. Antagonist antibodies may exert their activity by blocking receptor-ligand interactions.

[0148] Phenocopying of gelsolin, osteopontin, and Alzheimer's disease protective alleles There are three SNPs in the MS4A gene cluster that are associated with an increased risk of late-onset Alzheimer's disease. These 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, MS4A4A locus SNPs (rs2304933 and rs2304935) are associated with increased levels of MS4A4A and an increased risk of Alzheimer's disease, including late-onset Alzheimer's disease (LOAD) (Allen et al., 2012, Neurology, 79:221-228).

[0149] Genetic variants (SNPs) associated with Alzheimer's disease (AD) have been identified in the MS4A gene cluster. One of these variant alleles, rs1582763, is considered a protective allele because it is associated with increased CSF sTREM2 levels, a reduced risk of AD, and a delayed age of onset (Deming et al., 2018, bioRxiv, doi:dx doi org / 10.1101 / 352179). This rs1582763 protective allele is associated with reduced circulating MS4A4A mRNA levels. Furthermore, these findings suggest that the rs1582763 allele plays a protective role by reducing MS4A4A levels and thus the risk or severity of AD. This protective rs1582763 is also associated with increased osteopontin expression and increased gelsolin levels. The anti-MS4A4A antibodies of the present invention are effective in phenocopying protective alleles with respect to at least suppressing MS4A4A expression, increasing osteopontin expression, increasing gelsolin expression, and / or increasing sTREM levels. Some embodiments of the present disclosure provide anti-MS4A4A antibodies that phenocopy one or more alleles of MS4A associated with a reduced risk of Alzheimer's disease and / or a delayed age of onset of Alzheimer's disease.

[0150] The anti-MS4A4A antibodies of the present disclosure increased osteopontin (SPP1) mRNA levels and gelsolin (GSN) mRNA levels in human peripheral blood mononuclear cell-derived macrophages. In some embodiments, the anti-MS4A4A antibodies of the present disclosure increase osteopontin mRNA and / or protein levels. In some embodiments, the anti-MS4A4A antibodies of the present disclosure increase gelsolin mRNA and / or protein levels. In some embodiments, the anti-MS4A4A antibodies of the present disclosure increase osteopontin mRNA and / or protein levels in bone marrow 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%.

[0151] Anti-MS4A4A antibodies of the present disclosure increased osteopontin (SSP1) levels in non-human primates. In some embodiments, 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, anti-MS4A4A antibodies of the present disclosure increase osteopontin mRNA and / or protein levels in serum and / or CSF. In some embodiments, 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, anti-MS4A4A antibodies of the present disclosure increase osteopontin protein levels in the brain, hi some embodiments, anti-MS4A4A antibodies of the present disclosure increase osteopontin protein levels in the frontal lobe and / or hippocampus.

[0152] In some embodiments, the anti-MS4A4A antibodies of the present disclosure phenocopy the protective rs1582763 allele with respect to increased expression of osteopontin and gelsolin. In other aspects, the anti-MS4A4A antibodies of the present disclosure are effective in increasing expression of osteopontin, gelsolin, and / or sTREM2, and, like the protective rs1582763 allele, are biologically active in reducing the risk and / or severity of Alzheimer's disease. This data indicated that SPP1 and GSN are pharmacodynamic markers of the protective biological activity associated with the rs158273 allele.

[0153] CSFR1 and IL1RN expression Anti-MS4A4A antibodies of the present disclosure increased CSF1R levels in non-human primates. In some embodiments, 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, anti-MS4A4A antibodies of the present disclosure increase CSF1R mRNA and / or protein levels in serum and / or CSF. In some embodiments, 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, anti-MS4A4A antibodies of the present disclosure increase CSF1R protein levels in the brain, hi some embodiments, anti-MS4A4A antibodies of the present disclosure increase CSF1R protein levels in the frontal lobe and / or hippocampus.

[0154] CSF1R deficiency adversely 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 identified an association between mutations in the CSF1R gene and various disorders, such as adult-onset leukoencephalopathy with axonal spheroids and pigmented glia (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 antibody of the present disclosure inhibited cell death and sustained survival of human macrophages after CSF1R inhibition. Thus, in some embodiments, the anti-MS4A4A antibodies of the present disclosure are useful in treating individuals with a CSF1R-deficient disease or disorder, such as ALSP or HDLS.

[0155] Anti-MS4A4A antibodies of the present disclosure increased IL1RN levels in human peripheral blood mononuclear cell-derived macrophages. In some embodiments, anti-MS4A4A antibodies of the present disclosure increase IL1RN mRNA and / or protein levels. In some embodiments, anti-MS4A4A antibodies of the present disclosure increase IL1RN mRNA and / or protein levels. In some embodiments, 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%.

[0156] GPNMB expression GPNMB (glycoprotein nonmetastatic melanoma protein B) is a surface glycoprotein expressed in multiple cell types, including tissue macrophages and microglia. Several genetic variants are associated with the risk of Parkinson's disease (PD). GPNMB protein levels are elevated in the substantia nigra of PD patients, and GPNMB levels are elevated after lysosomal stress (Moloney et al., 2018, Neurobio Dis. 120:1-11). Additionally, increased GPNMB expression is associated with SNP rs199347, a risk SNP located within the GPNMB gene (Murthy et al., 2018). et al, Neurogenetics, 2017, 18:121-133).

[0157] The anti-MS4A4A antibodies of the present disclosure reduced the level of GPNMB cell surface protein in human primary macrophages. In some embodiments, the anti-MS4A4A antibodies of the present disclosure reduce the level of GPNMB cell surface protein in bone marrow 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, reducing GPNMB after adding an anti-MS4A4A antibody may provide a means of treating PD.

[0158] A. Exemplary Antibodies and Certain Other Antibody Embodiments In some embodiments, provided herein are: (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 sequence having at least about 95% identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 94, 108, 116, 146, 147, 308, and 311; (b) an HVR-H1 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. (c) an HVR-H2 comprising an amino acid sequence having at least about 95% identity to 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; (d) 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, or SEQ ID NOs: 100, 101, 102, 112, 123 , 124, 125, 126, 127, 128, 129, 154, 310, and 313; (d) 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, or 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. (e) an HVR-L1 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 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 sequence selected from the group consisting of SEQ ID NOs: 105, 106, 114, 139, 140, 141, 142, 143, 160, 161, 315, and 318;and (f) an 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 sequence having at least about 95% identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 107, 115, 144, 145, 163, 316, and 319;

[0159] In some embodiments, provided herein are: (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 sequence having at least about 95% identity 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 sequence having at least about 95% identity to an amino acid sequence 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 sequence having at least about 95% identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 100, 101, 102, and 310; (d) an HVR-H3 comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 103, and (f) an HVR-L3 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 107 and 316 or an amino acid sequence selected from the group consisting of SEQ ID NOs: 107 and 316.

[0160] In some embodiments, provided herein are: (a) an HVR-H1 comprising the amino acid sequence of SEQ ID NO: 108, or amino acids having at least about 95% homology to the amino acids 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 amino acids having at least about 95% homology to the amino acids of SEQ ID NO: 110 and 111; (c) an HVR-H3 comprising the amino acid sequence of SEQ ID NO: 112, or amino acids having at least about 95% homology to the amino acids of SEQ ID NO: 112; (d) an HVR-H4 comprising the amino acid sequence of SEQ ID NO: 112; Provided are anti-MS4A4A antibodies comprising at least one, two, three, four, five, or six HVRs selected from: (a) HVR-L1 comprising the amino acid sequence of SEQ ID NO: 113 or amino acids that are at least about 95% identical to the amino acid sequence of SEQ ID NO: 113; (b) HVR-L2 comprising the amino acid sequence of SEQ ID NO: 114 or amino acids that are at least about 95% identical 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 amino acids that are at least about 95% identical to the amino acid sequence of SEQ ID NO: 115.

[0161] In some embodiments, provided herein are: (a) an HVR-H1 comprising the amino acid sequence of SEQ ID NO: 116, or an amino acid sequence having at least about 95% identity to an amino acid sequence of SEQ ID NO: 116; (b) an amino acid sequence selected from the group consisting of SEQ ID NOs: 118, 119, 120, 121, and 122, or an amino acid sequence having at least about 95% identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 118, 119, 120, 121, and 122. (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 amino acids having at least about 95% identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 123, 124, 125, 126, 127, 128, and 129; (d) HVR-H4 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 130, 131, 132, 133, 134, 135, 136, 137, and 138; and (f) an HVR-L3 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 144 and 145, or an amino acid sequence that is at least about 95% identical to an amino acid sequence selected from the group consisting of SEQ ID NOs: 144 and 145.

[0162] In some embodiments, provided herein are: (a) an HVR-H1 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 146 and 147, or amino acids with at least about 95% homology to an amino acid sequence 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 amino acids with 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; (c) an HVR-H3 comprising an amino acid sequence of SEQ ID NO: 154, or amino acids with at least about 95% homology to an amino acid sequence of SEQ ID NO: 154; and (d) an HVR-H4 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 156, 157, and 158. and (f) an HVR-L3 comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 163 or amino acids that are at least about 95% identical to amino acids selected from the group consisting of SEQ ID NO: 163.

[0163] In some embodiments, provided herein are: (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; (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: 97; (c) HVR-L3 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: 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: (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: 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: 101; (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: 106; and (d) 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) an HVR-L3 comprising the amino acid sequence of SEQ ID NO: 107;

[0164] 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) 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.

[0165] In some embodiments, the present specification provides 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) 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.

[0166] In some embodiments, provided herein are: (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; (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: 119; (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: 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 -H3; (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: 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; (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; (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: 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; (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: 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; (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: 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: 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: 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: 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: 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: 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; (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) 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) the sequence (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-L3 comprising the amino acid sequence of SEQ ID NO: 134. (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. Provided are anti-MS4A4A antibodies comprising: (a) 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: 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.

[0167] In some embodiments, provided herein are: (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 -H3; (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;An anti-MS4A4A antibody is provided, comprising: (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: 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.

[0168] In some embodiments, provided herein is a HVR-H1 comprising at least one, at least two, or all three Vs selected from: (a) an HVR-H1 comprising the amino acid sequence of SEQ ID NO: 94, or an amino acid sequence having at least about 95% identity to the amino acid sequence of SEQ ID NO: 94; (b) an 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 having at least about 95% identity to the amino acid sequence selected from the group consisting of SEQ ID NOs: 96, 97, 98, and 99; and (c) an 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 having at least about 95% identity to the amino acid sequence selected from the group consisting of SEQ ID NOs: 100, 101, and 102. H Anti-MS4A4A antibodies containing HVR sequences are provided.

[0169] In some embodiments, provided herein are at least one, at least two, or all three VHs selected from: (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 308, or an amino acid sequence having at least about 95% identity 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 having at least about 95% identity 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 having at least about 95% identity to the amino acid sequence of SEQ ID NO: 310. H Anti-MS4A4A antibodies containing HVR sequences are provided.

[0170] In some embodiments, provided herein are at least one, at least two, or all three VHs selected from: (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 311, or an amino acid sequence having at least about 95% identity 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 having at least about 95% identity 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 having at least about 95% identity to the amino acid sequence of SEQ ID NO: 313. H Anti-MS4A4A antibodies containing HVR sequences are provided.

[0171] In some embodiments, provided herein is a HVR-L1 comprising at least one, at least two, or all three Vs selected from: (a) an 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% identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 103 and 104; (b) an 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% identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 105 and 106; and (c) an HVR-L3 comprising the amino acid sequence of SEQ ID NO: 107, or an amino acid sequence having at least about 95% identity to the amino acid sequence of SEQ ID NO: 107. L Anti-MS4A4A antibodies containing HVR sequences are provided.

[0172] In some embodiments, provided herein are at least one, at least two, or all three Vs selected from: (a) HVR-L1 comprising the amino acid sequence of SEQ ID NO: 314, or an amino acid sequence having at least about 95% identity to the amino acid sequence of SEQ ID NO: 314; (b) HVR-L2 comprising the amino acid sequence of SEQ ID NO: 315, or an amino acid sequence having at least about 95% identity to the amino acid sequence of SEQ ID NO: 315; and (c) HVR-L3 comprising the amino acid sequence of SEQ ID NO: 316, or an amino acid sequence having at least about 95% identity to the amino acid sequence of SEQ ID NO: 316. LAnti-MS4A4A antibodies containing HVR sequences are provided.

[0173] In some embodiments, provided herein are at least one, at least two, or all three Vs selected from: (a) HVR-L1 comprising the amino acid sequence of SEQ ID NO: 317, or an amino acid sequence having at least about 95% identity to the amino acid sequence of SEQ ID NO: 317; (b) HVR-L2 comprising the amino acid sequence of SEQ ID NO: 318, or an amino acid sequence having at least about 95% identity to the amino acid sequence of SEQ ID NO: 318; and (c) HVR-L3 comprising the amino acid sequence of SEQ ID NO: 319, or an amino acid sequence having at least about 95% identity to the amino acid sequence of SEQ ID NO: 319. L Anti-MS4A4A antibodies containing HVR sequences are provided.

[0174] In some embodiments, provided herein are at least one, at least two, or all three Vs selected from: (a) (i) an HVR-H1 comprising an amino acid sequence of SEQ ID NO: 94, or an amino acid sequence having at least about 95% identity to an amino acid sequence of SEQ ID NO: 94; (ii) an 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 having at least about 95% identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 96, 97, 98, and 99; and (iii) an 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 having at least about 95% identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 100, 101, and 102. H V containing HVR sequences Hand (b) at least one, at least two, or all three V domains selected from: (i) an 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% identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 103 and 104; (ii) an 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% identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 105 and 106; and (iii) an HVR-L3 comprising the amino acid sequence of SEQ ID NO: 107, or an amino acid sequence having at least about 95% identity to the amino acid sequence of SEQ ID NO: 107. L V containing HVR sequences L domain.

[0175] In some embodiments, provided herein are at least one, at least two, or all three VHs selected from: (a) (i) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 308, or an amino acid sequence having at least about 95% identity 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 having at least about 95% identity 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 having at least about 95% identity to the amino acid sequence of SEQ ID NO: 310. H V containing HVR sequences H domain, and (b) at least one, at least two, or all three V selected from: (i) an HVR-L1 comprising the amino acid sequence of SEQ ID NO: 314, or an amino acid sequence having at least about 95% identity to the amino acid sequence of SEQ ID NO: 314; (ii) an HVR-L2 comprising the amino acid sequence of SEQ ID NO: 315, or an amino acid sequence having at least about 95% identity to the amino acid sequence of SEQ ID NO: 315; and (iii) an HVR-L3 comprising the amino acid sequence of SEQ ID NO: 316, or an amino acid sequence having at least about 95% identity to the amino acid sequence of SEQ ID NO: 316. L V containing HVR sequences Ldomain.

[0176] In some embodiments, provided herein are at least one, at least two, or all three VHs selected from: (a) (i) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 311, or an amino acid sequence having at least about 95% identity 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 having at least about 95% identity 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 having at least about 95% identity to the amino acid sequence of SEQ ID NO: 313. H V containing HVR sequences H domain, and (b) at least one, at least two, or all three V selected from: (i) an HVR-L1 comprising the amino acid sequence of SEQ ID NO: 317, or an amino acid sequence having at least about 95% identity to the amino acid sequence of SEQ ID NO: 317; (ii) an HVR-L2 comprising the amino acid sequence of SEQ ID NO: 318, or an amino acid sequence having at least about 95% identity to the amino acid sequence of SEQ ID NO: 318; and (iii) an HVR-L3 comprising the amino acid sequence of SEQ ID NO: 316, or an amino acid sequence having at least about 95% identity to the amino acid sequence of SEQ ID NO: 316. L V containing HVR sequences L domain.

[0177] In some embodiments, provided herein are at least one, at least two, or all three Vs selected from: (a) an 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% identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 24, 25, 26, 27, 28, 29, and 30; (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 sequence having at least about 95% identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 110 and 111; and (c) an HVR-H3 comprising the amino acid sequence of SEQ ID NO: 112, or an amino acid sequence having at least about 95% identity to the amino acid sequence of SEQ ID NO: 112. H Anti-MS4A4A antibodies containing HVR sequences are provided.

[0178] In some embodiments, provided herein are at least one, at least two, or all three Vs selected from: (a) HVR-L1 comprising the amino acid sequence of SEQ ID NO: 113, or an amino acid sequence having at least about 95% identity 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 having at least about 95% identity 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 having at least about 95% identity to the amino acid sequence of SEQ ID NO: 115. L Anti-MS4A4A antibodies containing HVR sequences are provided.

[0179] In some embodiments, provided herein are at least one, at least two, or all three VHs selected from: (a) (i) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 108, or an amino acid sequence having at least about 95% identity 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 having at least about 95% identity to 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, or an amino acid sequence having at least about 95% identity to the amino acid sequence of SEQ ID NO: 112. H V containing HVR sequences H domain, and (b) at least one, at least two, or all three V selected from: (i) an HVR-L1 comprising the amino acid sequence of SEQ ID NO: 113, or an amino acid sequence having at least about 95% identity to the amino acid sequence of SEQ ID NO: 113; (ii) an HVR-L2 comprising the amino acid sequence of SEQ ID NO: 114, or an amino acid sequence having at least about 95% identity to the amino acid sequence of SEQ ID NO: 114; and (iii) an HVR-L3 comprising the amino acid sequence of SEQ ID NO: 115, or an amino acid sequence having at least about 95% identity to the amino acid sequence of SEQ ID NO: 115. L V containing HVR sequences L domain.

[0180] In some embodiments, provided herein are at least one, at least two, or all three Vs selected from: (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 116, or an amino acid sequence having at least about 95% identity to 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 NOs: 118, 119, 120, 121, and 122, or an amino acid sequence having at least about 95% identity 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 having at least about 95% identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 123, 124, 125, 126, 127, 128, and 129. H Anti-MS4A4A antibodies containing HVR sequences are provided.

[0181] In some embodiments, provided herein is an HVR-L1 comprising: (a) 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) an amino acid sequence selected from the group consisting of SEQ ID NOs: 139, 140, 141, 142, and 143. and (c) 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 sequence having at least about 95% identity to 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% identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 144 and 145. L Anti-MS4A4A antibodies containing HVR sequences are provided.

[0182] In some embodiments, provided herein are at least one, at least two, or all three Vs selected from: (a) (i) an HVR-H1 comprising an amino acid sequence of SEQ ID NO: 116, or an amino acid sequence having at least about 95% identity to the amino acid sequence of SEQ ID NO: 116; (ii) an 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 having at least about 95% identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 118, 119, 120, 121, and 122; and (iii) an 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 having at least about 95% identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 123, 124, 125, 126, 127, 128, and 129. H V containing HVR sequences H domain, and (b) (i) 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 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) 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% identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 139, 140, 141, 142, and 143; and (iii) an 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% identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 144 and 145. L V containing HVR sequences L domain.

[0183] In some embodiments, provided herein are at least one, at least two, or all three VHs selected from: (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 sequence having at least about 95% identity to an amino acid sequence 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 sequence having at least about 95% identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 149, 150, 151, 152, and 153; and (c) an HVR-H3 comprising the amino acid sequence of SEQ ID NO: 154, or an amino acid sequence having at least about 95% identity to the amino acid sequence of SEQ ID NO: 154. H Anti-MS4A4A antibodies containing HVR sequences are provided.

[0184] In some embodiments, provided herein are at least one, at least two, or all three Vs selected from: (a) an 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% identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 156, 157, and 158; (b) an 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% identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 160 and 161; and (c) an HVR-L3 comprising the amino acid sequence of SEQ ID NO: 163, or an amino acid sequence having at least about 95% identity to the amino acid sequence of SEQ ID NO: 163. L Anti-MS4A4A antibodies containing HVR sequences are provided.

[0185] In some embodiments, provided herein are at least one, at least two, or all three VHs selected from: (a) (i) an 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% identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 146 and 147; (ii) 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 sequence having at least about 95% identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 149, 150, 151, 152, and 153; and (iii) an HVR-H3 comprising the amino acid sequence of SEQ ID NO: 154, or an amino acid sequence having at least about 95% identity to the amino acid sequence of SEQ ID NO: 154. H V containing HVR sequences H and (b) at least one, at least two, or all three V domains selected from: (i) an 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% identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 156, 157, and 158; (ii) an 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% identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 160 and 161; and (iii) an HVR-L3 comprising the amino acid sequence of SEQ ID NO: 163, or an amino acid sequence having at least about 95% identity to the amino acid sequence of SEQ ID NO: 163. L V containing HVR sequences L domain.

[0186] 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 NOs: 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 In certain embodiments, the V 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: 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. HThe sequence may contain substitutions (e.g., conservative substitutions), insertions, or deletions compared to a reference sequence, but an anti-MS4A4A antibody comprising the sequence retains the ability to bind to MS4A4A. In certain embodiments, a total of 1 to 10 amino acids are 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 are 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, the substitutions, insertions, or deletions occur in regions outside the HVRs (i.e., FRs). Optionally, the anti-MS4A4A antibody comprises the V 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 In certain embodiments, V H comprises one, two, or three HVRs selected from: (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.

[0187] In another aspect, an anti-MS4A4A antibody is provided, the antibody comprising a light 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: 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, a V 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: 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 sequence may contain substitutions (e.g., conservative substitutions), insertions, or deletions compared to a reference sequence, but an anti-MS4A4A antibody comprising the sequence retains the ability to bind to MS4A4A. In some embodiments, a total of 1 to 10 amino acids are 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 are 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, the substitutions, insertions, or deletions occur in regions outside the HVRs (i.e., FRs). Optionally, the anti-MS4A4A antibody comprises the V 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 In certain embodiments, V L comprises one, two, or three HVRs selected from: (a) an 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) 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; and (c) an HVR-L3 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 107, 115, 144, 145, 163, 316, and 319.

[0188] In some embodiments, an anti-MS4A4A antibody is provided, which antibody comprises a V H and V in any of the previously provided embodiments. L In some embodiments, provided herein are anti-MS4A4A antibodies, which comprise any of the V of any of the previously provided embodiments. H and V in any of the previously provided embodiments. L In one embodiment, the antibody comprises V H and V L In one embodiment, the antibody comprises the sequences of SEQ ID NOs: 5 to 15 and SEQ ID NOs: 17 to 22, respectively, including post-translational modifications of these sequences. H and V L In one embodiment, the antibody comprises the sequences of SEQ ID NOs: 24-30 and SEQ ID NOs: 32-36, respectively, including post-translational modifications of these sequences. H and V L In one embodiment, the antibody comprises the sequences of SEQ ID NOs: 40 to 53 and SEQ ID NOs: 55 to 74, respectively, including post-translational modifications of these sequences. H and V L In one embodiment, the antibody comprises the sequences of SEQ ID NOs: 76-84 and 86-93, respectively, including post-translational modifications of these sequences. H and V L In one embodiment, the antibody comprises the sequences V H and V L The sequences are SEQ ID NO: 306 and SEQ ID NO: 307, respectively, including post-translational modifications of these sequences.

[0189] In some embodiments, as used herein, a heavy chain variable domain (V H ), and a light chain variable domain (V L ), and H and V LV 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:6 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: 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 Hand V comprising the amino acid sequence of SEQ ID NO: 307 L is selected from the group consisting of:

[0190] In some embodiments, as used herein, a heavy chain variable domain (V H ), and a light chain variable domain (V L ), and H and V L 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 Hand 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 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 is selected from the group consisting of:

[0191] In some embodiments, as used herein, a heavy chain variable domain (V H ), and a light chain variable domain (V L ), and H and V L 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: 57 L 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 Hand 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 H and 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 LV 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: 48 H 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 Hand 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 is selected from the group consisting of:

[0192] In some embodiments, as used herein, a heavy chain variable domain (V H ), and a light chain variable domain (V L ), and H and V L 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 Hand 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 is selected from the group consisting of:

[0193] 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 NOs: 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 304, and 306. H In certain embodiments, the V 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: 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 304, and 306. HThe sequence may contain substitutions (e.g., conservative substitutions), insertions, or deletions compared to a reference sequence, but an 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 the HVRs (i.e., FRs). Optionally, the anti-MS4A4A antibody comprises the V of SEQ ID NO: 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 304, or 306. H In certain embodiments, V H comprises one, two, or three HVRs selected from: (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.

[0194] In another aspect, an anti-MS4A4A antibody is provided, the antibody comprising a light 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: 17, 18, 19, 20, 21, 22, 305, and 307. L In certain embodiments, a V 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: 17, 18, 19, 20, 21, 22, 305, and 307. LThe sequence may contain substitutions (e.g., conservative substitutions), insertions, or deletions compared to a reference sequence, but an 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., FRs). Optionally, the anti-MS4A4A antibody may contain a VVR of SEQ ID NO: 17, 18, 19, 20, 21, 22, 305, or 307. L In certain embodiments, V L comprises one, two, or three HVRs selected from: (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.

[0195] 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 NOs: 24, 25, 26, 27, 28, 29, and 30. H In certain embodiments, a V 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: 24, 25, 26, 27, 28, 29, and 30. HThe sequence may contain substitutions (e.g., conservative substitutions), insertions, or deletions compared to a reference sequence, but an 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: 24, 25, 26, 27, 28, 29, or 30. In certain embodiments, a total of 1 to 5 amino acids have been substituted, inserted, and / or deleted in SEQ ID NO: 24, 25, 26, 27, 28, 29, or 30. In certain embodiments, the substitutions, insertions, or deletions occur in regions outside the HVRs (i.e., FRs). Optionally, the anti-MS4A4A antibody may contain a VVR of SEQ ID NO: 24, 25, 26, 27, 28, 29, or 30. H In certain embodiments, V H comprises one, two, or three HVRs selected from: (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.

[0196] In another aspect, an anti-MS4A4A antibody is provided, the antibody comprising a light 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: 32, 33, 34, 35, and 36. L In certain embodiments, a V 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: 32, 33, 34, 35, and 36. LThe sequence may contain substitutions (e.g., conservative substitutions), insertions, or deletions compared to a reference sequence, but an 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: 32, 33, 34, 35, or 36. In certain embodiments, a total of 1 to 5 amino acids have been substituted, inserted, and / or deleted in SEQ ID NO: 32, 33, 34, 35, or 36. In certain embodiments, the substitutions, insertions, or deletions occur in regions outside the HVRs (i.e., FRs). Optionally, the anti-MS4A4A antibody may contain a VVR of SEQ ID NO: 32, 33, 34, 35, or 36. L In certain embodiments, V L comprises one, two, or three HVRs selected from: (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.

[0197] 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 NOs: 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, and 53. H In certain embodiments, the V 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: 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, and 53. HThe sequence may contain substitutions (e.g., conservative substitutions), insertions, or deletions compared to a reference sequence, but an 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: 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 the HVRs (i.e., FRs). Optionally, the anti-MS4A4A antibody comprises the V of SEQ ID NO: 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, or 53 H In certain embodiments, V H comprises one, two, or three HVRs selected from: (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 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.

[0198] In another aspect, an anti-MS4A4A antibody is provided, the antibody comprising a light 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: 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, and 74. LIn certain embodiments, a V 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: 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, and 74. L The sequence may contain substitutions (e.g., conservative substitutions), insertions, or deletions compared to a reference sequence, but an 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: 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., FRs). Optionally, the anti-MS4A4A antibody comprises a VVR 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 In certain embodiments, V L comprises one, two, or three HVRs selected from: (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.

[0199] 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 NOs: 76, 77, 78, 79, 80, 81, 82, 83, and 84. H In certain embodiments, a V 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: 76, 77, 78, 79, 80, 81, 82, 83, and 84. H The sequence may contain substitutions (e.g., conservative substitutions), insertions, or deletions compared to a reference sequence, but an 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., FRs). Optionally, the anti-MS4A4A antibody may contain a VVR of SEQ ID NO: 76, 77, 78, 79, 80, 81, 82, 83, or 84. H In certain embodiments, V H comprises one, two, or three HVRs selected from: (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 the amino acid sequence of SEQ ID NO: 154.

[0200] In another aspect, an anti-MS4A4A antibody is provided, the antibody comprising a light 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: 86, 87, 88, 89, 90, 91, 92, and 93. L In certain embodiments, a V 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: 86, 87, 88, 89, 90, 91, 92, and 93. L The sequence may contain substitutions (e.g., conservative substitutions), insertions, or deletions compared to a reference sequence, but an 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: 86, 87, 88, 89, 90, 91, 92, or 93. In certain embodiments, a total of 1 to 5 amino acids have been substituted, inserted, and / or deleted in SEQ ID NO: 86, 87, 88, 89, 90, 91, 92, or 93. In certain embodiments, the substitutions, insertions, or deletions occur in regions outside the HVRs (i.e., FRs). Optionally, the anti-MS4A4A antibody may contain a VVR of SEQ ID NO: 86, 87, 88, 89, 90, 91, 92, or 93. L In certain embodiments, V L comprises one, two, or three HVRs selected from: (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 the amino acid sequence of SEQ ID NO: 163.

[0201] In another aspect, the anti-MS4A4A antibody comprises a full-length heavy chain amino acid sequence 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: 320-335 and 355-362. In certain embodiments, a full-length heavy chain amino acid sequence having 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 compared to the reference sequence, but an anti-MS4A4A antibody comprising that 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 to 335 and 355 to 362. In certain embodiments, a total of 1 to 5 amino acids have been substituted, inserted, and / or deleted in SEQ ID NOs: 320 to 335 and 355 to 362. In certain embodiments, the substitutions, insertions, or deletions occur in regions outside the HVRs (i.e., FRs).

[0202] In another aspect, the anti-MS4A4A antibody comprises a full-length light chain amino acid sequence 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: 363-365. In certain embodiments, a full-length heavy chain amino acid sequence having 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: 363-365 contains substitutions (e.g., conservative substitutions), insertions, or deletions compared to the reference sequence, but the anti-MS4A4A antibody comprising such sequence retains the ability to bind to MS4A4A. In certain embodiments, a total of 1 to 10 amino acids are 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 to 365. In certain embodiments, the substitutions, insertions, or deletions occur in regions outside the LVRs (i.e., FRs).

[0203] 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.

[0204] In some embodiments, an anti-MS4A4A antibody of the present disclosure competitively inhibits the binding of at least one reference antibody selected from 4A-202, 4A-301, 4A-302, 4A-330, 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 the 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, the anti-MS4A4A antibodies of the disclosure include 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-400, 4A-401, 4A-402, 4A-403, 4A-404, 4A-405, 4A-406, 4A-407, 4A-408, 4A-409, 4A-410, 4A-411, 4A-412, 4A-413, 4A-414, 4A-415, 4A-416, 4A-417, 4A-418, 4A-419, 4A-420, 4A-421, 4A-422, 4A-423, 4A-424, 4A-425, 4A-426, 4A-427, 4A-428, 4A-429, 4A-430, 4A-431, 4A-432 and 4A-381. In some embodiments, an anti-MS4A4A antibody of the present disclosure competitively inhibits the 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.

[0205] In some embodiments, the anti-MS4A4A antibodies of the disclosure include 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-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-34 9, 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, 4 and 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. Exemplary methods for mapping antibody-binding epitopes are described in detail in Morris (1996) "Epitope Mapping Protocols," in Methods in Molecular Biology, vol. 66 (Humana Press, Totowa, NJ).

[0206] In some embodiments, an anti-MS4A4A antibody of the present disclosure competitively inhibits the binding to MS4A4A 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, 4A-450, and any combination thereof. In some embodiments, an anti-MS4A4A antibody of the present disclosure competitively inhibits the binding to MS4A4A 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, 4A-331, and any combination thereof. In some embodiments, the anti-MS4A4A antibodies of the disclosure may be any of the following for binding to MS4A4A: 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-400, 4A-401, 4A-402, 4A-403, 4A-404, 4A-405, 4A-406, 4A-407, 4A-408, 4A-409, 4A-410, 4A-411, 4A-412, 4A-413, 4A-414, 4A-415, 4A-416, 4A-417, 4A-418, 4A-419, 4A-420, 4A-421, 4A-422, 4A-423, 4A-424, 4A-425, 4A-426, 4A-427, 4A-428, 4A-429, 4A-430, and 4A-381, and any combination thereof.In some embodiments, an anti-MS4A4A antibody of the present disclosure competitively inhibits the binding to MS4A4A 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.

[0207] In some embodiments, an anti-MS4A4A antibody of the present disclosure has an epitope on MS4A4A that is the same as or overlaps with 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 has an epitope on MS4A4A that is the same as or overlaps with 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, with respect to binding to MS4A4A. In some embodiments, the anti-MS4A4A antibodies of the disclosure may be 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-3 58, 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, with respect to binding to MS4A4A, an epitope on MS4A4A that is the same as or overlaps with at least one reference antibody selected from 4A-25, A-382, 4A-383, 4A-384, 4A-385, 4A-386, 4A-387, 4A-388, 4A-389, and 4A-390, and any combination thereof.

[0208] In some embodiments, anti-MS4A4A antibodies of the present disclosure bind to extracellular domain 1 (ECL1) of MS4A4A. In some embodiments, anti-MS4A4A antibodies of the present disclosure bind to one or more amino acids in the amino acid sequence CMASNTYGSNPIS (SEQ ID NO: 177) of SEQ ID NO: 1. In some embodiments, anti-MS4A4A antibodies of the present disclosure bind to extracellular domain 2 (ECL2) of MS4A4A. In some embodiments, anti-MS4A4A antibodies of the present disclosure bind to one or more amino acids in the amino acid sequence SFHHPYCNYYGNSNNCHGTMS (SEQ ID NO: 178) of SEQ ID NO: 1.

[0209] The anti-MS4A4A antibodies 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-420, 4A-421, 4A-422, 4A-423, 4A-424, 4A-425, 4A-426, 4A-427, 4A-428, 4A-429, 4A-430, 4A-431, 4A-432, 4A-433, 4A-434, 4A-435, 4A-436, 4A-437, 4A-438, 4A-439, 4A-440, 4A-441, 4A-442, 4A-443, 4A-444, 4A-445, 4A-446, 4A-447, 4A-448, 4A-449, 4A-500, 4A-501, 4A-502, 4A-503, 4A-504, 4A-505, 4A-506, 4A-507, 4A-508, 4A-509, 4A-601, 4A-610, 4A-61 -314, 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-3 28, 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, 4 A-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 competitive assay, immobilized MS4A4A or cells expressing MS4A4A on their cell surface are incubated with a solution containing a first labeled antibody (e.g., human or non-human primate) that binds to MS4A4A and a second unlabeled antibody to be tested for its ability to compete with the first antibody for binding to MS4A4A. The second antibody may be present in 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 that allow binding of the first antibody to MS4A4A, excess unbound antibody is removed, and the amount of label associated with the 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 compared to the control sample, this indicates that the second antibody competes with the first antibody for binding to MS4A4A. See Harlow and Lane (1988) Antibodies: A Laboratory Manual ch. 14 (Cold Spring Harbor Laboratory, Cold Spring Harbor, NY).

[0210] Further provided herein are anti-MS4A4A antibodies that competitively inhibit the binding of and / or compete for binding to an anti-MS4A4A antibody, the antibodies comprising: (a) a VVR comprising (i) an HVR-H1 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 94 and 308, (ii) an HVR-H2 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 96, 97, 98, 99, and 309, and (iii) an HVR-H3 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 100, 101, 102, and 310. Hdomain, and (b) a V domain comprising (i) an HVR-L1 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 103, 104, and 314, (ii) an HVR-L2 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 105, 106, and 315, and (iii) an HVR-L3 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 107 and 316. L In some embodiments, the antibody comprises a V H and V L The sequences are SEQ ID NOs: 5 to 15 and 304, and SEQ ID NOs: 17 to 22 and 305, respectively.

[0211] Provided herein are anti-MS4A4A antibodies that bind to an epitope on human MS4A4A that is the same as or overlaps with an epitope bound by an anti-MS4A4A antibody, and these antibodies comprise: (a) a VVR comprising (i) an HVR-H1 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 94 and 308, (ii) an HVR-H2 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 96, 97, 98, 99, and 309, and (iii) an HVR-H3 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 100, 101, 102, and 310. H domain, and (b) a V domain comprising (i) an HVR-L1 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 103, 104, and 314, (ii) an HVR-L2 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 105, 106, and 315, and (iii) an HVR-L3 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 107 and 316. L In some embodiments, the antibody comprises a V H and V L These include the sequences set forth in SEQ ID NOS: 5-15 and 304, and SEQ ID NOS: 17-22 and 305. In some embodiments, the epitope of human MS4A4A is the same epitope to which an anti-MS4A4A antibody binds.

[0212] Further provided herein are anti-MS4A4A antibodies that competitively inhibit the binding of and / or compete for binding to an anti-MS4A4A antibody, the antibodies comprising: (a) a VVR comprising (i) an HVR-H1 comprising the amino acid sequence of SEQ ID NO: 108, (ii) an HVR-H2 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 110 and 111, and (iii) an HVR-H3 comprising the amino acid sequence of SEQ ID NO: 112. H domain, and (b) a V domain comprising (i) an HVR-L1 comprising the amino acid sequence of SEQ ID NO: 113, (ii) an HVR-L2 comprising the amino acid sequence of SEQ ID NO: 114, and (iii) an HVR-L3 comprising the amino acid sequence of SEQ ID NO: 115. L In some embodiments, the antibody comprises a V H and V L The sequences are SEQ ID NOs: 24 to 30 and 32 to 36, respectively.

[0213] Provided herein are anti-MS4A4A antibodies that bind to an epitope in human MS4A4A that is the same as or overlaps with the epitope bound by an anti-MS4A4A antibody, and these antibodies comprise: (a) a VVR comprising (i) an HVR-H1 comprising the amino acid sequence of SEQ ID NO: 108, (ii) an HVR-H2 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 110 and 111, and (iii) an HVR-H3 comprising the amino acid sequence of SEQ ID NO: 112. H domain, and (b) a V domain comprising (i) an HVR-L1 comprising the amino acid sequence of SEQ ID NO: 113, (ii) an HVR-L2 comprising the amino acid sequence of SEQ ID NO: 114, and (iii) an HVR-L3 comprising the amino acid sequence of SEQ ID NO: 115. L In some embodiments, the antibody comprises a V H and V L These include the sequences set forth in SEQ ID NOs: 24 to 30 and SEQ ID NOs: 32 to 36, respectively. In some embodiments, the epitope of human MS4A4A is the same epitope to which an anti-MS4A4A antibody binds.

[0214] Further provided herein are anti-MS4A4A antibodies that competitively inhibit the binding of and / or compete for binding to an anti-MS4A4A antibody, the antibodies comprising: (a) a HVR-H1 comprising (i) an HVR-H1 having the amino acid sequence of SEQ ID NO: 116; (ii) an HVR-H2 having an amino acid sequence selected from the group consisting of SEQ ID NOs: 118, 119, 120, 121, and 122; and (iii) an HVR-H3 having an amino acid sequence selected from the group consisting of SEQ ID NOs: 123, 124, 125, 126, 127, 128, and 129. H and (b) a V domain comprising (i) 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, (ii) an HVR-L2 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 139, 140, 141, 142, and 143, and (iii) an HVR-L3 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 144 and 145. L In some embodiments, the antibody comprises a V H and V L The sequences are SEQ ID NOs: 40 to 53 and 55 to 74, respectively.

[0215] Provided herein are anti-MS4A4A antibodies that bind to an epitope on human MS4A4A that is the same as or overlaps with the epitope bound by an anti-MS4A4A antibody, and these antibodies comprise: (a) a VVR comprising (i) an HVR-H1 comprising the amino acid sequence of SEQ ID NO: 116, (ii) an HVR-H2 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 118, 119, 120, 121, and 122, and (iii) an HVR-H3 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 123, 124, 125, 126, 127, 128, and 129. Hand (b) a V domain comprising (i) 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, (ii) an HVR-L2 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 139, 140, 141, 142, and 143, and (iii) an HVR-L3 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 144 and 145. L In some embodiments, the antibody comprises a V H and V L These include the sequences set forth in SEQ ID NOs: 40 to 53 and SEQ ID NOs: 55 to 74, respectively. In some embodiments, the epitope of human MS4A4A is the same epitope to which an anti-MS4A4A antibody binds.

[0216] Further provided herein are anti-MS4A4A antibodies that competitively inhibit the binding of and / or compete for binding to an anti-MS4A4A antibody, the antibodies comprising: (a) a VVR comprising (i) an HVR-H1 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 146 and 147, (ii) an HVR-H2 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 149, 150, 151, 152, and 153, and (iii) an HVR-H3 comprising the amino acid sequence of SEQ ID NO: 154. H domain, and (b) a V domain comprising (i) an HVR-L1 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 156, 157, and 158, (ii) an HVR-L2 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 160 and 161, and (iii) an HVR-L3 comprising the amino acid sequence of SEQ ID NO: 163. L In some embodiments, the antibody comprises a V H and V L The sequences are SEQ ID NOs: 76 to 84 and 86 to 93, respectively.

[0217] Provided herein are anti-MS4A4A antibodies that bind to an epitope in human MS4A4A that is the same as or overlaps with the epitope bound by an anti-MS4A4A antibody, and these antibodies comprise: (a) a VVR comprising (i) an HVR-H1 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 146 and 147, (ii) an HVR-H2 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 149, 150, 151, 152, and 153, and (iii) an HVR-H3 comprising the amino acid sequence of SEQ ID NO: 154. H domain, and (b) a V domain comprising (i) an HVR-L1 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 156, 157, and 158, (ii) an HVR-L2 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 160 and 161, and (iii) an HVR-L3 comprising the amino acid sequence of SEQ ID NO: 163. L In some embodiments, the antibody comprises a V H and V L These include the sequences set forth in SEQ ID NOs: 76 to 84 and 86 to 93, respectively. In some embodiments, the epitope of human MS4A4A is the same epitope to which an anti-MS4A4A antibody binds.

[0218] In some embodiments, the anti-MS4A4A antibody of any of the preceding embodiments is a monoclonal antibody, including a humanized and / or human antibody. In some embodiments, the anti-MS4A4A antibody is an antibody fragment, e.g., an Fv, Fab, Fab', scFv, diabody, or F(ab')2 fragment. In some embodiments, the anti-MS4A4A antibody is a substantially full-length antibody, e.g., an IgG1 antibody, an IgG2a antibody, or other antibody class or isotype as defined herein.

[0219] In some embodiments, the anti-MS4A4A antibody of any of the preceding embodiments may incorporate any of the features, alone or in combination, as described in Sections 1-7 below.

[0220] (1) Anti-MS4A4A antibody binding affinity In some embodiments of any of the antibodies provided herein, the antibody has a cytotoxicity 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 less, e.g., 10 -8 M~10 -13 M, e.g., 10 -9 M~10 -13 The dissociation constant (Kd) can be determined by ELISA, surface plasmon resonance (SPR), biolayer interferometry (e.g., Octet from ForteBio), or other methods. Kd can be determined by any analytical technique, such as isothermal titration calorimetry (ITC), differential scanning calorimetry (DSC), circular dichroism (CD), stopped-flow analysis, and any biochemical or biophysical technique, such as colorimetric or fluorescent protein melting analysis (see, e.g., NIRS System), isothermal titration calorimetry (ITC), differential scanning calorimetry (DSC), circular dichroism (CD), stopped-flow analysis, and colorimetric or fluorescent protein melting analysis. In certain embodiments, Kd is measured by a radiolabeled antigen binding assay (RIA). In some embodiments, the RIA is performed using a Fab version of the antibody of interest or 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 using an immobilized antigen CM5 chip with approximately 10 response units (RU). In some embodiments, the 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). D is determined using a monovalent antibody (e.g., a Fab) or a full-length antibody. D is determined using the monovalent form of the full-length antibody.

[0221] (2) Antibody fragment 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 specific antibody fragments, see Hudson et al. Nat. Med. 9:129-134 (2003). For a review of scFv fragments, see, e.g., 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 that contain salvage receptor binding epitope residues and have extended in vivo half-lives, see U.S. Patent No. 5,869,046.

[0222] Diabodies are antibody fragments with two antigen-binding sites that can be bivalent or bispecific. See, e.g., EP 404097; WO 1993 / 01161; Hudson et al. Nat. Med. 9:129-134 (2003). Triabodies and tetrabodies are also described in Hudson et al. Nat. Med. 9:129-134 (2003). Single-domain antibodies are antibody fragments that contain all or part of the heavy chain variable domain or all or part of the light chain variable domain of an antibody. In certain embodiments, single-domain antibodies are human single-domain antibodies (see, e.g., U.S. Pat. No. 6,248,516).

[0223] Antibody fragments can be produced by a variety of techniques, including, but not limited to, proteolytic digestion of intact antibodies and production by recombinant host cells (e.g., E. coli or phage), as described herein.

[0224] (3) Chimeric and humanized antibodies In some embodiments of any of the antibodies provided herein, the antibody is a chimeric antibody. Particular chimeric antibodies are described, for example, in U.S. Patent No. 4,816,567. In some examples, chimeric antibodies contain 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 a further example, a chimeric antibody is a "class-switched" antibody in which the class or subclass has been changed from that of the parent antibody. Chimeric antibodies include antigen-binding fragments thereof.

[0225] In some embodiments of any antibody provided herein, the antibody is a humanized antibody. Generally, non-human antibodies are humanized to reduce immunogenicity in humans while retaining the specificity and affinity of the parent non-human antibody. In certain embodiments, humanized antibodies are substantially non-immunogenic in humans. In certain embodiments, humanized antibodies have substantially the same affinity for a target as an antibody from another species from which the humanized antibody is derived. See, e.g., U.S. Patent Nos. 5,530,101, 5,693,761, 5,693,762, and 5,585,089. In certain embodiments, amino acids in antibody variable domains are identified that can be modified to reduce immunogenicity without reducing the native affinity of the antigen-binding domain. See, e.g., U.S. Patent Nos. 5,766,886 and 5,869,619. Generally, humanized antibodies comprise one or more variable domains in which HVRs (or portions thereof) are derived from a non-human antibody and FRs (or portions thereof) are derived from human antibody sequences. A 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 (e.g., the antibody from which the HVR residues were derived), e.g., to restore or improve antibody specificity or affinity.

[0226] Humanized antibodies and methods for producing them are reviewed, for example, in Almagro et al. Front. Biosci. 13:1619-1633 (2008) and are also described, for example, in U.S. Pat. Nos. 5,821,337, 7,527,791, 6,982,321, and 7,087,409. Human framework regions that can be used for humanization include framework regions selected using the "best-fit" method (see, e.g., Sims et al. J. Immunol. 151:2296 (1993)); framework regions derived from consensus sequences 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 of FR libraries (see, e.g., Baca et al. al. J. Biol. Chem. 272:10678-10684 (1997) and Rosok et al. J. Biol. Chem. 271:22611-22618 (1996)).

[0227] (4) Human antibodies 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).

[0228] Human antibodies can be prepared by administering immunogens to intact human antibodies or transgenic animals modified to produce intact antibodies bearing human variable regions in response to antigen challenge. Mouse strains deficient in mouse antibody production can be engineered with large fragments of the human Ig loci, with the expectation that such mice will produce human antibodies in the absence of mouse antibodies. The large human Ig fragments allow for the diversity of large variable genes and the proper regulation of antibody production and expression. By exploiting the mouse machinery for antibody diversification and selection and the lack of immunological tolerance to human proteins, the recapitulated human antibody repertoire in these mouse strains can generate high-affinity, fully human antibodies against antigens of interest, including human antigens. Using hybridoma technology, antigen-specific human monoclonal antibodies with desired specificity can be produced and selected. Certain exemplary methods are described in U.S. Pat. No. 5,545,807, EP 546073, and EP 546073. See also, e.g., U.S. Patent Nos. 6,075,181 and 6,150,584, describing XENOMOUSE™ technology; U.S. Patent No. 5,770,429, describing HUMAB® technology; U.S. Patent No. 7,041,870, describing KM MOUSE® technology; and U.S. Patent Application Publication No. US2007 / 0061900, describing VELOCIMOUSE® technology. The human variable regions derived from intact antibodies produced by such animals can be further engineered, for example, by combining with different human constant regions.

[0229] Human antibodies can also be produced using hybridoma-based methods. Human myeloma and mouse-human heteromyeloma cell lines for the production of human monoclonal antibodies have been described (e.g., Kozbor J. Immunol. 133:3001 (1984) and Boerner et al. J. Immunol. 147:86 (1991)). Human antibodies generated via human B cell hybridoma technology are described in Li et al. Proc. Natl. Acad. Sci. USA, 103:3557-3562 (2006). Additional methods include those described in U.S. Pat. No. 7,189,826 (which describes the production of monoclonal human IgM antibodies from hybridoma cell lines). Human hybridoma technology (trioma technology) is 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 generated by isolating Fv clone variable domain sequences selected from a human-derived phage display library. Such variable domain sequences can then be combined with desired human constant domains. Techniques for selecting human antibodies from antibody libraries are described below.

[0230] In some embodiments of any of the antibodies provided herein, the antibody is a human antibody isolated by in vitro methods and / or by screening combinatorial libraries for antibodies possessing one or more desired activities. Suitable examples include, but are not limited to, phage display (CAT, Morphosys, Dyax, Biosite / Medarex, Xoma, Symphogen, Alexion (formerly Proliferon), Affimed), ribosome display (CAT), and yeast display (Adimab). In certain phage display methods, repertoires of VH and VL genes are separately cloned by polymerase chain reaction (PCR) and then randomly recombined into phage libraries as described in Winter et al. Ann. Rev. Immunol. 12:433-455 (1994). For example, various methods are known in the art for generating phage display libraries and screening such libraries for antibodies possessing desired binding characteristics. 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. See also Griffiths et al. J. Immunol. Methods 284(-2):119-132 (2004). Typically, phage display antibody fragments, either single-chain Fv (scFv) fragments or Fab fragments. Libraries derived from immunogens provide high-affinity antibodies to immunogens without the need to construct hybridomas. Alternatively, naive repertoires can be cloned (e.g., from humans) to provide a single source of antibodies against a wide range of non-self antigens as well as self antigens without immunization, as described in Griffiths et al. EMBO J. 12:725-734 (1993). Finally, a naive library was constructed by cloning unrearranged V gene segments from stem cells and encoding the highly variable HVR3 region using PCR primers containing random sequences, and in vitro rearrangement was achieved as described in Hoogenboom et al. J. Mol. Biol., 227:381-388, 1992. Patent documents describing human antibody phage libraries include, for example, U.S. Patent No. 5,750,373 and U.S. Patent Publication Nos. 2007 / 0292936 and 2009 / 0002360. Antibodies isolated from human antibody libraries are considered herein to be human antibodies or human antibody fragments.

[0231] (5) a constant region including an Fc region 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, IgM, or IgA class antibody.

[0232] In certain embodiments of any antibody provided herein, the antibody has an IgG2 isotype. In some embodiments, the antibody comprises a human IgG2 constant region. In some embodiments, the human IgG2 constant region comprises an Fc region. In some embodiments, the antibody induces one or more MS4A4A activities, or does so independent of binding 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).

[0233] In certain embodiments of any antibody provided herein, the antibody has an IgG1 isotype. In some embodiments, the antibody comprises a mouse IgG1 constant region. In some embodiments, the antibody comprises a human IgG1 constant region. In some embodiments, the human IgG1 constant region comprises 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).

[0234] In certain embodiments of any antibody provided herein, the antibody has an IgG4 isotype. In some embodiments, the antibody comprises a human IgG4 constant region. In some embodiments, the human IgG4 constant region comprises an Fc region. 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).

[0235] In certain embodiments of any of the antibodies provided herein, the antibody has a hybrid IgG2 / 4 isotype. In some embodiments, the antibody comprises an amino acid sequence comprising amino acids 118-260 of human IgG2 as defined by the EU numbering system and amino acids 261-447 of human IgG4 as defined by the EU numbering system (WO1997 / 11971; WO2007 / 106585).

[0236] In some embodiments, the Fc region enhances cluster formation without activating complement compared to a corresponding antibody comprising an Fc region that does not contain the amino acid substitutions. In some embodiments, the antibody induces one or more activities of a target to which the antibody specifically binds. In some embodiments, the antibody binds to MS4A4A.

[0237] It may also be desirable to modify the anti-MS4A4A antibodies of the present disclosure to modify effector function and / or increase the serum half-life of the antibody. For example, the Fc receptor binding site of the constant region may be modified or mutated to eliminate or reduce binding affinity to specific Fc receptors, such as FcγRI, FcγRII, and / or FcγRIII, thereby reducing antibody-dependent cellular cytotoxicity. In some embodiments, removing N-glycosylation in the Fc region of the antibody (e.g., in the CH2 domain of IgG) reduces the effector function. In some embodiments, modifications to regions such as residues 233-236, 297, and / or 327-331 of human IgG, as described in WO 99 / 58572 and Armour et al. Molecular Immunology 40:585-593 (2003); Reddy et al. J. Immunology 164:1925-1933 (2000), reduce effector function. In other embodiments, modifying the anti-MS4A4A antibodies of the present disclosure to modify effector function may also be desirable to enhance their selectivity for ITIM-containing FcgRIIb (CD32b), thereby enhancing clustering of the MS4A4A antibody with neighboring cells without activating humoral responses such as antibody-dependent cell-mediated cytotoxicity and antibody-dependent cellular phagocytosis.

[0238] To increase the serum half-life of the antibody, a salvage receptor binding epitope can be incorporated into the antibody (particularly an antibody fragment), for example, as described in U.S. Patent No. 5,739,277. As used herein, the term "salvage receptor binding epitope" refers to an epitope in the Fc region of an IgG molecule (e.g., 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.

[0239] (6) Multispecific antibodies A multispecific antibody is an antibody that has binding specificities for at least two different epitopes, such as epitopes of the same or different polypeptides (e.g., one or more MS4A4A polypeptides of the present disclosure). In some embodiments, the multispecific antibody may be a bispecific antibody. In some embodiments, the multispecific antibody may be a trispecific antibody. In some embodiments, the multispecific antibody may be a tetraspecific antibody. Such antibodies may be full-length antibodies or antibodies derived from 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.

[0240] Provided herein are multispecific antibodies comprising a first antigen-binding region comprising six HVRs of an antibody described herein that binds to MS4A4A, and a second antigen-binding region that binds to a second polypeptide. In some embodiments, the first antigen-binding region comprises six HVRs of an antibody described herein that binds to MS4A4A, and a second antigen-binding region that binds to a second polypeptide. H or V L Includes.

[0241] In some embodiments of any of the multispecific antibodies, the second polypeptide is selected from: (a) an antigen that promotes transport across the blood-brain barrier; (b) an antigen that promotes transport across the blood-brain barrier selected from transferrin receptor (TR), insulin receptor (HIR), insulin-like growth factor receptor (IGFR), low-density lipoprotein receptor-related proteins 1 and 2 (LPR-1 and 2), diphtheria toxin receptor, CRM197, a llama single domain antibody, TMEM30(A), a protein transduction domain, TAT, Syn-B, penetratin, a polyarginine peptide, angiopep peptide, and ANG1005; or (c) amyloid beta, oligomeric amyloid beta, amyloid beta plaque, amyloid precursor protein or fragments thereof, Tau, IA. PP, alpha-synuclein, TDP-43, FUS protein, C9orf72 (chromosome 9 open reading frame 72), c9RAN protein, prion protein, PrPSc, huntingtin, calcitonin, superoxide dismutase, ataxin, ataxin 1, ataxin 2, ataxin 3, ataxin 7, ataxin 8, ataxin 10, Lewy bodies, atrial natriuretic factor, islet amyloid polypeptide, insulin, apolipoprotein AI, serum amyloid A, medin, prolactin, transthyretin, lysozyme, beta-2 microglobulin, gelsolin, keratoepithelin, cystatin, immunoglobulin light chain AL, S-IBM protein, repeat-associated non-ATG (RAN) translation products, dipeptide repeats a disease-causing protein selected from 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;(d) a ligand and / or protein expressed on an immune cell, wherein the ligand and / or protein is 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) a protein, lipid, polysaccharide, or glycolipid expressed on one or more tumor cells, and any combination thereof;

[0242] Many antigens that facilitate transport across the blood-brain barrier are known in the art (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 2), diphtheria toxins such as CRM197 (a non-toxic variant of diphtheria toxin), llama single-domain antibodies such as TMEM30(A) (Flippase), protein transduction domains such as TAT, Syn-B, or penetratin, polyarginine or generally positively charged peptides, Angiopep peptides such as ANG1005 (see, e.g., Gabathuler, 2010), and other cell surface proteins that are enriched in blood-brain barrier endothelial cells (see, e.g., Daneman et al. PLoS One 5(10):e13741 (2010)).

[0243] The multivalent antibody may recognize the MS4A4A antigen as well as additional antigens, such as Aβ peptide, α-synuclein protein antigen, Tau protein antigen, TDP-43 protein antigen, prion protein antigen, huntingtin protein antigen, translation product antigens containing dipeptide repeats (DPR peptides) consisting of RAN, glycine-alanine (GA), glycine-proline (GP), glycine-arginine (GR), proline-alanine (PA), or proline-arginine (PR), insulin receptor, insulin-like growth factor receptor, etc. Transferrin receptor or other antigens that facilitate antibody translocation 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 alpha-synuclein.

[0244] The multivalent antibody comprises at least one polypeptide chain (and preferably two polypeptide chains), where one or more of the polypeptide chains comprises two or more variable domains. For example, one or more of the polypeptide chains may comprise VD1-(X1) n -VD2-(X2) n -Fc, where VD1 is a first variable domain, VD2 is a second variable domain, Fc is one polypeptide chain of an Fc region, X1 and X2 represent amino acids or polypeptides, and n is 0 or 1. Similarly, one or more polypeptide chains may be V H -C H 1. Flexible Linker-V H -C H 1—Fc region chain; or V H -C H 1-V H -C H1-Fc region chain. The multivalent antibodies herein preferably further comprise at least two (and preferably four) light chain variable domain polypeptides. The multivalent antibodies herein may comprise, for example, 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.

[0245] Techniques for generating 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 Nature 305:537 (1983), WO93 / 08829, and Traunecker et al. EMBO J. 10:3655 (1991)), and "knob-in-hole" genetic engineering (see, e.g., U.S. Pat. No. 5,731,168). See also WO2013 / 026833 (CrossMab). Multispecific antibodies can also be created by manipulating electrostatic steering effects to create antibody Fc-heterodimeric molecules (WO 2009 / 089004 A1); by cross-linking two or more antibodies (see, e.g., U.S. Pat. No. 4,676,980); using leucine; using "diabody" technology to create 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 by preparing triabodies as described, for example, in Tutt et al. J. Immunol. 147:60 (1991).

[0246] The present specification also includes genetically engineered antibodies with three or more functional antigen-binding sites, such as "octopus antibodies" (see, e.g., US2006 / 0025576). Antibodies herein also include "dual-acting FAbs" or "DAFs" that contain antigen-binding sites that bind to multiple MS4A4As (see, e.g., US2008 / 0069820).

[0247] (7) Antibody variants In some embodiments of any of the antibodies provided herein, amino acid sequence variants of the antibody are contemplated, e.g., where it is desirable to improve the binding affinity and / or other biological properties of the antibody.

[0248] (i) Substitution, insertion, and deletion variants Some embodiments of any of the antibodies provided herein provide antibody variants with one or more amino acid substitutions. Amino acid sequence variants of an antibody can be prepared by introducing appropriate modifications into the nucleotide sequence encoding the antibody, or by peptide synthesis. Such modifications can include, for example, deletions and / or insertions and / or substitutions of residues in the amino acid sequence of the antibody. [Table 2]

[0249] Substantial modification of the biological properties of the antibody is achieved by selecting substitutions that differ significantly in their effect on (a) the conformation of the polypeptide backbone in the area of ​​substitution, e.g., sheet or helix structure, (b) the charge or hydrophobicity of the molecule at the target site, or (c) maintaining side chain bulk. Naturally occurring residues are selected based on common side chain properties: (1) Hydrophobic: Norleucine, Met, Ala, Val, Leu, Ile; (2) Neutral hydrophilic: Cys, Ser, Thr, Asn, Gln; (3) Acidic: Asp, Glu; (4) basic: His, Lys, Arg; (5) residues that influence chain orientation: Gly, Pro; and (6) Aromatic: Divided into Trp, Tyr, and Phe.

[0250] For example, non-conservative substitutions involve exchanging a member of one of these classes for one of another class. Such substituted residues can be introduced, for example, into regions of the human antibody that are homologous with the non-human antibody, or into the non-homologous regions of the molecule.

[0251] When making modifications to the polypeptides or antibodies described herein, certain embodiments may take into account the hydropathic index of amino acids. Each amino acid is assigned a hydropathic index based on its hydrophobicity and charge characteristics. These include: isoleucine (+4.5); valine (+4.2); leucine (+3.8); phenylalanine (+2.8); cysteine / cysteine ​​(+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).

[0252] The importance of the hydropathic amino acid index in conferring interactive biological function to a protein is 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 having a similar hydropathic index or score and still retain similar biological activity. When making changes based on hydropathic index, certain embodiments include substitutions of amino acids whose hydropathic index is within ±2, certain embodiments include cases within ±1, and certain embodiments include cases within ±0.5.

[0253] It should also be understood that substitutions of like amino acids can be made effectively on the basis of hydrophilicity, particularly when the resulting biologically functional proteins or peptides are intended for use in immunological embodiments, as in the present case. In certain embodiments, the greatest local average hydrophilicity of a protein, as governed by the hydrophilicity of its adjacent amino acids, correlates with its immunogenicity and antigenicity, i.e., with a biological property of the protein.

[0254] These amino acid residues have been assigned hydrophilicity values: 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, certain embodiments include substitution of amino acids with hydrophilicity values ​​within ±2, certain embodiments include substitution of amino acids within ±1, and certain embodiments include substitution of amino acids within ±0.5. Epitopes can also be identified from primary amino acid sequences based on hydrophilicity. These regions are also referred to as "epitope core regions."

[0255] In certain embodiments, substitutions, insertions, or deletions may be made in one or more HVRs as long as such modifications do not substantially reduce the ability of the antibody to bind to the antigen. For example, conservative modifications (e.g., conservative substitutions provided herein) that do not substantially reduce binding affinity may be made in HVRs. Such modifications may, for example, be made outside the antigen-contacting residues of the HVRs. In certain embodiments of the foregoing mutant VH and VL sequences, each HVR is unaltered or contains no more than one, two, or three amino acid substitutions.

[0256] Amino acid sequence insertions include amino- and / or carboxyl-terminal fusions ranging from one residue to polypeptides of 100 or more residues in length, as well as intrasequence insertions of one or more amino acid residues. An example of a terminal insertion is an antibody with an N-terminal methionyl residue. Other insertional variants of antibody molecules include the fusion to the N- or C-terminus of the antibody (e.g., for ADEPT) or a polypeptide which increases the serum half-life of the antibody.

[0257] Cysteine ​​residues not involved in maintaining the proper conformation of the antibody also generally can be substituted with serine to improve the oxidative stability of the molecule and prevent aberrant crosslinking. Conversely, the addition of cysteine ​​bond(s) to an antibody can improve its stability, particularly where the antibody is an antibody fragment such as an Fv fragment.

[0258] (ii) Glycosylation variants In some embodiments of any of the antibodies provided herein, the antibody is modified to increase or decrease the extent to which the antibody is glycosylated. Addition or deletion of glycosylation sites to an antibody can be conveniently accomplished by altering the amino acid sequence to create or remove one or more glycosylation sites.

[0259] Glycosylation of antibodies is typically either N-linked or O-linked. N-linked refers to the attachment of the 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 recognition sequences for enzymatic attachment of the carbohydrate moiety to the asparagine side chain. Thus, 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, although 5-hydroxyproline or 5-hydroxylysine may also be used.

[0260] Addition of glycosylation sites to the antibody is conveniently accomplished by altering the amino acid sequence so that it contains one or more of the above-described tripeptide sequences (for N-linked glycosylation sites), or (for O-linked glycosylation sites) may be accomplished by adding or substituting one or more serine or threonine residues to the original antibody sequence.

[0261] If the antibody comprises an Fc region, the carbohydrate attached thereto can be modified. Native antibodies produced by mammalian cells typically contain branched, biantennary oligosaccharides that are typically N-linked to Asn297 (Kabat numbering) in the CH2 domain of the Fc region. These oligosaccharides can include various carbohydrates, such as mannose, N-acetylglucosamine (GlcNAc), galactose, and sialic acid, as well as fucose attached to the GlcNAc in the "stem" of the biantennary oligosaccharide structure. In some embodiments, oligosaccharide modifications on the antibodies of the present disclosure can be performed to generate antibody variants with specific improved properties.

[0262] In one embodiment, antibody variants are provided that have carbohydrate structures that lack fucose attached (directly or indirectly) to the Fc region. See, e.g., U.S. Patent Publication Nos. 2003 / 0157108 and 2004 / 0093621. Examples of publications relating to "defucosylated" or "fucosylated" antibody variants include: US2003 / 0157108; US2003 / 0115614; US2002 / 0164328; US2004 / 0093621; US2004 / 0132140; US2004 / 0110704; US2004 / 0110282; US2004 / 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, which are deficient in protein fucosylation (Ripka et al. Arch. Biochem. Biophys. 249:533-545 (1986); US2003 / 0157108), and knockout cell lines, such as alpha-1,6-fucosyltransferase gene, FUT8, knockout CHO cells (see, e.g., Yamane-Ohnuki et al. Biotech. Bioeng. 87:614 (2004) and Kanda et al., Biotechnol. Bioeng. 94(4):680-688 (2006)).

[0263] (iii) modified constant region In some embodiments of any of the antibodies provided herein, the antibody Fc is an antibody Fc isotype and / or modification, hi some embodiments, the antibody Fc isotype and / or modification is capable of binding to an Fc gamma receptor.

[0264] 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, one or more of the amino acid substitutions are selected from the group consisting of N297A (Bolt S et al. (1993) Eur J Immunol 23:403-411), D265A (Shields et al. (2001) RB 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), and the like, as defined by the EU numbering system. 57:1537-1543.31; 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.

[0265] In some embodiments of any IgG1 modified Fc, the Fc comprises an N297A mutation according to the EU numbering system. In some embodiments of any IgG1 modified Fc, the Fc comprises a D265A and N297A mutation according to the EU numbering system. In some embodiments of any IgG1 modified Fc, the Fc...

Claims

1. 1. An isolated antibody that binds to an MS4A4A protein, the antibody comprising a heavy chain variable region and a light chain variable region, a) 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; or b) 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. The antibody.

2. The antibody of claim 1, wherein the antibody comprises a heavy chain variable region and a light chain variable region, the heavy chain variable region comprising 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 comprising 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.

3. The antibody of claim 1, wherein the antibody comprises a heavy chain variable region and a light chain variable region, the heavy chain variable region comprising 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 comprising 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.

4. The antibody, 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. The antibody of claim 1 or 2, comprising:

5. The antibody, 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. The antibody of claim 1 or 2, comprising:

6. The antibody, 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. The antibody of claim 1 or 3, comprising:

7. The antibody, 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. The antibody of claim 1 or 3, comprising:

8. The antibody of any one of claims 1 to 7, wherein the antibody is human or humanized.

9. The antibody of any one of claims 1 to 7, wherein the antibody (a) reduces cell surface levels of MS4A4A or reduces intracellular levels of MS4A4A, (b) increases soluble TREM2 levels, increases membrane TREM2 levels, or increases soluble TREM2 and increases membrane TREM2 levels in bone marrow cells, and / or (c) inhibits expression of the M2 cell surface marker in bone marrow cells.

10. The antibody of claim 8, wherein the antibody inhibits the expression of CD200R, Dectin-1, or CD163 in bone marrow cells.

11. The antibody according to any one of claims 1 to 7, which enhances expression of gelsolin and / or osteopontin mRNA and / or protein in bone marrow cells.

12. The antibody according to any one of claims 1 to 11, wherein the antibody is a monoclonal antibody.

13. The antibody of any one of claims 1 to 12, wherein the antibody binds to human MS4A4A of SEQ ID NO: 1, or the antibody further binds to cynomolgus monkey MS4A4A of SEQ ID NO:

3.

14. The antibody of any one of claims 1 to 13, wherein the antibody belongs to the IgG class and has an IgG1, IgG2, or IgG4 isotype.

15. The antibody of claim 14, wherein the antibody has an IgG1 isotype.

16. The antibody of claim 15, wherein the IgG1 isotype comprises a modified Fc comprising a K322A mutation according to the EU numbering system.

17. 16. The antibody of claim 15, wherein the antibody comprises a modified Fc comprising the N325S and L328F mutations as defined by the EU numbering system.

18. The antibody according to any one of claims 1 to 17, wherein the antibody is an antibody fragment.

19. 19. The antibody of claim 18, wherein the antibody fragment is a Fab, Fab', Fab'-SH, F(ab')2, Fv, or scFv fragment.

20. The antibody according to any one of claims 1 to 19, which is a bispecific antibody that recognizes a first antigen and a second antigen.

21. The first antigen is MS4A4A, and the second antigen is (a) an antigen that facilitates 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 2), diphtheria toxin receptor, CRM197, llama single domain antibody, TMEM30(A), protein transduction domain, TAT, Syn-B, penetratin, polyarginine peptides, angiopeptides, basigin, Glut1, CD98hc, and ANG1005; (c) a pathogenic agent selected from the group consisting of 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 amyloid beta, oligomeric amyloid beta, amyloid beta plaque, 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, calcitonin, superoxide dismutase, ataxin, ataxin 1, ataxin 2, ataxin 3, ataxin 7, ataxin 8, ataxin 10, Lewy bodies, atrial natriuretic factor, islet amyloid polypeptide, insulin, apolipoprotein AI, serum amyloid A, medin, prolactin, transthyretin, lysozyme, beta 2 microglobulin, gelsolin, keratoepithelin, cystatin, immunoglobulin light chain AL, S-IBM protein, repeat-associated non-ATG (RAN) translation products, dipeptide repeats (Dipeptide repeats) the pathogenic agent is selected from the group consisting of dipeptidyl repeat (DPR) peptide, glycine-alanine (GA) repeat peptide, glycine-proline (GP) repeat peptide, glycine-arginine (GR) repeat peptide, proline-alanine (PA) repeat peptide, ubiquitin, and proline-arginine (PR) repeat peptide; (d) a ligand and / or protein expressed in an immune cell, the ligand and / or protein being 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 (e) a protein, lipid, polysaccharide, or glycolipid expressed by one or more tumor cells; The antibody of claim 20,

22. An isolated nucleic acid comprising a nucleic acid sequence encoding the antibody of any one of claims 1 to 21.

23. A vector comprising the nucleic acid of claim 22.

24. 23. An isolated host cell comprising the nucleic acid of claim 22.

25. 25. A method for producing an antibody that binds to human MS4A4A, comprising culturing the cell of claim 24 to produce the antibody.

26. 26. The method of claim 25, further comprising recovering the antibody produced by the cells.

27. A pharmaceutical composition comprising the antibody of any one of claims 1 to 21 and a pharma- ceutical acceptable carrier.

28. 22. A pharmaceutical composition for preventing, reducing the risk of, or treating an individual having a disease, disorder, or injury selected from the group consisting of Alzheimer's disease, late-onset Alzheimer's disease, and cognitive impairment, comprising a therapeutically effective amount of an antibody according to any one of claims 1 to 21.

29. A pharmaceutical composition for preventing, reducing the risk of, or treating an individual having a disease, disorder, condition, or injury caused by or associated with overexpression or increased activity of MS4A4A, the pharmaceutical composition comprising a therapeutically effective amount of an antibody described in any one of claims 1 to 21.

30. A pharmaceutical composition for treating an individual having a CSF1R deficient disease or disorder, said pharmaceutical composition comprising a therapeutically effective amount of an antibody according to any one of claims 1 to 21.

31. The pharmaceutical composition of claim 30, wherein the CSF1R deficiency disease or disorder is adult-onset leukoencephalopathy with axonal spheroids and pigmented glia (ALSP) or hereditary diffuse leukoencephalopathy (HDLS).

32. The pharmaceutical composition of any one of claims 28 to 31, wherein said preventing, reducing the risk of, or treating comprises detecting levels of osteopontin, gelsolin, membrane TREM2, and / or soluble TREM2 before and / or after administration of the pharmaceutical composition.

Images