Anti-trem-1 antibodies and uses thereof

Abstract

Provided herein are methods of identifying subjects suitable for treatment with an anti-TREM-1 antibody (i.e., an antagonistic anti-TREM-1 antibody), comprising measuring the expression level of a TREM-1 -associated gene. Also disclosed herein are methods of determining the efficacy of an anti-TREM-1 antibody, comprising measuring the expression level of a TREM-1 -associated gene. Also disclosed are methods of identifying non-responders to standard-of-care treatment and methods of treating a disease or disorder (e.g., inflammatory bowel disease) with an anti-TREM-1 antibody.

Description

[0001] Cross-references to related applications

[0002] This PCT application claims priority to U.S. Provisional Application No. 62 / 874,318, filed July 15, 2019, which is incorporated herein by reference in its entirety.

[0003] References to sequence listings submitted electronically via EFS-WEB

[0004] The contents of the sequence list submitted electronically in an ASCII text file (filename: 3338_1380000_SeqListing.txt; size: 486,499 bytes; creation date: July 14, 2019) submitted with this application are incorporated herein by reference in their entirety. Background Technology

[0005] TREM-1 is an activating receptor expressed on monocytes, macrophages, and neutrophils. By binding to its natural ligand, peptidoglycan recognition protein 1 (PGLYRP1), TREM-1 helps activate these cells, leading to the production of cytokines and other mediators that drive inflammation. Therefore, TREM-1 mRNA and protein expression are upregulated in many inflammatory diseases, including inflammatory bowel disease (IBD), and TREM-1-positive cells accumulate at sites of inflammation, which is correlated with disease severity. See Bouchon et al., Nature 410:1103-1107 (2001); and Schenk et al., Clin Invest 117:3097-3106 (2007).

[0006] Inflammatory bowel disease (IBD) (e.g., ulcerative colitis (UC) and Crohn's disease (CD)) is a chronic gastrointestinal disorder characterized by inflammation of the intestine or colon. Symptoms of IBD can vary, but typically include abdominal cramps, persistent diarrhea, and rectal bleeding. IBD can be debilitating and can lead to life-threatening complications if left untreated.

[0007] IBD has no known cure. Current treatment options include medications (e.g., anti-inflammatory agents, immunosuppressants, and antibiotics), nutritional supplements, and surgery. While such treatments can reduce signs and symptoms of the disease, they often have limited efficacy and / or adverse side effects. See, e.g., Martinez-Montiel, M.P., et al., Clin Exp Gastroenterol 8:257-269 (2015); Cunliffe, R.N., et al., Aliment Pharmacol Ther 16(4):647-662 (2002). Moreover, IBD is difficult to diagnose, and available diagnostic methods (e.g., blood / stool tests, X-rays, endoscopy (e.g., colonoscopy), and / or tissue biopsy) are often very invasive. Thus, IBD remains a major medical challenge worldwide, and there remains a need for safer and more effective new treatment and / or diagnostic options. SUMMARY

[0008] Provided herein is a method of identifying a subject having a disease or disorder amenable to treatment with an antagonistic anti-TREM-1 antibody. In certain embodiments, the method comprises measuring the level of expression of a TREM-1 -related gene in a sample of the subject, wherein the TREM-1 -related gene comprises nicotinamide phosphoribosyltransferase (NAMPT); dehydrogenase / reductase 9 (DHRS9); cyclin-dependent kinase inhibitor 1A (CDKN1A); CD52 molecule (CD52); myotubularin-related protein 11 (MTMR11); EH domain-containing 1 (EHD1); solute carrier family 27 member 3 (SLC27A3); interleukin 24 (IL24); Pim-2 proto-oncogene, serine / threonine kinase (PIM2); chitinase 3-like 1 (CHI3L1); polypeptide N-acetylgalactosaminyltransferase 6 (GALNT6); acyl-CoA thioesterase 7 (ACOT7); cytokine inducible SH2 containing protein (CISH); sequence similarity family 129 member A (FAM129A); polo-like kinase 3 (PLK3); major facilitator superfamily domain containing 12 (MFSD12); StAR-related lipid transfer domain containing 4 (STARD4); C-type lectin domain family 12 member A (CLEC12A); CD55 molecule (Cromer blood group) (CD55); interferon lambda receptor 1 (IFNLR1), or a combination thereof.

[0009] In some embodiments, the method further comprises administering to a subject exhibiting an increased level of expression of the TREM-1 -associated gene compared to a reference a therapeutically effective dose of the antagonistic anti-TREM-1 antibody, wherein the reference comprises a subject that does not have the disease or disorder (e.g., a healthy subject).

[0010] Also provided herein is a method of treating a disease or disorder in a subject in need thereof, comprising administering to the subject a therapeutically effective dose of an antagonistic anti-TREM-1 antibody, wherein the subject exhibits an increased level of expression of a TREM-1 -associated gene, wherein the TREM-1 -associated gene comprises nicotinamide phosphoribosyltransferase (NAMPT); Dehydrogenase / reductase 9 (DHRS9); Cyclin-dependent kinase inhibitor 1A (CDKN1A); CD52 molecule (CD52); Myotubularin-related protein 11 (MTMR11); EH domain-containing 1 (EHD1); Solute carrier family 27 member 3 (SLC27A3); Interleukin 24 (IL24); Pim-2 proto-oncogene, serine / threonine kinase (PIM2); Chitinase 3-like 1 (CHI3L1); Polypeptide N-acetylgalactosaminyltransferase 6 (GALNT6); Acyl-CoA thioesterase 7 (ACOT7); Cytokine inducible SH2 containing protein (CISH); Sequence similarity family 129 member A (FAM129A); Polo-like kinase 3 (PLK3); Major facilitator superfamily domain containing 12 (MFSD12); StAR-related lipid transfer domain containing 4 (STARD4); C-type lectin domain family 12 member A (CLEC12A); CD55 molecule (Cromer blood group) (CD55); Interferon lambda receptor 1 (IFNLR1), or a combination thereof.

[0011] In some embodiments, the subject was previously treated with a standard of care treatment for the disease or disorder and did not respond to the treatment.

[0012] In some embodiments, the standard of care treatment comprises an anti-TNF-a antibody. In certain embodiments, the anti-TNF-a antibody comprises infliximab certolizumab pegol etanercept adalimumab golimumab or a combination thereof.

[0013] The present disclosure also provides a method of identifying a non-responder to a standard of care treatment for a disease or disorder, comprising measuring the level of expression of a TREM-1 -related gene in a sample of a subject who has received the standard of care treatment, wherein the subject exhibits an increased level of expression of the TREM-1 -related gene, and wherein the TREM-1 -related gene comprises nicotinamide phosphoribosyltransferase (NAMPT); dehydrogenase / reductase 9 (DHRS9); cyclin-dependent kinase inhibitor 1A (CDKN1A); CD52 molecule (CD52); myotubularin-related protein 11 (MTMR11); EH domain-containing 1 (EHD1); solute carrier family 27 member 3 (SLC27A3); interleukin 24 (IL24); Pim-2 proto-oncogene, serine / threonine kinase (PIM2); chitinase 3-like 1 (CHI3L1); polypeptide N-acetylgalactosaminyltransferase 6 (GALNT6); acyl-CoA thioesterase 7 (ACOT7); cytokine inducible SH2 containing protein (CISH); sequence similarity family 129 member A (FAM129A); polo-like kinase 3 (PLK3); major facilitator superfamily domain containing 12 (MFSD12); StAR-related lipid transfer domain containing 4 (STARD4); C-type lectin domain family 12 member A (CLEC12A); CD55 molecule (Cromer blood group) (CD55); interferon lambda receptor 1 (IFNLR1), or a combination thereof.

[0014] In some embodiments, the standard of care treatment comprises an anti-TNF-a antibody (e.g., ).

[0015] In some embodiments, the method of identifying a non-responder to a standard of care treatment for a disease or disorder further comprises administering an additional therapeutic agent to a subject who has been identified as a non-responder to the standard of care treatment. In certain embodiments, the additional therapeutic agent comprises an antagonistic anti-TREM-1 antibody (e.g., those disclosed herein).

[0016] Provided herein is a method of determining the efficacy of an antagonistic anti-TREM-1 antibody in treating a disease or disorder in a subject in need thereof, comprising administering the antagonistic anti-TREM-1 antibody to the subject and measuring the expression level of a TREM-1 -related gene in a sample of the subject, wherein the subject exhibits a decrease in the expression level of the TREM-1 -related gene after the administration, wherein the TREM-1 -related gene comprises nicotinamide phosphoribosyltransferase (NAMPT); Dehydrogenase / reductase 9 (DHRS9); Cyclin-dependent kinase inhibitor 1A (CDKN1A); CD52 molecule (CD52); Myotubularin-related protein 11 (MTMR11); EH domain-containing 1 (EHD1); Solute carrier family 27 member 3 (SLC27A3); Interleukin 24 (IL24); Pim-2 proto-oncogene, serine / threonine kinase (PIM2); Chitinase 3-like 1 (CHI3L1); Polypeptide N-acetylgalactosaminyltransferase 6 (GALNT6); Acyl-CoA thioesterase 7 (ACOT7); Cytokine inducible SH2 containing protein (CISH); Family with sequence similarity 129, member A (FAM129A); Polo-like kinase 3 (PLK3); Major facilitator superfamily domain containing 12 (MFSD12); StAR-related lipid transfer domain containing 4 (STARD4); C-type lectin domain family 12 member A (CLEC12A); CD55 molecule (Cromer blood group) (CD55); Interferon lambda receptor 1 (IFNLR1), or a combination thereof. In some embodiments, the subject continues treatment with the antagonistic anti-TREM-1 antibody.

[0017] In some embodiments, any of the methods disclosed herein further comprises measuring one or more scores prior to, concurrently with, or after measuring the expression level of the TREM-1 -related gene and / or administering the antagonistic anti-TREM-1 antibody, the scores comprising a baseline Mayo score, a grade 2B lamina propria neutrophil infiltration score, and a fecal calprotectin level.

[0018] In some embodiments, a subject (as applicable to any of the methods disclosed herein) exhibits one or more of an increase in baseline Mayo score, an increase in grade 2B lamina propria neutrophil infiltration score, and an increase in fecal calprotectin level prior to administration of the antagonistic anti-TREM-1 antibody.

[0019] In some embodiments, a subject exhibits an increase in baseline Mayo score of at least about 5%, at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, or more, as compared to the reference.

[0020] In some embodiments, the subject exhibits a baseline Mayo score greater than about 6, 7, 8, 9, 10, 11, or 12 prior to administration.

[0021] In some embodiments, the subject exhibits an increase in grade 2B lamina propria neutrophil infiltration score of at least about 5%, at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, or more compared to the reference.

[0022] In some embodiments, the subject exhibits a grade 2B lamina propria neutrophil infiltration score greater than about 0, about 0.1, about 0.2, or about 0.3.

[0023] In some embodiments, the subject exhibits an increase in fecal calprotectin level of at least about 5%, at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, or more compared to the reference.

[0024] In some embodiments, the subject exhibits a fecal calprotectin level (pg / g of feces) greater than about 1.5 logio, greater than about 2.0 logio, greater than about 2.5 logio, greater than about 3.0 logio, or greater than about 3.5 logio.

[0025] In some embodiments, administration of an antagonistic anti-TREM-1 antibody (e.g., those disclosed herein) reduces expression of the TREM-1 -associated gene in the subject.

[0026] In some embodiments, administration of an antagonistic anti-TREM-1 antibody reduces the baseline Mayo score, grade 2B lamina propria neutrophil infiltration score, and / or fecal calprotectin level of the subject. In certain embodiments, the baseline Mayo score is reduced by at least about 5%, at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, or more. In some embodiments, the grade 2B lamina propria neutrophil infiltration score is reduced by at least about 5%, at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, or more. In further embodiments, the fecal calprotectin level is reduced by at least about 5%, at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, or more.

[0027] In some embodiments, the expression level of the TREM-1 -related gene is increased in the presence of a natural ligand for TREM-1 but is not increased in the presence of an agonistic anti-TREM-1 antibody.

[0028] In some embodiments, the sample comprises a tissue, blood, serum, plasma, saliva, urine, or a combination thereof.

[0029] In some embodiments, the disease or disorder (as applicable to any of the methods disclosed herein) is associated with increased degranulation, reactive oxygen species formation, and / or proinflammatory cytokine release by neutrophils. In some embodiments, the disease or disorder is associated with activation of monocytes and / or increased inflammatory cytokine and chemokine production by monocytes. In certain embodiments, the disease or disorder is associated with hypoxia. In further embodiments, the disease or disorder is associated with increased cell surface TREM-1 protein expression and / or increased levels of soluble TREM-1 protein.

[0030] In some embodiments, the disease or condition (as applicable to any of the methods disclosed herein) comprises inflammatory bowel disease (IBD), Crohn's disease (CD), ulcerative colitis (UC), irritable bowel syndrome, rheumatoid arthritis (RA), psoriasis, psoriatic arthritis, systemic lupus erythematosus (SLE), lupus nephritis, vasculitis, sepsis, systemic inflammatory response syndrome (SIRS), type I diabetes, Graves' disease, multiple sclerosis (MS), autoimmune myocarditis, Kawasaki disease, coronary artery disease, chronic obstructive pulmonary disease, interstitial lung disease, autoimmune thyroiditis, scleroderma, systemic sclerosis, osteoarthritis, atopic dermatitis, vitiligo, graft versus host disease, Sjogren's syndrome, autoimmune nephritis, Goodpasture's syndrome, chronic inflammatory demyelinating polyneuropathy, allergy, asthma, other autoimmune diseases resulting from acute or chronic inflammation, chronic kidney disease, or a combination thereof. In certain embodiments, the disease or disorder is inflammatory bowel disease. In some embodiments, the inflammatory bowel disease comprises Crohn's disease and ulcerative colitis.

[0031] In some embodiments, an anti-TREM-1 antibody (e.g., those disclosed herein) comprises a heavy chain CDR1, CDR2, and CDR3 and a light chain CDR1, CDR2, and CDR3, wherein the heavy chain CDR3 comprises DMGIRRQFAY (SEQ ID NO: 19) or DMGIRRQFAY (SEQ ID NO: 19) but with one or two substitutions. In certain embodiments, the heavy chain CDR3 comprises DQGIRRQFAY (SEQ ID NO: 72).

[0032] In some embodiments, the anti-TREM-1 antibody comprises a heavy chain CDR1, CDR2, and CDR3 and a light chain CDR1, CDR2, and CDR3, wherein the heavy chain CDR2 comprises RIRTKSSNYATYYAASVKG (SEQ ID NO: 18) or RIRTKSSNYATYYAASVKG (SEQ ID NO: 18), but with one or two substitutions.

[0033] In some embodiments, the anti-TREM-1 antibody comprises a heavy chain CDR1, CDR2, and CDR3 and a light chain CDR1, CDR2, and CDR3, wherein the heavy chain CDR1 comprises TYAMH (SEQ ID NO: 17) or TYAMH (SEQ ID NO: 17), but with one or two substitutions.

[0034] In some embodiments, the anti-TREM-1 antibody comprises a heavy chain CDR1, CDR2, and CDR3 and a light chain CDR1, CDR2, and CDR3, wherein the light chain CDR1 comprises RASQSVDTFDYSFLH (SEQ ID NO: 24) or RASQSVDTFDYSFLH (SEQ ID NO: 24), but with one or two substitutions.

[0035] In some embodiments, the anti-TREM-1 antibody comprises a heavy chain CDR1, CDR2, and CDR3 and a light chain CDR1, CDR2, and CDR3, wherein the light chain CDR2 comprises RASNLES (SEQ ID NO: 21) or RASNLES (SEQ ID NO: 21), but with one or two substitutions.

[0036] In some embodiments, the anti-TREM-1 antibody comprises a heavy chain CDR1, CDR2, and CDR3 and a light chain CDR1, CDR2, and CDR3, wherein the light chain CDR3 comprises QQSNQDPYT (SEQ ID NO: 25) or QQSNQDPYT (SEQ ID NO: 25), but with one or two substitutions.

[0037] In some embodiments, the anti-TREM-1 antibody comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein the VH comprises an amino acid sequence as set forth in SEQ ID NO: 15 or 26-29, and the VL comprises an amino acid sequence as set forth in SEQ ID NO: 23.

[0038] In some embodiments, an anti-TREM-1 antibody comprises a heavy chain (HC) and a light chain (LC), wherein the HC comprises an amino acid sequence as set forth in SEQ ID NO: 30, 31, 32, or 33. In certain embodiments, the LC comprises an amino acid sequence as set forth in SEQ ID NO: 34.

[0039] In some embodiments, an anti-TREM-1 antibody comprises a heavy chain CDR1, CDR2, and CDR3 and a light chain CDR1, CDR2, and CDR3, wherein (a) the heavy chain CDR1, CDR2, and CDR3 comprise the amino acid sequences as set forth in SEQ ID NOs: 61, 62, and 63, respectively, and the light chain CDR1, CDR2, and CDR3 comprise the amino acid sequences as set forth in SEQ ID NOs: 64, 65, and 66, respectively; (b) the heavy chain CDR1, CDR2, and CDR3 comprise the amino acid sequences as set forth in SEQ ID NOs: 67, 68, and 69, respectively, and the light chain CDR1, CDR2, and CDR3 comprise the amino acid sequences as set forth in SEQ ID NOs: 70, 71, and 72, respectively; (c) the heavy chain CDR1, CDR2, and CDR3 comprise the amino acid sequences as set forth in SEQ ID NOs: 67, 68, and 69, respectively, and the light chain CDR1, CDR2, and CDR3 comprise the amino acid sequences as set forth in SEQ ID NOs: 64, 65, and 73, respectively; (d) the heavy chain CDR1, CDR2, and CDR3 comprise the amino acid sequences as set forth in SEQ ID NOs: 74, 75, and 76, respectively, and the light chain CDR1, CDR2, and CDR3 comprise the amino acid sequences as set forth in SEQ ID NOs: 70, 77, and 78, respectively; (e) the heavy chain CDR1, CDR2, and CDR3 comprise the amino acid sequences as set forth in SEQ ID NOs: 79, 80, and 81, respectively, and the light chain CDR1, CDR2, and CDR3 comprise the amino acid sequences as set forth in SEQ ID NOs: 70, 71, and 72, respectively; (f) the heavy chain CDR1, CDR2, and CDR3 comprise the amino acid sequences as set forth in SEQ ID NOs: 159, 160, and 161, respectively, and the light chain CDR1, CDR2, and CDR3 comprise the amino acid sequences as set forth in SEQ ID NOs: 70, 71, and 162, respectively; or (g) the heavy chain CDR1, CDR2, and CDR3 comprise the amino acid sequences as set forth in SEQ ID NOs: 159, 160, and 161, respectively, and the light chain CDR1, CDR2, and CDR3 comprise the amino acid sequences as set forth in SEQ ID NOs: 70, 71, and 133, respectively.

[0040] In some embodiments, the anti-TREM-1 antibody for use in the methods disclosed herein comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein the VH comprises an amino acid sequence as set forth in SEQ ID NO: 53, 55, 58, 60, or 153, and wherein the VL comprises an amino acid sequence as set forth in SEQ ID NO: 54, 56, 57, 59, 154, or 155.

[0041] In some embodiments, the anti-TREM-1 antibody further comprises a heavy chain (HC) constant region and a light chain (LC) constant region, wherein the HC constant region comprises an amino acid sequence as set forth in SEQ ID NO: 48, SEQ ID NO: 47, SEQ ID NO: 11, or SEQ ID NO: 12. In certain embodiments, the LC constant region comprises an amino acid sequence as set forth in SEQ ID NO: 35.

[0042] In some embodiments, the anti-TREM-1 antibody disclosed herein comprises a heavy chain CDR1, CDR2, and CDR3, and a light chain CDR1, CDR2, and CDR3, wherein (a) the heavy chain CDR1 comprises amino acids 31 to 35 of SEQ ID NO: 13 (TYAMH); (b) the heavy chain CDR2 comprises amino acids 50 to 68 of SEQ ID NO: 13 (RIRTKSSNYATYYAASVKG); (c) the heavy chain CDR3 comprises amino acids 101 to 110 of SEQ ID NO: 13 (DMGQRRQFAY); (d) the light chain CDR1 comprises amino acids 24 to 38 of SEQ ID NO: 14 (RASESVDTFDYSFLH); (e) the light chain CDR2 comprises amino acids 54 to 60 of SEQ ID NO: 14 (RASNLES); and / or (f) the light chain CDR3 comprises amino acids 93 to 101 of SEQ ID NO: 14 (QQSNEDPYT).

[0043] In some embodiments, the anti-TREM-1 antibody comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein the VH comprises amino acids 1 to 121 of SEQ ID NO: 13, and wherein the VL comprises amino acids 1 to 111 of SEQ ID NO: 14.

[0044] In some embodiments, the anti-TREM-1 antibody comprises a heavy chain (HC) and a light chain (LC), wherein the HC comprises an amino acid sequence as set forth in SEQ ID NO: 13, and wherein the LC comprises an amino acid sequence as set forth in SEQ ID NO: 14. BRIEF DESCRIPTION OF DRAWINGS

[0045] Figures 1A-1D A comparison of TNF-a production by human monocytes following stimulation with Peptidoglycan Recognition Protein-1 (PGLYRP1) and / or peptidoglycan (PGN) is provided. The different stimulation conditions were as follows: (i) unstimulated (“Unstim”), (ii) PGLYRP1 alone (“PGRP”), (iii) PGN alone (“PGN”), and (iv) both PGLYRP1 and PGN (“PGRP+PGN”). In Figure 1A , 1B In 1A, 1B, and 1C, the PGN was derived from Staphylococcus aureus (PGN-SA), Escherichia coli (PGN-EK), and Bacillus subtilus (PGN-BS), respectively. In Figure 1D In 1D, PGN lacking Toll-like Receptor 2 (TLR2) binding (PGN-ECndss) was used to stimulate the monocytes. In Figures 1A-1D In each of 1A-1D, the TNF-a values are provided as mean ± s.e.m.

[0046] Figure 2 A principal component analysis plot is provided showing the correlation of all monocyte samples following 24 hour stimulation under six different conditions. The stimulation conditions were as follows: (i) unstimulated (squares), (ii) PGN-ECndss alone (“PGN-EC”; circles), (iii) PGN-ECndss+PGLYRP1 (“PGN+PGRP”; triangles), (iv) PGN-ECndss+PGLYRP1+isotype antibody (“PGN+PGRP+isotype”; crosses), (v) PGN-ECndss+PGLYRP1+anti-TREM1 blocking antibody (“PGN+PGRP+Trem1”; stars), and (vi) PGLYRP1 alone (“PL”; diamonds).

[0047] Figure 3A scatter plot is provided that illustrates the specificity of genes induced upon TREM-1 ligand stimulation to the TREM-1 signaling pathway. The x-axis shows the log2 fold change in gene expression upon stimulation with PGLYRP1 + PGN-ECndss vs PGN-ECndss ("P+L vs P"). The y-axis shows the log2 fold change in gene expression upon anti-TREM-1 inhibition with anti-TREM-1 antibody vs isotype control ("P+L + aTREM vs P+L + iso"). The diagonal line represents the reference line that indicates where points should fall if they came from a population with the same distribution. Genes that are increased in expression upon PGN-ECndss + PGLYRP1 ("P+L") treatment compared to PGN-ECndss alone ("P") are shown in dark grey. Genes that are decreased in expression upon treatment with PGN-ECndss + PGLYRP1 + anti-TREM1 blocking antibody ("P+L + aTREM") compared to PGN-ECndss + PGLYRP1 + isotype antibody control ("P+L + Iso") are shown in light grey.

[0048] Figure 4A and 4B A principal component analysis plot is provided that shows the correlation of neutrophils cultured under different stimulation conditions. Figure 4A and 4B The stimulation conditions for both are as follows: (i) unstimulated (squares), (ii) PGN-ECndss alone ("PGN-EC"; circles), (iii) PGN-ECndss + PGLYRP1 ("PGN + PGRP"; triangles), (iv) PGN-ECndss + PGLYRP1 + isotype control antibody ("PGN + PGRP + isotype"; crosses), (v) PGN-ECndss + PGLYRP1 + anti-TREM1 blocking antibody ("PGN + PGRP + Trem1"; stars), and (vi) PGLYRP1 alone ("PL"; diamonds). Figure 4A Results after 6 hours of stimulation are provided. Figure 4B Results from 4 different donors are provided: D249 (squares), D254 (circles), D274 (triangles), and D299 (diamonds).

[0049] Figure 5 A comparison of the overlap of gene expression profiles in monocytes stimulated with different TREM-1 agonists is provided. Figure 5A scatter plot showing the log2 fold change of the TREM-1 natural ligand (PGLYRP1 + PGN-ECndss vs PGN-ECndss alone, x-axis) vs the log2 fold change of the agonistic anti-TREM1 antibody MAB1278 (agTREM1 vs isotype antibody). Dark grey points represent genes that changed expression level upon stimulation with the TREM-1 natural ligand. Light grey points represent genes that changed expression level upon stimulation with the agonistic anti-TREM1 antibody. The black straight line represents the linear regression of the genes that changed expression level for each of the TREM-1 agonists.

[0050] Figures 6A-6F A comparison of the levels of cytokines produced by monocytes upon TREM-1 stimulation is provided. TREM-1 expressing monocytes were stimulated with PGN alone (“PGN”) or PGN in combination with PGLYRP1 (“PGRP+PGN”). Unstimulated monocytes (“Unstim”) were used as a negative control. To confirm that the produced cytokines are specific to TREM-1 activation, some monocytes were stimulated with PGN and PGLYRP1 in combination with an antagonistic anti-TREM-1 antibody (“PGRP+PGN+anti-TREM1”). Figure 6A 、 6B Figures 6A, 6B, 6C, 6D, 6E, and 6F, respectively, show the protein levels of CCL20, IL-1 b, IL-12p40, IL-23p19, IL-6, and TNFa produced by monocytes. Figure 6D 、 6E Figures 6A, 6B, 6C, 6D, 6E, and 6F, respectively, show the protein levels of CCL20, IL-1 b, IL-12p40, IL-23p19, IL-6, and TNFa produced by monocytes. Figures 6D-6F In Figure 6F, gene expression levels are shown as fold increase compared to the expression level of unstimulated monocytes. Data are shown as mean ± s.e.m.

[0051] Figure 7 Figure 7 shows the expression of the TREM-1 gene signature in lesion (“DIS”) or non-lesion (“NOR”) colon biopsies of patients with ulcerative colitis. The TREM-1 gene signature is shown as ssGSEA score, which is a ranking-based score that summarizes the collective expression enrichment of all genes in the TREM-1 module. See Example 2. Ulcerative colitis patients are from a clinical phase 2 trial to evaluate efficacy of an anti-IP10 antibody (ClinicalTrials.gov Identifier NCT00656890). Both data are shown separately and as mean ± s.e.m. Boxplot height represents the 25% and 75% quantile and the median in the center.

[0052] Figure 8The correlation between the TREM-1 gene signature and TREM-1 mRNA expression levels in colonic biopsies of patients with ulcerative colitis ("DIS") or without ("NOR") is shown. The TREM-1 gene signature is shown as ssGSEA score, which is a rank-based score that summarizes the collective expression enrichment of all genes in the TREM-1 module. See Example 2. TREM-1 mRNA expression levels are shown on the X-axis and the TREM-1 signature score (based on the TREM 180 gene module from monocytes, see Example 2) is shown on the Y-axis. The diagonal line represents the best fit linear regression.

[0053] Figure 9 The TREM-1 gene signature in patients with ulcerative colitis who received or did not receive prior standard-of-care treatment (i.e., anti-TNF therapy or oral corticosteroid use) is shown. Data were derived from colonic biopsies of patients with ulcerative colitis in a clinical phase 2 trial to evaluate efficacy of an anti-IP10 antibody (ClinicalTrials.gov Identifier NCT00656890). Patients with a history of anti-TNF therapy were considered anti-TNF inadequate responders / non-responders ("anti-TNF IR / NR"). Patients with no anti-TNF record were considered anti-TNF naive ("anti-TNF naive"). In each of the anti-TNF therapy groups, patients who received oral corticosteroids ("YES") and patients who did not receive oral corticosteroids ("NO") are shown. The TREM-1 gene signature is shown as ssGSEA score.

[0054] Figure 10 The TREM-1 gene signature in patients with ulcerative colitis (UC) or Crohn's disease (CD) after treatment with infliximab is shown. Data were derived from a public dataset (GSE16879) of colonic biopsies from patients with inflammatory bowel disease at baseline and 4-6 weeks after treatment with infliximab compared to non-inflammatory bowel disease colonic biopsies. For each patient, the ssGSEA score was calculated, which is a rank-based score that summarizes the collective expression enrichment of all genes in the TREM-1 module (shown on the y-axis). In both the UC and CD groups, patients were classified as responders ("TNF responders") or non-responders ("TNF non-responders") to infliximab. Then, the ssGSEA scores of patients before (light gray) and after (dark gray) treatment are shown. Both data are shown separately and as mean ± s.e.m. Boxplot height represents the 25% and 75% quantile and the median in the center.

[0055] Figure 11A and 11BA heat map showing the ulcerative colitis (UC)-specific TREM-1 gene signature within each individual gene in the UC-specific TREM-1 signature from lesion biopsies from different UC patients. The Y-axis shows the individual genes and the x-axis shows each patient. The baseline partial Mayo and Geboes Global JS scores are provided for each patient. The brackets show the hierarchical clustering of the patients genes and how the patients are clustered together according to the way the 38 genes are expressed. Figure 11A A heat map showing the ulcerative colitis (UC)-specific TREM-1 gene signature within each individual gene in the UC-specific TREM-1 signature from lesion biopsies from different UC patients. The Y-axis shows the individual genes and the x-axis shows each patient. The baseline partial Mayo and Geboes Global JS scores are provided for each patient. The brackets show the hierarchical clustering of the patients genes and how the patients are clustered together according to the way the 38 genes are expressed. Figure 11B A histogram showing the distribution of the UC-specific TREM-1 gene signature score in UC patients (shown on the x-axis). The black line represents the best fit curve.

[0056] Figures 12A-12C A heat map showing the ulcerative colitis (UC)-specific TREM-1 gene signature within each individual gene in the UC-specific TREM-1 signature from lesion biopsies from different UC patients. The Y-axis shows the individual genes and the x-axis shows each patient. The baseline partial Mayo and Geboes Global JS scores are provided for each patient. The brackets show the hierarchical clustering of the patients genes and how the patients are clustered together according to the way the 38 genes are expressed. Figure 12A A comparison of the UC-specific TREM-1 signature score (y-axis) with the baseline Mayo score (x-axis) for the patients. Figure 12B A comparison of the TREM-1 signature score (y-axis) with the Geboes 2B grade: lamina propria (LP) neutrophil infiltration (a measure of the Geboes grading system) score (x-axis). For the LP neutrophil infiltration score, the scores shown are as follows: 0.0 - no increase; 0.1 - mild but definite increase; 0.2 - moderate increase; 0.3 - marked increase). Figure 12C A comparison of the TREM-1 signature score (y-axis) with the fecal calprotectin level (shown as log10, x-axis). In Figure 12A and 12C In and the diagonal represents the best fit linear regression. DETAILED DESCRIPTION

[0057] I. DEFINITIONS

[0058] For easier understanding of the present description, certain terms are first defined. Additional definitions are set forth throughout the DETAILED DESCRIPTION.

[0059] It is important to note that the term "a" or "an" entity refers to one or more than one of that entity; for example, "a nucleotide sequence" is understood to mean one or more nucleotide sequences. As such, the terms "a" (or "an"), "one or more", and "at least one" can be used interchangeably herein.

[0060] Also, as used herein, "and / or," where ever used, is to be interpreted as specific disclosure of each of the two specified features or components individually as well as a disclosure of both features or components together. For example, the terms "comprises", "comprising", "includes", "including" and the like are to be interpreted expansively and quantitatively to mean "including at least" so that when

[0061] It should be understood that wherever aspects are described herein with the wording "comprising" other aspects are also provided which are "consisting of and / or "consisting essentially of the described aspects.

[0062] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure is related. For example, the Concise Dictionary of Biomedicine and Molecular Biology, Juo, Pei-Show, 2nd ed., 2002, CRC Press; The Dictionary of Cell and Molecular Biology, 3rd ed., 1999, Academic Press; and the Oxford Dictionary Of Biochemistry And Molecular Biology, Revised, 2000, Oxford University Press, provide one of skill with a general dictionary of many of the terms used in this disclosure.

[0063] Units, prefixes, and symbols are denoted in their Systeme International de Unites (SI) accepted form. Numeric ranges are inclusive of the numbers defining the range. Unless otherwise indicated, nucleotide sequences are written left to right in 5' to 3' orientation. Amino acid sequences are written left to right in amino-to-carboxy orientation. The headings provided herein are not limitations of the various aspects of the disclosure which can be had by reference to the specification as a whole. Accordingly, the terms defined immediately below are to be given their broadest interpretation as is typical in the patent arts.

[0064] The term“about” is used herein to mean approximately, roughly, or in the range of ±10%, ±9%, ±8%, ±7%, ±6%, ±5%, ±4%, ±3%, ±2%, ±1%, or less, as long as this does not exceed the precision of the numerical value. The term“about” can modify the numerical values such as the aforementioned values more than and less than the stated values by a variance of, for example, 10%, or more or less.

[0065] The term“triggering receptor expressed on myeloid cells 1” (also known as TREM1, TREM-1, and CD354) refers to a receptor expressed on monocytes, macrophages, and neutrophils. The primary ligand for TREM-1 includes Peptidoglycan Recognition Protein 1 (PGLYRP1), which belongs to the family of Peptidoglycan (PGN)-binding proteins (PGRPs). When activated, TREM-1 associates with the ITAM-containing signaling adaptor protein DAP12. Downstream signaling can include activation of NFAT transcription factors, leading to upregulation of proinflammatory cytokine production. As used herein, the term“TREM-1” includes any variant or isoform of TREM-1.

[0066] Three isoforms of human TREM-1 have been identified. Isoform 1 (Accession No. NP_061113.1; SEQ ID NO: 1) consists of 234 amino acids and represents the canonical sequence. Isoform 2 (Accession No. NP_001229518.1; SEQ ID NO: 2) consists of 225 amino acids and differs from the canonical sequence at amino acid residues 201-234. Amino acid residues encode the transmembrane domain and a portion of the cytoplasmic domain. Isoform 3 (Accession No. NP_001229519; SEQ ID NO: 3) consists of 150 amino acids and is soluble. It lacks amino acid residues 151-234, which encode the transmembrane domain, the cytoplasmic domain, and a portion of the extracellular domain. Amino acid residues 138-150 also differ from the canonical sequence described above.

[0067] The following are the amino acid sequences of the three known isoforms of human TREM-1.

[0068] (A) Human TREM-1 Isoform 1 (Accession No. NP_061113.1; SEQ ID NO: 1; encoded by the nucleotide sequence having Accession No. NM_018643; SEQ ID NO: 4):

[0069] MRKTRLWGLLWMLFVSELRAATKLTEEKYELKEGQTLDVKCDYTLEKFASSQKAWQIIRDGEMPKTLACTERPSKNSHPVQVGRIILEDYHDHGLLRVRMVNLQVEDSGLYQCVIYQPPKEPHMLFDRIRLVVTKGFSGTPGSNENSTQNVYKIPPTTTKALCPLYTSPRTVTQAPPKSTADVSTPDSEINLTNVTDIIRVPVFNIVILLAGGFLSKSLVFSVLFAVTLRSFVP (signal sequence is underlined);

[0070] (B) Human TREM-1 isoform 2 (Accession No. NP_001229518.1; SEQ ID NO: 2; encoded by nucleotide sequence having Accession No. NM_001242589; SEQ ID NO: 5):

[0071] MRKTRLWGLLWMLFVSELRA ATKLTEEKYELKEGQTLDVKCDYTLEKFASSQKAWQIIRDGEMPKTLACTERPSKNSHPVQVGRIILEDYHDHGLLRVRMVNLQVEDSGLYQCVIYQPPKEPHMLFDRIRLVVTKGFSGTPGSNENSTQNVYKIPPTTTKALCPLYTSPRTVTQAPPKSTADVSTPDSEINLTNVTDIIRYSFQVPGPLVWTLSPLFPSLCAERM (signal sequence is underlined);

[0072] (C) Human TREM-1 isoform 3 (Accession No. NP_001229519; SEQ ID NO: 3; encoded by nucleotide sequence having Accession No. NM_001242590; SEQ ID NO: 6):

[0073] MRKTRLWGLLWMLFVSELRA ATKLTEEKYELKEGQTLDVKCDYTLEKFASSQKAWQIIRDGEMPKTLACTERPSKNSHPVQVGRIILEDYHDHGLLRVRMVNLQVEDSGLYQCVIYQPPKEPHMLFDRIRLVVTKGFRCSTLSFSWLVDS (signal sequence is underlined).

[0074] The Macaca fascicularis TREM-1 protein (Accession No. XP_001082517; SEQ ID NO: 7) is predicted to have the following amino acid sequence:

[0075] ATKLTEEKYELKEGQTLDVKCDYTLEKFASSQKAWQIIRDGEMPKTLACTERPSKNSHPVQVGRIILEDYHDHGLLRVRMVNLQVEDSGLYQCVIYQPPKEPHMLFDRIRLVVTKGFRCSTLSFSWLVDS (signal sequence is underlined).

[0074] The Macaca fascicularis TREM-1 protein (Accession No. XP_001082517; SEQ ID NO: 7) is predicted to have the following amino acid sequence:

[0075] MRKTRLWGLLWMLFVSELRA TTELTEEKYEYKEGQTLEVKCDYALEKYANSRKAWQKMEGKMPKILAKTERPSENSHPVQVGRITLEDYPDHGLLQVQMTNLQVEDSGLYQCVIYQHPKESHVLFNPICLVVTKGSSGTPGSSENSTQNVYRTPSTTAKALGPRYTSPRTVTQAPPESTVVVSTPGSEINLTNVTDIIRVPVFNIVIIVAGGFLSKSLVFSVLFAVTLRSFGP (signal sequence is underlined).

[0076] As used herein, the terms“peptidoglycan recognition protein 1” and“PGLYRP1” refer to a natural ligand of the TREM-1 protein. PGLYRP1 is a highly conserved protein of 196 amino acids in length, consisting of a signal peptide and a peptidoglycan binding domain, expressed in neutrophils and released upon activation. The amino acid sequence of PGLYRP1 (Accession No. NP_005082.1; SEQ ID NO: 8) is provided below:

[0077] MSRRSMLLAWALPSLLRLGAA QETEDPACCSPIVPRNEWKALASECAQHLSLPLRYVVVSHTAGSSCNTPASCQQQARNVQHYHMKTLGWCDVGYNFLIGEDGLVYEGRGWNFTGAHSGHLWNPMSIGISFMGNYMDRVPTPQAIRAAQGLLACGVAQGALRSNYVLKGHRDVQRTLSPGNQLYHLIQNWPHYRSP (signal sequence is underlined).

[0078] As used herein, the term“inflammatory bowel disease” or“IBD” refers to a group of disorders that cause inflammation of the intestines and / or colon, often manifested in symptoms including, but not limited to, abdominal cramping and pain, diarrhea, weight loss, and intestinal bleeding. The main forms of IBD are ulcerative colitis (UC) and Crohn’s disease (CD).

[0079] “Ulcerative colitis” is a chronic, episodic, inflammatory disease of the large intestine and rectum characterized by bloody diarrhea. Ulcerative colitis is characterized by chronic inflammation of the colonic mucosa and can be classified according to the site: “proctitis” involves only the rectum, “proctosigmoiditis” affects the rectum and sigmoid colon, “left-sided colitis” encompasses the entire left side of the large intestine, and “pancolitis” inflames the entire colon.

[0080] “Crohn’s disease” (also known as “regional enteritis”) is a chronic autoimmune disease that can affect any part of the gastrointestinal tract, but most commonly the ileum (the region where the small and large intestines meet). In contrast to ulcerative colitis, Crohn’s disease is characterized by chronic inflammation extending to all layers of the intestinal wall and involving the mesentery and regional lymph nodes. The basic pathological process is the same whether or not the small intestine or colon is involved.

[0081] The severity of IBD in a subject can be determined according to various methods known in the art, which generally rely on a combination of patient characteristics. Non-limiting examples of these methods include Disease Activity Index (DAI) / Mayo score, Geboes score, Truelove and Witts severity index, St Mark’s index, Clinical Activity Index (CAI), Activity Index (AI), Simple Clinical Colitis

[0082] As used herein, the term “Mayo score” or “Mayo scoring system” refers to a 12-point composite index consisting of input from the patient and a person (e.g., a physician) who treats the patient. See US20160324919A1 and Schroeder et al., N Engl J Med 317(26): 1625-29 (1987). Each subscore of the Mayo system ranges from 0 to 3, depending on severity. The sum of the individual subscores provides the total Mayo score. See Table 1 (below).

[0083] Table 1. Mayo Scoring System

[0084]

[0085] a “Normal” stool frequency refers to the average number of bowel movements per day when the patient is in remission.

[0086] b The physician global assessment is based on rectal bleeding, stool frequency, mucosal appearance, patient-reported abdominal pain, patient’s overall sense of well-being, and physical examination findings.

[0087] As used herein, the term Geboes score refers to a histopathological scoring system that measures disease activity based on architectural changes, chronic inflammatory infiltrate, neutrophils and eosinophils in the lamina propria, neutrophils in the epithelium, crypt destruction, erosion, and ulceration using a 6-point grading system (0-5). See WO2017095875A1 and Geboes, K. et al., Gut 47(3):404-9 (2000), which are hereby incorporated in their entireties. Higher grades indicate more severe disease activity. See Table 2 (below).

[0088] Table 2. Geboes Scoring System

[0089]

[0090]

[0091] As used herein, the term “Grade 2B lamina propria neutrophilic infiltration score” refers to a grade in the Geboes scoring system (see Table 2 above).

[0092] As used herein, the term “fecal calprotectin” refers to a biochemical measurement of the protein calprotectin in stool. Calprotectin is a member of the S100 calcium-binding protein family and exists as a heterodimer of S100A8 and S100A9 proteins. Calprotectin is primarily produced by neutrophils, and elevated levels of calprotectin have been used as a diagnostic marker for diseases such as IBD, celiac disease, infectious colitis, necrotizing enterocolitis, intestinal cystic fibrosis, and colorectal cancer. See Konikoff, M. R. et al., Inflamm Bowel Dis 12(6):524-34 (2006). In some embodiments, reference fecal calprotectin levels (in pg / g of stool) are as follows: (i) normal (≤ 50.0), (ii) borderline (50.1-120.0), and (iii) abnormal (≥ 120.1). Fecal calprotectin levels can be determined by any method known in the art (e.g., ELISA, immunofluorescence assay). See Labaere, D. et al., United European Gastroenterol J 2(1):30-37 (2014).

[0093] As used herein, the term "antibody" refers to a protein derived from a germline immunoglobulin sequence that is capable of specifically binding to an antigen (TREM-1) or a portion thereof. The term includes full-length antibodies of any class or isotype (i.e., IgA, IgE, IgG, IgM, and / or IgY), as well as any single chains or fragments thereof. An antibody that specifically binds to an antigen or a portion thereof can bind exclusively to that antigen or portion thereof, or it can bind to a limited number of cognate antigens or portions thereof. Full-length antibodies typically comprise at least four polypeptide chains: two heavy (H) chains and two light (L) chains interconnected by disulfide bonds. One immunoglobulin subclass of particular pharmaceutical interest is the IgG family. In humans, the IgG class can be subdivided into 4 subclasses based on the sequence of their heavy chain constant regions: IgGl, IgG2, IgG3, and IgG4. The light chains can be classified into two types, kappa and lambda, based on their sequence composition differences. IgG molecules are composed of two heavy chains and two light chains, the heavy chains being interconnected by two or more disulfide bonds, each light chain being linked to a heavy chain by a disulfide bond. The heavy chains can comprise a heavy chain variable region (VH) and up to three heavy chain constant (CH) regions: CHI, CH2, and CH3. The light chains can comprise a light chain variable region (VL) and a light chain constant region (CL). The VHand VLregions can be further subdivided into regions of hypervariability, termed complementarity determining regions (CDRs), interspersed with regions that are more conserved, termed framework regions (FRs). The VHand VLregions typically comprise three CDRs and four FRs, arranged from amino-terminus to carboxy-terminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. The hypervariable regions of the heavy and light chains form a binding domain that is capable of interacting with an antigen, while the constant regions of the antibodies can mediate the binding of the immunoglobulin to host tissues or factors, including, but not limited to, various cells of the immune system (effector cells), Fc receptors, and the first component (Clq) of the classical complement system. The antibodies of the present invention can be isolated. The term "isolated antibody" refers to an antibody that has been separated and / or recovered from one or more other components of the environment in which the antibody is produced, and / or has been purified from a mixture of components present in the environment in which the antibody is produced. Certain antigen-binding fragments of antibodies can be suitable for use in the context of the present invention, as it has been shown that the antigen-binding function of an antibody can be performed by fragments of a full-length antibody.

[0094] As described herein, the term "antigen-binding portion" of an antibody refers to one or more fragments of an antibody that retain the ability to specifically bind to an antigen, such as TREM-1. Examples of antigen-binding fragments include Fab, Fab', F(ab)2, F(ab')2, F(ab)S, Fv (typically the VL and VH domains of a single arm of an antibody), single chain Fv (scFv; see, e.g., Bird et al., Science 242: 42S-426 (1988); Huston et al., PNAS 85: 5879-5883 (1988)), dsFv, Fd (typically the VH and CHI domains), and dAb (typically the VH domain) fragments; VH, VL, VhH, and V-NAR domains; monovalent molecules comprising a single VH and a single VL chain; minibodies, diabodies, triabodies, tetrabodies, and kappa bodies (see, e.g., Ill et al., Protein Eng 10: 949-57 (1997)); camel IgG; IgNAR; and one or more isolated CDRs or paratopes, where the isolated CDRs or antigen-binding residues or polypeptides can be bound or linked together to form a functional antibody fragment. Various types of antibody fragments have been described or reviewed in, e.g., Holliger and Hudson, Nat Biotechnol 23: 1126-1136 (2005); International Publication No. WO 2005 / 040219 and U.S. Publication Nos. 2005 / 0238646 and 2002 / 0161201. These antibody fragments can be obtained using conventional techniques known to those with skill in the art, and the fragments can be screened as described herein for use in the same manner as whole antibodies.

[0095] A "human" antibody (HuMAb) refers to an antibody having variable regions in which both the framework and CDR regions are derived from human germline immunoglobulin sequences. Furthermore, if the antibody contains a constant region, the constant region also is derived from human germline immunoglobulin sequences. The anti-TREM-1 antibodies described herein can include amino acid residues not encoded by human germline immunoglobulin sequences (e.g., mutations introduced by random or site-specific mutagenesis in vitro or by somatic mutation in vivo). However, the term "human antibody", as used herein, is not intended to include antibodies in which CDR sequences derived from the germline of another mammalian species, such as a mouse, have been grafted onto human framework sequences. The terms "human" and "fully human" antibodies are used synonymously.

[0096] A "humanized" antibody refers to a human / non-human chimeric antibody that contains one or more sequences derived from a non-human immunoglobulin (CDR regions or portions thereof). Thus, a humanized antibody is a human immunoglobulin (recipient antibody) in which at least residues from a hypervariable region of the recipient are replaced by residues from a hypervariable region of an antibody (donor antibody) of a different species (such as mouse, rat, rabbit, or non-human primate) such that the desired specificity, affinity, sequence composition, and function are achieved. In some instances, FR residues of the human immunoglobulin are replaced by corresponding non-human residues. One example of such a modification is the introduction of one or more so-called back mutations, which are typically amino acid residues derived from the donor antibody. Humanization of antibodies can be performed using recombinant techniques known to those of skill in the art (see, e.g., Antibody Engineering, Methods in Molecular Biology, Vol. 248, Benny K.C. Lo, ed.). Suitable human recipient frameworks for both light and heavy chain variable domains can be identified, for example, by sequence or structural homology. Alternatively, fixed recipient frameworks can be used, for example, based on knowledge of structural, biophysical, and biochemical properties. The recipient framework can be germlined or derived from a mature antibody sequence. CDR regions from the donor antibody can be transferred by CDR grafting. CDR-grafted humanized antibodies can be further optimized, for example, for affinity, function, and biophysical properties, by identifying key framework positions where re-introduction of amino acid residues from the donor antibody (back mutations) have a beneficial effect on the properties of the humanized antibody. In addition to back mutations from the donor antibody, the humanized antibody can be engineered by introducing germline residues in the CDR or framework regions, eliminating immunogenic epitopes, site-directed mutagenesis, affinity maturation, and the like.

[0097] Furthermore, humanized antibodies can comprise residues that are not found in the recipient antibody or in the donor antibody. These modifications are made to further improve antibody performance. In general, a humanized antibody will comprise substantially all of at least one (and typically two) variable domains, in which all or substantially all of the CDR regions correspond to those of a non-human immunoglobulin and all or substantially all of the FR residues are those of a human immunoglobulin sequence. The humanized antibody optionally also can comprise at least a portion of an immunoglobulin constant region (Fc), typically that of a human immunoglobulin. The term "humanized antibody derivative" refers to any modified form of a humanized antibody, such as a conjugate of the antibody with another agent or antibody.

[0098] As used herein, the term "recombinant human antibody" includes all human antibodies that are prepared, expressed, created or isolated by recombinant means, such as (a) antibodies isolated from an animal (e.g., a mouse) that is transgenic or transchromosomal for human immunoglobulin genes or a hybridoma prepared therefrom, (b) antibodies isolated from a host cell transformed to express the antibody, e.g., from a transfectoma, (c) antibodies isolated from a recombinant, combinatorial human antibody library and (d) antibodies prepared, expressed, created or isolated by any other means that involve splicing of human immunoglobulin gene sequences to other DNA sequences. Such recombinant human antibodies comprise variable and constant regions, which are of specific human germline immunoglobulin sequences, but which contain subsequent rearrangements and mutations that occur, for example, during maturation of the antibody. As is known in the art (see, e.g., Lonberg Nature Biotech. 23(9): 1117-1125 (2005)), the variable region contains the antigen binding domain, which is encoded by various genes that rearrange to form a specific antibody to a foreign antigen. In addition to rearrangement, the variable region can be further modified by changes in individual amino acids (termed somatic mutation or hypermutation), which increase the affinity of the antibody to the foreign antigen. The constant region will change in response to the antigen (i.e., isotype switching). Thus, the rearranged and somatically mutated nucleic acid molecules encoding the light and heavy chain immunoglobulin polypeptides in response to an antigen cannot have sequence identity with the original nucleic acid molecules, but are substantially identical or similar (i.e., at least 80% identical).

[0099] A "chimeric antibody" refers to an antibody in which the variable region is derived from one species and the constant region is derived from another species, such as an antibody in which the variable region is derived from a mouse antibody and the constant region is derived from a human antibody.

[0100] As used herein, "isotype" refers to the antibody class (e.g., IgGl, IgG2, IgG3, IgG4, IgM, IgAl, IgA2, IgD, and IgE antibodies) that is encoded by heavy chain constant region genes.

[0101] “Allotype” refers to naturally occurring variants within a particular isotype group that differ by several amino acids (see, e.g., Jefferis et al., mAbs 1 : 1 (2009)). The anti-TREM-1 antibodies described herein can be any allotype. In some embodiments, the anti-TREM-1 antibodies of the present disclosure belong to the “IgG1.3f” allotype, which comprises one or more amino acid substitutions selected from the group consisting of L234A, L235E, and G237A according to EU numbering, as compared to a wild-type IgG1 isotype (e.g., SEQ ID NO: 9). In other embodiments, the anti-TREM-1 belongs to the “IgG1.1f” allotype, which comprises one or more amino acid substitutions selected from the group consisting of L234A, L235E, G237A, A330S, and P331S according to EU numbering, as compared to a wild-type IgG1 isotype (e.g., SEQ ID NO: 9). In further embodiments, the anti-TREM-1 antibodies disclosed herein belong to the “IgG1-Aba” allotype, which comprises one or more amino acid substitutions selected from the group consisting of K214R, C226S, C229S, and P238S according to EU numbering, as compared to a wild-type IgG1 isotype (e.g., SEQ ID NO: 9). In some embodiments, the anti-TREM-1 antibodies disclosed herein belong to the “IgG4-Aba” allotype, which comprises the CH1 domain of a wild-type IgG4 isotype (e.g., SEQ ID NO: 10) and the CH2 and CH3 domains of IgG1. In certain embodiments, the IgG4-Aba allotype antibody comprises one or more amino acid substitutions selected from the group consisting of S131C, K133R, G137E, G138S, Q196K, I199T, N203D, K214R, C226S, C229S, and P238S according to EU numbering, as compared to a wild-type IgG1 isotype (e.g., SEQ ID NO: 9).

[0102] The phrases“an antibody recognizing an antigen” and“an antibody specific for an antigen” are used interchangeably herein with the term“an antibody that specifically binds to an antigen.”

[0103] As used herein, “isolated antibody” is intended to refer to an antibody that has been separated and / or recovered from one or more other components of the environment in which the antibody is produced, and / or has been purified from a mixture of components present in the environment in which the antibody is produced.

[0104] “Effector function” refers to the interaction of an antibody Fc region with an Fc receptor or ligand, or the biochemical event that results therefrom. Exemplary “effector functions” include Clq binding, complement dependent cytotoxicity (CDC), Fc receptor binding, FcyR-mediated effector functions such as ADCC and antibody-dependent cell-mediated phagocytosis (ADCP), and down regulation of cell surface receptors (e.g., B cell receptor; BCR). Such effector functions generally require the combination of an Fc region with a binding domain (e.g., an antibody variable domain). In one embodiment, the anti-TREM-1 antibodies of the present disclosure comprise an Fc region that does not bind to one or more FcyRs and thus lack effector function (i.e., an effectorless).

[0105] “Fc receptor” or “FcR” is a receptor that binds to the Fc region of an immunoglobulin. FcRs that bind to IgG antibodies include the receptors of the FcyR family, including allelic variants and alternatively spliced forms of these receptors. The FcyR family is composed of three activating (Fc gamma Rl, Fc gamma RIII, and Fc gamma RIV in mice; Fc gamma RIA, Fc gamma RIIA, and Fc gamma RIIIA in humans) and one inhibitory (Fc gamma RIB) receptor. Various properties of human FcyRs are known in the art. Most innate effector cell types co-express one or more activating FcyRs and the inhibitory FcyRIB, whereas natural killer (NK) cells selectively express one activating Fc receptor (Fc gamma RIII in mice and Fc gamma RIIIA in humans) but not the inhibitory FcyRIB in both mice and humans. Human IgGl binds to most human Fc receptors and is considered equivalent to murine IgG2a in terms of the type of activating Fc receptor it binds to.

[0106] “Fc region” (fragment, crystallizable region) or “Fc domain” or “Fc” refers to the C-terminal region of the heavy chain of an antibody that mediates the binding of the immunoglobulin to host tissues or factors, including binding to Fc receptors (FcRs) located on various cells of the immune system (e.g., effector cells) or to the first component (Clq) of the classical complement system. Thus, the Fc region includes the constant region of an antibody except the first constant region immunoglobulin domain (e.g., CHI or CL).

[0107] In IgG, the Fc region comprises the immunoglobulin domains CH2 and CH3 and the hinge between the CH1 and CH2 domains. While the definition of the boundaries of the Fc region of an immunoglobulin heavy chain might vary slightly, as defined herein, the human IgG heavy chain Fc region is defined to stretch from an amino acid residue D221 of IgG1, V222 of IgG2, L221 of IgG3, and P224 of IgG4, to the carboxy-terminus of the heavy chain, wherein the numbering is according to the EU index as in Kabat. The CH2 domain of a human IgG Fc region stretches from amino acid 237 to amino acid 340, and the CH3 domain is located C-terminally to the CH2 domain in the Fc region, i.e. it stretches from amino acid 341 to amino acid 447 or 446 (if the C-terminal lysine residue is not present) or 445 (if the C-terminal glycine and lysine residues are not present) of IgG. As used herein, the Fc region can be a native sequence Fc, including any allotype variant, or a variant Fc (e.g., non-naturally occurring Fc). Fc can also refer to this region in isolation or in the context of an Fc-containing protein polypeptide, such as a "binding protein comprising an Fc region", also referred to as an "Fc fusion protein" (e.g., an antibody or immunoadhesion).

[0108] A "native sequence Fc region" or "native sequence Fc" comprises an amino acid sequence that is identical to the amino acid sequence of a Fc region found in nature. Native sequence human Fc regions include a native sequence human IgGl Fc region; a native sequence human IgG2 Fc region; a native sequence human IgG3 Fc region; and a native sequence human IgG4 Fc region as well as naturally occurring variants thereof. Native sequence Fc includes Fc of various allotypes (see, e.g., Jefferis et al., mAbs 1 : 1 (2009)).

[0109] A "variant sequence Fc region" or "non-naturally occurring Fc" comprises modifications that generally alter one or more functional properties thereof, such as serum half-life, complement fixation, Fc receptor binding, protein stability, and / or antigen-dependent cellular cytotoxicity, or lack of these properties, among others. In some embodiments, the anti-TREM-1 antibodies of the present disclosure can be chemically modified (e.g., one or more chemical moieties can be attached to the antibody) or modified to alter their glycosylation, additionally altering one or more functional properties of the antibody. In one embodiment, the anti-TREM-1 antibody is of the IgGl isotype and carries a modified Fc domain comprising one or more and possibly all of the following mutations that will result in reduced affinity for certain Fc receptors (L234A, L235E, and G237A) and reduced Clq-mediated complement fixation (A330S and P331S) (residues numbered according to EU index).

[0110] The terms "hinge," "hinge domain," "hinge region," and "antibody hinge region" refer to the domain that connects the CH1 domain to the heavy chain constant region of the CH2 domain, and includes the upper, middle, and lower portions of the hinge (Roux et al., J Immunol 161 :4083 (1998)). The hinge provides varying degrees of flexibility between the binding and effector regions of an antibody, and also provides a site for intermolecular disulfide bonding between the two heavy chain constant regions. As used herein, the hinge begins at Glu216 and ends at Gly237 of all IgG isotypes (Roux et al., J Immunol 161 :4083 (1998)). The sequences of wild-type IgGl, IgG2, IgG3, and IgG4 hinges are known in the art (e.g., International PCT Publication No. WO 2017 / 087678). In one embodiment, the hinge region of CH1 of an anti-TREM-1 antibody is modified such that the number of cysteine residues in the hinge region is altered, e.g., increased or decreased. This approach is further described, for example, in U.S. Patent No. 5,677,425.

[0111] The constant region can be modified to stabilize the antibody, for example, to reduce the risk of a bivalent antibody being split into two monovalent VH-VL fragments. For example, in an IgG4 constant region, residue S228 (residue numbered according to the EU index) can be mutated to a proline (P) residue to stabilize the inter-heavy chain disulfide bridge formation at the hinge (see, e.g., Angal et al., Mol Immunol. 30:105-8 (1995)). Antibodies or fragments thereof can also be defined in terms of their complementarity determining regions (CDRs). As used herein, the term “complementarity determining region” or “hypervariable region” refers to a region of an antibody that specifies the amino acid residues involved in antigen binding. The regions of hypervariable or CDRs can be identified as regions of highest variability in an amino acid alignment of antibody variable domains. Databases can be used for CDR identification, such as the Kabat database, CDRs are defined, for example, as comprising amino acid residues 24-34 (CDR1), 50-59 (CDR2), and 89-97 (CDR3) in the light chain variable domain and 31-35 (CDR1), 50-65 (CDR2), and 95-102 (CDR3) in the heavy chain variable domain; (Kabat et al., 1991; Sequences of Proteins of Immunological Interest, Fifth Edition, U.S. Department of Health and Human Services, NIH Publication No. 91-3242) or, CDRs can be defined as those residues from “hypervariable loops” (residues 26-33 (LI), 50-52 (L2), and 91-96 (L3) in the light chain variable domain and 26-32 (HI), 53-55 (H2), and 96-101 (H3) in the heavy chain variable domain) (Chothia and Lesk, J. Mol. Biol 196:901-917 (1987)). Generally, the numbering of amino acid residues in this region is performed by the method described in Kabat et al., supra. Phrases such as “Kabat position,” “Kabat residue,” and “according to Kabat” refer herein to this numbering system of the heavy chain variable domain or light chain variable domain. Using the Kabat numbering system, the actual linear amino acid sequence of a peptide can contain fewer or additional amino acids corresponding to a shortening of, or insertion into, a framework (FR) or CDR of the variable domain. For example, a heavy chain variable domain can include amino acid insertions following residue 52 of CDR H2 (residues 52a, 52b, and 52c according to Kabat) and insertions of residues following heavy chain FR residue 82 (e.g., residues 82a, 82b, and 82c, etc. according to Kabat). The Kabat numbering of residues in a given antibody can be determined by alignment of the “standard” Kabat numbered sequences with the sequence of the antibody.

[0112] The term "epitope" or "antigenic determinant" refers to the site on an antigen, e.g., TREM-1, to which an immunoglobulin or antibody binds specifically, e.g., as defined by the particular method used to identify it. Epitopes can be formed both by contiguous amino acids (usually linear epitopes) or by amino acids that are disposed contiguously in three-dimensional space (usually conformational epitopes). Epitopes formed from contiguous amino acids are typically but not always maintained in the same spatial conformation after denaturing solvent treatment, while epitopes formed by amino acids that are disposed contiguously in three-dimensional space can not be maintained in the same spatial conformation after denaturing solvent treatment. Epitopes typically include at least 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 amino acids in a unique spatial conformation. Methods of determining the epitope bound by a given antibody (i.e., epitope mapping) are well known in the art and include, e.g., immunoblotting and immunoprecipitation assays in which overlapping or contiguous peptides from, e.g., TREM-1, are tested for reactivity with a given antibody, e.g., an anti-TREM-1 antibody. Methods of determining the spatial conformation of an epitope include techniques in the art and described herein, e.g., X-ray crystallography, antigenic mutation analysis, 2-dimensional nuclear magnetic resonance, and HDX-MS (see, e.g., Epitope Mapping Protocols in Methods in Molecular Biology, Vol. 66, G. E. Morris, ed. (1996)).

[0113] The term "bind to the same epitope" with respect to two or more antibodies means that the antibodies bind to the same segment of amino acid residues, as determined by a given method. Techniques for determining whether antibodies bind to the "same epitope on TREM-1" of the antibodies described herein include, e.g., epitope mapping methods such as crystal X-ray analysis of antigen: antibody complexes, which provides atomic resolution of the epitope, and hydrogen / deuterium exchange mass spectrometry (HDX-MS). Other methods monitor binding of antibodies to antigen fragments or antigenic mutant variants, where loss of binding due to modification of an amino acid residue within the antigen sequence is generally taken as an indication of an epitope component. In addition, computational combinatorial methods for epitope mapping can also be used. These methods rely on the ability of an antibody of interest to affinity isolate a particular short peptide from a combinatorial phage display peptide library. Antibodies having the same VH and VL or the same CDR1, 2, and 3 sequences are expected to bind to the same epitope.

[0114] An antibody that "competes with another antibody for binding to a target" is an antibody that inhibits (partially or fully) binding of the other antibody to the target. Whether two antibodies compete with each other for binding to a target, i.e., whether one antibody inhibits binding of the other antibody to the target and to what extent, can be determined using known competition experiments, e.g., Surface plasmon resonance (SPR) analysis. In certain embodiments, an antibody competes with and inhibits the binding of another antibody to a target by at least 50%, 60%, 70%, 80%, 90%, or 100%. The level of inhibition or competition can vary depending on which antibody is the "blocking antibody" (i.e., the cold antibody that is incubated first with the target). Competition assays can be performed as described in, for example, Ed Harlow and David Lane, Cold Spring Harb Protoc; 2006; doi: 10.1101 / pdb.prot4277 or Chapter 11 of "Using Antibodies", Ed Harlow and David Lane, eds., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, USA 1999. Two antibodies "cross-compete" if they block each other by at least 50%, i.e., no matter which antibody is contacted first with the antigen in the competition experiment.

[0115] As used herein, the terms "specifically binds," "selectively binds," "selectively binds," and "binds specifically" mean that an antibody binds to an epitope on a predetermined antigen. Typically, (i) an antibody binds to a predetermined antigen with an affinity that is at least two-fold greater than its affinity for binding to a non-specific antigen (e.g., BSA, casein) (other than the predetermined antigen or a closely related antigen), and (ii) an antibody binds to the predetermined antigen with an equilibrium dissociation constant (KD) of about less than 10 2000 instrument using a predetermined antigen (e.g., recombinant human TREM-1) as the analyte and the antibody as the ligand, or a Scatchard analysis of the binding of the antibody to antigen-positive cells. - 7 M, 10 -8 M, 10 -10 M or even lower, and (ii) an antibody binds to the predetermined antigen with an affinity that is at least two-fold greater than its affinity for binding to a non-specific antigen (e.g., BSA, casein) (other than the predetermined antigen or a closely related antigen). Thus, an antibody that "binds specifically to human TREM-1" refers to an antibody that binds to soluble or human TREM-1 with a K -7 M or less, such as about less than 10 -8 M, 10 -9 M or 10 -10 M or even lower, and (ii) an antibody binds to the predetermined antigen with an affinity that is at least two-fold greater than its affinity for binding to a non-specific antigen (e.g., BSA, casein) (other than the predetermined antigen or a closely related antigen). Thus, an antibody that "binds specifically to human TREM-1" refers to an antibody that binds to soluble or human TREM-1 with a K D M or less, such as about less than 10 -7 M, 10 -8 M or 10 -9 M or 10 -10 M or even lower, and (ii) an antibody binds to the predetermined antigen with an affinity that is at least two-fold greater than its affinity for binding to a non-specific antigen (e.g., BSA, casein) (other than the predetermined antigen or a closely related antigen). Thus, an antibody that "binds specifically to human TREM-1" refers to an antibody that binds to soluble or human TREM-1 with a KD Antibodies that bind to cynomolgus TREM-1. In certain embodiments, such antibodies that do not cross-react with TREM-1 from non-human species exhibit essentially no detectable binding to these proteins in standard binding assays.

[0116] The term "anti-TREM-1 antibody", as used herein, refers to an antibody (including fragments thereof) that specifically binds to human TREM-1. Unless indicated otherwise, the anti-TREM-1 antibodies disclosed herein are antagonistic antibodies (i.e., inhibit or suppress the activity of TREM-1 on cells (e.g., monocytes, macrophages or neutrophils) (i.e., do not agonize when bound)).

[0117] The term "binding specificity" refers herein to the interaction of a molecule, such as an antibody or fragment thereof, with a single, specific antigen or a limited number of highly homologous antigens (or epitopes). In contrast, an antibody that is capable of specifically binding to TREM-1 is not capable of binding to a different molecule. An antibody according to the application can not be capable of binding to the natural killer cell p44-related protein Nkp44.

[0118] The specificity of the interaction and the value of the equilibrium binding constant can be determined directly by well-known methods. Standard assays to assess the ability of a ligand, such as an antibody, to bind its target are known in the art and include, for example, ELISA, Western blot, RIA, and flow cytometry. The binding kinetics and binding affinity of an antibody can also be assessed by standard assays known in the art, such as SPR.

[0119] A competitive binding assay to determine whether two antibodies are competing for binding or cross-competing for binding includes: competing for binding to bone marrow cells expressing TREM-1, for example, as determined by flow cytometry such as described in the Examples. Other methods include: SPR (e.g., Biacore®), BIAcore®, and Biacore® analysis. ), solid phase direct or indirect radioimmunoassay (RIA), solid phase direct or indirect enzyme immunoassay (EIA), sandwich competition assay (see Stahli et al., Methods in Enzymology 9:242 (1983)); solid phase direct biotin-streptavidin EIA (see Kirkland et al., J. Immunol. 137:3614 (1986)); solid phase direct labeled assay, solid phase direct labeled sandwich assay (see Harlow and Lane, Antibodies: A Laboratory Manual, Cold Spring Harbor Press (1988)); solid phase direct labeled RIA using 1-125 labeling (see Morel et al., Mol. Immunol. 25(1):7 (1988)); solid phase direct biotin-streptavidin EIA (Cheung et al., Virology 176:546 (1990)); and direct labeled RIA. (Moldenhauer et al., Scand. J. Immunol. 32:77 (1990)).

[0120] As used herein, the term "bin" is defined using reference antibodies. A second antibody is said to belong to the same "bin" as a reference antibody if the second antibody cannot bind to the antigen simultaneously with the reference antibody. In this case, the reference antibody and the second antibody compete for binding to the same part of the antigen and are referred to as "competing antibodies." A second antibody is said to belong to a separate "bin" if the second antibody can bind to the antigen simultaneously with the reference antibody. In this case, the reference antibody and the second antibody do not compete for binding to the same part of the antigen and are referred to as "non-competing antibodies."

[0121] The antibody "bin" does not provide direct information about the epitope. Competing antibodies, i.e., antibodies belonging to the same "bin," can have the same epitope, overlapping epitopes, or even separate epitopes. The latter is the case if the reference antibody bound to an epitope on the antigen occupies the space needed for a second antibody to contact an epitope on its antigen ("steric hindrance"). Non-competing antibodies typically have separate epitopes.

[0122] The term "binding affinity" refers herein to a measure of the strength of the non-covalent interaction between two molecules, e.g., an antibody or fragment thereof, and an antigen. The term "binding affinity" is used to describe monovalent interactions (intrinsic activity).

[0123] The binding affinity of a monovalent interaction between two molecules, e.g., an antibody or fragment thereof, and an antigen, can be quantified by determining the equilibrium dissociation constant (K D ). In turn, K DThe association and dissociation rate constants corresponding to the association and dissociation of the monovalent complex can be determined by measuring the kinetics of the complex formation and dissociation, for example, using the SPR method. The rate constants corresponding to the association and dissociation of the monovalent complex are called the association rate constants k. a (or k) on and dissociation rate constant k d (or k) 0ff K D Through equation K D =k d / k a With k a and k d Related. According to the above definition, binding affinity related to different molecular interactions, such as comparing the binding affinity of different antibodies for a given antigen, can be achieved by comparing the K0 of individual antibody / antigen complexes. D Use values ​​to compare.

[0124] As used in this article, the term "high affinity" for IgG antibodies refers to an affinity of 10 for the target antigen. -8 M or smaller, 10 - 9 M or smaller or 10 -10 M or smaller K D Antibodies. However, "high affinity" binding can vary depending on the antibody isotype. For example, "high affinity" binding to the IgM isotype refers to an antibody having a binding affinity of 10... -10 M or smaller, or 10 -8 M or smaller K D .

[0125] In the context of in vitro or in vivo assays using antibodies or their antigen-binding fragments, the term "EC50" refers to the concentration of the antibody or its antigen-binding fragment that induces a response, which is 50% of the maximum response, i.e., the midpoint between the maximum response and the baseline.

[0126] As used herein, the term "naturally occurring" refers to the fact that an object can exist in nature. For example, polypeptide or polynucleotide sequences that exist in organisms (including viruses), can be isolated from natural sources, and have not been intentionally modified artificially in a laboratory are naturally occurring.

[0127] A polypeptide is a chain containing at least two consecutively linked amino acid residues, with no upper limit on chain length. One or more amino acid residues in a protein may contain modifications, such as, but not limited to, glycosylation, phosphorylation, or disulfide bond formation. A protein may contain one or more polypeptides.

[0128] As used herein, the term "nucleic acid molecule" is intended to include both DNA and RNA molecules. Nucleic acid molecules can be single-stranded or double-stranded and can be cDNA.

[0129] A "conservative amino acid substitution" refers to the replacement of an amino acid residue by another residue having similar side chain. Families of amino acid residues having similar side chains have been defined in the art. These families include amino acids with basic side chains (e.g., lysine, arginine, histidine), acidic side chains (e.g., aspartic acid, glutamic acid), uncharged polar side chains (e.g., glycine, asparagine, glutamine, serine, threonine, tyrosine, cysteine, tryptophan), nonpolar side chains (e.g., alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine), beta-branched side chains (e.g., threonine, valine, isoleucine), and aromatic side chains (e.g., tyrosine, phenylalanine, tryptophan, histidine). In certain embodiments, a predicted non-essential amino acid residue in an anti-TREM-1 antibody is replaced with another amino acid residue from the same side chain family. Methods of identifying nucleotide and amino acid conservative substitutions that do not eliminate antigen binding are well known in the art (see, e.g., Brummell et al., Biochem. 32:1180-1187 (1993); Kobayashi et al., Protein Eng. 12(10):879-884 (1999); and Burks et al., Proc. Natl. Acad. Sci. USA 94:412-417 (1997)).

[0130] For nucleic acids, the term "substantial homology" means that two nucleic acids, or specified sequences thereof, are identical with, or have a specified percentage of, nucleotides in optimally aligning and comparing sequences, with appropriate gaps and allowances for nucleotide insertions or deletions. Alternatively, substantial homology exists when a fragment of a selected sequence hybridizes to the complement of a strand under selective hybridization conditions.

[0131] For polypeptides, the term "substantial homology" means that two polypeptides, or specified sequences thereof, are identical with, or have a specified percentage of, amino acids in optimally aligning and comparing sequences, with appropriate gaps and allowances for amino acid insertions or deletions.

[0132] The percent identity between two sequences is the number of identical positions shared by the sequences (i.e., % homology = # identical positions / total positions x 100), taking into account the number and length of gaps introduced to achieve optimal alignment of the two sequences. The comparison of sequences and determination of percent identity between two sequences can be accomplished using a mathematical algorithm, as described in the non-limiting examples below.

[0133] The percent identity between two nucleotide sequences can be determined using the GAP program in the GCG software package (available at worldwideweb.gcg.com), using a NWSgapdna.CMP matrix and a 40, 50, 60, 70, or 80 penalty for gap creation and a 1, 2, 3, 4, 5, or 6 penalty for gap extension. The percent identity between two amino acid sequences can be determined using the GAP program in the GCG software package (available at worldwideweb.gcg.com), using a PAM120 weight residue table, a gap creation penalty of 12, and a gap extension penalty of 4. In addition, the percent identity between two amino acid sequences can be determined using the Needleman and Wunsch (J. Mol. Biol. (48):444-453 (1970)) algorithm which has been incorporated into the GAP program in the GCG software package (available at worldwideweb.gcg.com), using either a Blossum 62 matrix or a PAM250 matrix, and a gap penalty of 16, 14, 12, 10, 8, 6, or 4, and the gap length penalty of 1, 2, 3, 4, 5, or 6.

[0134] The nucleic acid and protein sequences described herein can further be used as a "query sequence" to perform a search against public databases to, e.g., identify related sequences. Such searches can be performed using the NBLAST and XBLAST programs (version 2.0) of Altschul et al. (1990) J. Mol. Biol. 215:403-10. BLAST nucleotide searches can be performed with the NBLAST program, score = 100, wordlength = 12 to obtain nucleotide sequences homologous to nucleic acid molecules described herein. BLAST protein searches can be performed with the XBLAST program, score = 50, wordlength = 3 to obtain amino acid sequences homologous to protein molecules described herein. To obtain gapped alignments for comparison purposes, Gapped BLAST (Altschul et al., (1997) Nucleic Acids Res. 25(17):3389-3402) can be utilized as described below. When utilizing BLAST and Gapped BLAST programs, the default parameters of the respective programs (e.g., XBLAST and NBLAST) can be used. See worldwideweb.ncbi.nlm.nih.gov.

[0135] Nucleic acids can be present in intact cells, in a cell lysate, or in a partially purified or substantially pure form. A nucleic acid is "isolated" or "rendered substantially pure" when purified away from other cellular components or other contaminants, such as other cellular nucleic acids (e.g., other portions of a chromosome) or proteins, by standard techniques, including alkaline / SDS treatment, CsCl banding, column chromatography, agarose gel electrophoresis, and others well known in the art. See, F. Ausubel et al., eds. Current Protocols in Molecular Biology, Greene Publishing and Wiley Interscience, New York (1987).

[0136] Nucleic acids, such as cDNAs, can be mutated according to standard techniques to provide gene sequences. For coding sequences, these mutations can affect the amino acid sequence as desired. In particular, DNA sequences that are substantially homologous to or derived from (where "derived" means identical to or modified from another sequence) the native V, D, J, constant, switch, and other such sequences described herein are contemplated.

[0137] As used herein, the term "vector" is intended to refer to a nucleic acid molecule capable of transporting another nucleic acid to which it has been linked. One type of vector is a "plasmid", which refers to a circular double stranded DNA loop into which additional DNA segments can be ligated. Another type of vector is a viral vector, wherein additional DNA segments can be ligated into the viral genome. Certain vectors are capable of autonomous replication in a host cell into which they are introduced (e.g., bacterial vectors having a bacterial origin of replication and episomal mammalian vectors). Other vectors (e.g., non-episomal mammalian vectors) can be integrated into the genome of a host cell upon introduction into the host cell, and thereby are replicated along with the host genome. Moreover, certain vectors are capable of directing the expression of genes to which they are operatively linked. Such vectors are referred to herein as "recombinant expression vectors" (or simply, "expression vectors"). In general, expression vectors of utility in recombinant DNA technologies are often in the form of plasmids. In this context, the term "plasmid" and "vector" can be used interchangeably, as the plasmid is the most commonly used form of vector. However, other forms of expression vectors are also included, such as viral vectors (e.g., replication defective retroviruses, adenoviruses, and adeno-associated viruses), which serve equivalent functions.

[0138] As used herein, the term "recombinant host cell" (or simply "host cell") is intended to refer to a cell that comprises a nucleic acid that does not naturally occur in the cell, and can be a cell into which a recombinant expression vector has been introduced. It should be understood that such terms are intended to refer not only to the particular subject cell but to the progeny of such a cell. Because certain modifications can occur in succeeding generations due to either mutation or environmental influences, such progeny can not, in fact, be identical to the parent cell, but are still included within the scope of the term "host cell" as used herein.

[0139] As used herein, the term "linked" refers to the association of two or more molecules. The linkage can be covalent or non-covalent. The linkage can also be genetic (i.e., recombinant fusion). Such linkage can be achieved using a variety of techniques well known in the art, such as chemical conjugation and recombinant protein production.

[0140] As used herein, "administration" refers to physical introduction of a composition comprising a therapeutic agent into a subject using any of a variety of methods and delivery systems known to those of skill in the art. Different routes of administration of the anti-TREM-1 antibodies described herein include intravenous, intraperitoneal, intramuscular, subcutaneous, spinal or other parenteral routes of administration, e.g., by injection or infusion. As used herein, the phrase "parenteral administration" refers to modes of administration other than enteral and topical administration, usually by injection, including, but not limited to, intravenous, intraperitoneal, intramuscular, intraarterial, intrathecal, intralymphatic, intralesional, intracapsular, intraorbital, intracardiac, intradermal, transtracheal, subcutaneous, subcuticular, intraepidermal, intrarticular, subcapsular, subarachnoid, intraspinally, epidural, and intrasternal injection and infusion, as well as in vivo electroporation. Alternatively, the antibodies described herein can be administered by a non-parenteral route, such as a topical, epidermal, or mucosal route of administration, e.g., intranasal, oral, vaginal, rectal, sublingual, or topical. Administration can also be, e.g., one, multiple, and / or one or more extended periods of time.

[0141] As used herein, the terms "inhibit" or "block" (e.g., referring to inhibition / blocking of binding of a TREM-1 ligand to TREM-1 on a cell) are used interchangeably and encompass partial and complete inhibition / blocking. In some embodiments, an anti-TREM-1 antibody inhibits binding of a TREM-1 ligand to TREM-1 by at least about 50%, e.g., about 60%, 70%, 80%, 90%, 95%, 99%, or 100%, e.g., as determined as described further herein. In some embodiments, an anti-TREM-1 antibody inhibits binding of a TREM-1 ligand to TREM-1 by no more than 50%, e.g., about 40%, 30%, 20%, 10%, 5%, or 1%, e.g., as determined as described further herein.

[0142] The terms "treat," "treating," and "treatment" as used herein refer to any type of intervention or process performed on a subject with the objective of reversing, alleviating, improving, inhibiting or slowing down or preventing the progression, development, severity or recurrence of a symptom, complication, disorder or biochemical indicia associated with a disease, or enhancing overall survival. Treatment can be performed on a subject who has a disease or a subject who does not have a disease (e.g., for prophylaxis).

[0143] The terms "effective dose" or "effective amount" are defined as an amount that is sufficient to achieve or at least partially achieve a desired effect. A "therapeutically effective amount" or "therapeutically effective dose" of a drug or therapeutic agent is any amount of a drug that, when used alone or in combination with another therapeutic agent, promotes disease regression as evidenced by a decrease in severity, frequency, or duration of symptoms, an increase in frequency and duration of disease symptom-free periods, or a prevention of impairment or disability due to the disease affliction. A therapeutically effective amount or dose of a drug includes a "prophylactically effective amount" or "prophylactically effective dose," which is any amount of a drug that, when administered alone or in combination with another therapeutic agent to a subject at risk of developing a disease or condition, inhibits the development or progression of the disease. The ability of a therapeutic agent to promote disease regression or inhibit disease development or progression can be assessed using a variety of methods known to the skilled artisan, such as in human subjects during clinical trials, in animal model systems predictive of efficacy in humans, or by determining the activity of the agent in in vitro assays.

[0144] The term "patient" includes human and other mammalian subjects who receive prophylactic or therapeutic treatment.

[0145] The term "subject" as used herein includes any human or non-human animal. For example, the methods and compositions described herein can be used to treat a subject having a cancer. The term "non-human animal" includes all vertebrates, e.g., mammals and non-mammals, such as non-human primates, sheep, dogs, cows, chickens, amphibians, reptiles, etc.

[0146] The terms "ug" and "uM" as used herein are used interchangeably with "pg" and "pM," respectively.

[0147] The various aspects described herein are described in more detail in the subsections below.

[0148] II. Methods of the Disclosure

[0149] Methods of identifying a subject suitable for treatment

[0150] Disclosed herein are methods of identifying a subject having a disease or disorder amenable to treatment with an anti-TREM-1 antibody (i.e., an antagonistic anti-TREM-1 antibody). In some embodiments, the methods disclosed herein comprise measuring the expression level of a TREM-1 -associated gene in a sample of the subject. In some embodiments, the TREM-1 -associated gene comprises one or more of the genes listed in Table 3 (below). In some embodiments, the expression level of a TREM-1 -associated gene (e.g., disclosed herein) is increased when a natural TREM-1 ligand (i.e., PGLYRP1) binds to TREM-1, but is not increased when an agonistic anti-TREM-1 antibody binds to TREM-1.

[0151] Table 3. TREM-1 -associated genes

[0152]

[0153]

[0154] In some embodiments, a subject amenable to treatment with an anti-TREM-1 antibody exhibits an increased expression level of a TREM-1 -associated gene compared to a reference (e.g., a subject not having a disease or disorder, e.g., a healthy subject). In certain embodiments, the expression level of a TREM-1 -associated gene (e.g., disclosed herein) in the subject is increased by at least about 5%, at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, or more, compared to a reference (e.g., a subject not having a disease or disorder, e.g., a healthy subject).

[0155] In some embodiments, an increased expression level of a TREM-1 -associated gene disclosed herein is associated with an increase in one or more other biomarkers. In some embodiments, the one or more other biomarkers comprise a baseline Mayo score, a grade 2B lamina propria neutrophil infiltration score, a fecal calprotectin level, or a combination thereof. Thus, in certain embodiments, a subject amenable to treatment with an anti-TREM-1 antibody exhibits an increased baseline Mayo score, an increased grade 2B lamina propria neutrophil infiltration score, and / or an increased fecal calprotectin level compared to a reference (e.g., a subject not having a disease or disorder, e.g., a healthy subject). In some embodiments, a method of identifying a subject amenable to treatment with an anti-TREM-1 antibody comprises determining the baseline Mayo score, the grade 2B lamina propria neutrophil infiltration score, and / or the fecal calprotectin level in a sample of the subject.

[0156] In some embodiments, the subject has an increase in baseline Mayo score of at least about 5%, at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, or more, compared to a reference (e.g., a subject who does not have the disease or disorder, e.g., a healthy subject). In some embodiments, the subject has a baseline Mayo score of greater than about 6, about 7, about 8, about 9, about 10, about 11, or about 12.

[0157] In some embodiments, the subject has an increase in grade 2B lamina propria neutrophil infiltration score of at least about 5%, at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, or more, compared to a reference (e.g., a subject who does not have the disease or disorder, e.g., a healthy subject). In some embodiments, the subject has a grade 2B lamina propria neutrophil infiltration score of greater than about 0, about 0.1, about 0.2, or about 0.3.

[0158] In some embodiments, the subject has an increase in fecal calprotectin level of at least about 5%, at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, or more, compared to a reference (e.g., a subject who does not have the disease or disorder, e.g., a healthy subject). In some embodiments, the fecal calprotectin level (pg / g) is greater than about 1.5 log10, about 2.0 log10, about 2.5 log10, about 3.0 log10, or about 3.5 log10.

[0159] In some embodiments, the method of identifying a subject suitable for treatment with an anti-TREM-1 antibody disclosed herein further comprises administering an effective dose of the anti-TREM-1 antibody to the subject.

[0160] Methods of determining treatment efficacy

[0161] Also disclosed herein are methods of determining the efficacy of an anti-TREM-1 antibody (i.e., an antagonistic anti-TREM-1 antibody) in treating a disease or disorder in a subject in need thereof. In some embodiments, the methods disclosed herein comprise administering an anti-TREM-1 antibody to a subject and measuring the level of expression of a TREM-1 -associated gene in a sample of the subject. In some embodiments, the TREM-1 -associated gene comprises one or more of the genes listed in Table 3 (above). In some embodiments, the TREM-1 -associated gene (e.g., disclosed herein) is increased when a natural TREM-1 ligand (i.e., PGLYRP1) binds to TREM-1, but is not increased when an agonistic anti-TREM-1 antibody binds to TREM-1.

[0162] In some embodiments, the subject exhibits a decrease in the level of expression of a TREM-1 -associated gene after administration of an anti-TREM-1 antibody compared to a reference (e.g., the corresponding value in the subject prior to administration). In certain embodiments, the level of expression of a TREM-1 -associated gene in the subject is decreased by at least about 5%, at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, or more after administration compared to a reference (e.g., the corresponding value in the subject prior to administration). In some embodiments, a decrease in the level of expression of a TREM-1 -associated gene after administration indicates that the anti-TREM-1 antibody is effective in the subject (e.g., reduces and / or prevents one or more symptoms associated with the disease or disorder).

[0163] In some embodiments, a decrease in the level of expression of a TREM-1 -associated gene is associated with a decrease in one or more other biomarkers. In some embodiments, the one or more other biomarkers comprise a baseline Mayo score, a grade 2B lamina propria neutrophil infiltration score, a fecal calprotectin level, or a combination thereof. Accordingly, in some embodiments, the methods of determining the efficacy of an anti-TREM-1 antibody comprise administering an anti-TREM-1 antibody to a subject and determining the baseline Mayo score, the grade 2B lamina propria neutrophil infiltration score, and / or the fecal calprotectin level in a sample of the subject. In some embodiments, the anti-TREM-1 antibody is effective (e.g., reduces and / or prevents one or more symptoms associated with the disease or disorder) when the baseline Mayo score, the grade 2B lamina propria neutrophil infiltration score, and / or the fecal calprotectin level is decreased compared to a reference (e.g., the corresponding value in the subject prior to administration of the anti-TREM-1 antibody).

[0164] In some embodiments, the baseline Mayo score is reduced by at least about 5%, at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, or more, compared to a reference (e.g., the corresponding value in the subject prior to administration). In certain embodiments, the grade 2B lamina propria neutrophil infiltration score is reduced by at least about 5%, at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, or more, compared to a reference (e.g., the corresponding value in the subject prior to administration). In some embodiments, the fecal calprotectin level is reduced by at least about 5%, at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, or more, compared to a reference (e.g., the corresponding value in the subject prior to administration).

[0165] In some embodiments, the subject has a baseline Mayo score of at least about 4, at least about 5, at least about 6, at least about 7, at least about 8, at least about 9, at least about 10, at least about 11, or at least about 12, prior to administration of the anti-TREM-1 antibody. In some embodiments, the subject has a grade 2B lamina propria neutrophil infiltration score of greater than about 0, greater than about 0.1, greater than about 0.2, or greater than about 0.3, prior to administration of the anti-TREM-1 antibody. In some embodiments, the subject has a fecal calprotectin level of greater than about 1.5 logio, greater than about 2.0 logio, greater than about 2.5 logio, greater than about 3.0 logio, or greater than about 3.5 logio, prior to administration of the anti-TREM-1 antibody.

[0166] In some embodiments, measuring the expression level of the TREM-1 -related gene occurs 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 2 weeks, at least 3 weeks, or at least 4 weeks or more, after the anti-TREM-1 antibody is administered to the subject. In some embodiments, determining the baseline Mayo score, the grade 2B lamina propria neutrophil infiltration score, and / or the fecal calprotectin level occurs 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 2 weeks, at least 3 weeks, or at least 4 weeks or more, after the anti-TREM-1 antibody is administered to the subject. In certain embodiments, measuring the expression level of the TREM-1 -related gene and / or determining the baseline Mayo score, the grade 2B lamina propria neutrophil infiltration score, and / or the fecal calprotectin level occurs at multiple time points after administration of the anti-TREM-1 antibody.

[0167] In some embodiments, the subject continues to receive an anti-TREM-1 antibody, wherein the anti-TREM-1 antibody treatment is determined to be ineffective for the subject. In some embodiments, the subject receives an adjusted dose of the anti-TREM-1 antibody, wherein the initial dose of the anti-TREM-1 antibody is determined to be ineffective for the subject (e.g., the subject’s expression level of a TREM-1 -associated gene, Mayo score, grade 2B lamina propria neutrophil infiltration score, and / or fecal calprotectin level is not reduced after administration).

[0168] Methods of identifying non-responders to standard-of-care treatments

[0169] The present disclosure also provides methods of identifying non-responders to a standard-of-care treatment for a disease or disorder. As used herein, the term “non-responder” refers to a subject who does not exhibit an improvement in one or more symptoms associated with the disease or disorder. As used herein, the term “standard-of-care treatment” refers to a treatment accepted by medical experts as an appropriate treatment for a certain type of disease and widely used by health care professionals. This term is used interchangeably with any of the following terms: “best practice,” “standard medical care,” and “standard therapy.” In some embodiments, the disease or disorder comprises inflammatory bowel disease (e.g., ulcerative colitis or Crohn’s disease), and the standard-of-care treatment comprises drugs (e.g., anti-inflammatory agents, immunosuppressants, and antibiotics), nutritional supplements, and surgery. In certain embodiments, the standard-of-care treatment comprises an anti-TNF-a antibody. In some embodiments, the anti-TNF-a antibody comprises infliximab Certolizumab pegol Etanercept Adalimumab Golimumab or combinations thereof. In other embodiments, the standard-of-care treatment comprises an oral corticosteroid. In some embodiments, the standard-of-care treatment comprises an anti-IP-10 antibody.

[0170] In some embodiments, the method of identifying non-responders to a standard-of-care treatment comprises measuring the expression level of a TREM-1 -associated gene in a sample of a subject who has previously received the standard-of-care treatment. In some embodiments, the TREM-1 -associated gene comprises one or more genes listed in Table 3 (above). In some embodiments, the TREM-1 -associated gene (e.g., disclosed herein) is increased when a natural TREM-1 ligand (i.e., PGLYRP1) binds to TREM-1, but is not increased when an agonistic anti-TREM-1 antibody binds to TREM-1.

[0171] In some embodiments, a subject is a non-responder if the expression level of a TREM-1 -associated gene in the subject is not decreased by at least about 5%, at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, or more, as compared to a reference (e.g., a subject prior to standard-of-care treatment administration). In certain embodiments, a non-responder has a baseline Mayo score that is increased by at least 5%, 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 more, as compared to a reference (e.g., a subject prior to standard-of-care treatment administration).

[0172] In some embodiments, a subject is a non-responder if the expression level of a TREM-1 -associated gene in the subject is not decreased by at least about 5%, at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, or more, as compared to a reference (e.g., a subject prior to standard-of-care treatment administration). In certain embodiments, a non-responder has a baseline Mayo score that is increased by at least 5%, 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 more, as compared to a reference (e.g., a subject prior to standard-of-care treatment administration).

[0173] As described above, in some embodiments, the expression level of a TREM-1 -associated gene (e.g., disclosed herein) is correlated with one or more other biomarkers, such as baseline Mayo score, grade 2B lamina propria neutrophil infiltration score, and / or fecal calprotectin level. Accordingly, in some embodiments, a method of identifying a non-responder to a standard-of-care treatment for a disease or disorder comprises determining the baseline Mayo score, grade 2B lamina propria neutrophil infiltration score, and / or fecal calprotectin level in a sample of a subject who has previously received a standard-of-care treatment.

[0174] In some embodiments, a subject is a non-responder if the baseline Mayo score of the subject is increased as compared to a reference (e.g., a subject not suffering from a disease or disorder, e.g., a healthy subject). In certain embodiments, the baseline Mayo score of a non-responder is increased by at least 5%, 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 more, as compared to a reference (e.g., a subject not suffering from a disease or disorder, e.g., a healthy subject). In certain embodiments, a non-responder to a standard-of-care treatment has a baseline Mayo score of at least about 4, at least about 5, at least about 6, at least about 7, at least about 8, at least about 9, at least about 10, at least about 11, or at least about 12.

[0175] In some embodiments, a subject is a non-responder if the subject has an increased 2B grade lamina propria neutrophil infiltration score compared to a reference (e.g., a subject that does not have the disease or disorder, e.g., a healthy subject). In some embodiments, a non-responder has an increased 2B grade lamina propria neutrophil infiltration score of at least 5%, 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 more, compared to a reference (e.g., a subject that does not have the disease or disorder, e.g., a healthy subject). In certain embodiments, the 2B grade lamina propria neutrophil infiltration score is greater than about 0, greater than about 0.1, greater than about 0.2, or greater than about 0.3.

[0176] In some embodiments, a subject is a non-responder if the subject has an increased fecal calprotectin level compared to a reference (e.g., a subject that does not have the disease or disorder, e.g., a healthy subject). In some embodiments, a non-responder disclosed herein has an increased fecal calprotectin level of at least 5%, 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 more, compared to a reference (e.g., a subject that does not have the disease or disorder, e.g., a healthy subject). In some embodiments, a non-responder has a fecal calprotectin level that is greater than about 1.5 logio, greater than about 2.0 logio, greater than about 2.5 logio, greater than about 3.0 logio, or greater than about 3.5 logio.

[0177] In some embodiments, a subject is a non-responder to a standard of care treatment if the subject does not have a decreased Mayo score after receiving the standard of care treatment. In certain embodiments, a subject does not have a decreased Mayo score of at least about 5%, at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, or more, compared to a reference (e.g., the subject prior to administration of the standard of care treatment).

[0178] In some embodiments, a subject is a non-responder if the subject has an increased 2B grade lamina propria neutrophil infiltration score compared to a reference (e.g., a subject that does not have the disease or disorder, e.g., a healthy subject). In some embodiments, a non-responder has an increased 2B grade lamina propria neutrophil infiltration score of at least 5%, 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 more, compared to a reference (e.g., a subject that does not have the disease or disorder, e.g., a healthy subject). In certain embodiments, the 2B grade lamina propria neutrophil infiltration score is greater than about 0, greater than about 0.1, greater than about 0.2, or greater than about 0.3.

[0179] In some embodiments, a subject is a non-responder if the subject's fecal calprotectin level does not decrease after receiving standard-of-care treatment. In some embodiments, the subject's fecal calprotectin level does not decrease by at least about 5%, at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, or more, compared to a reference (e.g., the subject prior to administration of the standard-of-care treatment).

[0180] In some embodiments, a non-responsive subject has a baseline Mayo score of at least about 4, at least about 5, at least about 6, at least about 7, at least about 8, at least about 9, at least about 10, at least about 11, or at least about 12 prior to administration of the standard-of-care treatment. In some embodiments, a non-responsive subject has a 2B grade lamina propria neutrophil infiltration score of greater than about 0, greater than about 0.1, greater than about 0.2, or greater than about 0.3 prior to administration of the standard-of-care treatment. In some embodiments, a non-responsive subject has a fecal calprotectin level of greater than about 1.5 logio, greater than about 2.0 logio, greater than about 2.5 logio, greater than about 3.0 logio, or greater than about 3.5 logio prior to administration of the standard-of-care treatment.

[0181] In some embodiments, measuring the expression level of a TREM-1 -associated gene occurs 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 2 weeks, at least 3 weeks, or at least 4 weeks or more after administering an anti-TREM-1 antibody to the subject. In some embodiments, determining the baseline Mayo score, 2B grade lamina propria neutrophil infiltration score, and / or fecal calprotectin level occurs 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 2 weeks, at least 3 weeks, or at least 4 weeks or more after administering an anti-TREM-1 antibody to the subject.

[0182] In some embodiments, the methods disclosed herein further comprise administering an additional therapeutic agent to a subject who has been identified as a non-responder to a standard-of-care treatment. In certain embodiments, the additional therapeutic agent comprises an anti-TREM-1 antibody.

[0183] Methods of treating a disease or disorder

[0184] Disclosed herein are methods of treating a disease or disorder in a subject in need thereof, comprising administering to the subject an effective dose of an anti-TREM-1 antibody (i.e., an antagonistic anti-TREM-1 antibody), wherein the subject exhibits an increased level of expression of a TREM-1 -associated gene compared to a reference (e.g., a subject that does not have the disease or disorder, e.g., a healthy subject). In some embodiments, the TREM-1 -associated gene comprises one or more genes listed in Table 3 (above). In some embodiments, the TREM-1 -associated gene (e.g., disclosed herein) is increased when a natural TREM-1 ligand (i.e., PGLYRP1) binds to TREM-1, but is not increased when an agonistic anti-TREM-1 antibody binds to TREM-1.

[0185] In some embodiments, the level of expression of a TREM-1 -associated gene in the subject is increased by at least about 5%, at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, or more, compared to a reference (e.g., a subject that does not have the disease or disorder, e.g., a healthy subject).

[0186] In some embodiments, administration of the anti-TREM-1 antibody to the subject decreases the level of expression of a TREM-1 -associated gene compared to a reference (e.g., the corresponding value in the subject prior to administration). In certain embodiments, the level of expression of a TREM-1 -associated gene is decreased by at least about 5%, at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, or more, after administration.

[0187] In some embodiments, the subject exhibits an increased baseline Mayo score, an increased grade 2B lamina propria neutrophil infiltration score, and / or an increased fecal calprotectin level compared to a reference (e.g., a subject that does not have the disease or disorder, e.g., a healthy subject) prior to administration of the anti-TREM-1 antibody.

[0188] In some embodiments, the baseline Mayo score of the subject is increased by at least about 5%, at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, or more, compared to a reference (e.g., a subject that does not have the disease or disorder, e.g., a healthy subject). In some embodiments, the baseline Mayo score of the subject is greater than about 6, greater than about 7, greater than about 8, greater than about 9, greater than about 10, greater than about 11, or greater than about 12, prior to administration of the anti-TREM-1 antibody.

[0189] In some embodiments, the subject has an increased fecal calprotectin level of at least about 5%, at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, or more, compared to a reference (e.g., a subject who does not have the disease or disorder, e.g., a healthy subject). In some embodiments, the fecal calprotectin level (pg / g) is greater than about 1.5 logio, greater than about 2.0 logio, greater than about 2.5 logio, greater than about 3.0 logio, or greater than about 3.5 logio prior to administration.

[0190] In some embodiments, the subject has an increased fecal calprotectin level of at least about 5%, at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, or more, compared to a reference (e.g., a subject who does not have the disease or disorder, e.g., a healthy subject). In some embodiments, the fecal calprotectin level (pg / g) is greater than about 1.5 logio, greater than about 2.0 logio, greater than about 2.5 logio, greater than about 3.0 logio, or greater than about 3.5 logio prior to administration.

[0191] In some embodiments, the subject has an increased fecal calprotectin level of at least about 5%, at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, or more, compared to a reference (e.g., a subject who does not have the disease or disorder, e.g., a healthy subject). In some embodiments, the fecal calprotectin level (pg / g) is greater than about 1.5 logio, greater than about 2.0 logio, greater than about 2.5 logio, greater than about 3.0 logio, or greater than about 3.5 logio prior to administration.

[0192] In some embodiments, the method of treating a disease or disorder in a subject in need thereof further comprises measuring the expression level of a TREM-1 -related gene and / or determining the baseline Mayo score, 2B-grade lamina propria neutrophil infiltration score, and / or fecal calprotectin level prior to administering the anti-TREM-1 antibody to the subject.

[0193] In some embodiments, the methods of treating a disease or disorder disclosed herein comprise measuring the expression level of a TREM-1 -associated gene and / or determining the baseline Mayo score, grade 2B lamina propria neutrophil infiltration score, and / or fecal calprotectin level after administering an anti-TREM-1 antibody to the subject. In certain embodiments, measuring the expression level of a TREM-1 -associated gene and / or determining the baseline Mayo score, grade 2B lamina propria neutrophil infiltration score, and / or fecal calprotectin level occurs 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 2 weeks, at least 3 weeks, or at least 4 weeks or more after administering the anti-TREM-1 antibody.

[0194] The methods and compositions disclosed herein can be used to treat (e.g., treat) a variety of diseases or disorders, wherein the disease or disorder is associated with an increase in TREM-1 activity. In some embodiments, the disease or disorder is associated with increased degranulation, reactive oxygen species formation, and / or proinflammatory cytokine release by neutrophils. In certain embodiments, the disease or disorder is associated with activation of monocytes and / or increased inflammatory cytokine and chemokine production by monocytes. In some embodiments, the disease or disorder is associated with hypoxia. In some embodiments, the disease or disorder is associated with an increase in cell surface TREM-1 protein expression and / or an increase in soluble TREM-1 protein levels.

[0195] Non-limiting examples of such diseases include inflammatory bowel disease (IBD), Crohn’s disease (CD), ulcerative colitis (UC), irritable bowel syndrome, rheumatoid arthritis (RA), psoriasis, psoriatic arthritis, systemic lupus erythematosus (SLE), lupus nephritis, type I diabetes, Graves’ disease, multiple sclerosis (MS), autoimmune myocarditis, Kawasaki disease, coronary artery disease, chronic obstructive pulmonary disease, interstitial lung disease, autoimmune thyroiditis, scleroderma, systemic sclerosis, osteoarthritis, atopic dermatitis, vitiligo, graft-versus-host disease, Sjogren’s syndrome, autoimmune nephritis, Goodpasture’s syndrome, chronic inflammatory demyelinating polyneuropathy, allergy, asthma, and other autoimmune diseases caused by acute or chronic inflammation. In some embodiments, the disease or disorder is inflammatory bowel disease. In certain embodiments, the inflammatory bowel disease comprises Crohn’s disease and ulcerative colitis.

[0196] In some embodiments, the sample of the subject in which the expression level of a TREM-1 related gene is measured comprises tissue, blood, serum, plasma, saliva, urine, or a combination thereof. In some embodiments, the sample of the subject in which the baseline Mayo score, grade 2B intestinal submucosal neutrophil infiltration score, or calprotectin level is determined comprises tissue, blood, serum, plasma, saliva, urine, or a combination thereof.

[0197] III. Anti-TREM-1 Antibodies

[0198] Certain aspects of the present disclosure include administering a therapeutically effective amount of an anti-TREM-1 antibody (i.e., an antagonistic anti-TREM-1 antibody) to a subject in need thereof. Anti-TREM-1 antibodies (or VH / VL domains derived therefrom) suitable for use in the present disclosure can be produced using methods known in the art. Alternatively, anti-TREM-1 antibodies well known in the art can be used. See, e.g., WO 2016 / 009086 and WO 2017 / 152102, each of which is incorporated by reference herein in its entirety.

[0199] Anti-TREM-1 antibodies (e.g., fully human monoclonal antibodies) useful in the methods disclosed herein are characterized by specific functional features or properties, which are provided throughout the DETAILED DESCRIPTION. In some embodiments, anti-TREM-1 antibodies that can be used with the present methods exhibit one or more of the following properties:

[0200] (a) bind to soluble and / or membrane-bound human TREM-1 (e.g., at a site on the extracellular domain that is bound by a TREM-1 ligand (e.g., PGLYRP1));

[0201] (b) cross-react with TREM-1 from one or more non-human primates (e.g., cynomolgus monkey TREM-1);

[0202] (c) block or inhibit binding of PGLYRP1 to TREM-1;

[0203] (d) block or inhibit production of inflammatory cytokines (e.g., IL-6, TNF-a, IL-8, IL-1b, IL-12, and combinations thereof) by cells (e.g., macrophages, dendritic cells, neutrophils) upon activation;

[0204] (e) does not induce release of proinflammatory cytokines by myeloid cells (e.g., dendritic cells);

[0205] (f) does not bind to one or more FcyRs;

[0206] (g) has a viscosity profile of less than about 5 cP at a concentration of 80 mg / mL, or less than about 10 cP at a concentration of 130 mg / mL; and / or

[0207] (h) reduces or prevents the occurrence of an inflammatory cytokine storm upon administration to a subject.

[0208] In some embodiments, the anti-TREM-1 antibodies described herein bind to human TREM-1 with high affinity, for example, as determined by BIACORE TM Kdof 10-7M or less, 10 D 为 10 -7 M or less, 10 -8 M or less, 10 -9 M (1 nM) or less, 10 -10 M or less, 10 -11 M or less, 10 -12 M or less, 10 -12 M to 10 -7 M, 10 -11 M to 10 -7 M, 10 -10 M to 10 -7 M or 10 -9 M to 10-7M. In some embodiments, the anti-TREM-1 antibodies described herein bind to cynomolgus monkey TREM-1, for example, as determined by BIACORE TM Kdof 10-7M or less, 10 例如 Kdof 10-7M or less, 10 D为 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 M or less, 10 -12 M to 10 -7 M, 10 -11 M to 10 -7 M, 10 -10 M to 10 -7 M or 10-9M to 10-7M.

[0209] In some embodiments, the anti-TREM-1 antibody cross-competes for binding to human TREM-1 with mAb 0170 and / or mAb 0318. In certain embodiments, the anti-TREM-1 antibody also cross-competes for binding to cynomolgus monkey TREM-1 with mAb 0170 and / or mAb 0318. In other words, in certain embodiments, the anti-TREM-1 antibody that can be used with the present methods belongs to the same "bin" as mAb 0170 and / or mAb 0318.

[0210] The mAb 0170 antibody has a heavy chain variable region (VH) and a light chain variable region (VL), wherein the VH comprises amino acids 1 to 121 of SEQ ID NO: 13, and wherein the VL comprises amino acids 1 to 111 of SEQ ID NO: 14. The mAb 0170 antibody also has a heavy chain CDR1, CDR2, and CDR3, and a light chain CDR1, CDR2, and CDR3, wherein (a) the heavy chain CDR1 comprises amino acids 31 to 35 of SEQ ID NO: 13; (b) the heavy chain CDR2 comprises amino acids 50 to 68 of SEQ ID NO: 13; (c) the heavy chain CDR3 comprises amino acids 101 to 110 of SEQ ID NO: 13; (d) the light chain CDR1 comprises amino acids 24 to 38 of SEQ ID NO: 14; (e) the light chain CDR2 comprises amino acids 54 to 60 of SEQ ID NO: 14; and (f) the light chain CDR3 comprises amino acids 93 to 101 of SEQ ID NO: 14. See WO 2016 / 009086, which is incorporated by reference herein in its entirety.

[0211] Accordingly, in some embodiments, an anti-TREM-1 antibody useful for the present disclosure comprises a VH and a VL, wherein the VH comprises an amino acid sequence as set forth in SEQ ID NO: 15 (i.e., amino acids 1 to 121 of SEQ ID NO: 13), and wherein the VL comprises an amino acid sequence as set forth in SEQ ID NO: 16 (i.e., amino acids 1 to 111 of SEQ ID NO: 14). In some embodiments, the VH of the anti-TREM-1 antibody comprises a CDR1 sequence as set forth in SEQ ID NO: 17 (TYAMH), a CDR2 sequence as set forth in SEQ ID NO: 18 (RIRTKSSNYATYYAASVKG), and a CDR3 sequence as set forth in SEQ ID NO: 19 (DMGIRRQFAY). In some embodiments, the VL of the anti-TREM-1 antibody comprises a CDR1 sequence as set forth in SEQ ID NO: 20 (RASESVDTFDYSFLH), a CDR2 sequence as set forth in SEQ ID NO: 21 (RASNLES), and a CDR3 sequence as set forth in SEQ ID NO: 22 (QQSNEDPYT).

[0212] The mAb 0318 antibody has a heavy chain variable region (VH) comprising SEQ ID NO: 15 and a light chain variable region (VL) comprising SEQ ID NO: 23. See International Publication No. 2016 / 009086. The mAb 0318 also has heavy chain CDR1, CDR2, and CDR3, which correspond to amino acids 31-35, 50-68, and 101-110 of SEQ ID NO: 15, respectively. The light chain CDR1, CDR2, and CDR3 of the mAb 0318 antibody correspond to amino acids 24-38, 54-60, and 93-101 of SEQ ID NO: 23.

[0213] Accordingly, in some embodiments, an anti-TREM-1 antibody comprises a VH and a VL of SEQ ID NOs: 15 and 23, respectively. In some embodiments, the VH of the anti-TREM-1 antibody comprises a CDR1 sequence of amino acids 31-35 of SEQ ID NO: 15 (TYAMH), wherein one of the amino acids can be substituted with a different amino acid. In certain embodiments, the VH of the anti-TREM-1 antibody comprises a CDR2 sequence of amino acids 50-68 of SEQ ID NO: 15 (RIRTKSSNYATYYAASVKG), wherein one, two, or three of the amino acids can be substituted with a different amino acid. In some embodiments, the VH of the anti-TREM-1 antibody comprises a CDR3 sequence of amino acids 101-110 of SEQ ID NO: 15 (DMGIRRQFAY), wherein one, two, or three of the amino acids can be substituted with a different amino acid.

[0214] In some embodiments, the VL of the anti-TREM-1 antibody comprises the CDR1 sequence of amino acids 24-38 (RASQSVDTFDYSFLH) of SEQ ID NO: 23, wherein one, two, or three of the amino acids can be substituted with a different amino acid. In other embodiments, the VL of the anti-TREM-1 antibody comprises the CDR2 sequence of amino acids 54-60 (RASNLES) of SEQ ID NO: 23, wherein one or two of the amino acids can be substituted with a different amino acid. In some embodiments, the VL of the anti-TREM-1 antibody comprises the CDR3 sequence of amino acids 93-101 (QQSNQDPYT) of SEQ ID NO: 23, wherein one or two of the amino acids can be substituted with a different amino acid.

[0215] Methionine residues in the CDRs of antibodies can be oxidized, leading to potential chemical degradation and thus reduced potency of the antibody. Accordingly, one or more of the methionine residues in the heavy and / or light chain CDRs of the anti-TREM-1 antibodies disclosed herein can be replaced with an amino acid residue that does not undergo oxidative degradation. In some embodiments, the methionine residues within the heavy chain CDR1 and CDR3 are replaced with an amino acid residue that does not undergo oxidative degradation (e.g., glutamine or leucine). Accordingly, in some embodiments, the VH of the anti-TREM-1 antibody comprises the CDR3 sequence of amino acids 101-110 (DQGIRRQFAY) of SEQ ID NO: 26 or amino acids 101-110 (DLGIRRQFAY) of SEQ ID NO: 27. In other embodiments, the VH of the anti-TREM-1 antibody comprises the CDR1 sequence of amino acids 31-35 (TYAQH) of SEQ ID NO: 28 or amino acids 31-35 (TYALH) of SEQ ID NO: 29. Similarly, in some embodiments, deamidation sites can be removed from the anti-TREM-1 antibodies, particularly in the CDRs.

[0216] In some embodiments, the VH and VL of the anti-TREM-1 antibody comprise the VH and VL sequences of an anti-TREM-1 antibody disclosed in International Publication No. WO 2017 / 152102 A2, which is incorporated by reference herein in its entirety. In some embodiments, the VL of the anti-TREM-1 antibody comprises a CDR1 sequence selected from the group consisting of SEQ ID NOs:9-27 of WO 2017 / 152102, a CDR2 sequence selected from the group consisting of SEQ ID NOs:28-40 of WO 2017 / 152102, and / or a CDR3 sequence selected from the group consisting of SEQ ID NOs:41-119 of WO 2017 / 152102. In one embodiment, the VH of the anti-TREM-1 antibody comprises a CDR1 sequence selected from the group consisting of SEQ ID NOs: 120-143 of WO 2017 / 152102, a CDR2 sequence selected from the group consisting of SEQ ID NOs: 144-172 of WO 2017 / 152102, and / or a CDR3 sequence selected from the group consisting of SEQ ID NOs: 173-247 of WO 2017 / 152102.

[0217] In some embodiments, the anti-TREM-1 antibody of the present disclosure comprises CDR and / or variable region sequences that are at least 80% identical (e.g., at least 85%, at least 95%, at least 95%, or at least 99% identical) to the CDR and / or variable region sequences of the mAb 0318 antibody.

[0218] In some embodiments, the anti-TREM-1 antibody of the present disclosure comprises a heavy chain variable region (VH) selected from the group consisting of SEQ ID NOs:396-475 of WO 2017 / 152102 and / or a light chain variable region (VL) selected from the group consisting of SEQ ID NOs:316-395 of WO 2017 / 152102.

[0219] In some embodiments, the anti-TREM-1 antibody comprises a heavy chain (HC) and a light chain (LC), wherein the HC comprises SEQ ID NO:50, SEQ ID NO:51, SEQ ID NO:52, or SEQ ID NO:53. In some embodiments, the LC comprises SEQ ID NO:54.

[0220] In some embodiments, an anti-TREM-1 antibody comprises a heavy chain and a light chain, wherein the heavy chain and the light chain comprise the amino acid sequences as shown in Table 7. In some embodiments, an anti-TREM-1 antibody comprises a heavy chain and a light chain, wherein the heavy chain comprises the amino acid sequence as shown in SEQ ID NO: 30, and the light chain comprises the amino acid sequence as shown in SEQ ID NO: 34. In some embodiments, an anti-TREM-1 antibody comprises a heavy chain and a light chain, wherein the heavy chain comprises the amino acid sequence as shown in SEQ ID NO: 31, and the light chain comprises the amino acid sequence as shown in SEQ ID NO: 34. In some embodiments, an anti-TREM-1 antibody comprises a heavy chain and a light chain, wherein the heavy chain comprises the amino acid sequence as shown in SEQ ID NO: 32, and the light chain comprises the amino acid sequence as shown in SEQ ID NO: 34. In some embodiments, an anti-TREM-1 antibody comprises a heavy chain and a light chain, wherein the heavy chain comprises the amino acid sequence as shown in SEQ ID NO: 33, and the light chain comprises the amino acid sequence as shown in SEQ ID NO: 34.

[0221] Heavy chains and light chains comprising an amino acid sequence that is at least 99%, 98%, 97%, 96%, 95%, 90%, 85%, or 80% identical to any of the heavy chains or light chains shown herein (e.g., SEQ ID NOs: 30-34) can be used to form an anti-TREM-1 antibody having the desired characteristics, such as those described further herein.

[0222] In some embodiments, an anti-TREM-1 antibody of the present disclosure comprises a heavy chain and a light chain, wherein the heavy chain comprises an amino acid sequence that is at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to the amino acid sequence shown in SEQ ID NO: 30, 31, 32, or 33, and wherein the light chain comprises an amino acid sequence that is at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to the amino acid sequence shown in SEQ ID NO: 34.

[0223] In some embodiments, an anti-TREM-1 antibody that can be used with the present methods is epitope-directed. As used herein, the term “epitope-directed” refers to an anti-TREM-1 antibody that is selected to bind to an epitope other than D38 to L45, E46 to Q56, and / or Y90 to L96 of human TREM-1 (SEQ ID NO: 1). In some embodiments, an epitope-directed anti-TREM-1 antibody binds to one or more epitopes selected from the group consisting of: (1)27 EKYELKEGQTL 37 (SEQ ID NO: 50), (2) 88 EDYHDHGLLRVRM 100 (SEQ ID NO: 51), (3) 120 KEPHMLFDR 128 (SEQ ID NO: 52), and any combination thereof.

[0224] Epitope-directed anti-TREM-1 antibodies described herein can be produced by any method known in the art, such as those described in the Examples. In some embodiments, epitope-directed anti-TREM-1 antibodies can be produced by immunizing an animal (e.g., a mouse) with a human TREM-1 polypeptide comprising a mutation at one of the epitopes described above (e.g., amino acid residues 38-48 of SEQ ID NO: 1). Following immunization, the produced antibodies can be further characterized for binding to human TREM-1. In some embodiments, a synthetic peptide comprising the epitope of interest can be synthesized and used to immunize an animal (e.g., a mouse). In some embodiments, an alternative scaffold comprising the epitope of interest (e.g., the tenth human fibronectin type III domain, 10Fn3; or a3D, a highly thermostable three-helical bundle protein) can be used.

[0225] In some embodiments, the anti-TREM-1 antibodies (i.e., epitope-directed) do not cross-compete with mAb 0170 and / or mAb 0318 for binding to TREM-1 (e.g., human or cynomolgus monkey). In other words, in certain embodiments, the anti-TREM-1 antibodies disclosed herein belong to a different "bin" than mAb 0170 and / or mAb 0318.

[0226] In some embodiments, the epitope-directed anti-TREM-1 antibodies comprise a VH and a VL, wherein:

[0227] (a) the VH and VL comprise the amino acid sequences set forth in SEQ ID NOs: 53 and 54, respectively;

[0228] (b) the VH and VL comprise the amino acid sequences set forth in SEQ ID NOs: 55 and 56, respectively;

[0229] (c) the VH and VL comprise the amino acid sequences set forth in SEQ ID NOs: 55 and 57, respectively;

[0230] (d) the VH and VL comprise the amino acid sequences set forth in SEQ ID NOs: 58 and 59, respectively;

[0231] (e) the VH and VL comprise the amino acid sequences set forth in SEQ ID NOs: 60 and 56, respectively;

[0232] (f) the VH and VL comprise the amino acid sequences set forth in SEQ ID NOs: 153 and 154, respectively; or

[0233] (g) the VH and VL comprise the amino acid sequences set forth in SEQ ID NOs: 153 and 155, respectively.

[0234] In some embodiments, the epitope-directed anti-TREM-1 antibodies disclosed herein comprise the CDRs of the heavy chain variable region selected from the group consisting of SEQ ID NOs: 53, 55, 58, 60, and 153. In some embodiments, the epitope-directed anti-TREM-1 antibodies disclosed herein comprise the CDRs of the light chain variable region selected from the group consisting of SEQ ID NOs: 54, 56, 57, 59, 154, and 155.

[0235] In some embodiments, the epitope-directed anti-TREM-1 antibodies that can be used with the present methods comprise a heavy chain variable region (VH) CDR1, CDR2, and CDR3, and a light chain variable region (VL) CDR1, CDR2, and CDR3, wherein:

[0236] (a) the VH CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 61, the VH CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 62, the VH CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 63, the VL CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 64, the VL CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 65, and the VL CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 66;

[0237] (b) the VH CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 67, the VH CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 68, the VH CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 69, the VL CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 70, the VL CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 71, and the VL CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 72;

[0238] (c) the VH CDR1 comprises an amino acid sequence as set forth in SEQ ID NO: 67, the VH CDR2 comprises an amino acid sequence as set forth in SEQ ID NO: 68, the VH CDR3 comprises an amino acid sequence as set forth in SEQ ID NO: 69, the VL CDR1 comprises an amino acid sequence as set forth in SEQ ID NO: 64, the VL CDR2 comprises an amino acid sequence as set forth in SEQ ID NO: 65, and the VL CDR3 comprises an amino acid sequence as set forth in SEQ ID NO: 38;

[0239] (d) the VH CDR1 comprises an amino acid sequence as set forth in SEQ ID NO: 74, the VH CDR2 comprises an amino acid sequence as set forth in SEQ ID NO: 75, the VH CDR3 comprises an amino acid sequence as set forth in SEQ ID NO: 76, the VL CDR1 comprises an amino acid sequence as set forth in SEQ ID NO: 70, the VL CDR2 comprises an amino acid sequence as set forth in SEQ ID NO: 77, and the VL CDR3 comprises an amino acid sequence as set forth in SEQ ID NO: 78;

[0240] (e) the VH CDR1 comprises an amino acid sequence as set forth in SEQ ID NO: 79, the VH CDR2 comprises an amino acid sequence as set forth in SEQ ID NO: 80, the VH CDR3 comprises an amino acid sequence as set forth in SEQ ID NO: 81, the VL CDR1 comprises an amino acid sequence as set forth in SEQ ID NO: 70, the VL CDR2 comprises an amino acid sequence as set forth in SEQ ID NO: 71, and the VL CDR3 comprises an amino acid sequence as set forth in SEQ ID NO: 72;

[0241] (f) the VH CDR1 comprises an amino acid sequence as set forth in SEQ ID NO: 159, the VH CDR2 comprises an amino acid sequence as set forth in SEQ ID NO: 160, the VH CDR3 comprises an amino acid sequence as set forth in SEQ ID NO: 161, the VL CDR1 comprises an amino acid sequence as set forth in SEQ ID NO: 70, the VL CDR2 comprises an amino acid sequence as set forth in SEQ ID NO: 71, and the VL CDR3 comprises an amino acid sequence as set forth in SEQ ID NO: 162; or

[0242] (g) the VH CDR1 comprises an amino acid sequence as set forth in SEQ ID NO: 159, the VH CDR2 comprises an amino acid sequence as set forth in SEQ ID NO: 160, the VH CDR3 comprises an amino acid sequence as set forth in SEQ ID NO: 161, the VL CDR1 comprises an amino acid sequence as set forth in SEQ ID NO: 70, the VL CDR2 comprises an amino acid sequence as set forth in SEQ ID NO: 71, and the VL CDR3 comprises an amino acid sequence as set forth in SEQ ID NO: 133.

[0243] In some embodiments, an epitope-directed anti-TREM-1 antibody that can be used with the present methods comprises a heavy chain variable region (VH) CDR1, CDR2, and CDR3 and a light chain variable region (VL) CDR1, CDR2, and CDR3, wherein one or more CDRs comprise one or more amino acid mutations (e.g., substitutions or deletions) relative to one or more of the anti-TREM-1 antibodies disclosed herein. Thus, in certain embodiments, an epitope-directed anti-TREM-1 antibody comprises a VH CDR1 comprising X1, X2, X3, X4, and X5, wherein X1 is S or N; X2 is S, Y, or E; X3 is Y, G, or A; X4 is W, M, or I; and X5 is S, T, H, or N. In some embodiments, an epitope-directed anti-TREM-1 antibody comprises a VH CDR2 comprising X1, X2, X3, X4, X5, X6, X7, X8, X9, X10, X11, X12, X13, X14, X15, X16, and X17, wherein X1 is Y, V, or G; X2 is T or I; X3 is W, I, or absent; X4 is H, Y, or P; X5 is Y, D, or I; X6 is S, G, or F; X7 is G, S, or D; X8 is I, Y, N, or T; X9 is S, T, or K; X10 is N or Y; X11 is Y or G; X12 is N or A; X13 is P, D, or Q; X14 is S or K; X15 is L, V, or F; X16 is K or Q; and X17 is S or G. In some embodiments, an epitope-directed anti-TREM-1 antibody comprises a VH CDR3 comprising X1, X2, X3, X4, X5, X6, X7, X8, X9, X10, X11, X12, G, X13, X14, X15, X16, X17, X18, D, and X19, wherein X1 is E, D, M, T, or absent; X2 is G, V, or Y; X3 is Y, R, or absent; X4 is D, H, G, or absent; X5 is I, Y, or absent; X6 is L, Y, or absent; X7 is T, G, N, or absent; X8 is G, S, Y, or absent; X9 is Y, V, T, F, or H; X10 is E, L, S, or Y; X11 is Y, W, F, or H; X12 is Y or F; X13 is E or absent; X14 is L or absent; X15 is L or absent; X16 is P or absent; X17 is L or absent; X18 is M or L; and X19 is V or Y. In certain embodiments, an epitope-directed anti-TREM-1 antibody comprises a VL CDR1 comprising R, A, S, Q, X1, X2, X3, S, S, X4, L, and A, wherein X1 is S or G; X2 is V or I; X3 is S or absent; and X4 is Y or A. In some embodiments, an epitope-directed anti-TREM-1 antibody comprises a VL CDR2 comprising X1, A, S, S, X2, X3, and X4, wherein X1 is G, D, or A; X2 is R or L; X3 is A, E, or Q; and X4 is T or S.In certain embodiments, the epitope-directed anti-TREM-1 antibody comprises a VL CDR3 comprising Q, Q, X1, X2, S, X3, P, X4, and T, wherein X1 is Y or F; X2 is G or N; X3 is S or Y; and X4 is L, Y, I, or is absent.

[0244] In some embodiments, the anti-TREM-1 antibody useful for the present disclosure comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein

[0245] (a) the VH comprises an amino acid sequence as set forth in SEQ ID NO: 82, and the VL comprises an amino acid sequence as set forth in SEQ ID NO: 83;

[0246] (b) the VH comprises an amino acid sequence as set forth in SEQ ID NO: 84, and the VL comprises an amino acid sequence as set forth in SEQ ID NO: 85;

[0247] (c) the VH comprises an amino acid sequence as set forth in SEQ ID NO: 86, and the VL comprises an amino acid sequence as set forth in SEQ ID NO: 87;

[0248] (d) the VH comprises an amino acid sequence as set forth in SEQ ID NO: 88, and the VL comprises an amino acid sequence as set forth in SEQ ID NO: 89;

[0249] (e) the VH comprises an amino acid sequence as set forth in SEQ ID NO: 88, and the VL comprises an amino acid sequence as set forth in SEQ ID NO: 90;

[0250] (f) the VH comprises an amino acid sequence as set forth in SEQ ID NO: 88, and the VL comprises an amino acid sequence as set forth in SEQ ID NO: 83;

[0251] (g) the VH comprises an amino acid sequence as set forth in SEQ ID NO: 91, and the VL comprises an amino acid sequence as set forth in SEQ ID NO: 90;

[0252] (h) the VH comprises an amino acid sequence as set forth in SEQ ID NO: 88, and the VL comprises an amino acid sequence as set forth in SEQ ID NO: 92;

[0253] (i) the VH comprises an amino acid sequence as set forth in SEQ ID NO: 93, and the VL comprises an amino acid sequence as set forth in SEQ ID NO: 94;

[0254] (j) the VH comprises an amino acid sequence as set forth in SEQ ID NO: 95, and the VL comprises an amino acid sequence as set forth in SEQ ID NO: 96;

[0255] (k) the VH comprises an amino acid sequence as set forth in SEQ ID NO: 97, and the VL comprises an amino acid sequence as set forth in SEQ ID NO: 83;

[0256] (l) the VH comprises an amino acid sequence as set forth in SEQ ID NO: 97, and the VL comprises an amino acid sequence as set forth in SEQ ID NO: 98;

[0257] (m) the VH comprises an amino acid sequence as set forth in SEQ ID NO: 97, and the VL comprises an amino acid sequence as set forth in SEQ ID NO: 99;

[0258] (n) the VH comprises an amino acid sequence as set forth in SEQ ID NO: 97, and the VL comprises an amino acid sequence as set forth in SEQ ID NO: 100;

[0259] (o) the VH comprises an amino acid sequence as set forth in SEQ ID NO: 97, and the VL comprises an amino acid sequence as set forth in SEQ ID NO: 101;

[0260] (p) the VH comprises an amino acid sequence as set forth in SEQ ID NO: 97, and the VL comprises an amino acid sequence as set forth in SEQ ID NO: 89;

[0261] (q) the VH comprises an amino acid sequence as set forth in SEQ ID NO: 102, and the VL comprises an amino acid sequence as set forth in SEQ ID NO: 83;

[0262] (r) the VH comprises an amino acid sequence as set forth in SEQ ID NO: 102, and the VL comprises an amino acid sequence as set forth in SEQ ID NO: 92;

[0263] (s) the VH comprises an amino acid sequence as set forth in SEQ ID NO: 103, and the VL comprises an amino acid sequence as set forth in SEQ ID NO: 83;

[0264] (t) the VH comprises an amino acid sequence as set forth in SEQ ID NO: 104, and the VL comprises an amino acid sequence as set forth in SEQ ID NO: 83;

[0265] (u) the VH comprises an amino acid sequence as set forth in SEQ ID NO: 105, and the VL comprises an amino acid sequence as set forth in SEQ ID NO: 106;

[0266] (v) the VH comprises an amino acid sequence as set forth in SEQ ID NO: 107, and the VL comprises an amino acid sequence as set forth in SEQ ID NO: 108;

[0267] (w) the VH comprises an amino acid sequence as set forth in SEQ ID NO: 109, and the VL comprises an amino acid sequence as set forth in SEQ ID NO: 83;

[0268] (x) the VH comprises an amino acid sequence as set forth in SEQ ID NO: 110, and the VL comprises an amino acid sequence as set forth in SEQ ID NO: 111;

[0269] (y) the VH comprises an amino acid sequence as set forth in SEQ ID NO: 112, and the VL comprises an amino acid sequence as set forth in SEQ ID NO: 89;

[0270] (z) the VH comprises an amino acid sequence as set forth in SEQ ID NO: 156, and the VL comprises an amino acid sequence as set forth in SEQ ID NO: 157;

[0271] (aa) the VH comprises an amino acid sequence as set forth in SEQ ID NO: 156, and the VL comprises an amino acid sequence as set forth in SEQ ID NO: 83; or

[0272] (bb) the VH comprises an amino acid sequence as set forth in SEQ ID NO: 88, and the VL comprises an amino acid sequence as set forth in SEQ ID NO: 158.

[0273] In some embodiments, an anti-TREM-1 antibody for use in the present disclosure is non-epitope directed (i.e., can cross-compete with mAb0170 and / or mAb 0318 for binding to TREM-1 (human or cynomolgus monkey)). In some embodiments, a non-epitope directed anti-TREM-1 antibody comprises the CDRs of a heavy chain variable region selected from the group consisting of SEQ ID NOs: 82, 84, 86, 88, 91, 93, 95, 97, 102, 103, 104, 105, 107, 109, 110, 112, and 156. In some embodiments, a non-epitope directed anti-TREM-1 antibody comprises the CDRs of a light chain variable region selected from the group consisting of 83, 85, 87, 89, 90, 92, 94, 96, 98, 99, 100, 101, 106, 108, 111, 157, and 158.

[0274] In some embodiments, a non-epitope directed anti-TREM-1 antibody of the present disclosure comprises a heavy chain variable region (VH) CDR1, CDR2, and CDR3 and a light chain variable region (VL) CDR1, CDR2, and CDR3, wherein

[0275] (a) the VH CDR1 comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 74, 113, 118, 122, 128, 136, 139, 142, and 163;

[0276] (b) the VH CDR2 comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 114, 119, 123, 126, 127, 129, 131, 134, 137, 140, 143, 146, 149, and 164;

[0277] (c) the VH CDR3 comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 115, 120, 124, 130, 135, 138, 141, 144, 145, 147, 150, and 165;

[0278] (d) the VL CDR1 comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 116 and 42;

[0279] (e) the VL CDR2 comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 77 and 65; and / or

[0280] (f) the VL CDR3 comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 73, 78, 117, 121, 125, 133, 148, and 166.

[0281] In some embodiments, the anti-TREM-1 antibody (non-epitope directed) comprises VH CDR1, CDR2, and CDR3, and VL CDR1, CDR2, and CDR3, wherein:

[0282] (a) the VH CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 113, the VH CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 114, the VH CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 115, the VL CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 116, the VL CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 77, and the VL CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 117;

[0283] (b) the VH CDR1 comprises an amino acid sequence as set forth in SEQ ID NO: 118, the VH CDR2 comprises an amino acid sequence as set forth in SEQ ID NO: 119, the VH CDR3 comprises an amino acid sequence as set forth in SEQ ID NO: 120, the VL CDR1 comprises an amino acid sequence as set forth in SEQ ID NO: 116, the VL CDR2 comprises an amino acid sequence as set forth in SEQ ID NO: 77, and the VL CDR3 comprises an amino acid sequence as set forth in SEQ ID NO: 121 ;

[0284] (c) the VH CDR1 comprises an amino acid sequence as set forth in SEQ ID NO: 122, the VH CDR2 comprises an amino acid sequence as set forth in SEQ ID NO: 123, the VH CDR3 comprises an amino acid sequence as set forth in SEQ ID NO: 124, the VL CDR1 comprises an amino acid sequence as set forth in SEQ ID NO: 116, the VL CDR2 comprises an amino acid sequence as set forth in SEQ ID NO: 77, and the VL CDR3 comprises an amino acid sequence as set forth in SEQ ID NO: 125;

[0285] (d) the VH CDR1 comprises an amino acid sequence as set forth in SEQ ID NO: 122, the VH CDR2 comprises an amino acid sequence as set forth in SEQ ID NO: 126, the VH CDR3 comprises an amino acid sequence as set forth in SEQ ID NO: 124, the VL CDR1 comprises an amino acid sequence as set forth in SEQ ID NO: 116, the VL CDR2 comprises an amino acid sequence as set forth in SEQ ID NO: 77, and the VL CDR3 comprises an amino acid sequence as set forth in SEQ ID NO: 78;

[0286] (e) the VH CDR1 comprises an amino acid sequence as set forth in SEQ ID NO: 122, the VH CDR2 comprises an amino acid sequence as set forth in SEQ ID NO: 126, the VH CDR3 comprises an amino acid sequence as set forth in SEQ ID NO: 124, the VL CDR1 comprises an amino acid sequence as set forth in SEQ ID NO: 116, the VL CDR2 comprises an amino acid sequence as set forth in SEQ ID NO: 77, and the VL CDR3 comprises an amino acid sequence as set forth in SEQ ID NO: 117;

[0287] (f) the VH CDR1 comprises an amino acid sequence as set forth in SEQ ID NO: 122, the VH CDR2 comprises an amino acid sequence as set forth in SEQ ID NO: 127, the VH CDR3 comprises an amino acid sequence as set forth in SEQ ID NO: 124, the VL CDR1 comprises an amino acid sequence as set forth in SEQ ID NO: 116, the VL CDR2 comprises an amino acid sequence as set forth in SEQ ID NO: 77, and the VL CDR3 comprises an amino acid sequence as set forth in SEQ ID NO: 78;

[0288] (g) the VH CDR1 comprises an amino acid sequence as set forth in SEQ ID NO: 128, the VH CDR2 comprises an amino acid sequence as set forth in SEQ ID NO: 129, the VH CDR3 comprises an amino acid sequence as set forth in SEQ ID NO: 130, the VL CDR1 comprises an amino acid sequence as set forth in SEQ ID NO: 116, the VL CDR2 comprises an amino acid sequence as set forth in SEQ ID NO: 77, and the VL CDR3 comprises an amino acid sequence as set forth in SEQ ID NO: 78;

[0289] (h) the VH CDR1 comprises an amino acid sequence as set forth in SEQ ID NO: 128, the VH CDR2 comprises an amino acid sequence as set forth in SEQ ID NO: 131, the VH CDR3 comprises an amino acid sequence as set forth in SEQ ID NO: 132, the VL CDR1 comprises an amino acid sequence as set forth in SEQ ID NO: 116, the VL CDR2 comprises an amino acid sequence as set forth in SEQ ID NO: 77, and the VL CDR3 comprises an amino acid sequence as set forth in SEQ ID NO: 117;

[0290] (i) the VH CDR1 comprises an amino acid sequence as set forth in SEQ ID NO: 128, the VH CDR2 comprises an amino acid sequence as set forth in SEQ ID NO: 131, the VH CDR3 comprises an amino acid sequence as set forth in SEQ ID NO: 132, the VL CDR1 comprises an amino acid sequence as set forth in SEQ ID NO: 116, the VL CDR2 comprises an amino acid sequence as set forth in SEQ ID NO: 77, and the VL CDR3 comprises an amino acid sequence as set forth in SEQ ID NO: 133;

[0291] (j) VH CDR1 comprises an amino acid sequence as set forth in SEQ ID NO: 128, VH CDR2 comprises an amino acid sequence as set forth in SEQ ID NO: 131, VH CDR3 comprises an amino acid sequence as set forth in SEQ ID NO: 132, VL CDR1 comprises an amino acid sequence as set forth in SEQ ID NO: 116, VL CDR2 comprises an amino acid sequence as set forth in SEQ ID NO: 77, and VL CDR3 comprises an amino acid sequence as set forth in SEQ ID NO: 78;

[0292] (k) VH CDR1 comprises an amino acid sequence as set forth in SEQ ID NO: 122, VH CDR2 comprises an amino acid sequence as set forth in SEQ ID NO: 131, VH CDR3 comprises an amino acid sequence as set forth in SEQ ID NO: 124, VL CDR1 comprises an amino acid sequence as set forth in SEQ ID NO: 116, VL CDR2 comprises an amino acid sequence as set forth in SEQ ID NO: 77, and VL CDR3 comprises an amino acid sequence as set forth in SEQ ID NO: 117;

[0293] (l) VH CDR1 comprises an amino acid sequence as set forth in SEQ ID NO: 74, VH CDR2 comprises an amino acid sequence as set forth in SEQ ID NO: 134, VH CDR3 comprises an amino acid sequence as set forth in SEQ ID NO: 135, VL CDR1 comprises an amino acid sequence as set forth in SEQ ID NO: 116, VL CDR2 comprises an amino acid sequence as set forth in SEQ ID NO: 77, and VL CDR3 comprises an amino acid sequence as set forth in SEQ ID NO: 117;

[0294] (m) VH CDR1 comprises an amino acid sequence as set forth in SEQ ID NO: 136, VH CDR2 comprises an amino acid sequence as set forth in SEQ ID NO: 137, VH CDR3 comprises an amino acid sequence as set forth in SEQ ID NO: 138, VL CDR1 comprises an amino acid sequence as set forth in SEQ ID NO: 116, VL CDR2 comprises an amino acid sequence as set forth in SEQ ID NO: 77, and VL CDR3 comprises an amino acid sequence as set forth in SEQ ID NO: 117;

[0295] (n) the VH CDR1 comprises an amino acid sequence as set forth in SEQ ID NO: 139, the VH CDR2 comprises an amino acid sequence as set forth in SEQ ID NO: 140, the VH CDR3 comprises an amino acid sequence as set forth in SEQ ID NO: 141, the VL CDR1 comprises an amino acid sequence as set forth in SEQ ID NO: 116, the VL CDR2 comprises an amino acid sequence as set forth in SEQ ID NO: 77, and the VL CDR3 comprises an amino acid sequence as set forth in SEQ ID NO: 78;

[0296] (o) the VH CDR1 comprises an amino acid sequence as set forth in SEQ ID NO: 142, the VH CDR2 comprises an amino acid sequence as set forth in SEQ ID NO: 143, the VH CDR3 comprises an amino acid sequence as set forth in SEQ ID NO: 144, the VL CDR1 comprises an amino acid sequence as set forth in SEQ ID NO: 116, the VL CDR2 comprises an amino acid sequence as set forth in SEQ ID NO: 77, and the VL CDR3 comprises an amino acid sequence as set forth in SEQ ID NO: 125;

[0297] (p) the VH CDR1 comprises an amino acid sequence as set forth in SEQ ID NO: 142, the VH CDR2 comprises an amino acid sequence as set forth in SEQ ID NO: 143, the VH CDR3 comprises an amino acid sequence as set forth in SEQ ID NO: 145, the VL CDR1 comprises an amino acid sequence as set forth in SEQ ID NO: 116, the VL CDR2 comprises an amino acid sequence as set forth in SEQ ID NO: 77, and the VL CDR3 comprises an amino acid sequence as set forth in SEQ ID NO: 117;

[0298] (q) the VH CDR1 comprises an amino acid sequence as set forth in SEQ ID NO: 74, the VH CDR2 comprises an amino acid sequence as set forth in SEQ ID NO: 146, the VH CDR3 comprises an amino acid sequence as set forth in SEQ ID NO: 147, the VL CDR1 comprises an amino acid sequence as set forth in SEQ ID NO: 116, the VL CDR2 comprises an amino acid sequence as set forth in SEQ ID NO: 77, and the VL CDR3 comprises an amino acid sequence as set forth in SEQ ID NO: 148;

[0299] (r) VH CDR1 comprises an amino acid sequence as set forth in SEQ ID NO: 74, VH CDR2 comprises an amino acid sequence as set forth in SEQ ID NO: 149, VH CDR3 comprises an amino acid sequence as set forth in SEQ ID NO: 150, VL CDR1 comprises an amino acid sequence as set forth in SEQ ID NO: 116, VL CDR2 comprises an amino acid sequence as set forth in SEQ ID NO: 77, and VL CDR3 comprises an amino acid sequence as set forth in SEQ ID NO: 78;

[0300] (s) VH CDR1 comprises an amino acid sequence as set forth in SEQ ID NO: 163, VH CDR2 comprises an amino acid sequence as set forth in SEQ ID NO: 164, VH CDR3 comprises an amino acid sequence as set forth in SEQ ID NO: 165, VL CDR1 comprises an amino acid sequence as set forth in SEQ ID NO: 116, VL CDR2 comprises an amino acid sequence as set forth in SEQ ID NO: 77, and VL CDR3 comprises an amino acid sequence as set forth in SEQ ID NO: 166;

[0301] (t) VH CDR1 comprises an amino acid sequence as set forth in SEQ ID NO: 163, VH CDR2 comprises an amino acid sequence as set forth in SEQ ID NO: 164, VH CDR3 comprises an amino acid sequence as set forth in SEQ ID NO: 165, VL CDR1 comprises an amino acid sequence as set forth in SEQ ID NO: 116, VL CDR2 comprises an amino acid sequence as set forth in SEQ ID NO: 77, and VL CDR3 comprises an amino acid sequence as set forth in SEQ ID NO: 117; or

[0302] (t) VH CDR1 comprises an amino acid sequence as set forth in SEQ ID NO: 122, VH CDR2 comprises an amino acid sequence as set forth in SEQ ID NO: 126, VH CDR3 comprises an amino acid sequence as set forth in SEQ ID NO: 124, VL CDR1 comprises an amino acid sequence as set forth in SEQ ID NO: 167, VL CDR2 comprises an amino acid sequence as set forth in SEQ ID NO: 65, and VL CDR3 comprises an amino acid sequence as set forth in SEQ ID NO: 73.

[0303] In some embodiments, anti-TREM-1 antibodies that can be used with the present methods comprise CDR and / or variable region sequences that are at least 80% identical (e.g., at least 85%, at least 95%, at least 95%, or at least 99% identical) to the CDR and / or variable region sequences disclosed herein (e.g., Table 9).

[0304] In some embodiments, an anti-TREM-1 antibody comprises a heavy chain and a light chain, wherein the heavy chain comprises a VH domain disclosed herein (e.g., those provided in Table 9) fused to a heavy chain constant region described herein (e.g., SEQ ID NO: 47, 48, 11, or 12). In some embodiments, an anti-TREM-1 antibody disclosed herein comprises a heavy chain and a light chain, wherein the light chain comprises a VL domain disclosed herein (e.g., those provided in Table 9) fused to a light chain constant region described herein (e.g., SEQ ID NO: 35).

[0305] In some embodiments, an anti-TREM-1 antibody comprises a heavy chain and a light chain, wherein the heavy chain comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 168-202, and / or wherein the light chain comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 203-210.

[0306] Heavy chains and light chains comprising an amino acid sequence that is at least 99%, 98%, 97%, 96%, 95%, 90%, 85%, or 80% identical to any of the heavy chains or light chains described herein can be used to form an anti-TREM-1 antibody having the desired characteristics, e.g., those further described herein.

[0307] In some embodiments, an anti-TREM1 antibody comprises a heavy chain constant region, wherein the heavy chain constant region comprises one or more amino acid substitutions selected from the group consisting of K214R, L234A, L235E, G237A, D356E, L358M, and any combination thereof (according to EU numbering). In some embodiments, an anti-TREM-1 antibody comprises a heavy chain constant region, wherein the heavy chain constant region comprises one or more amino acid substitutions selected from the group consisting of K214R, L234A, L235E, G237A, A330S, P331S, D356E, L358M, and any combination thereof (according to EU numbering). In some embodiments, an anti-TREM-1 antibody comprises a heavy chain constant region, wherein the heavy chain constant region comprises one or more amino acid substitutions selected from the group consisting of K214R, C226S, C229S, P238S, and any combination thereof (according to EU numbering). In some embodiments, an anti-TREM-1 antibody comprises a heavy chain constant region, wherein the heavy chain constant region comprises one or more amino acid substitutions selected from the group consisting of S131C, K133R, G137E, G138S, Q196K, I199T, N203D, K214R, C226S, C229S, P238S, and any combination thereof (according to EU numbering).

[0308] In some embodiments, the antibodies disclosed herein bind to anti-TREM-1 at one or more epitopes that are the same as the mAb 0318 antibody. In some embodiments, the anti-TREM-1 antibody is capable of specifically binding to (i) at least one amino acid residue selected from the group consisting of A21, T22, K23, L24, T25, E26, and any combination thereof of human TREM-1 (e.g., isoform 1, SEQ ID NO: 1), and (ii) at least one amino acid residue selected from the group consisting of A49, S50, S51, Q52, K53, A54, W55, Q56, 157, 158, R59, D60, G61, E62, M63, P64, K65, T66, L67, A68, C69, T70, E71, R72, P73, S74, K75, N76, S77, H78, P79, V80, Q81, V82, G83, R84, 185, and any combination thereof, and (iii) at least one amino acid residue selected from the group consisting of C113, V114, 1115, Y116, Q117, P118, P119, and any combination thereof. See WO 2016 / 009086.

[0309] In some embodiments, the anti-TREM-1 antibody is capable of specifically binding to amino acids D38 to F48 of SEQ ID NO: 1 (human TREM-1), as determined using, e.g., HX-MS or X-ray diffraction. In some embodiments, the anti-TREM-1 antibody has an epitope comprising one, two, three, four, five, six, seven, or all of the amino acid residues D38, V39, K40, C41, D42, Y43, T44, and L45 of SEQ ID NO: 1 (human TREM-1), and one, two, or all of the amino acid residues selected from the group consisting of E46, K47, and F48 of SEQ ID NO: 1 (human TREM-1), as determined using, e.g., HX-MS or X-ray diffraction. In certain embodiments, the anti-TREM-1 antibody has an epitope comprising one, two, three, or all of the amino acid residues selected from the group consisting of D42, E46, D92, and H93 of SEQ ID NO: 1 (human TREM-1), as determined using a variant of TREM-1 and surface plasmon resonance.

[0310] In some embodiments, the anti-TREM-1 antibodies of the present disclosure have an epitope comprising at least amino acid residues E46 and / or D92 of SEQ ID NO: 1 (human TREM-1), as determined using variants of TREM-1 and surface plasmon resonance. In some embodiments, the anti-TREM-1 antibodies comprise one, two, or all of the amino acid residues selected from the group consisting of L31, 186, and V101 of SEQ ID NO: 1 (human TREM-1). In certain embodiments, the anti-TREM-1 antibodies are capable of specifically binding to a polypeptide comprising amino acid residues E19 to L26 of Macaca fascicularis TREM-1 (SEQ ID NO: 7), as determined using, for example, HX-MS or X-ray diffraction.

[0311] In some embodiments, the anti-TREM-1 antibodies are capable of specifically binding to human TREM-1, wherein the epitope of the antibody comprises one, two, three, four, five, six, seven, eight, nine, or all of the amino acid residues selected from the group consisting of V39, K40, C41, D42, Y43, L45, E46, K47, F48, and A49 of SEQ ID NO: 1.

[0312] In some embodiments, the anti-TREM-1 antibodies are capable of specifically binding to human TREM-1, wherein the epitope of the antibody comprises D42 of SEQ ID NO: 1. In other embodiments, the anti-TREM-1 antibodies are capable of specifically binding to human TREM-1, wherein the epitope of the antibody comprises E46 of SEQ ID NO: 1. In some embodiments, the epitope of the antibody can comprise V39, C41, D42, Y43, L45 of SEQ ID NO: 1. In further embodiments, the epitope of the antibody can comprise E46, K47, and A49 of SEQ ID NO: 1. In one particular embodiment, the epitope of the anti-TREM-1 antibody can also comprise F48 of SEQ ID NO: 1.

[0313] In some embodiments, the anti-TREM-1 antibodies of the present disclosure comprise mutations in which one or more negatively charged residues in the light chain CDR1 and CDR3 regions of the antibody are replaced with uncharged residues. In some embodiments, the anti-TREM-1 antibodies comprise a substitution at one or more of amino acid residues D1, D30, D33, D74, D98, E27, and E97 of SEQ ID NO: 23 with an amino acid residue selected from the group consisting of glycine, alanine, serine, asparagine, glutamine, threonine, cysteine, and tyrosine. These mutations are referred to herein as “charge- compensating” mutations.

[0314] In some embodiments, the anti-TREM-1 antibodies of the present disclosure comprise mutations in the Fab-Fab interaction region of SEQ ID NO: 15 to reduce Fab-Fab dimerization. Previous mAb0318 antibody suggested that, due to the antibody comprising two Fabs, multimerization can affect viscosity. These mutations are referred to as“Fab-Fab interaction” mutations. In certain embodiments, the anti-TREM-1 antibodies comprise a mutation at any one of residues Y32, R52, S55, S56, N57, A59, M102, 1104, and R106 of SEQ ID NO: 15 or residues F32, D33, Y34, Y53, R54, and D98 of SEQ ID NO: 23 to an amino acid residue selected from the group consisting of glycine, alanine, serine, asparagine, glutamine, threonine, cysteine, lysine, arginine, tryptophan, histidine, and tyrosine.

[0315] In some embodiments, an anti-TREM-1 antibody as disclosed herein comprises a mutation at position 32 of SEQ ID NO: 23, wherein the phenylalanine is mutated to an amino acid selected from the group consisting of glycine, serine, threonine, cysteine, alanine, valine, leucine, isoleucine, and methionine. This mutation is based on the observation that an Ala substitution at position Y90 of SEQ ID NO: 1 improved the affinity of SEQ ID NO: 3 to TREM-1. It was found that Y90 interacts with the phenylalanine residue of SEQ ID NO: 23. Mutations made to SEQ ID NO: 23 to improve Fab-TREM-1 interaction are referred to as “Fab-TREM-1 interaction” mutations. Provided herein are anti-TREM-1 antibodies whose variable regions are linked (e.g., covalently linked or fused) to an Fc, e.g., an IgGl, IgG2, IgG3, or IgG4 Fc, which can be any allotype or alloallotype, e.g., for IgGl: G1m, G1m1(a), G1m2(x), G1m3(f), G1m17(z); for IgG2: G2m, G2m23(n); for IgG3: G3m, G3m21(g1), G3m28(g5), G3m11(b0), G3m5(b1), G3m13(b3), G3m14(b4), G3m10(b5), G3m15(s), G3m16(t), G3m6(c3), G3m24(c5), G3m26(u), G3m27(v); and for K: Km, Km1, Km2, Km3 (see, e.g., Jeffries et al. (2009) mAbs 1:1). In some embodiments, the variable regions of an anti-TREM-1 antibody disclosed herein are linked to an effectorless or largely effectorless Fc, e.g., IgGl. In some embodiments, the variable regions of an anti-TREM-1 antibody are linked to an Fc that has reduced or no binding to one or more FcyRs.

[0316] In some embodiments, the VH domain of an anti-TREM-1 antibody described herein can be fused to a constant domain of a human IgG (i.e., Fc), e.g., IgGl, IgG2, IgG3, or IgG4, which is naturally occurring or modified, e.g., as further described herein. For example, the VH domain can comprise the amino acid sequence of any of the VH domains described herein fused to a human IgG (e.g., IgGl) constant region, such as the following wild-type human IgGl constant domain amino acid sequence:

[0317] ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPA VLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPC PAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVH NAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKG QPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPV LDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK (SEQ ID NO: 9) or an allotype variant of the amino acid sequence of SEQ ID NO: 9, and having the following amino acid sequence:

[0318] (SEQ ID NO: 46; allotype-specific amino acid residues are in bold and underlined).

[0319] In some embodiments, the VH domain of an anti-TREM-1 antibody described herein can comprise the amino acid sequence of any of the VH domains described herein fused to an effectorless constant region, such as the following effectorless human IgGl constant domain amino acid sequence

[0320]

[0321] DKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK “IgGl.lf”, comprising substitutions L234A, L235E, G237A, A330S, and P331S, according to EU numbering, which are underlined)

[0322] or

[0323]

[0324]

[0325] (SEQ ID NO: 48; “IgGl.3f”, comprising substitutions L234A, L235E, and G237A, according to EU numbering, which are underlined).

[0326] For example, the allotype variant of IgGl comprises K97R, D239E, and / or L241M (indicated below by underlining and boldface) and is numbered according to the numbering in SEQ ID NOs: 46-48. Within the full-length heavy chain region, these amino acid substitutions are numbered as K214R, D356E, and L358M according to the EU numbering. In some embodiments, the constant region of the anti-TREM-1 antibody further comprises one or more mutations or substitutions at amino acids L117, A118, G120, A213, and P214 (indicated below by underlining) as numbered in SEQ ID NOs: 46-48 or L234, A235, G237, A330, and P331 according to the EU numbering. In further embodiments, the constant region of the anti-TREM-1 antibody comprises one or more mutations or substitutions at amino acids L117A, A118E, G120A, A213S, and P214S of SEQ ID NOs: 46-48 or L234A, L235E, G237A, A330S, and P331S according to the EU numbering. The constant region of the anti-TREM-1 antibody can further comprise one or more mutations or substitutions of L117A, A118E, and G120A of SEQ ID NO: 9 or L234A, L235E, and G237A according to the EU numbering.

[0327] In some embodiments, the VH domain of an anti-TREM-1 antibody described herein comprises the amino acid sequence of any of the VH domains described herein fused to an IgGl constant domain comprising the following amino acid sequence:

[0328]

[0329] (SEQ ID NO: 11; "IgGl-Aba", comprising substitutions K214R, C226S, C229S, and P238S, which are indicated below by underlining, according to the EU numbering); or

[0330] (SEQ ID NO: 12; "IgG4-Aba", comprising substitutions S131C, K133R, G137E, G138S, Q196K, I199T, N203D, K214R, C226S, C229S, and P238S, which are indicated below by underlining, according to the EU numbering).

[0331] The VL domains described herein can be fused to a constant domain of a human kappa or lambda light chain. For example, the VL domain of an anti-TREM-1 antibody can comprise the amino acid sequence of any of the VL domains described herein fused to the following human IgGl kappa light chain amino acid sequence:

[0332] RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 35)

[0333] In certain embodiments, the heavy chain constant region comprises a lysine or another amino acid at the C-terminus, e.g., it comprises the following last amino acids: LSPGK (SEQ ID NO: 151) in the heavy chain. In certain embodiments, the heavy chain constant region lacks one or more amino acids at the C-terminus and has, e.g., the C-terminal sequence LSPG (SEQ ID NO: 152) or LSP.

[0334] In some embodiments, the variable region of the anti-TREM-1 antibody is linked to an effectorless or predominantly effectorless Fc. In certain embodiments, the variable region of the anti-TREM-1 antibody is linked to an Fc selected from the group consisting of IgG1.1f, IgG1.3f, IgG1-Aba, and IgG4-Aba, as described herein.

[0335] In general, the variable regions described herein can be linked to an Fc comprising one or more modifications that generally alter one or more functional properties of the antibody, such as Fc receptor binding, inflammatory cytokine release, serum half-life, complement fixation, and / or antigen-dependent cellular cytotoxicity. In addition, the antibodies described herein can be chemically modified (e.g., one or more chemical moieties can be attached to the antibody) or modified to alter their glycosylation, thereby altering one or more functional properties of the antibody. Each of these embodiments is described in more detail below. Residue numbering in the Fc region is that of the EU index of Kabat.

[0336] An Fc region encompasses a domain derived from a constant region of an immunoglobulin (e.g., IgG1, IgG2, IgG3, IgG4, and other classes, such as IgA, IgD, IgE, and IgM), including fragments, analogs, variants, mutants, or derivatives of the constant region. A constant region of an immunoglobulin is defined as a naturally occurring or synthetically produced polypeptide that is homologous to the C-terminal region of an immunoglobulin and can include a CH1 domain, a hinge, a CH2 domain, a CH3 domain, or a CH4 domain, alone or in combination.

[0337] Ig molecules interact with various classes of cellular receptors. For example, IgG molecules interact with three classes of Fc receptors (FcγR) that are specific for IgG class antibodies, i.e., FcγRI, FcγRII, and FcγRIII. The important sequences for IgG binding to FcγR receptors have been mapped to the CH2 and CH3 domains. The serum half-life of an antibody is affected by the ability of the antibody to bind to Fc receptors (FcR).

[0338] In some embodiments, the Fc region of an anti-TREM-1 antibody is a variant Fc region, e.g., an Fc sequence that has been modified (e.g., by amino acid substitution, deletion, and / or insertion) relative to a parent Fc sequence (e.g., an unmodified Fc polypeptide from which the variant is subsequently derived) to provide a desired structural feature and / or biological activity.

[0339] For example, modifications can be made in the Fc region to generate an Fc variant that (a) increases or decreases antibody-dependent cell-mediated cytotoxicity (ADCC), (b) increases or decreases complement-mediated cytotoxicity (CDC), (c) increases or decreases affinity for Clq, and / or (d) increases or decreases affinity for an Fc receptor, relative to a parent Fc. Such Fe region variants typically comprise at least one amino acid modification in the Fe region. Combinatorial amino acid modifications are considered particularly desirable. For example, a variant Fc region can comprise two, three, four, five, etc. substitutions, e.g., of particular Fc region positions identified herein.

[0340] A variant Fc region can also comprise sequence alterations in which amino acids involved in disulfide bond formation are removed or replaced with other amino acids. Such removal can avoid reaction with other cysteine-containing proteins present in the host cell used to produce the anti-TREM-1 antibodies described herein. Even if the cysteine residues are removed, a single chain Fc domain can still form a dimeric Fc domain that is non-covalently bound together. In other embodiments, the Fc region can be modified to make it more compatible with a selected host cell. For example, the PA sequence near the N-terminus of a typical native Fc region, which can be recognized by a digestive enzyme in E. coli such as proline imino peptidase, can be removed. In other embodiments, one or more glycosylation sites within the Fc domain can be removed. Residues that are typically glycosylated (e.g., asparagine) can be deleted or replaced with a non-glycosylated residue (e.g., alanine). In other embodiments, sites involved in interaction with complement, such as the Clq binding site, can be removed from the Fc region. For example, the EKK sequence of human IgGl can be deleted or replaced. In certain embodiments, sites that affect binding to Fc receptors, preferably other than the salvage receptor binding site, can be removed. In other embodiments, the Fc region can be modified to remove ADCC sites. ADCC sites are known in the art; see, e.g., Sarmay et al., Molec. Immunol. 29(5):633-9 (1992) for ADCC sites in IgGl. Specific examples of variant Fc domains are disclosed in, e.g., WO 97 / 34631 and WO 96 / 32478.

[0341] In some embodiments, the hinge region of the Fc is modified such that the number of cysteine residues in the hinge region is altered, e.g., increased or decreased. This approach is further described in U.S. Patent No. 5,677,425 to Bodmer et al. The number of cysteine residues in the hinge region of the Fc is altered to, e.g., facilitate assembly of the light and heavy chains or to increase or decrease the stability of the antibody. In one embodiment, the Fc hinge region of the antibody is mutated to decrease the biological half-life of the antibody. More specifically, one or more amino acid mutations are introduced into the CH2-CH3 domain interface region of the Fc-hinge fragment such that the antibody has impaired binding to staphylococcal protein A (SpA) relative to native Fc-hinge domain SpA binding. This approach is described in greater detail in U.S. Patent No. 6,165,745 to Ward et al.

[0342] In yet other embodiments, the Fc region is altered by replacing at least one amino acid residue with a different amino acid residue to alter one or more effector functions of the antibody. For example, one or more amino acids selected from amino acid residues 234, 235, 236, 237, 297, 318, 320, 322, 330, and / or 331 can be replaced with a different amino acid residue such that the affinity of the antibody for an effector ligand is altered, but the antigen binding ability of the parent antibody is preserved. The effector ligand to which affinity is altered can be, for example, an Fc receptor or the Cl component of complement. This approach is described in greater detail in U.S. Patent Nos. 5,624,821 and 5,648,260 to Winter et al.

[0343] In some embodiments, one or more amino acids selected from amino acid residues 329, 331, and 322 can be replaced with a different amino acid residue such that Clq binding and / or complement dependent cytotoxicity (CDC) of the antibody is decreased or eliminated. This approach is described in greater detail in U.S. Patent No. 6,194,551 to Idusogie et al.

[0344] In some embodiments, one or more amino acid residues within amino acid positions 231 and 239 are altered to thereby alter the ability of the antibody to fix complement. This approach is further described in PCT Publication WO 94 / 29351 to Bodmer et al.

[0345] In some embodiments, the Fc region can be modified by modifying one or more amino acids at positions 234, 235, 236, 238, 239, 240, 241, 243, 244, 245, 247, 248, 249, 252, 254, 255, 256, 258, 262, 263, 264, 265, 267, 268, 269, 270, 272, 276, 278, 280, 283, 285, 286, 289, 290, 292, 293, 294, 295, 296, 298, 299, 301, 303, 305, 307, 309, 312, 313, 315, 320, 322, 324, 325, 326, 327, 329, 330, 331, 332, 333, 334, 335, 337, 338, 340, 360, 373, 376, 378, 382, 388, 389, 398, 414, 416, 419, 430, 433, 434, 435, 436, 437, 438, or 439 to reduce antibody-dependent cellular cytotoxicity (ADCC) and / or reduce affinity for Fcy receptors. Exemplary substitutions include 236A, 239D, 239E, 268D, 267E, 268E, 268F, 324T, 332D, and 332E. Exemplary variants include 239D / 332E, 236A / 332E, 236A / 239D / 332E, 268F / 324T, 267E / 268F, 267E / 324T, and 267E / 268F / 324T. Other modifications to enhance FcyR and complement interactions include, but are not limited to, substitutions 298A, 333A, 334A, 326A, 2471, 339D, 339Q, 280H, 290S, 298D, 298V, 243L, 292P, 300L, 396L, 3051, and 396L. These and other modifications are reviewed in Strohl, 2009, Current Opinion in Biotechnology 20:685-691.

[0346] Other Fc modifications that can be made to the Fc are those that serve to reduce or eliminate binding to FcyR and / or complement proteins, thereby reducing or eliminating Fc- mediated effector functions such as ADCC, ADCP, and CDC. Exemplary modifications include, but are not limited to, substitutions, insertions, and deletions at positions 234, 235, 236, 237, 267, 269, 325, 328, 330, and / or 331 (e.g., 330 and 331), where numbering is according to the EU index. Exemplary substitutions include, but are not limited to, 234A, 235E, 236R, 237A, 267R, 269R, 325L, 328R, 330S, and 331S (e.g., 330S and 331S), where numbering is according to the EU index. Fc variants can comprise 236R / 328R. Other modifications that reduce FcyR and complement interactions include substitutions 297A, 234A, 235A, 237A, 318A, 228P, 236E, 268Q, 309L, 330S, 331S, 220S, 226S, 229S, 238S, 233P, and 234V, and removal of glycosylation at position 297 by mutation or enzymatic means or by production in organisms that do not glycosylate proteins, such as bacteria. These and other modifications are reviewed in Strohl, 2009, Current Opinion in Biotechnology 20:685-691.

[0347] Optionally, the Fc region can comprise non-naturally occurring amino acid residues at additional and / or alternative positions known to those of skill in the art (see, e.g., U.S. Patent Nos. 5,624,821; 6,277,375; 6,737,056; 6,194,551; 7,317,091; 8,101,720; International Publication Nos. WO 00 / 42072; WO 01 / 58957; WO 02 / 06919; WO 04 / 016750; WO 04 / 029207; WO 04 / 035752; WO 04 / 074455; WO 04 / 099249; WO 04 / 063351; WO 05 / 070963; WO 05 / 040217, WO 05 / 092925, and WO 06 / 020114).

[0348] The affinity and binding properties of the Fc region for its ligands can be determined by a variety of in vitro assay methods (biochemical or immunological-based assays) known in the art, including but not limited to equilibrium methods (e.g., enzyme-linked immunosorbent assay (ELISA) or radioimmunoassay (RIA)) or kinetics (e.g., BIACORE analysis) and other methods such as indirect binding assays, competitive inhibition assays, fluorescence resonance energy transfer (FRET), gel electrophoresis, and chromatography (e.g., gel filtration). These and other methods can employ labeling of one or more of the components under examination and / or employ a variety of detection methods including, but not limited to, colorimetric, fluorescent, luminescent, or isotopic labels. Detailed descriptions of binding affinities and kinetics can be found in Paul, W. E., ed., Fundamental immunology, 4th Ed., Lippincott-Raven, Philadelphia (1999), which focuses on antibody-immunogen interactions.

[0349] In certain embodiments, the anti-TREM-1 antibodies of the present disclosure comprise an Fc with reduced or no ability to bind to FcyR. In some embodiments, the anti-TREM-1 antibody has reduced binding affinity for FcyRI (CD64), FcyRIIA (CD32), FcyRIIB (CD32), FcyRIIIA (CD16a), FcyRIIIB (CD16b), or any combination thereof, compared to an antibody comprising a heavy chain consisting of the amino acid sequence set forth in SEQ ID NO: 30 and a light chain consisting of the amino acid sequence set forth in SEQ ID NO: 34. In some embodiments, the anti-TREM-1 antibody has at least two-fold, at least three-fold, at least four-fold, at least five-fold, at least six-fold, at least seven-fold, at least eight-fold, at least nine-fold, or at least 10-fold reduced binding affinity for FcyRI (CD64) compared to an antibody comprising a heavy chain consisting of the amino acid sequence set forth in SEQ ID NO: 30 and a light chain consisting of the amino acid sequence set forth in SEQ ID NO: 34.

[0350] In some embodiments, the anti-TREM-1 antibody comprises an IgGl Fc variant comprising: (a) one or more amino acid substitutions selected from the group consisting of L234A, L235E, G237A, and any combination thereof according to EU numbering; (b) one or more amino acid substitutions selected from the group consisting of L234A, L235E, G237A, A330S, P331S, and any combination thereof; (c) one or more amino acid substitutions selected from the group consisting of K214R, C226S, C229S, P238S, and any combination thereof; (d) one or more amino acid substitutions selected from the group consisting of S131C, K133R, G137E, G138S, Q196K, I199T, N203D, K214R, C226S, C229S, P238S, and any combination thereof.

[0351] In some embodiments, an anti-TREM-1 antibody as disclosed herein has (a) an IgGl isotype and comprises one or more amino acid substitutions at amino acid residues in the Fc region selected from the group consisting of N297A, N297Q, D270A, D265A, L234A, L235A, C226S, C229S, P238S, E233P, L234V, P238A, A327Q, A327G, P329A, K322A, L234F, L235E, P331S, T394D, A330L, M252Y, S254T, T256E, L328E, P238D, S267E, L328F, E233D, G237D, H268D, P271G, A330R, and any combination thereof, wherein the numbering of residues is according to the EU or Kabat numbering, or comprises an amino acid deletion at a position in the Fc region corresponding to glycine 236; (b) an IgG2 isotype and comprises one or more amino acid substitutions at amino acid residues in the Fc region selected from the group consisting of P238S, V234A, G237A, H268A, H268Q, H268E, V309L, N297A, N297Q, A330S, P331S, C232S, C233S, M252Y, S254T, T256E, and any combination thereof, wherein the numbering of residues is according to the EU or Kabat numbering; or (c) an IgG4 isotype and comprises one or more amino acid substitutions at amino acid residues in the Fc region selected from the group consisting of E233P, F234V, L234A / F234A, L235A, G237A, E318A, S228P, L236E, S241P, L248E, T394D, M252Y, S254T, T256E, N297A, N297Q, and any combination thereof, wherein the numbering of residues is according to the EU or Kabat numbering. In some embodiments, (a) the Fc region further comprises one or more additional amino acid substitutions at amino acid residues selected from the group consisting of A330L, L234F; L235E, P331S, and any combination thereof, wherein the numbering of residues is according to the EU or Kabat numbering; (b) the Fc region further comprises one or more additional amino acid substitutions at positions selected from the group consisting of M252Y, S254T, T256E, and any combination thereof, wherein the numbering of residues is according to the EU or Kabat numbering; or (c) the Fc region further comprises a S228P amino acid substitution according to the EU or Kabat numbering. See WO2017 / 152102.

[0352] In certain embodiments, an Fc with reduced complement fixation is selected. An exemplary Fc with reduced complement fixation, e.g., an IgGl Fc, has the following two amino acid substitutions: A330S and P331S.

[0353] In certain embodiments, an Fc with substantially no effector function is selected, i.e., it has reduced binding to FcyRs and reduced complement fixation. An exemplary Fc with no effector function, e.g., an IgGl Fc, comprises the following five mutations: L234A, L235E, G237A, A330S, and P331S.

[0354] IV. Nucleic Acids, Vectors, and Cells

[0355] Another aspect described herein relates to nucleic acid molecules encoding the anti-TREM-1 antibodies described herein. The nucleic acids can be present in intact cells, in cell lysates, or in a partially purified or substantially pure form. The nucleic acids are "isolated" or "rendered substantially pure" when purified to a degree that removes other cellular materials and other contaminants, e.g., by standard techniques that include alkaline / SDS treatment, CsCl banding, column chromatography, restriction enzyme digestion, agarose gel electrophoresis, and others well known in the art. See, F. Ausubel et al. eds. (1987) Current Protocols in Molecular Biology, Greene Publishing and Wiley Interscience, New York. The nucleic acids described herein can be, e.g., DNA or RNA and can or can not contain intronic sequences. In some embodiments, the nucleic acids are cDNA molecules.

[0356] The nucleic acids described herein can be obtained using standard molecular biology techniques. For antibodies expressed by hybridomas (e.g., hybridomas prepared from transgenic mice carrying human immunoglobulin genes, as described further below), the cDNAs encoding the light and heavy chains of the antibodies produced by the hybridomas can be obtained by standard PCR amplification or cDNA cloning techniques. For antibodies obtained from an immunoglobulin gene library (e.g., using phage display techniques), the nucleic acid encoding the antibody can be recovered from the library.

[0357] In some embodiments, the nucleic acids described herein are those encoding the VH and VL sequences of the anti-TREM-1 antibodies of the disclosure. Exemplary DNA sequences encoding the VH and VL sequences are set forth in SEQ ID NOs: 36-39, 226-260 and 40, 261-295, respectively.

[0358] Methods for making anti-TREM-1 antibodies as disclosed herein can include expressing the heavy and light chains in a cell line comprising nucleotide sequences encoding the heavy and light chains and signal peptides (e.g., SEQ ID NOs: 36-39, 226-260, and 40, 261-295, respectively). Host cells comprising these nucleotide sequences are encompassed herein.

[0359] Once DNA fragments encoding the VH and VL segments are obtained, these DNA fragments can be further manipulated by standard recombinant DNA techniques, for example, to convert the variable region genes to full-length antibody chain genes, Fab fragment genes or scFv genes. In these manipulations, the DNA fragments encoding the VL or VH are operatively linked to another DNA segment via recombinant means. The term "operatively linked" is intended to mean that the two DNA fragments are connected in such a way as to permit them to be transcribed together into mRNA.

[0360] An isolated DNA encoding a VH region can be converted to a full-length heavy chain gene by operatively linking the VH-encoding DNA to another DNA molecule encoding a heavy chain constant region (hinge, CHI, CH2, and / or CH3). The sequences of human heavy chain constant region genes are known in the art (see, e.g., Kabat, E. A. et al. (1991) Sequences of Proteins of Immunological Interest, Fifth Edition, U.S. Department of Health and Human Services, NIH Publication No. 91-3242) and DNA fragments

[0361] An isolated DNA encoding a VL region can be converted to a full-length light chain gene (as well as a Fab light chain gene) by operatively linking the DNA encoding the VL to another DNA molecule encoding a light chain constant region, CL. Sequences of human light chain constant region genes are known in the art (see, e.g., Kabat, E. A. et al. (1991) Sequences of Proteins of Immunological Interest, Fifth Edition, U.S. Department of Health and Human Services, NIH Publication No. 91-3242) and DNA fragments encompassing these regions can be obtained by standard PCR amplification. The light chain constant region can be a kappa or lambda constant region.

[0362] Another aspect described herein relates to cells (e.g., host cells) expressing (e.g., recombinantly expressing) the anti-TREM-1 antibodies described herein and related polynucleotides and expression vectors. Also provided herein are vectors comprising a polynucleotide comprising a nucleotide sequence encoding an anti-TREM-1 antibody or fragment thereof. In some embodiments, the vectors can be used for recombinantly expressing the anti-TREM-1 antibodies described herein in host cells (e.g., mammalian cells). In some embodiments, the vectors can be used for gene therapy.

[0363] Suitable vectors for use in the present disclosure include expression vectors, viral vectors, and plasmid vectors. In some embodiments, the vector is a viral vector.

[0364] As used herein, an expression vector refers to any nucleic acid construct that contains the elements needed for transcription and translation of an inserted coding sequence, or, in the case of RNA viral vectors, the elements needed for replication and translation when introduced into an appropriate host cell. Expression vectors can include plasmids, phagemids, viruses, and their derivatives.

[0365] An expression vector of the present disclosure can include a polynucleotide encoding an antibody or antigen-binding portion thereof described herein. In some embodiments, the coding sequence for the antibody or antigen-binding portion thereof is operably linked to an expression control sequence. As used herein, two nucleic acid sequences are operably linked if they are covalently linked in a manner which allows each component nucleic acid sequence to retain its functional ability. Coding sequences and gene expression control sequences are said to be operably linked when they are covalently linked in a manner which places expression or transcription and / or translation of the coding sequence under the influence or control of the gene expression control sequence. Two DNA sequences are said to be operably linked if induction of a promoter in the 5' gene expression sequence results in transcription of the coding sequence, and if the nature of the linkage between the two DNA sequences does not (1) cause the introduction of a frame-shift mutation, (2) interfere with the ability of the promoter region to direct transcription of the coding sequence, or (3) interfere with the ability of the corresponding RNA transcript to be translated into a protein. Thus, a gene expression sequence will be operably linked to a coding nucleic acid sequence if the gene expression sequence is capable of effecting transcription of the coding nucleic acid sequence in such a manner that the resulting transcript is translated into the desired antibody or antigen-binding portion thereof.

[0366] Viral vectors include, but are not limited to, nucleic acid sequences from retroviruses, such as Moloney murine leukemia virus, Harvey murine sarcoma virus, murine mammary tumor virus, and Rous sarcoma virus; lentivirus; adenovirus; adeno-associated virus; SV40-type viruses; polyoma virus; Epstein-Barr virus; papilloma virus; herpes virus; vaccinia virus; polio virus; and RNA viruses such as retroviruses. Other vectors well known in the art can be readily employed. Certain viral vectors are based on noncytopathic eukaryotic viruses in which a nonessential gene has been replaced by the gene of interest. Noncytopathic viruses include retroviruses, whose life cycle involves reverse transcription of genomic viral RNA to DNA, followed by integration of the provirus into host cell DNA. Retroviruses have been approved for use in human gene therapy trials. Most useful are those that are replication defective (i.e., capable of directing the synthesis of the desired protein, but not of manufacturing infectious particles). Such genetically altered retroviral expression vectors have general utility for efficient transduction of genes in vivo. Standard protocols for producing replication defective retroviruses, including the steps of incorporating foreign genetic material into a plasmid, transfecting a packaging cell line with the plasmid, producing recombinant retroviruses by the packaging cell line, collecting viral particles from tissue culture medium, and infecting target cells with the viral particles, are provided in Kriegler, M., Gene Transfer and Expression, A Laboratory Manual, W. H. Freeman Co., New York (1990) and Murry, E. J., Methods in Molecular Biology, Vol. 7, Humana Press, Inc., Cliffton, N. J. (1991).

[0367] In some embodiments, the virus is an adeno-associated virus, a double-stranded DNA virus. Adeno-associated virus can be engineered to be replication defective and is capable of infecting a wide range of cell types and species. It also has advantages such as heat and lipid solvent stability; high transduction frequency in different lineages of cells, including hematopoietic cells; and lack of superinfection inhibition allowing multiple rounds of transduction. Adeno-associated virus has been reported to integrate into human cell DNA in a site-specific manner, minimizing the potential for insertional mutagenesis and variability in expression of inserted genes characteristic of retroviral infection. Furthermore, wild-type adeno-associated virus infection has been passed in tissue culture for over 100 passages in the absence of selective pressure, meaning that adeno-associated virus genome integration is a relatively stable event. Adeno-associated virus can also function in an extrachromosomal manner.

[0368] V. Immunoconjugates

[0369] The present disclosure also provides immunoconjugates comprising any of the anti-TREM-1 antibodies disclosed herein. In some embodiments, the immunoconjugate comprises an antibody or antigen-binding portion linked to an agent. In some embodiments, the immunoconjugate comprises a bispecific molecule disclosed herein linked to an agent (e.g., as a therapeutic or diagnostic agent).

[0370] For diagnostic purposes, suitable agents are detectable labels for whole body imaging, including radioisotopes, as well as radioisotopes, enzymes, fluorescent labels, and other suitable antibody tags for sample testing. Detectable labels that can be attached to any of the anti-TREM-1 antibodies described herein can be any of a variety of types currently used in the in vitro diagnostics field, including particulate labels, including metal sols such as colloidal gold, isotopes such as, for example, N 2 S 2 , N3S or N 4型 peptide chelators provide I 125 or Tc99), chromophores (including fluorescent labels, luminescent labels, phosphorescent labels, and the like), as well as enzymatic labels that convert a given substrate to a detectable label upon contact, and polynucleotide tags that are revealed upon amplification such as by polymerase chain reaction. Suitable enzymatic labels include horseradish peroxidase, alkaline phosphatase, and the like. For example, the label can be alkaline phosphatase detected by the presence or formation of chemiluminescence upon conversion of a 1,2 dioxetane substrate such as adamantlymethyloxyphosphoryloxyphenyl dioxetane (AMPPD), 3-(4-(methoxyspiro{1,2-dioxetane-3,2'-(5'-chloro)tricyclo{3.3.1.13,7}decane}-4-yl)phenyl phosphate disodium (CSPD)), and CDP- and CDP-Star®. Or other luminescent substrates well known to those skilled in the art, such as chelates of suitable lanthanides such as terbium (III) and europium (III). The means of detection is determined by the label chosen. The appearance of the label or its reaction product can be observed using the naked eye (in the case of a particulate label and accumulation at appropriate levels), or using an instrument such as a spectrophotometer, luminometer, fluorometer, and the like, all in accordance with standard protocols. or other luminescent substrates well known to those skilled in the art, such as chelates of suitable lanthanides such as terbium (III) and europium (III). The means of detection is determined by the label chosen. The appearance of the label or its reaction product can be observed using the naked eye (in the case of a particulate label and accumulation at appropriate levels), or using an instrument such as a spectrophotometer, luminometer, fluorometer, and the like, all in accordance with standard protocols.

[0371] In some embodiments, the conjugation method results in a substantially (or nearly) non-immunogenic linkage, such as a peptide bond (i.e., amide bond), a thio bond, a (steric hindered), disulfide bond, a hydrazone bond, and an ether bond. These linkages are nearly non-immunogenic and show reasonable stability in serum (see, e.g., Senter, P.D., Curr. Opin. Chem. Biol. 13 (2009) 235-244; WO 2009 / 059278; WO 95 / 17886).

[0372] Depending on the part and the biochemical properties of the antibody, different conjugation strategies can be employed. In case the part is naturally occurring or a recombinant of 50 to 500 amino acids, standard procedures for the synthetic chemistry of protein conjugates are described in textbooks and can easily be followed by the skilled person (see, e.g., Hackenberger, C. P. R., and Schwarzer, D., Angew. Chem. Int. Ed. Engl. 47 (2008) 10030-10074). In some embodiments, the reaction of a maleimido moiety with a cysteine residue within the antibody or the part is used. This is particularly suitable coupling chemistry in case, e.g., Fab or Fab'-fragments of antibodies are used. Alternatively, in some embodiments, coupling to the C-terminus of the antibody or the part is performed. C-terminal modification of proteins, e.g., Fab-fragments, can be performed as described (Sunbul, M., and Yin, J., Org. Biomol. Chem. 7 (2009) 3361-3371).

[0373] Generally, site-specific reactions and covalent coupling are based on the conversion of natural amino acids into amino acids with a reactivity that is orthogonal to the reactivity of the other functional groups present. For example, a specific cysteine in a rare sequence context can be enzymatically converted into an aldehyde (see Frese, M. A., and Dierks, T., ChemBioChem. 10 (2009) 425-427). Desired amino acid modifications can also be obtained by employing the specific enzymatic reactivity of certain enzymes with natural amino acids in a given sequence context (see, e.g., Taki, M., et al., Prot. Eng. Des. Sel. 17 (2004) 119-126; Gautier, A., et al. Chem. Biol. 15 (2008) 128-136; and Protease-catalyzed formation of C—N bonds is used by Bordusa, F., Highlights in Bioorganic Chemistry (2004) 389-403). Site-specific reactions and covalent coupling can also be achieved by selective reaction of terminal amino acids with appropriate modification reagents.

[0374] The reactivity of N-terminal cysteines with benzonitriles (see Ren, H., et al., Angew. Chem. Int. Ed. Engl. 48 (2009) 9658-9662) can be used to achieve site-specific covalent coupling.

[0375] Natural chemical ligation can also rely on C-terminal cysteine residues (Taylor, E. Vogel; Imperiali, B, Nucleic Acids and Molecular Biology (2009), 22 (Protein Engineering), 65-96).

[0376] US6437095 B1 describes a conjugation method based on the faster reaction of a cysteine in a stretch of negatively charged amino acids with a cysteine positioned in a stretch of positively charged amino acids.

[0377] This moiety can also be a synthetic peptide or a peptidomimetic. Insofar as the polypeptide is chemically synthesized, amino acids with orthogonal chemical reactivity can be incorporated during such synthesis (see, e.g., de Graaf, A. J. et al., Bioconjug. Chem. 20 (2009) 1281-1295). As various orthogonal functional groups are key and can be introduced into synthetic peptides, conjugation of such peptides to linkers is standard chemistry.

[0378] To obtain singly labeled polypeptides, the 1 : 1 stoichiometric conjugate can be separated from other conjugation byproducts by chromatography. This process can be facilitated by using dye-labeled binding pair members and charged linkers. By using this type of labeled and highly negatively charged binding pair members, singly conjugated polypeptides are easily separated from unlabeled polypeptides and polypeptides carrying more than one linker due to differences in charge and molecular weight that can be used for separation. Fluorescent dyes can be used to purify the complex from unbound components as labeled monovalent binders.

[0379] In some embodiments, the moiety linked to the anti-TREM-1 antibody is selected from the group consisting of a binding moiety, a labeling moiety, and a biologically active moiety.

[0380] The anti-TREM-1 antibodies described herein can also be conjugated to a therapeutic agent to form an immunoconjugate, such as an antibody-drug conjugate (ADC). Suitable therapeutic agents include antimetabolites, alkylating agents, DNA minor groove binders, DNA intercalators, DNA cross-linking agents, histone deacetylase inhibitors, nuclear export inhibitors, proteasome inhibitors, topoisomerase I or II inhibitors, heat shock protein inhibitors, tyrosine kinase inhibitors, antibiotics, and antimitotic agents. In ADCs, the antibody and therapeutic agent are preferably conjugated through a cleavable linker, such as a peptidyl, disulfide, or hydrazone linker. In some embodiments, the linker is a peptidyl linker, such as Val-Cit, Ala-Val, Val-Ala-Val, Lys-Lys, Pro-Val-Gly-Val-Val (SEQ ID NO: 49), Ala-Asn-Val, Val-Leu-Lys, Ala-Ala-Asn, Cit-Cit, Val-Lys, Lys, Cit, Ser, or Glu. ADCs can be prepared as described in U.S. Patent Nos. 7,087,600; 6,989,452; and 7,129,261; PCT publications WO 02 / 096910; WO 07 / 038658; WO 07 / 051081; WO 07 / 059404; WO 08 / 083312; and WO 08 / 103693; U.S. Patent Publications 20060024317; 20060004081; and 20060247295.

[0381] Anti-TREM-1 antibodies (e.g., those described herein) can also be used to detect TREM-1, such as human TREM-1, e.g., human TREM-1, in a tissue or tissue sample. The antibodies can be used, for example, in an ELISA assay or flow cytometry. In some embodiments, the anti-TREM-1 antibody is contacted with the cells (e.g., cells in a tissue) for a time suitable for specific binding to occur, and then a reagent (e.g., an antibody that detects the anti-TREM-1 antibody) is added. Exemplary assays are provided in the Examples. The anti-TREM-1 antibody can be a fully human antibody, or it can be a chimeric antibody, such as an antibody having human variable regions and murine constant regions or portions thereof. Exemplary methods for detecting TREM-1 (e.g., human TREM-1) in a sample (a cell or tissue sample) include (i) contacting the sample with an anti-TREM-1 antibody for a time sufficient to allow the anti-TREM-1 antibody to specifically bind to TREM-1 in the sample, and (2) contacting the sample with a detection reagent, e.g., an antibody, that specifically binds to the anti-TREM-1 antibody (such as to the Fc region of the anti-TREM-1 antibody), thereby detecting TREM-1 to which the anti-TREM-1 antibody is bound. A washing step can be included after incubation with the antibody and / or detection reagent. The anti-TREM-1 antibody used in these methods need not be linked to a label or detection reagent, as a separate detection reagent can be used.

[0382] Other uses of anti-TREM-1 antibodies, e.g., as monotherapy or in combination therapy, are provided elsewhere herein, e.g., in the section relating to combination therapy.

[0383] VI. Bispecific molecules

[0384] The anti-TREM-1 antibodies described herein can be used to form bispecific molecules. The anti-TREM-1 antibodies, or antigen-binding portions thereof, can be derivatized or linked to another functional molecule, such as another peptide or protein (e.g., a ligand for another antibody or receptor), to produce a bispecific molecule that binds to at least two different binding sites or target molecules. For example, an anti-TREM-1 antibody can be linked to an antibody or scFv that specifically binds to any protein that can serve as a potential target for a combination therapy, such as a protein described herein (e.g., an antibody to IP-10 or TNF-a). The antibodies described herein can in fact be derivatized or linked to more than one other functional molecule to produce a multispecific molecule that binds to more than two different binding sites and / or target molecules; such multispecific molecules are also intended to be encompassed by the term "bispecific molecule" as used herein. To produce the bispecific molecules described herein, the antibodies described herein can be functionally linked (e.g., by chemical coupling, genetic fusion, noncovalent

[0385] Accordingly, provided herein are bispecific molecules comprising at least a first binding specificity for TREM-1 and a second binding specificity for a second target epitope. In some embodiments described herein where the bispecific molecule is multispecific, the molecule can further comprise a third binding specificity.

[0386] In some embodiments, the bispecific molecules described herein comprise at least one antibody, or antibody fragment thereof, including, for example, a Fab, Fab', F(ab')2, Fv, or a single chain Fv (scFv), as a binding specificity. The antibody can also be a light chain or heavy chain dimer, or any minimal fragment thereof such as Fv or a single chain construct, as described in Ladner et al. U.S. Patent No. 4,946,778.

[0387] While human monoclonal antibodies are preferred, other antibodies that can be used in the bispecific molecules described herein are murine, chimeric, and humanized monoclonal antibodies.

[0388] The bispecific molecules described herein can be prepared by conjugating the binding specificities using methods known in the art. For example, each binding specificity of the bispecific molecule can be produced separately and then conjugated to one another. When the binding specificities are proteins or peptides, a variety of coupling or cross-linking agents can be used for covalent conjugation. Examples of cross-linking agents include protein A, carbodiimide, N-succinimidyl-acetyl-thioacetate (SATA), 5,5'-dithiobis(2-nitrobenzoic acid) (DTNB), o-phenylenedimaleimide (oPDM), N-3-(2-pyridyldithio)propionic acid succinimidyl ester (SPDP), and 4-(N-maleimidomethyl) cyclohexane-l-carboxylate sulfosuccinimidyl ester (sulfo-SMCC) (see, e.g., Karpovsky et al. (1984) J. Exp. Med. 160: 1686; Liu, MA et al. (1985) Proc. Natl. Acad. Sci. USA 82: 8648). Other methods include those described in Paulus (1985) Behring Ins. Mitt. No. 78, 118-132; Brennan et al. (1985) Science 229: 81-83; and Glennie et al. (1987) J. Immunol. 139: 2367-2375. Some conjugating agents are SATA and sulfo-SMCC, both of which are available from Pierce Chemical Co. (Rockford, IL).

[0389] When the binding specificities are antibodies, they can be conjugated by the sulfhydryl groups of the C-terminal hinge regions of the two heavy chains. In some embodiments, the hinge regions are modified to contain an odd number of sulfhydryl residues, preferably one sulfhydryl residue, prior to conjugation.

[0390] Alternatively, the two binding specificities can be encoded in the same vector and expressed and assembled in the same host cell. This approach is particularly useful when the bispecific molecule is a mAb x mAb, mAb x Fab, mAb x (scFv)2, Fab x F(ab')2, or ligand x Fab fusion protein. The bispecific antibody can comprise an antibody containing a scFv at the C-terminus of each heavy chain. The bispecific molecules described herein can be a single chain molecule comprising one single chain antibody and a binding determinant, or a single chain bispecific molecule comprising two binding determinants. The bispecific molecule can comprise at least two single chain molecules. Methods of making bispecific molecules are described, for example, in U.S. Patent No. 5,260,203; U.S. Patent No. 5,455,030; U.S. Patent No. 4,881,175; U.S. Patent No. 5,132,405; U.S. Patent No. 5,091,513; U.S. Patent No. 5,476,786; U.S. Patent No. 5,013,653; U.S. Patent No. 5,258,498; and U.S. Patent No. 5,482,858.

[0391] Binding of the bispecific molecules to their particular targets can be confirmed using art- recognized methods such as enzyme-linked immunosorbent assay (ELISA), radioimmunoassay (RIA), FACS analysis, bioassay (e.g., growth inhibition), or Western blot assay. Each of these assays generally detects the presence of a protein-antibody complex of particular interest by employing a labeled reagent (e.g., an antibody) specific for the complex of interest.

[0392] VII. Kits

[0393] Provided herein are kits comprising one or more of the anti-TREM-1 antibodies, or antigen binding portions thereof, bispecific molecules, or immunoconjugates thereof described herein. In some embodiments, provided herein is a pharmaceutical pack or kit comprising one or more containers filled with one or more of the components of a pharmaceutical composition described herein, such as one or more of the antibodies or antigen binding portions thereof provided herein, optionally with instructions for use. In some embodiments, the kit contains a pharmaceutical composition described herein and any prophylactic or therapeutic agents, such as those described herein.

[0394] VIII. Compositions and Formulations

[0395] Also provided herein are compositions (e.g., pharmaceutical compositions) and formulations comprising one or more of the anti-TREM-1 antibodies disclosed herein (including polynucleotides, vectors, and cells that encode and / or express the anti-TREM-1 antibodies). For example, in one embodiment, the present disclosure provides a pharmaceutical composition comprising one or more of the anti-TREM-1 antibodies disclosed herein formulated together with a pharmaceutically acceptable carrier.

[0396] As used herein, “pharmaceutically acceptable carrier” includes any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like that are physiologically compatible. In some embodiments, the carrier is suitable for intravenous, intramuscular, subcutaneous, parenteral, spinal, or topical administration (e.g., by injection or infusion). Depending on the route of administration, the active compound (i.e., the antibody, immunoconjugate, or bispecific molecule) can be coated in a material to protect the compound from the action of acids and other natural conditions that can inactivate the compound.

[0397] Accordingly, it is an object of the present disclosure to provide a pharmaceutical formulation that improves the stability of anti-TREM-1 antibodies and thus allows for their long-term storage. In some embodiments, the pharmaceutical formulations disclosed herein comprise: (a) an anti-TREM-1 antibody; (b) a buffer; (c) a stabilizer; (d) a salt; (e) a bulking agent; and / or (f) a surfactant. In some embodiments, the pharmaceutical formulation is stable for at least 1 month, at least 2 months, at least 3 months, at least 6 months, at least 1 year, at least 2 years, at least 3 years, at least 5 years, or more. In some embodiments, the formulation is stable when stored at 4°C, 25°C, or 40°C.

[0398] Buffer

[0399] The buffers useful in the present application can be weak acids or bases that serve to maintain the acidity (pH) of a solution near a selected value upon the addition of another acid or base. Suitable buffers can maximize the stability of the pharmaceutical formulation by maintaining the pH control of the formulation. Suitable buffers can also ensure physiologic compatibility or optimize solubility. Rheology, viscosity, and other properties can also depend on the pH of the formulation. Common buffers include, but are not limited to, histidine, citrate, succinate, acetate, and phosphate. In some embodiments, the buffer comprises histidine (e.g., L-histidine) and an isotonic agent, and the pH can be adjusted with acids or bases known in the art. In certain embodiments, the buffer is L-histidine. In certain embodiments, the pH of the formulation is maintained between about 2 and about 10, or between about 4 and about 8.

[0400] Stabilizers

[0401] Stabilizing agents are added to pharmaceutical products to stabilize the product. Such agents can stabilize proteins in a variety of different ways. Common stabilizing agents include, but are not limited to, amino acids such as glycine, alanine, lysine, arginine, or threonine, carbohydrates such as glucose, sucrose, trehalose, raffmose, or maltose, polyols such as glycerol, mannitol, sorbitol, cyclodextrin, or any type and molecular weight of defrming or PEG. In one aspect of the application, the stabilizing agent is selected to maximize the stability of the FIX polypeptide in the lyophilized preparation. In certain embodiments, the stabilizing agent is sucrose and / or arginine.

[0402] Fillers

[0403] Fillers can be added to pharmaceutical products to increase the bulk and mass of the product, thus facilitating their accurate metering and handling. Common fillers include, but are not limited to, lactose, sucrose, glucose, mannitol, sorbitol, calcium carbonate, or magnesium stearate.

[0404] Surfactants

[0405] Surfactants are amphiphilic substances that have a hydrophilic and a hydrophobic group. Surfactants can be anionic, cationic, zwitterionic, or non-ionic surfactants. Examples of non-ionic surfactants include, but are not limited to, alkyl ethoxylates, nonylphenol ethoxylates, amine ethoxylates, polyethylene oxide, polypropylene oxide, fatty alcohols such as cetyl alcohol or oleyl alcohol, cocamide MEA, cocamide DEA, polysorbate, or dodecyldimethylamine oxide. In some embodiments, the surfactant is polysorbate 20 or polysorbate 80.

[0406] In some embodiments, the pharmaceutical formulation of the present disclosure comprises:

[0407] (a) about 0.25 mg / mL to 250 mg / mL (e.g., 10 to 200 mg / mL) of an anti-TREM-1 antibody;

[0408] (b) about 20 mM histidine;

[0409] (c) about 150 mM sucrose;

[0410] (d) about 25 mM arginine; and

[0411] (e) about 50 mM NaCl.

[0412] The formulations can also contain one or more of a buffering system, a preservative, a tonicity agent, a chelating agent, a stabilizer, and / or a surfactant, as well as various combinations thereof. The use of preservatives, isotonic agents, chelating agents, stabilizers, and surfactants in pharmaceutical compositions is well known to the skilled artisan. Reference can be made to Remington: The Science and Practice of Pharmacy, 20thEdition, 1995. 19

[0413] In some embodiments, the pharmaceutical formulation is an aqueous formulation. Such formulations are typically solutions or suspensions, but can also include gels, dispersions, emulsions, and multiple phase materials. The term "aqueous formulation" is defined as a formulation comprising at least 50% w / w water. Likewise, the term "aqueous solution" is defined as a solution comprising at least 50% w / w water, and the term "aqueous suspension" is defined as a suspension comprising at least 50% w / w water.

[0414] In some embodiments, the pharmaceutical formulation is a lyophilized formulation to which a solvent and / or diluent is added by the physician or patient prior to use.

[0415] The pharmaceutical compositions described herein can also be administered in combination therapy, i.e., combined with other agents. For example, the combination therapy can include an anti-TREM-1 antibody described herein in combination with at least one other therapeutic agent. Examples of therapeutic agents that can be used in combination therapy can include other compounds, drugs, and / or agents used in the treatment of a disease or disorder (e.g., an inflammatory disorder). Such compounds, drugs, and / or agents can include, for example, anti-inflammatory drugs or antibodies that block or reduce inflammatory cytokine production. In some embodiments, the therapeutic agent can include an anti-IP-10 antibody, an anti-TNF-a antibody (e.g., adalimumab golimumab infliximab certolizumab pegol interferon beta-1a (e.g., AVONEX®), interferon beta-1b (e.g., BETASERON®), glatiramer acetate (e.g., COPAXONE®), mitoxantrone (e.g., NOVANTRONE®), non-steroidal anti-inflammatory drugs (NSAIDs), analgesics, corticosteroids, and combinations thereof.

[0416] ​The pharmaceutical compounds described herein can include one or more pharmaceutically acceptable salts. "Pharmaceutically acceptable salt" refers to salts that retain the desired biological activity of the parent compound and do not impart undesired toxicological effects (see, e.g., Berge, S.M. et al. (1977) J. Pharm. Sci. 66: 1-19). Examples of such salts include acid addition salts and base addition salts. Acid addition salts include those derived from nontoxic inorganic acids, such as hydrochloric, nitric, phosphoric, sulfuric, hydrobromic, hydroiodic, phosphorous, and the like, as well as those derived from nontoxic organic acids, such as aliphatic mono- and dicarboxylic acids, phenyl-substituted alkanoic acids, hydroxy alkanoic acids, aromatic acids, aliphatic and

[0417] The pharmaceutical compositions described herein can also include a pharmaceutically acceptable antioxidant. Examples of pharmaceutically acceptable antioxidants include: (1) water-soluble antioxidants, such as ascorbic acid, cysteine hydrochloride, sodium bisulfate, sodium metabisulfite, sodium sulfite, and the like; (2) oil-soluble antioxidants, such as ascorbyl palmitate, butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), lecithin, propyl gallate, alpha-tocopherol, and the like; (3) metal chelating agents, such as citric acid, ethylenediamine tetraacetic acid (EDTA), sorbitol, tartaric acid, phosphoric acid, and the like.

[0418] Examples of suitable aqueous and nonaqueous carriers that can be employed in the pharmaceutical compositions described herein include water, ethanol, polyols (such as glycerol, propylene glycol, polyethylene glycol, and the like), and suitable mixtures thereof, vegetable oils, such as olive oil, and injectable organic esters, such as ethyl oleate. Proper fluidity can be maintained, for example, by the use of coating materials, such as lecithin, by the maintenance of the required particle size in the case of dispersion, and by the use of surfactants.

[0419] These compositions can also contain adjuvants such as preservatives, wetting agents, emulsifying agents, and dispersing agents. Prevention of presence of microorganisms can be ensured both by sterilization procedures, supra, and by the inclusion of various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol sorbic acid, and the like. It can also be desirable to include isotonic agents, for example, sugars, sodium chloride, and the like into the compositions. In addition, prolonged absorption of the injectable pharmaceutical form can be brought about by the inclusion of agents which delay absorption, for example, aluminum monostearate and gelatin.

[0420] Pharmaceutically acceptable carriers include sterile aqueous solutions or dispersions and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersions. The use of such media and agents for pharmaceutically active substances is known in the art. Except insofar as any conventional media or agent is incompatible with the active compound, its use in the pharmaceutical compositions described herein is contemplated. The pharmaceutical compositions can contain preservatives or can be preservative-free. Supplementary active compounds can be incorporated into the compositions.

[0421] Therapeutic compositions generally must be sterile to prevent infection of the subject to be treated. The compositions can be formulated to be stable under the conditions of manufacture and storage, yet to be delivered to the subject in a form suitable for use. Sterile injectable forms can be aqueous or non-aqueous solutions, suspensions, emulsions, or other like form. Examples of non-aqueous

[0422] Sterile injectable solutions can be prepared by incorporating the active compound in the required amount in an appropriate solvent with one or a combination of ingredients enumerated above, as required, followed by sterilization microfiltration. Generally, dispersions are prepared by incorporating the active compound into a sterile vehicle that contains a basic dispersion medium and the required other ingredients from those enumerated above. In the case of sterile powders for the preparation of sterile injectable solutions, some methods of preparation are vacuum drying and freeze-drying (lyophilization) that yield a powder of the active ingredient plus any additional desired ingredient from the previously sterile-filtered solution thereof.

[0423] The amount of active ingredient that can be combined with the carrier materials to produce a single dosage form will vary depending upon the subject being treated and the particular mode of administration. The amount of active ingredient that can be combined with a carrier material to produce a single dosage form will generally be that amount of the composition that produces a therapeutic effect. Generally, out of 100%, this amount will range from about 0.01% to about 99%, about 0.1% to about 70%, or about 1% to about 30% of the active ingredient, combined with a pharmaceutically acceptable carrier.

[0424] Dosage regimens are adjusted to provide the optimum desired response (e.g., a therapeutic response). For example, a single dose can be administered, several divided doses can be administered over time or the dose can be proportionally reduced or increased as indicated by the exigencies of the therapeutic situation. It is especially advantageous to formulate parenteral compositions in dosage unit form for ease of administration and uniformity of dosage. Dosage unit form as used herein refers to physically discrete units suited as unitary dosages for the subjects to be treated; each unit containing a predetermined quantity of active compound calculated to produce the desired therapeutic effect in association with the required pharmaceutical carrier. The specification for the dosage unit forms described herein are dictated by and directly dependent on (a) the unique characteristics of the active compound and the particular therapeutic effect to be achieved, and (b) the limitations inherent in the art of compounding such an active compound for the treatment of an individual.

[0425] For administration of, e.g., an anti-TREM-1 antibody described herein, the dosage ranges from about 0.0001 to 100 mg / kg, and more usually 0.01 to 5 or 10 mg / kg, of the host body weight. For example dosages can be 0.3 mg / kg, 1 mg / kg, 3 mg / kg, 5 mg / kg or 10 mg / kg of body weight or within the range of 1-10 mg / kg. An exemplary treatment regimen entails administration once per week, once every two weeks, once every three weeks, once every four weeks, once a month, once every 3 months, or once every three to 6 months. Exemplary dosage regimens for an anti-TREM-1 antibody described herein include 1 mg / kg or 3 mg / kg of body weight via intravenous administration, with the antibody being given using one of the following dosing schedules: (i) six doses every four weeks, then once every three months; (ii) every three weeks; (iii) 3 mg / kg once, then 1 mg / kg every three weeks.

[0426] In some embodiments, an anti-TREM-1 antibody is administered at a flat dose (flat dosing regimen). In other embodiments, an anti-TREM-1 antibody is administered at a fixed dose with another antibody. In certain embodiments, an anti-TREM-1 antibody is administered at a weight-based dose.

[0427] In some methods, two or more monoclonal antibodies with different binding specificities are administered simultaneously, in which case the dose of each antibody to be administered falls within the ranges indicated. The antibodies will usually be administered in a variety of combinations. The interval between single doses can be, for example, weekly, monthly, every three months or yearly. The interval can also be irregular, as indicated by the blood levels of the antibodies against the target antigens in the patient. In some methods, the dose is adjusted to achieve a plasma antibody concentration of about 1-1000 pg / ml, and in some methods, about 25-300 pg / ml.

[0428] The antibodies can be administered as sustained-release formulations, in which case less frequent administration is required. Dosage and frequency of administration will vary depending on the half-life of the antibody in the patient. In general, human antibodies have the longest half-life, followed by humanized antibodies, chimeric antibodies and non-human antibodies. The dosage and frequency of administration can vary depending on whether the treatment is prophylactic or therapeutic. In prophylactic applications, a relatively low dosage is administered at relatively infrequent intervals over a long period of time. Some patients continue to receive treatment for the rest of their lives. In therapeutic applications, the relatively low dosage can be administered at relatively frequent intervals up until the disease progresses to a point where the treatment is no longer effective. At such time, the patient can be given a more intensive treatment in an attempt to check the progress of the disease. Subsequently, the patient can be given a prophylactic treatment.

[0429] Actual dosage levels of the active ingredients in the pharmaceutical compositions described herein can be varied so as to obtain an amount of the active ingredient which is effective to achieve the desired therapeutic response for a particular patient, composition, and mode of administration, without being toxic to the patient. The selected dosage level will depend on a variety of pharmacokinetic factors including the activity of the particular compositions described herein, or an ester, salt or amide thereof, employed, the route of administration, the time of administration, the rate of excretion of the particular compound being employed, the duration of the treatment, other drugs, compounds and / or materials used in combination with the particular compositions employed, the age, sex, weight, condition, general health and prior medical history of the patient being treated, and like factors well known in the medical arts.

[0430] The compositions described herein can be administered using one or more routes of administration using one or more of a variety of methods known in the art. The skilled artisan will appreciate that the route and / or means of administration will vary depending on the desired results. Routes of administration of the anti-TREM-1 antibodies described herein can include intravenous, intramuscular, intradermal, intraperitoneal, subcutaneous, spinal, or other parenteral routes of administration, such as by injection or infusion. As used herein, the phrase “parenteral administration” refers to modes of administration other than enteral and topical administration, usually by injection, including, but not limited to, intravenous, intramuscular, intraarterial, intrathecal, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subcuticular, intraarticular, subcapsular, subarachnoid, intraspinally, epidural, and intrastemal injection and infusion.

[0431] Alternatively, the antibodies described herein can potentially be administered by non-parenteral routes, such as topical, epidermal or mucosal routes of administration, for example, intranasally, orally, vaginally, rectally, sublingually or topically.

[0432] The active compounds can be prepared with carriers that protect the compound against rapid release, such as a controlled release formulation, including implants, transdermal patches, and microencapsulated delivery systems. Biodegradable, biocompatible polymers can be used, such as ethylene vinyl acetate, polyanhydrides, polyglycolic acid, collagen, polyorthoesters, and polylactic acid. Many methods for the preparation of such formulations are patented or well known to those skilled in the art. See, e.g., Sustained and Controlled Release Drug Delivery Systems, J.R. Robinson, ed., Marcel Dekker, Inc., New York, 1978.

[0433] The therapeutic compositions can be administered with medical devices known in the art. For example, in one particular embodiment, the therapeutic compositions described herein can be administered with a needleless hypodermic injection device, such as the devices disclosed in U.S. Patent Nos. 5,399,163; 5,383,851; 5,312,335; 5,064,413; 4,941,880; 4,790,824; or 4,596,556. Examples of well-known implants and modules that can be used with the anti-TREM-1 antibodies described herein include: U.S. Patent No. 4,487,603, which discloses an implantable micro-infusion pump for dispensing medication at a controlled rate; U.S. Patent No. 4,486,194, which discloses a therapy device for administering medication through the skin; U.S. Patent No. 4,447,233, which discloses a medication infusion pump for delivering medication at a precise infusion rate; U.S. Patent No. 4,447,224, which discloses a variable flow implantable infusion apparatus for continuous drug delivery; U.S. Patent No. 4,439,196, which discloses an osmotic drug delivery system having multi-chamber compartments; and U.S. Patent No. 4,475,196, which discloses an osmotic drug delivery system. These patents are incorporated herein by reference. Numerous other such implants, delivery systems, and modules are known to those skilled in the art.

[0434] In some embodiments, the anti-TREM-1 antibodies described herein can be formulated to ensure proper in vivo distribution. For example, the blood-brain barrier (BBB) excludes many highly hydrophilic compounds. To ensure that the therapeutic compounds described herein cross the BBB, if desired, e.g., for brain cancer, they can be formulated, e.g., in a liposome. For methods of manufacturing liposomes, see, e.g., U.S. Patents 4,522,811; 5,374,548; and 5,399,331. Liposomes can comprise one or more moieties which are selectively transported into specific cells or organs, thus enhancing targeted drug delivery (see, e.g., V. V. Ranade (1989) J. Clin. Pharmacol. 29:685). Exemplary targeting moieties include folate or biotin (see, e.g., U.S. Patent 5,416,016 to Low et al.); a mannose glycoside (Umezawa et al. (1988) Biochem. Biophys. Res. Commun. 153:1038); an antibody (P.G. Bloeman et al. (1995) FEBS Lett. 357:140; M. Owais et al. (1995) Antimicrob. Agents Chemother. 39:180); a surfactant protein A receptor (Briscoe et al. (1995) Am. J. Physiol. 1233:134); pl20 (Schreier et al. (1994) J. Biol. Chem. 269:9090); see also K. Keinanen; M.L. Laukkanen (1994) FEBS Lett. 346:123; J.J. Killion; I.J. Fidler (1994) Immunomethods 4:273.

[0435] The following examples are offered by way of illustration and not by way of limitation. The contents of all references cited throughout this application are expressly incorporated herein by reference.

[0436] EXAMPLE

[0437] Example 1: Comparison of TREM-1 activation with PGLYRP1 alone or in combination with different PGNs

[0438] By identifying PGLYRP1 as a ligand for TREM-1, the differential expression of genes upon TREM-1 receptor ligation was assessed in monocytes and neutrophils. Monocytes and neutrophils were isolated from peripheral blood mononuclear cells (PBMCs) of whole blood from healthy human donors. To isolate the cells, whole blood from healthy donors was layered on a ficoll gradient. Monocytes were extracted from the peripheral blood mononuclear cell (PBMC) layer, while neutrophils were isolated from the red blood cell (RBC) containing layer. The neutrophil layer was then resuspended in HETASEP TM solution (Stem Cell Technologies) and incubated at 37°C for 1 hour to precipitate RBCs. Isolated neutrophils were then resuspended in sterile water for 30 seconds, followed by the addition of 0.6 M KCl. For monocyte isolation, ficoll purified PBMCs were washed and then purified using EASYSEP TM Human Monocyte Enrichment Kit (Stem Cell Technologies) to purify monocytes.

[0439] Once isolated, monocytes and neutrophils (1 x 10 6 cells / well) were plated in 24-well plates. Cells were then stimulated by the following conditions: (i) no stimulation, (ii) PGLYRP1 (soluble), (iii) PGLYRP1 (plated) or (iv) PGLYRP1 + PGN. Previous studies have shown that PGLYRP1 in combination with the bacterial component peptidoglycan (PGN) can effectively induce TREM-1 signaling. PGN derived from S. aureus (PGN-SA), E. coli (PGN-EK), B. subtilis (PGN-BS) or lacking TLR2 activity (PGN-ECndss) were used. Recombinant PGRP was plated at 5 pg / ml O / N on Nunc Maxisorp plates. Plates were washed and PGN with purified monocytes and neutrophils were added to a final concentration of 10 pg / ml. Cultures were incubated overnight at 37°C, 5% C02 and media was removed to measure cytokine production. To measure TREM-1 activity, TNF-a levels produced by monocytes and neutrophils were measured by AlphaLISA assay.

[0440] Soluble PGLYRP1 was unable to stimulate TNF-a expression in purified human monocytes (data not shown), while plate- immobilized PGLYRP1 induced TNF-a production Figure 1A ). PGN derived from S. aureus (PGN-SA) induced TNF-a secretion from human monocytes to similar levels as PGLYRP1 plated, while the addition of PGLYRP1 and PGN-SA demonstrated a synergistic effect, asFigure 1A PGN extracted from E. coli (PGN-EK) and B. subtilis (PGN-BS) were observed to have similar PGN-dependent monocyte activation through TLR2 receptor engagement. See Figure 1B and 1C PGN-ECndss (i.e., lacking TLR2 activity) was unable to induce TNF-a production alone, but effectively increased the PGLYRP1 -mediated response Figure 1D ). Thus, to reduce background from PGN signaling, PGN-ECndss was used in subsequent examples.

[0441] Example 2: Evaluation of TREM-1 gene signatures following TREM-1 engagement

[0442] To generate robust TREM-1 gene signatures by gene expression analysis, the transcriptomic profiling pattern following TREM-1 engagement was evaluated. Briefly, peripheral blood mononuclear cells and neutrophils were isolated from healthy donors and plated onto 24-well plates as described in Example 1 above. Isolated mononuclear cells and neutrophils were stimulated for 6 or 24 hours with the following conditions: (i) no stimulation, (ii) PGLYRP1 alone (PGRP), (iii) PGN-ECndss alone (PGN), or (iv) a combination of PGLYRP1 and PGN-ECndss (PGN+PGRP). PGN-ECndss, PGN-EK, PGN-SA, and PGN-BS were obtained through Invivogen. To ensure that any induced genes were specific to TREM-1 pathway engagement, some mononuclear cells and neutrophils were stimulated with PGLYRP1 + PGN-ECndss in the presence of a TREM-1 blocking antibody or isotype antibody control. Following stimulation, RNA was isolated from cells using the RNeasy Micro Kit (Qiagen, Valencia, CA). RNA quality was monitored using the Agilent 2100 Bioanalyzer (Agilent Technologies, Palo Alto, CA) and RNA quantity was measured using the NanoDrop (NanoDrop Technologies). Fifty nanograms of RNA was amplified and labeled with the Nugen WT-Pico Ovation System and Encore Biotin Module Assay (NuGEN Technologies Inc.). Labeled cRNA / cDNA was hybridized on the Affymetrix GeneChip Human Genome U219 Array Plate (Affymetrix) and processed according to the manufacturer’s recommended protocol.

[0443] Custom CDF BrainArray was used to annotate Affy-based mRNA expression data (.cel files from U219 and U133Plus platforms) in units of Entrez Gene IDs. Log2RMA was used to normalize expression values. To identify differentially expressed genes and construct the TREM-1 module, a linear mixed model was fitted with ligand stimulation or antibody treatment as fixed effects and donors as random effects. Single-sample gene set enrichment (ssGSEA) was used to apply gene modules to tissue or blood mRNA profiling data from patients and derive quantitative scores representing gene module enrichment. Bioinformatics and statistical analyses (e.g. multiple regression to identify TREM-1 signature-associated biomarkers) were performed in R using relevant Bioconductor packages (e.g. LIMMA) and Omicsoft ArrayStudio. Visualizations were performed in R ggplot framework and Omicsoft ArrayStudio.

[0444] As shown in Figure 2 , principal component analysis showed that PGN-ECndss alone (media with P) did not significantly enhance or suppress global gene expression patterns in monocytes at 24 hours compared to unstimulated. However, PGN-ECndss with PGLYRP1 (PGN+PGRP) induced a large number of genes that formed a separate cluster shown in the PCA analysis (upper right quadrant, Figure 2 ). PGLYRP1 alone enhanced a subset of genes that was further increased when PGN-ECndss was added. Addition of anti-TREM-1 blocking antibody (but not isotype control antibody) suppressed the genes induced by PGLYRP1 + PGN-ECndss treatment, confirming that the induced genes were specific to the TREM-1 pathway (lower left quadrant, Figure 2 ; see also Figure 3 ). In neutrophils, we observed weaker expression pattern changes under PGLYRP1 + PGN-ECndss stimulation at 6 hours and greater donor-to-donor differences, as shown in Figure 4A and 4B . Therefore, gene expression in monocytes after 24 hours stimulation was used as the primary source to generate the TREM-1 module.

[0445] Gene selection for the TREM-1 module was based on the following points: 1) PGN-ECndss+PGLYRP1 stimulation significantly upregulated compared to PGN-ECndss alone (P+L vs P, fold change >4, FDR <0.05, 292 genes in total); 2) PGN-ECndss+PGLYRP1+TREM-1 antibody significantly downregulated compared to PGN-ECndss+PGLYRP1 (P+L+TREM-1 vs P+L, fold change <-2, FDR <0.05, 286 out of 292 genes in 1), and 3) no significant differential expression in PGN-ECndss stimulation (P vs media, fold FDR >0.05, 180 out of 286 genes in 2) and no significant differential expression in PGN-ECndss+PGLYRP1 vs PGN-ECndss+isotype control (P+L vs P+L+iso, FDR >0.05, all 180 genes).

[0446] A total of 180 genes passed the defined selection criteria. GO functional annotation of genes in the TREM-1 module showed enrichment in extracellular space and plasma membrane-localized proteins (Table 4 below) (LOD = log odds; pVal = probability value; Pcor = probability of correlation). Metacore pathway enrichment analysis of the TREM-1 module included gene networks involved in chemotaxis, inflammatory response (TH17 derived and innate inflammation), and cell proliferation (Table 5 below) (FDR = false discovery rate). The top 20 genes in the 180 gene module ordered by the magnitude of PGLYRP1 specific stimulation are listed in Table 6 (below).

[0447] Table 4.

[0448] Item Description LOD Pval Pcor GO:0005615 extracellular space 34.8 1.51E-35 3.94E-33 GO:0016020 membrane 18.1 2.27E-19 5.91E-17 GO:0044444 cytoplasmic part 15.8 1.95E-16 5.08E-14 GO:0009986 cell surface 11.4 4.76E-12 1.24E-09 GO:0031012 extracellular matrix 10.2 6.56E-11 1.71E-08 GO:0031982 vesicle 9.12 1.07E-09 2.79E-07 GO:0012505 endomembrane system 6.81 2.33E-07 6.05E-05 GO:0044422 organelle part 5.84 2.62E-06 6.81E-04 GO:0005634 nucleus 5.19 1.17E-05 0.00303 GO:0031252 cell front 3.88 0.000158 0.041

[0449] Table 5.

[0450]

[0451] Table 6.

[0452]

[0453]

[0454] Example 3: Comparison of TREM-1 gene signature using different TREM-1 agonists

[0455] To understand the overlap in gene expression between agonistic anti-TREM-1 antibodies and the natural TREM-1 ligand (PGLYRP1), the gene modules generated upon ligand (891 genes, P+L vs P, fold change >2, FDR <0.05) and agonistic antibody (331 genes, agTREM-1 vs isotype, fold change >2, FDR <0.05) stimulation were evaluated. Agonistic antibodies are described in Dower K et al., Journal of immunology 180:3520-3534 (2008).

[0456] As shown in Figure 5 , there is only a small subset of genes that overlap between the two TREM-1 agonists.

[0457] Example 4: Evaluation of TREM-1 gene signature in peripheral blood mononuclear cells (PBMCs)

[0458] One of the key elements for clinical development is the development of a robust pharmacodynamic (PD) assay that can be measured in the clinical trial setting. To this end, it was evaluated whether the TREM-1 specific gene changes could be reproduced in PBMCs, which are more easily accessible in the clinic than monocytes. Briefly, PBMCs from healthy human donors were isolated by ficoll purification. PBMCs were then plated onto 24-well plates (1 x 10 6 cells / well) and stimulated for 24 hours using the following conditions: (i) medium alone (i.e., no stimulation), (ii) PGN-ECndss alone, (iii) PGN-ECndss + PGRP, and (iv) PGN-ECndss + PGRP + agonistic anti-TREM-1 antibody. Then, the expression of several genes (i.e., CCL20, IL-1 beta, IL-12 p40, and IL-23 beta) that are highly induced (and inhibited by TREM-1 blocking antibodies) were selected from a 180 gene signature and their expression pattern was evaluated using RT-PCR and / or cytokine expression analysis.

[0459] Similar to purified monocytes (see Figures 1A-1D ), PGN-ECndss induced very little CCL20, IL1 beta, and IL12 p40 expression (at the protein and mRNA level). See Figures 6A-6F . In contrast, PGLYRP1 and PGN-ECndss strongly induced mRNA and protein expression of these cytokines and chemokines, which was blocked by TREM-1 blocking antibodies Figures 6A-6F . These results confirmed the finding that TREM-1 induced changes can be blocked with anti-TREM-1 antibodies in monocytes can also be extended to PBMCs and have the potential to be used as a PD marker in clinical trials.

[0460] Example 5: Distribution of TREM-1 marker scores in IBD biopsy expression profiling datasets at baseline and post-treatment

[0461] The expression patterns of the TREM-1 gene module generated in colon biopsies from IBD patients in the above embodiments were evaluated. Gene expression data from a phase 2 trial evaluating the efficacy of the anti-IP10 antibody in UC patients (ClinicalTrials.gov identifier NCT00656890) was used as the primary data source. At baseline (D1), matched lesion and non-lesion biopsies and whole blood samples from 78 UC patients were analyzed using the Affymetrix U219 platform. As a secondary data source, the public dataset (GSE16879) was used to investigate gene expression before and after infliximab (IFX) treatment in CD and UC patients. This dataset contains gene expression profiles from baseline and 4–6 weeks after infliximab treatment from 61 IBD patients (24 UC, 19 CD colon, and 18 CD ileum) (along with clinical annotations and treatment responses) and 12 non-IBD controls (6 colon, 6 ileum) (Affymetrix platform). For each patient, the ssGSEA score was calculated using the GSVABioconductor package in R, which is a sort-based score that summarizes the collective expression enrichment of all genes in the TREM-1 module.

[0462] like Figure 7 As shown, in the anti-IP10 trial dataset, we found that the TREM-1 module score in lesions was increased compared to non-lesion biopsies at baseline (P < 0.001). The TREM-1 module score was also positively correlated with TREM-1 expression in UC lesion biopsies (Rho = 0.85, P < 0.001). Figure 8 This indicates TREM-1 activation at baseline in this patient population.

[0463] Then, the standard of care (SOC) was assessed, such as how oral steroids or the use of anti-TNF drugs affected changes in TREM-1 module scores in tissues. All patients with a history of anti-IP10 trials and anti-TNF therapy were considered non-responders / inadequate responders (NR / IR). Patients without a history of anti-TNF use were considered anti-TNF initiators.

[0464] Regardless of the use of oral corticosteroids, TREM-1 gene marker scores showed no statistically significant difference between anti-TNF naïve and anti-TNF NR / IR IBD patients. Figure 9). Analysis of the GSE16879 dataset showed that in those patients considered as treatment non-responders, the TREM-1 module score remained elevated after treatment with IFX, but decreased in TNF responders ( Figure 10 ) Furthermore, at baseline, IFX non-responders had significantly higher TREM-1 module score compared to IFX responders ( Figure 10 ) These findings applied to UC and CD patients with colonic involvement. Altogether, these data suggest that the TREM-1 pathway is more active and remains elevated in the TNF non-responder population.

[0465] Example 6: Application of the TREM-1 gene module as potential blood pharmacodynamic biomarker candidate

[0466] As the genes in the TREM-1 gene module reflect the transcriptomic changes upon TREM-1 pathway activation, it was evaluated whether these genes could be used as potential PD biomarker candidates. To select blood-based PD marker candidates, genes in the TREM-1 gene module were filtered according to the following criteria: a) expression in UC baseline blood (mean log2 RMA > 5); b) small change in UC baseline blood (IQR < 0.7); c) expression in extracellular space (GO functional annotation). The percentage of inhibition upon anti-TREM-1 treatment was calculated by dividing the magnitude of fold change in PGLYRP1 + PGN-ECndss by the fold change in PGLYRP1 + PGN-ECndss stimulation with PGN-ECndss stimulation. TREM-1 ligand stimulation is the log2 fold change in PGLYRP1 + PGN-ECndss stimulation and these candidates are ranked in descending order. Table 7 shows the top 20 blood PD candidates with mean and IQR of these mRNAs in IM129-005 UC baseline blood and internal healthy and UC blood. These genes will be evaluated in future clinical trials.

[0467] Table 7.

[0468]

[0469]

[0470] Example 7: Application of the TREM-1 module to generate a UC-specific TREM-1 signature

[0471] Using the TREM-1 gene modules generated in the above examples, the following criteria were used to generate a UC-specific TREM-1 signature: a) associated with the TREM-1 pathway (genes from the TREM-1 module); b) UC-relevant expression in lesion biopsies (mean log2 RMA > 4); c) UC-relevant upregulation in lesion biopsies (lesion vs. non-lesion FDR < 0.05); d) different expression in UC lesion biopsy samples (IQR > 1). This filtering resulted in a signature with 38 genes that represent potential patient stratification biomarker candidates. We then investigated the expression pattern of these 38 genes in baseline UC colon biopsies from the anti-IP10 trial Figure 11A

[0472] As shown in Figure 11B , while there is significant heterogeneity in the expression of these genes between patients, two distinct patient clusters were identified. This bimodal distribution suggests the presence of a UC patient population with high TREM-1 module scores (> 0.33; Figure 12B ) that represents the majority of the patient population in this dataset. To further support the heterogeneity of the TREM-1 module in UC patients, matched non-lesion tissue was used as a reference. Using the TREM-1 module score to predict lesion or non-lesion tissue, the score that reaches the highest AUC (0.25) can be an optimal value to separate patient populations with positive or negative TREM-1 signature.

[0473] Example 8: Application of TREM-1 gene signature to identify alternative biomarkers for patient stratification through correlation analysis with clinical parameters

[0474] Assuming that the TREM-1 gene signature in lesion colon biopsies reflects pathway activation in IBD patients, the correlation of the TREM-1 gene signature in UC colon biopsies with other potential biomarkers was investigated. Since measuring the TREM-1 gene signature requires a lesion biopsy in disease tissue (e.g. colon) and either a dedicated gene panel or global RNASeq analysis, this study allowed the identification of potential biomarkers as alternatives to TREM-1 pathway activation. These biomarkers can be clinically easier to measure.

[0475] Regression models were fitted to investigate the association of multiple clinical and biomarker factors with the TREM-1 module score from patient stratification candidates (Table 8).

[0476] Table 8.

[0477]

[0478] As shown in Figures 12A-12C ​As shown, baseline Mayo score, grade 2B intrinsic layer neutrophil infiltration score (one component of the Geboes score), and fecal calprotectin were the factors significantly associated with the TREM-1 gene signature in UC patients. The positive correlation signal after correction with other factors suggests the presence of a UC patient population with high fecal calprotectin (FC) and high TREM-1 score. In other words, most UC patients with high FC levels can have high TREM-1 pathway activation. Since FC is more clinically accessible, it can serve as a surrogate biomarker for the TREM-1 gene signature.

[0479] Table 9. Exemplary anti-TREM-1 antibody sequences

[0480]

[0481]

[0482]

[0483]

[0484]

[0485]

[0486]

[0487]

[0488]

[0489]

[0490]

[0491] SEQUENCE LISTING <110> BRISTOL-MYERS SQUIBB COMPANY <120> Anti-TREM-1 antibodies and uses thereof <130> 3338.1380000 / ELE / C-K / DKC <140> US 62 / 874,318 <141> 2019-07-15 <160> 295 <170> PatentIn 3.5 version <210> 1 <211> 234 <212> PRT <213> Artificial sequence <220> <223> TREM-1, isoform 1 <400> 1 Met Arg Lys Thr Arg Leu Trp Gly Leu Leu Trp Met Leu Phe Val Ser 1 5 10 15 Glu Leu Arg Ala Ala Thr Lys Leu Thr Glu Glu Lys Tyr Glu Leu Lys 20 25 30 Glu Gly Gln Thr Leu Asp Val Lys Cys Asp Tyr Thr Leu Glu Lys Phe 35 40 45 Ala Ser Ser Gln Lys Ala Trp Gln Ile Ile Arg Asp Gly Glu Met Pro 50 55 60 Lys Thr Leu Ala Cys Thr Glu Arg Pro Ser Lys Asn Ser His Pro Val 65 70 75 80 Gln Val Gly Arg Ile Ile Leu Glu Asp Tyr His Asp His Gly Leu Leu 85 90 95 Arg Val Arg Met Val Asn Leu Gln Val Glu Asp Ser Gly Leu Tyr Gln 100 105 110 Cys Val Ile Tyr Gln Pro Pro Lys Glu Pro His Met Leu Phe Asp Arg 115 120 125 Ile Arg Leu Val Val Thr Lys Gly Phe Ser Gly Thr Pro Gly Ser Asn 130 135 140 Glu Asn Ser Thr Gin Asn Val Tyr Lys lie Pro Pro Thr Thr Thr Lys 145 150 155 160 Ala Leu Cys Pro Leu Tyr Thr Ser Pro Arg Thr Val Thr Gin Ala Pro 165 170 175 Pro Lys Ser Thr Ala Asp Val Ser Thr Pro Asp Ser Glu lie Asn Leu 180 185 190 Thr Asn Val Thr Asp lie lie Arg Val Pro Val Phe Asn lie Val lie 195 200 205 Leu Leu Ala Gly Gly Phe Leu Ser Lys Ser Leu Val Phe Ser Val Leu 210 215 220 Phe Ala Val Thr Leu Arg Ser Phe Val Pro 225 230 <210> 2 <211> 225 <212> PRT <213> Artificial Sequence <220> <223> TREM-1, isoform 2 <400> 2 Met Arg Lys Thr Arg Leu Trp Gly Leu Leu Trp Met Leu Phe Val Ser 1 5 10 15 Glu Leu Arg Ala Ala Thr Lys Leu Thr Glu Glu Lys Tyr Glu Leu Lys 20 25 30 Glu Gly Gin Thr Leu Asp Val Lys Cys Asp Tyr Thr Leu Glu Lys Phe 35 40 45 Ala Ser Ser Gin Lys Ala Trp Gin He He Arg Asp Gly Glu Met Pro 50 55 60 Lys Thr Leu Ala Cys Thr Glu Arg Pro Ser Lys Asn Ser His Pro Val 65 70 75 80 Gln Val Gly Arg He He Leu Glu Asp Tyr His Asp His Gly Leu Leu 85 90 95 Arg Val Arg Met Val Asn Leu Gin Val Glu Asp Ser Gly Leu Tyr Gin 100 105 110 Cys Val He Tyr Gin Pro Pro Lys Glu Pro His Met Leu Phe Asp Arg 115 120 125 He Arg Leu Val Val Thr Lys Gly Phe Ser Gly Thr Pro Gly Ser Asn 130 135 140 Glu Asn Ser Thr Gin Asn Val Tyr Lys He Pro Pro Thr Thr Thr Lys 145 150 155 160 Ala Leu Cys Pro Leu Tyr Thr Ser Pro Arg Thr Val Thr Gin Ala Pro 165 170 175 Pro Lys Ser Thr Ala Asp Val Ser Thr Pro Asp Ser Glu He Asn Leu 180 185 190 Thr Asn Val Thr Asp lie lie Arg Tyr Ser Phe Gin Val Pro Gly Pro 195 200 205 Leu Val Trp Thr Leu Ser Pro Leu Phe Pro Ser Leu Cys Ala Glu Arg 210 215 220 Met 225 <210> 3 <211> 150 <212> PRT <213> Artificial Sequence <220> <223> TREM-1, Isoform 3 <400> 3 Met Arg Lys Thr Arg Leu Trp Gly Leu Leu Trp Met Leu Phe Val Ser 1 5 10 15 Glu Leu Arg Ala Ala Thr Lys Leu Thr Glu Glu Lys Tyr Glu Leu Lys 20 25 30 Glu Gly Gin Thr Leu Asp Val Lys Cys Asp Tyr Thr Leu Glu Lys Phe 35 40 45 Ala Ser Ser Gin Lys Ala Trp Gin lie lie Arg Asp Gly Glu Met Pro 50 55 60 Lys Thr Leu Ala Cys Thr Glu Arg Pro Ser Lys Asn Ser His Pro Val 65 70 75 80 Gln Val Gly Arg lie lie Leu Glu Asp Tyr His Asp His Gly Leu Leu 85 90 95 Arg Val Arg Met Val Asn Leu Gin Val Glu Asp Ser Gly Leu Tyr Gin 100 105 110 Cys Val He Tyr Gin Pro Pro Lys Glu Pro His Met Leu Phe Asp Arg 115 120 125 He Arg Leu Val Val Thr Lys Gly Phe Arg Cys Ser Thr Leu Ser Phe 130 135 140 Ser Trp Leu Val Asp Ser 145 150 <210> 4 <211> 3252 <212> DNA <213> Artificial Sequence <220> <223> TREM-1, Isoform 1 <400> 4 agcagttgga gctggtgcac aggaaggatg aggaagacca ggctctgggg gctgctgtgg 60 atgctctttg tctcagaact ccgagctgca actaaattaa ctgaggaaaa gtatgaactg 120 aaagaggggc agaccctgga tgtgaaatgt gactacacgc tagagaagtt tgccagcagc 180 cagaaagctt ggcagataat aagggacgga gagatgccca agaccctggc atgcacagag 240 aggccttcaa agaattccca tccagtccaa gtggggagga tcatactaga agactaccat 300 gatcatggtt tactgcgcgt ccgaatggtc aaccttcaag tggaagattc tggactgtat 360 cagtgtgtga tctaccagcc tcccaaggag cctcacatgc tgttcgatcg catccgcttg 420 gtggtgacca agggtttttc agggacccct ggctccaatg agaattctac ccagaatgtg 480 tataagattc ctcctaccac cactaaggcc ttgtgcccac tctataccag ccccagaact 540 gtgacccaag ctccacccaa gtcaactgcc gatgtctcca ctcctgactc tgaaatcaac 600 cttacaaatg tgacagatat catcagggtt ccggtgttca acattgtcat tctcctggct 660 ggtggattcc tgagtaagag cctggtcttc tctgtcctgt ttgctgtcac gctgaggtca 720 tttgtaccct aggcccacga acccacgaga atgtcctctg acttccagcc acatccatct 780 ggcagttgtg ccaagggagg agggaggagg taaaaggcag ggagttaata acatgaatta 840 aatctgtaat caccggctat ttctaaagtc agcgtctcac cttcctgccc actgccctcg 900 ttcctctaat aatcttgggt gggcatttgt gcctcagaaa agaagttaca gccccaaaca 960 tgcttggtcc ttcattccac cagccactlg gggttggcat gaaatacaga cagctcaatg 1020 cttttcaccg taattctctt gtgggggctg tgacatgcag aaggcacacc tgatacttct 1080 cctgctcagt tttgccctgg accatacaat tttggcctga cctggacaga gctcccacta 1140 cagaagcatc ctgctcgccc catgctggga cttcctcttt ctagcatcag acacttgggt 1200 ttcatgctta tgtgtggttc tttccaacac tcccagaaaa gggtgttgaa gattgtggaa 1260 cctggagaaa taagacatcg tggtgagaaa gtgcatcctt ctcagagaaa agagttaaac 1320 tgagtatctt cttctgggga aatactggca ggccgagatg ggatccatag gagagcaaca 1380 acagaccatg tcagacatcc tgtgtgcatt tatcgctgga tcctgaaaat agccccgtga 1440 aggcagaaat gtatgtgact agaacgaggc cacatgaata agccactgcc cactggcagg 1500 agtgaaaact gaagcgctcc ttacctgaag gaccccaaaa ccatatagaa tagaataacc 1560 aggagttccg cctgtgtcta aatgcctctt ttcctgtatc acacaagggt cagggatggt 1620 ggagtaaaag ctctccccct gggaggcttc tggaggctgt ccccatgtgc ttgcctagtt 1680 ccccactctg ccctcctcct cttctctcag tctgctcctg gaacacctgc ctcagtttcc 1740 atgctctctc cagtgccctc cccggtgaag caggtaggtg ttcaggccac cacagagaca 1800 atctctgtgg gagattgtct tgcaatctcc cacagatttc aatcaggatt ttgttatttc 1860 ctactttgag ctttaaaggg aaatgggcct catgggtggg gaaaggatgg tgggtccttc 1920 cagcccaatt tagtgatgcc cagggcagat attatcctca gttcccaaga gcaatcatac 1980 ttttccacac ataccgtgtg tctcatgtta ggtaaatgta tttttacaat gagcaccact 2040 tctgtggaaa aagttccctg cacggggagg tccagcttcc agactgctcc atcgcataag 2100 gacttcccca ttcccctaaa tgctgctctg tcagaacctg cccaggtaat ggtaatgacc 2160 ctagagagat gatttctgaa ccgcaatttt gagcccatta gaaggtgtgt ggtgggcatt 2220 tatttcatcc tgatgctctg gtgagaatct ttgcagacgc actagatcca gaagctgtta 2280 atcttggtgc atttattttc ctacctaaaa gaaccaagca gctcagaggc agtgactgta 2340 caggatgcag tgtttataat aatgctgagc ttgctggtct ggaaccccac actttagcaa 2400 tcccagcatt gttcctgttt atgaagttga caaagtgacc agggcaaggg ggtattatca 2460 ttaaatacac tctaggagag gcagaacaca tgagggcaat gtttttcaga ggtctttagg 2520 ccaccgcatc agattctcct ggagcataaa gcaaatgctt tatgagtcca gggcccctgc 2580 agacctactg tatactagta tacagctccc tcttagtgga tctcaagctt gtttccaaaa 2640 agtcattaca ctccttacca aagcccatga cacattcata cagattcatc cagacataac 2700 ccactgcatg gtccagtgca tgcttgtgtg cttaacttat tatagatcaa gtgttattta 2760 agtccaacat attaaacgtg actgaaatat tatatgcact aaatcggaag acaaatggcc 2820 gtgctggata ctctcccatt tgcacttaca ggccacgcca tgctccatcc tccccatcct 2880 gcctgggaac ctgaccaatg tggctaaacc agcaggctcc ctggctgcca gcttctggct 2940 ggttgagcca atgggaagca tgagaggaga gccgagggtg gggagacagt cagatcaggg 3000 tttctgtcct cagggctccc tccctggcag gtggtggcag gagtggctgc attcccctct 3060 gaaggctcct tcaagcctct cagcaaacag ctcccatctc caagtccagc cacctgttcc 3120 atcctctcct ctgtaggccc agcagtggga atgacccacc actattgcca gcctcagcat 3180 actgaagcac cccttactgg attccctaaaa ttctatgcac atgtttatta aatgctcctc 3240 aattacccag tt 3252 <210> 5 <211> 2128 <212> DNA <213> Artificial Sequence <220> <223> TREM-1, Isoform 2 <400> 5 gtgcaacttt ccgaagcctc taggtcattg tggtgccttg tagctgtccc gggagccctc 60 agcagcagtt ggagctggtg cacaggaagg atgaggaaga ccaggctctg ggggctgctg 120 tggatgctct ttgtctcaga actccgagct gcaactaaat taactgagga aaagtatgaa 180 ctgaaagagg ggcagaccct ggatgtgaaa tgtgactaca cgctagagaa gtttgccagc 240 agccagaaag cttggcagat aataagggac ggagagatgc ccaagaccct ggcatgcaca 300 gagaggcctt caaagaattc ccatccagtc caagtgggga ggatcatact agaagactac 360 catgatcatg gtttactgcg cgtccgaatg gtcaaccttc aagtggaaga ttctggactg 420 tatcagtgtg tgatctacca gcctcccaag gagcctcaca tgctgttcga tcgcatccgc 480 ttggtggtga ccaagggttt ttcagggacc cctggctcca atgagaattc tacccagaat 540 gtgtataaga ttcctcctac caccactaag gccttgtgcc cactctatac cagccccaga 600 actgtgaccc aagctccacc caagtcaact gccgatgtct ccactcctga ctctgaaatc 660 aaccttacaa atgtgacaga tatcatcagg tatagtttcc aggtccctgg gcccctggtt 720 tggacactga gccctttgtt tcccagtctg tgtgctgaga ggatgtgaaa gtgagggaaa 780 ggggagggtg gggcaggaga agacttgagt cacattagtc tgggtagaaa tgtccagggg 840 aagaaggaag tggtgatgga gaatagggga ggctctcagc caggctgtcc ttctccccag 900 ttcaccttct ttgtttcctt gcaacctgag tattaaagag agggaaatgg catcttcccc 960 aagttccagt ggagctcatc caaccccagg ggcctgatgg gcagtgggaa agcactctga 1020 gtgaggggcc ctggatctag tgttggcctg actaactgaa tgtgactttg ggtgagtcag 1080 gaaccctctc tggctttagc ttctttgaca attcagtagg catggtagaa acccaaagct 1140 ggaagacatt gtccacctaa taactctcag caggagctgg agctggggct aaatgcagta 1200 ttggtttttg ccttattgtt ttttaaataa cattgttgca tgtgcccaat tatagataaa 1260 tagaatcaga atttttgcag atgaggtccc tctcagtatt tttaataaga tgtttaggag 1320 atttctaatg tgtaggcaaa cttaagaacc actaacttag caatttcaca cctctttaca 1380 gttattaaat gttgatttat attagaatgt gtttatcatt aaatactgaa ttatcaggat 1440 ggaaatactt ttccacatca ccactagtct aacatgtct tttctcata ttctctcta 1500 gtctttattc atgtttatac atattttaat atggcagtaa gtctattat acatgttttt 1560 attctttcc ttgctgaat tattctttga tgctgctacc taaatttcat aaatataatt 1620 tcaatgagtg catatttttc tatcaatttgg gaagccata acttacataa ccattggcct 1680 gttcttagac attgatatag tgttgataac tctgagataa acatattctt gcagatatat 1740 ttttctctt ttaaatatt aaccgaaat ttccatgaga taatttaaac actgatatca 1800 ttttgtatac aaatagcatc taccaatggt cttccaaag attggaaca atttgcagtg 1860 tgatccatta agcataata aagcagtgtc atggcaggct tacagacacg gatttagtct 1920 tttaaatcat taaatagtgt gagttaaatc acatcctgc taaaacaat gtgaactggt 1980 gcctgctcta atttctctat gagtgtagac tccacctcca tatggtagt ggcagtgcct 2040 ttttccccat tatgttgttt ggggaacaaa gtgctcatta aacttctgtg gaataaatca 2100 aacgaatgat caaaaaaaaa aaaaaaaa 2128 <210> 6 <211> 3140 <212> DNA <213> Artificial Sequence <220> <223> TREM-1, Isoform 3 <400> 6 gtgcaacttt ccgaagcctc taggtcattg tggtgccttg tagctgtccc gggagccctc 60 agcagcagtt ggagctggtg cacaggaagg atgaggaaga ccaggctctg ggggctgctg 120 tggatgctct ttgtctcaga actccgagct gcaactaaat taactgagga aaagtatgaa 180 ctgaaagagg ggcagaccct ggatgtgaaa tgtgactaca cgctagagaa gtttgccagc 240 agccagaaag cttggcagat aataagggac ggagagatgc ccaagaccct ggcatgcaca 300 gagaggcctt caaagaattc ccatccagtc caagtgggga ggatcatact agaagactac 360 catgatcatg gtttactgcg cgtccgaatg gtcaaccttc aagtggaaga ttctggactg 420 tatcagtgtg tgatctacca gcctcccaag gagcctcaca tgctgttcga tcgcatccgc 480 tatcagtgtg tgatctacca gcctcccaag gagcctcaca tgctgttcga tcgcatccgc 480ttggtggtga ccaaggggtt ccggtgttca acattgtcat tctcctggct ggtggattcc 540 tgagtaagag cctggtcttc tctgtcctgt ttgctgtcac gctgaggtca tttgtaccct 600 aggcccacga acccacgaga atgtcctctg acttccagcc acatccatct ggcagttgtg 660 ccaagggagg agggaggagg taaaaggcag ggagttaata acatgaatta aatctgtaat 720 caccggctat ttctaaagtc agcgtctcac cttcctgccc actgccctcg ttcctctaat 780 aatcttgggt gggcatttgt gcctcagaaa agaagttaca gccccaaaca tgcttggtcc 840 ttcattccac cagccactlg gggttggcat gaaatacaga cagctcaatg cttttcaccg 900 taattctctt gtgggggctg tgacatgcag aaggcacacc tgatacttct cctgctcagt 960 tttgccctgg accatacaat tttggcctga cctggacaga gctcccacta cagaagcatc 1020 ctgctcgccc catgctggga cttcctcttt ctagcatcag acacttgggt ttcatgctta 1080 tgtgtggttc tttccaacac tcccagaaaa gggtgttgaa gattgtggaa cctggagaaa 1140 taagacatcg tggtgagaaa gtgcatcctt ctcagagaaa agagttaaac tgagtatctt 1200 cttctgggga aatactggca ggccgagatg ggatccatag gagagcaaca acagaccatg 1260 tcagacatcc tgtgtgcatt tatcgctgga tcctgaaaat agccccgtga aggcagaaat 1320 gtatgtgact agaacgaggc cacatgaata agccactgcc cactggcagg agtgaaaact 1380 gaagcgctcc ttacctgaag gaccccaaaa ccatatagaa tagaataacc aggagttccg 1440 cctgtgtcta aatgcctctt ttcctgtatc acacaagggt cagggatggt ggagtaaaag 1500 ctctccccct gggaggcttc tggaggctgt ccccatgtgc ttgcctagtt ccccactctg 1560 ccctcctcct cttctctcag tctgctcctg gaacacctgc ctcagtttcc atgctctctc 1620 cagtgccctc cccggtgaag caggtaggtg ttcaggccac cacagagaca atctctgtgg 1680 gagattgtct tgcaatctcc cacagatttc aatcaggatt ttgttatttc ctactttgag 1740 ctttaaaggg aaatgggcct catgggtggg gaaaggatgg tgggtccttc cagcccaatt 1800 tagtgatgcc cagggcagat attatcctca gttcccaaga gcaatcatac ttttccacac 1860 ataccgtgtg tctcatgtta ggtaaatgta tttttacaat gagcaccact tctgtggaaa 1920 aagttccctg cacggggagg tccagcttcc agactgctcc atcgcataag gacttcccca 1980 ttcccctaaa tgctgctctg tcagaacctg cccaggtaat ggtaatgacc ctagagagat 2040 gatttctgaa ccgcaatttt gagcccatta gaaggtgtgt ggtgggcatt tatttcatcc 2100 tgatgctctg gtgagaatct ttgcagacgc actagatcca gaagctgtta atcttggtgc 2160 atttattttc ctacctaaaa gaaccaagca gctcagaggc agtgactgta caggatgcag 2220 tgtttataat aatgctgagc ttgctggtct ggaaccccac actttagcaa tcccagcatt 2280 gttcctgttt atgaagttga caaagtgacc agggcaaggg ggtattatca ttaaatacac 2340 tctaggagag gcagaacaca tgagggcaat gtttttcaga ggtctttagg ccaccgcatc 2400 agattctcct ggagcataaa gcaaatgctt tatgagtcca gggcccctgc agacctactg 2460 tatactagta tacagctccc tcttagtgga tctcaagctt gtttccaaaa agtcattaca 2520 ctccttacca aagcccatga cacattcata cagattcatc cagacataac ccactgcatg 2580 gtccagtgca tgcttgtgtg cttaacttat tatagatcaa gtgttattta agtccaacat 2640 attaaacgtg actgaaatat tatatgcact aaatcggaag acaaatggcc gtgctggata 2700 ctctcccatt tgcacttaca ggccacgcca tgctccatcc tccccatcct gcctgggaac 2760 ctgaccaatg tggctaaacc agcaggctcc ctggctgcca gcttctggct ggttgagcca 2820 atgggaagca tgagaggaga gccgagggtg gggagacagt cagatcaggg tttctgtcct 2880 cagggctccc tccctggcag gtggtggcag gagtggctgc attcccctct gaaggctcct 2940 tcaagcctct cagcaaacag ctcccatctc caagtccagc cacctgttcc atcctctcct 3000 ctgtaggccc agcagtggga atgacccacc actattgcca gcctcagcat actgaagcac 3060 cccttactgg attccctaaa ttctatgcac atgtttatta aatgctcctc aattacccag 3120 ttaaaaaaaa aaaaaaaaaa 3140 <210> 7 <211> 233 <212> PRT <213> Artificial Sequence <220> <223> TREM-1 (predicted) <400> 7 Met Arg Lys Thr Arg Leu Trp Gly Leu Leu Trp Met Leu Phe Val Ser 1 5 10 15 Glu Leu Arg Ala Thr Thr Glu Leu Thr Glu Glu Lys Tyr Glu Tyr Lys 20 25 30 Glu Gly Gln Thr Leu Glu Val Lys Cys Asp Tyr Ala Leu Glu Lys Tyr 35 40 45 Ala Asn Ser Arg Lys Ala Trp Gln Lys Met Glu Gly Lys Met Pro Lys 50 55 60 Ile Leu Ala Lys Thr Glu Arg Pro Ser Glu Asn Ser His Pro Val Gln 65 70 75 80 Val Gly Arg Ile Thr Leu Glu Asp Tyr Pro Asp His Gly Leu Leu Gln 85 90 95 Val Gln Met Thr Asn Leu Gln Val Glu Asp Ser Gly Leu Tyr Gln Cys 100 105 110 Val Ile Tyr Gln His Pro Lys Glu Ser His Val Leu Phe Asn Pro Ile 115 120 125 Cys Leu Val Val Thr Lys Gly Ser Ser Gly Thr Pro Gly Ser Ser Glu 130 135 140 Asn Ser Thr Gln Asn Val Tyr Arg Thr Pro Ser Thr Thr Ala Lys Ala 145 150 155 160 Leu Gly Pro Arg Tyr Thr Ser Pro Arg Thr Val Thr Gln Ala Pro Pro 165 170 175 Glu Ser Thr Val Val Val Ser Thr Pro Gly Ser Glu lie Asn Leu Thr 180 185 190 Asn Val Thr Asp lie lie Arg Val Pro Val Phe Asn lie Val lie lie 195 200 205 Val Ala Gly Gly Phe Leu Ser Lys Ser Leu Val Phe Ser Val Leu Phe 210 215 220 Ala Val Thr Leu Arg Ser Phe Gly Pro 225 230 <210> 8 <211> 196 <212> PRT <213> Artificial Sequence <220> <223> Peptidoglycan recognition protein 1 <400> 8 Met Ser Arg Arg Ser Met Leu Leu Ala Trp Ala Leu Pro Ser Leu Leu 1 5 10 15 Arg Leu Gly Ala Ala Gin Glu Thr Glu Asp Pro Ala Cys Cys Ser Pro 20 25 30 lie Val Pro Arg Asn Glu Trp Lys Ala Leu Ala Ser Glu Cys Ala Gin 35 40 45 His Leu Ser Leu Pro Leu Arg Tyr Val Val Val Ser His Thr Ala Gly 50 55 60 Ser Ser Cys Asn Thr Pro Ala Ser Cys Gin Gin Gin Ala Arg Asn Val 65 70 75 80 Gln His Tyr His Met Lys Thr Leu Gly Trp Cys Asp Val Gly Tyr Asn 85 90 95 Phe Leu Ile Gly Glu Asp Gly Leu Val Tyr Glu Gly Arg Gly Trp Asn 100 105 110 Phe Thr Gly Ala His Ser Gly His Leu Trp Asn Pro Met Ser Ile Gly 115 120 125 Ile Ser Phe Met Gly Asn Tyr Met Asp Arg Val Pro Thr Pro Gln Ala 130 135 140 Ile Arg Ala Ala Gln Gly Leu Leu Ala Cys Gly Val Ala Gln Gly Ala 145 150 155 160 Leu Arg Ser Asn Tyr Val Leu Lys Gly His Arg Asp Val Gln Arg Thr 165 170 175 Leu Ser Pro Gly Asn Gln Leu Tyr His Leu Ile Gln Asn Trp Pro His 180 185 190 Tyr Arg Ser Pro 195 <210> 9 <211> 330 <212> PRT <213> Artificial Sequence <220> <223> Wild-type IgG1 Heavy Chain Constant Region <400> 9 Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys 1 5 10 15 Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr 20 25 30 Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser 35 40 45 Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser 50 55 60 Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr 65 70 75 80 Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys 85 90 95 Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys 100 105 110 Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro 115 120 125 Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys 130 135 140 Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp 145 150 155 160 Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu 165 170 175 Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu 180 185 190 His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn 195 200 205 Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly 210 215 220 Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu 225 230 235 240 Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr 245 250 255 Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn 260 265 270 Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe 275 280 285 Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn 290 295 300 Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr 305 310 315 320 Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 325 330 <210> 10 <211> 327 <212> PRT <213> Artificial Sequence <220> [[ID=​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​130 135 140 Asp Val Ser Gin Glu Asp Pro Glu Val Gin Phe Asn Trp Tyr Val Asp 145 150 155 160 Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gin Phe 165 170 175 Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gin Asp 180 185 190 Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu 195 200 205 Pro Ser Ser Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gin Pro Arg 210 215 220 Glu Pro Gin Val Tyr Thr Leu Pro Pro Ser Gin Glu Glu Met Thr Lys 225 230 235 240 Asn Gin Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp 245 250 255 Ile Ala Val Glu Trp Glu Ser Asn Gly Gin Pro Glu Asn Asn Tyr Lys 260 265 270 Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser 275 280 285 Arg Leu Thr Val Asp Lys Ser Arg Trp Gin Glu Gly Asn Val Phe Ser 290 295 300 Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser 305 310 315 320 Leu Ser Leu Ser Leu Gly Lys 325 <210> 11 <211> 330 <212> PRT <213> Artificial Sequence <220> <223> IgG1 -Aba heavy chain constant region <400> 11 Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys 1 5 10 15 Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr 20 25 30 Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser 35 40 45 Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser 50 55 60 Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr 65 70 75 80 Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys 85 90 95 Arg Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Ser Pro Pro Ser 100 105 110 Pro Ala Pro Glu Leu Leu Gly Gly Ser Ser Val Phe Leu Phe Pro Pro 115 120 125 Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys 130 135 140 Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp 145 150 155 160 Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu 165 170 175 Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu 180 185 190 His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn 195 200 205 Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly 210 215 220 Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu 225 230 235 240 Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr 245 250 255 Pro Ser Asp lie Ala Val Glu Trp Glu Ser Asn Gly Gin Pro Glu Asn 260 265 270 Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe 275 280 285 Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gin Gin Gly Asn 290 295 300 Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr 305 310 315 320 Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 325 330 <210> 12 <211> 330 <212> PRT <213> Artificial Sequence <220> <223> IgG4-Aba Heavy Chain Constant Region <400> 12 Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Cys Ser Arg 1 5 10 15 Ser Thr Ser Glu Ser Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr 20 25 30 Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser 35 40 45 Gly Val His Thr Phe Pro Ala Val Leu Gin Ser Ser Gly Leu Tyr Ser 50 55 60 Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Lys Thr 65 70 75 80 Tyr Thr Cys Asn Val Asp His Lys Pro Ser Asn Thr Lys Val Asp Lys 85 90 95 Arg Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Ser Pro Pro Ser 100 105 110 Pro Ala Pro Glu Leu Leu Gly Gly Ser Ser Val Phe Leu Phe Pro Pro 115 120 125 Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys 130 135 140 Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp 145 150 155 160 Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu 165 170 175 Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu 180 185 190 His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn 195 200 205 Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly 210 215 220 Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu 225 230 235 240 Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr 245 250 255 Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn 260 265 270 Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe 275 280 285 Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn 290 295 300 Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr 305 310 315 320 Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 325 330 <210> 13 <211> 448 <212> PRT <213> Artificial Sequence <220> <223> HC of mAb 0170 <400> 13 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Lys Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Thr Tyr 20 25 30 Ala Met His Trp Val Arg Gin Ala Ser Gly Lys Gly Leu Glu Trp Val 35 40 45 Gly Arg He Arg Thr Lys Ser Ser Asn Tyr Ala Thr Tyr Tyr Ala Ala 50 55 60 Ser Val Lys Gly Arg Phe Thr He Ser Arg Asp Asp Ser Lys Asn Thr 65 70 75 80 Ala Tyr Leu Gin Met Asn Ser Leu Lys Thr Glu Asp Thr Ala Val Tyr 85 90 95 Tyr Cys Thr Arg Asp Met Gly He Arg Arg Gin Phe Ala Tyr Trp Gly 100 105 110 Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser 115 120 125 Val Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr Ala 130 135 140 Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val 145 150 155 160 Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala 165 170 175 Val Leu Gin Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val 180 185 190 Pro Ser Ser Ser Leu Gly Thr Lys Thr Tyr Thr Cys Asn Val Asp His 195 200 205 Lys Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Ser Lys Tyr Gly 210 215 220 Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe Leu Gly Gly Pro Ser 225 230 235 240 Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg 245 250 255 Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gin Glu Asp Pro 260 265 270 Glu Val Gin Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala 275 280 285 Lys Thr Lys Pro Arg Glu Glu Gin Phe Asn Ser Thr Tyr Arg Val Val 290 295 300 Ser Val Leu Thr Val Leu His Gin Asp Trp Leu Asn Gly Lys Glu Tyr 305 310 315 320 Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys Thr 325 330 335 Ile Ser Lys Ala Lys Gly Gin Pro Arg Glu Pro Gin Val Tyr Thr Leu 340 345 350 Pro Pro Ser Gin Glu Glu Met Thr Lys Asn Gin Val Ser Leu Thr Cys 355 360 365 Leu Val Lys Gly Phe Tyr Pro Ser Asp He Ala Val Glu Trp Glu Ser 370 375 380 Asn Gly Gin Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp 385 390 395 400 Ser Asp Gly Ser Phe Phe Leu Tyr Ser Arg Leu Thr Val Asp Lys Ser 405 410 415 Arg Trp Gin Glu Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala 420 425 430 Leu His Asn His Tyr Thr Gin Lys Ser Leu Ser Leu Ser Leu Gly Lys 435 440 445 <210> 14 <211> 218 <212> PRT <213> Artificial Sequence <220> <223> LC of mAb 0170 <400> 14 Asp He Val Leu Thr Gin Ser Pro Asp Ser Leu Ala Val Ser Leu Gly 1 5 10 15 Glu Arg Ala Thr lie Asn Cys Arg Ala Ser Glu Ser Val Asp Thr Phe 20 25 30 Asp Tyr Ser Phe Leu His Trp Tyr Gin Gin Lys Pro Gly Gin Pro Pro 35 40 45 Lys Leu Leu lie Tyr Arg Ala Ser Asn Leu Glu Ser Gly Val Pro Asp 50 55 60 Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr lie Ser 65 70 75 80 Ser Leu Gin Ala Glu Asp Val Ala Val Tyr Tyr Cys Gin Gin Ser Asn 85 90 95 Glu Asp Pro Tyr Thr Phe Gly Gin Gly Thr Lys Leu Glu lie Lys Arg 100 105 110 Thr Val Ala Ala Pro Ser Val Phe lie Phe Pro Pro Ser Asp Glu Gin 115 120 125 Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr 130 135 140 Pro Arg Glu Ala Lys Val Gin Trp Lys Val Asp Asn Ala Leu Gin Ser 145 150 155 160 Gly Asn Ser Gin Glu Ser Val Thr Glu Gin Asp Ser Lys Asp Ser Thr 165 170 175 Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys 180 185 190 His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro 195 200 205 Val Thr Lys Ser Phe Asn Arg Gly Glu Cys 210 215 <210> 15 <211> 121 <212> PRT <213> Artificial Sequence <220> <223> VH of mAb 0170, 0318 <400> 15 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Lys Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Thr Tyr 20 25 30 Ala Met His Trp Val Arg Gln Ala Ser Gly Lys Gly Leu Glu Trp Val 35 40 45 Gly Arg Ile Arg Thr Lys Ser Ser Asn Tyr Ala Thr Tyr Tyr Ala Ala 50 55 60 Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asp Ser Lys Asn Thr 65 70 75 80 Ala Tyr Leu Gln Met Asn Ser Leu Lys Thr Glu Asp Thr Ala Val Tyr 85 90 95 Tyr Cys Thr Arg Asp Met Gly Ile Arg Arg Gin Phe Ala Tyr Trp Gly 100 105 110 Gln Gly Thr Leu Val Thr Val Ser Ser 115 120 <210> 16 <211> 111 <212> PRT <213> Artificial Sequence <220> <223> VL of mAb 0170 <400> 16 Asp Ile Val Leu Thr Gin Ser Pro Asp Ser Leu Ala Val Ser Leu Gly 1 5 10 15 Glu Arg Ala Thr Ile Asn Cys Arg Ala Ser Glu Ser Val Asp Thr Phe 20 25 30 Asp Tyr Ser Phe Leu His Trp Tyr Gin Gin Lys Pro Gly Gin Pro Pro 35 40 45 Lys Leu Leu Ile Tyr Arg Ala Ser Asn Leu Glu Ser Gly Val Pro Asp 50 55 60 Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser 65 70 75 80 Ser Leu Gin Ala Glu Asp Val Ala Val Tyr Tyr Cys Gin Gin Ser Asn 85 90 95 Glu Asp Pro Tyr Thr Phe Gly Gin Gly Thr Lys Leu Glu Ile Lys 100 105 110 <210> 17 <211> 5 <212> PRT <213> Artificial Sequence <220> <223> CDR1 VH of mAb 0170, 0318 <400> 17 Thr Tyr Ala Met His 1 5 <210> 18 <211> 19 <212> PRT <213> Artificial Sequence <220> <223> CDR2 VH of mAb 0170, 0318 <400> 18 Arg Ile Arg Thr Lys Ser Ser Asn Tyr Ala Thr Tyr Tyr Ala Ala Ser 1 5 10 15 Val Lys Gly <210> 19 <211> 10 <212> PRT <213> Artificial Sequence <220> <223> CDR3 VH of mAb 0170, 0318 <400> 19 Asp Met Gly Ile Arg Arg Gln Phe Ala Tyr 1 5 10 <210> 20 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> CDR1 VL of mAb 0170 <400> 20 Arg Ala Ser Glu Ser Val Asp Thr Phe Asp Tyr Ser Phe Leu His 1 5 10 15 <210> 21 <211> 7 <212> PRT <213> Artificial Sequence <220> <223> CDR2 VL of mAb 0170, 0318 <400> 21 Arg Ala Ser Asn Leu Glu Ser 1 5 <210> 22 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> CDR3 VL of mAb 0170 <400> 22 Gln Gln Ser Asn Glu Asp Pro Tyr Thr 1 5 <210> 23 <211> 111 <212> PRT <213> Artificial Sequence <220> <223> VL of mAb 0318 <400> 23 Asp Ile Val Leu Thr Gln Ser Pro Asp Ser Leu Ala Val Ser Leu Gly 1 5 10 15 Glu Arg Ala Thr Ile Asn Cys Arg Ala Ser Gln Ser Val Asp Thr Phe 20 25 30 Asp Tyr Ser Phe Leu His Trp Tyr Gln Gln Lys Pro Gly Gln Pro Pro 35 40 45 Lys Leu Leu Ile Tyr Arg Ala Ser Asn Leu Glu Ser Gly Val Pro Asp 50 55 60 Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser 65 70 75 80 Ser Leu Gln Ala Glu Asp Val Ala Val Tyr Tyr Cys Gln Gln Ser Asn 85 90 95 Gln Asp Pro Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys 100 105 110 <210> twenty four <211> 15 <212> PRT <213> Artificial sequence <220> <223> CDR1 VL of mAb 0318 <400> twenty four Arg Ala Ser Gln Ser Val Asp Thr Phe Asp Tyr Ser Phe Leu His 1 5 10 15 <210> 25 <211> 9 <212> PRT <213> Artificial sequence <220> <223> CDR3 VL of mAb 0318 <400> 25 Gln Gln Ser Asn Gln Asp Pro Tyr Thr 1 5 <210> 26 <211> 121 <212> PRT <213> Artificial sequence <220> <223> VH of 318 Ab-mutant #1 <400> 26 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Lys Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Thr Tyr 20 25 30 Ala Met His Trp Val Arg Gln Ala Ser Gly Lys Gly Leu Glu Trp Val 35 40 45 Gly Arg Ile Arg Thr Lys Ser Ser Asn Tyr Ala Thr Tyr Tyr Ala Ala 50 55 60 Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asp Ser Lys Asn Thr 65 70 75 80 Ala Tyr Leu Gln Met Asn Ser Leu Lys Thr Glu Asp Thr Ala Val Tyr 85 90 95 Tyr Cys Thr Arg Asp Gln Gly Ile Arg Arg Gln Phe Ala Tyr Trp Gly 100 105 110 Gln Gly Thr Leu Val Thr Val Ser Ser 115 120 <210> 27 <211> 121 <212> PRT <213> Artificial Sequence <220> <223> VH of 318 Ab - Mutant #2 <400> 27 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Lys Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Thr Tyr 20 25 30 Ala Met His Trp Val Arg Gln Ala Ser Gly Lys Gly Leu Glu Trp Val 35 40 45 Gly Arg Ile Arg Thr Lys Ser Ser Asn Tyr Ala Thr Tyr Tyr Ala Ala 50 55 60 Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asp Ser Lys Asn Thr 65 70 75 80 Ala Tyr Leu Gln Met Asn Ser Leu Lys Thr Glu Asp Thr Ala Val Tyr 85 90 95 Tyr Cys Thr Arg Asp Leu Gly Ile Arg Arg Gln Phe Ala Tyr Trp Gly 100 105 110 Gln Gly Thr Leu Val Thr Val Ser Ser 115 120 <210> 28 <211> 121 <212> PRT <213> Artificial Sequence <220> <223> 318 Ab - Mutant #3 VH <400> 28 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Lys Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Thr Tyr 20 25 30 Ala Gln His Trp Val Arg Gln Ala Ser Gly Lys Gly Leu Glu Trp Val 35 40 45 Gly Arg Ile Arg Thr Lys Ser Ser Asn Tyr Ala Thr Tyr Tyr Ala Ala 50 55 60 Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asp Ser Lys Asn Thr 65 70 75 80 Ala Tyr Leu Gln Met Asn Ser Leu Lys Thr Glu Asp Thr Ala Val Tyr 85 90 95 Tyr Cys Thr Arg Asp Met Gly Ile Arg Arg Gln Phe Ala Tyr Trp Gly 100 105 110 Gln Gly Thr Leu Val Thr Val Ser Ser 115 120 <210> 29 <211> 121 <212> PRT <213> Artificial Sequence <220> <223> 318 Ab - mutant #4 VH <400> 29 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Lys Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Thr Tyr 20 25 30 Ala Leu His Trp Val Arg Gln Ala Ser Gly Lys Gly Leu Glu Trp Val 35 40 45 Gly Arg Ile Arg Thr Lys Ser Ser Asn Tyr Ala Thr Tyr Tyr Ala Ala 50 55 60 Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asp Ser Lys Asn Thr 65 70 75 80 Ala Tyr Leu Gln Met Asn Ser Leu Lys Thr Glu Asp Thr Ala Val Tyr 85 90 95 Tyr Cys Thr Arg Asp Met Gly Ile Arg Arg Gln Phe Ala Tyr Trp Gly 100 105 110 Gln Gly Thr Leu Val Thr Val Ser Ser 115 120 <210> 30 <211> 451 <212> PRT <213> Artificial Sequence <220> <223> 0318-IgG1.3f HC <400> 30 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Lys Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Thr Tyr 20 25 30 Ala Met His Trp Val Arg Gln Ala Ser Gly Lys Gly Leu Glu Trp Val 35 40 45 Gly Arg lie Arg Thr Lys Ser Ser Asn Tyr Ala Thr Tyr Tyr Ala Ala 50 55 60 Ser Val Lys Gly Arg Phe Thr lie Ser Arg Asp Asp Ser Lys Asn Thr 65 70 75 80 Ala Tyr Leu Gin Met Asn Ser Leu Lys Thr Glu Asp Thr Ala Val Tyr 85 90 95 Tyr Cys Thr Arg Asp Met Gly lie Arg Arg Gin Phe Ala Tyr Trp Gly 100 105 110 Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser 115 120 125 Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala 130 135 140 Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Gin Pro Val Thr Val 145 150 155 160 Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala 165 170 175 Val Leu Gin Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val 180 185 190 Pro Ser Ser Ser Leu Gly Thr Gin Thr Tyr lie Cys Asn Val Asn His 195 200 205 Lys Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Pro Lys Ser Cys 210 215 220 Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Glu Gly 225 230 235 240 Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met 245 250 255 Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His 260 265 270 Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val 275 280 285 His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr 290 295 300 Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly 305 310 315 320 Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile 325 330 335 Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val 340 345 350 Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser 355 360 365 Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp lie Ala Val Glu 370 375 380 Trp Glu Ser Asn Gly Gin Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro 385 390 395 400 Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val 405 410 415 Asp Lys Ser Arg Trp Gin Gin Gly Asn Val Phe Ser Cys Ser Val Met 420 425 430 His Glu Ala Leu His Asn His Tyr Thr Gin Lys Ser Leu Ser Leu Ser 435 440 445 Pro Gly Lys 450 <210> 31 <211> 450 <212> PRT <213> Artificial Sequence <220> <223> 0318-IgG1.1f HC <400> 31 Glu Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly 1 5 10 15 Ser Leu Lys Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Thr Tyr 20 25 30 Ala Met His Trp Val Arg Gin Ala Ser Gly Lys Gly Leu Glu Trp Val 35 40 45 Gly Arg Ile Arg Thr Lys Ser Ser Asn Tyr Ala Thr Tyr Tyr Ala Ala 50 55 60 Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asp Ser Lys Asn Thr 65 70 75 80 Ala Tyr Leu Gln Met Asn Ser Leu Lys Thr Glu Asp Thr Ala Val Tyr 85 90 95 Tyr Cys Thr Arg Asp Met Gly Ile Arg Arg Gln Phe Ala Tyr Trp Gly 100 105 110 Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser 115 120 125 Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala 130 135 140 Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val 145 150 155 160 Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala 165 170 175 Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val 180 185 190 Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His 195 200 205 Lys Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Pro Lys Ser Cys 210 215 220 Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Glu Gly 225 230 235 240 Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met 245 250 255 Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His 260 265 270 Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val 275 280 285 His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr 290 295 300 Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly 305 310 315 320 Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ser Ser Ile 325 330 335 Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val 340 345 350 Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser 355 360 365 Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu 370 375 380 Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro 385 390 395 400 Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val 405 410 415 Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met 420 425 430 His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser 435 440 445 Pro Gly 450 <210> 32[[ID=2……此处原文似乎缺失部分内容,请补充完整以便准确翻译]] <211> 451 <212> PRT <213> Artificial Sequence <220> <223> 0318 - IgG1 - Aba HC <400> 32 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Lys Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Thr Tyr 20 25 30 Ala Met His Trp Val Arg Gln Ala Ser Gly Lys Gly Leu Glu Trp Val 35 40 45 Gly Arg Ile Arg Thr Lys Ser Ser Asn Tyr Ala Thr Tyr Tyr Ala Ala 50 55 60 Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asp Ser Lys Asn Thr 65 70 75 80 Ala Tyr Leu Gln Met Asn Ser Leu Lys Thr Glu Asp Thr Ala Val Tyr 85 90 95 Tyr Cys Thr Arg Asp Met Gly Ile Arg Arg Gln Phe Ala Tyr Trp Gly 100 105 110 Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser 115 120 125 Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala 130 135 140 Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val 145 150 155 160 Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala 165 170 175 Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val 180 185 190 Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His 195 200 205 Lys Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Pro Lys Ser Cys 210 215 220 Asp Lys Thr His Thr Ser Pro Pro Ser Pro Ala Pro Glu Leu Leu Gly 225 230 235 240 Gly Ser Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met 245 250 255 Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His 260 265 270 Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val 275 280 285 His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr 290 295 300 Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly 305 310 315 320 Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile 325 330 335 Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val 340 345 350 Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser 355 360 365 Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu 370 375 380 Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro 385 390 395 400 Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val 405 410 415 Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met 420 425 430 His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser 435 440 445 Pro Gly Lys 450 <210> 33 <211> 451 <212> PRT <213> Artificial Sequence <220> <223> 0318-IgG4-Aba HC <400> 33 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Lys Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Thr Tyr 20 25 30 Ala Met His Trp Val Arg Gln Ala Ser Gly Lys Gly Leu Glu Trp Val 35 40 45 Gly Arg Ile Arg Thr Lys Ser Ser Asn Tyr Ala Thr Tyr Tyr Ala Ala 50 55 60 Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asp Ser Lys Asn Thr 65 70 75 80 Ala Tyr Leu Gln Met Asn Ser Leu Lys Thr Glu Asp Thr Ala Val Tyr 85 90 95 Tyr Cys Thr Arg Asp Met Gly Ile Arg Arg Gln Phe Ala Tyr Trp Gly 100 105 110 Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser 115 120 125 Val Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr Ala 130 135 140 Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val 145 150 155 160 Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala 165 170 175 Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val 180 185 190 Pro Ser Ser Ser Leu Gly Thr Lys Thr Tyr Thr Cys Asn Val Asp His 195 200 205 Lys Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Pro Lys Ser Cys 210 215 220 Asp Lys Thr His Thr Ser Pro Pro Ser Pro Ala Pro Glu Leu Leu Gly 225 230 235 240 Gly Ser Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met 245 250 255 Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His 260 265 270 Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val 275 280 285 His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr 290 295 300 Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly 305 310 315 320 Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile 325 330 335 Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val 340 345 350 Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser 355 360 365 Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu 370 375 380 Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro 385 390 395 400 Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val 405 410 415 Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met 420 425 430 His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser 435 440 445 Pro Gly Lys 450 <210> 34 <211> 218 <212> PRT <213> Artificial Sequence <220> <223> 0318-IgG1.3f LC; 0318-IgG1.1f LC; 0318-IgG1-Aba LC; 0318-IgG4-Aba LC <400> 34 Asp Ile Val Leu Thr Gln Ser Pro Asp Ser Leu Ala Val Ser Leu Gly 1 5 10 15 Glu Arg Ala Thr lie Asn Cys Arg Ala Ser Gin Ser Val Asp Thr Phe 20 25 30 Asp Tyr Ser Phe Leu His Trp Tyr Gin Gin Lys Pro Gly Gin Pro Pro 35 40 45 Lys Leu Leu lie Tyr Arg Ala Ser Asn Leu Glu Ser Gly Val Pro Asp 50 55 60 Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr lie Ser 65 70 75 80 Ser Leu Gin Ala Glu Asp Val Ala Val Tyr Tyr Cys Gin Gin Ser Asn 85 90 95 Gln Asp Pro Tyr Thr Phe Gly Gin Gly Thr Lys Leu Glu lie Lys Arg 100 105 110 Thr Val Ala Ala Pro Ser Val Phe lie Phe Pro Pro Ser Asp Glu Gin 115 120 125 Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr 130 135 140 Pro Arg Glu Ala Lys Val Gin Trp Lys Val Asp Asn Ala Leu Gin Ser 145 150 155 160 Gly Asn Ser Gin Glu Ser Val Thr Glu Gin Asp Ser Lys Asp Ser Thr 165 170 175 Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys 180 185 190 His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro 195 200 205 Val Thr Lys Ser Phe Asn Arg Gly Glu Cys 210 215 <210> 35 <211> 107 <212> PRT <213> Artificial Sequence <220> <223> IgG1 kappa light chain <400> 35 Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu 1 5 10 15 Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe 20 25 30 Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln 35 40 45 Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser 50 55 60 Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu 65 70 75 80 Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser 85 90 95 Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys 100 105 <210> 36 <211> 1407 <212> PRT <213> Artificial Sequence <220> <223> 0318-IgG1.3f HC <400> 36 Ala Thr Gly Ala Gly Gly Gly Cys Thr Thr Gly Gly Ala Thr Cys Thr 1 5 10 15 Thr Cys Thr Thr Thr Cys Thr Gly Cys Thr Cys Thr Gly Cys Cys Thr 20 25 30 Gly Gly Cys Cys Gly Gly Gly Cys Gly Cys Gly Cys Cys Thr Thr Gly 35 40 45 Gly Cys Cys Gly Ala Ala Gly Thr Gly Cys Ala Gly Cys Thr Cys Gly 50 55 60 Thr Gly Gly Ala Gly Thr Cys Cys Gly Gly Cys Gly Gly Ala Gly Gly 65 70 75 80 Ala Cys Thr Gly Gly Thr Cys Cys Ala Ala Cys Cys Thr Gly Gly Cys 85 90 95 Gly Gly Cys Thr Cys Cys Cys Thr Gly Ala Ala Gly Cys Thr Gly Thr 100 105 110 Cys Cys Thr Gly Cys Gly Cys Thr Gly Cys Cys Thr Cys Cys Gly Gly 115 120 125 Cys Thr Thr Cys Ala Cys Cys Thr Thr Cys Thr Cys Cys Ala Cys Cys 130 135 140 Thr Ala Cys Gly Cys Cys Ala Thr Gly Cys Ala Cys Thr Gly Gly Gly 145 150 155 160 Thr Cys Ala Gly Gly Cys Ala Gly Gly Cys Cys Thr Cys Cys Gly Gly 165 170 175 Ala Ala Ala Gly Gly Gly Cys Cys Thr Gly Gly Ala Ala Thr Gly Gly 180 185 190 Gly Thr Cys Gly Gly Cys Cys Gly Gly Ala Thr Thr Ala Gly Gly Ala 195 200 205 Cys Cys Ala Ala Gly Thr Cys Cys Ala Gly Cys Ala Ala Cys Thr Ala 210 215 220 Cys Gly Cys Thr Ala Cys Cys Thr Ala Cys Thr Ala Cys Gly Cys Cys 225 230 235 240 Gly Cys Cys Ala Gly Cys Gly Thr Cys Ala Ala Gly Gly Gly Cys Cys 245 250 255 Gly Gly Thr Thr Cys Ala Cys Ala Ala Thr Cys Thr Cys Cys Cys Gly 260 265 270 Gly Gly Ala Cys Gly Ala Cys Thr Cys Cys Ala Ala Gly Ala Ala Cys 275 280 285 Ala Cys Cys Gly Cys Cys Thr Ala Thr Cys Thr Cys Cys Ala Gly Ala 290 295 300 Thr Gly Ala Ala Cys Ala Gly Cys Cys Thr Gly Ala Ala Gly Ala Cys 305 310 315 320 Ala Gly Ala Gly Gly Ala Cys Ala Cys Cys Gly Cys Cys Gly Thr Gly 325 330 335 Thr Ala Cys Thr Ala Thr Thr Gly Cys Ala Cys Cys Cys Gly Gly Gly 340 345 350 Ala Thr Ala Thr Gly Gly Gly Cys Ala Thr Thr Cys Gly Gly Ala Gly 355 360 365 Gly Cys Ala Gly Thr Thr Cys Gly Cys Cys Thr Ala Thr Thr Gly Gly 370 375 380 Gly Gly Cys Cys Ala Gly Gly Gly Cys Ala Cys Cys Cys Thr Gly Gly 385 390 395 400 Thr Gly Ala Cys Ala Gly Thr Cys Ala Gly Cys Thr Cys Cys Gly Cys 405 410 415 Cys Ala Gly Cys Ala Cys Ala Ala Ala Ala Gly Gly Ala Cys Cys Thr 420 425 430 Ala Gly Cys Gly Thr Gly Thr Thr Cys Cys Cys Cys Cys Thr Gly Gly 435 440 445 Cys Cys Cys Cys Thr Ala Gly Cys Ala Gly Cys Ala Ala Gly Thr Cys 450 455 460 Cys Ala Cys Ala Ala Gly Cys Gly Gly Cys Gly Gly Cys Ala Cys Cys 465 470 475 480 Gly Cys Thr Gly Cys Cys Cys Thr Gly Gly Gly Cys Thr Gly Thr Cys 485 490 495 Thr Gly Gly Thr Gly Ala Ala Ala Gly Ala Cys Thr Ala Cys Thr Thr 500 505 510 Thr Cys Cys Cys Gly Ala Gly Cys Cys Cys Gly Thr Gly Ala Cys Ala 515 520 525 Gly Thr Gly Ala Gly Cys Thr Gly Gly Ala Ala Cys Thr Cys Thr Gly 530 535 540 Gly Cys Gly Cys Cys Cys Thr Gly Ala Cys Ala Thr Cys Cys Gly Gly 545 550 555 560 Ala Gly Thr Gly Cys Ala Cys Ala Cys Cys Thr Thr Cys Cys Cys Thr 565 570 575 Gly Cys Cys Gly Thr Gly Cys Thr Cys Cys Ala Gly Thr Cys Cys Ala 580 585 590 Gly Cys Gly Gly Cys Cys Thr Gly Thr Ala Cys Ala Gly Cys Cys Thr 595 600 605 Gly Ala Gly Cys Ala Gly Cys Gly Thr Cys Gly Thr Gly Ala Cys Cys 610 615 620 Gly Thr Cys Cys Cys Thr Ala Gly Cys Ala Gly Cys Ala Gly Cys Cys 625 630 635 640 Thr Gly Gly Gly Ala Ala Cys Cys Cys Ala Gly Ala Cys Cys Thr Ala 645 650 655 Cys Ala Thr Cys Thr Gly Cys Ala Ala Cys Gly Thr Gly Ala Ala Cys 660 665 670 Cys Ala Cys Ala Ala Gly Cys Cys Cys Thr Cys Cys Ala Ala Cys Ala 675 680 685 Cys Cys Ala Ala Gly Gly Thr Gly Gly Ala Cys Ala Ala Gly Ala Gly 690 695 700 Gly Gly Thr Gly Gly Ala Gly Cys Cys Cys Ala Ala Gly Thr Cys Cys 705 710 715 720 Thr Gly Thr Gly Ala Cys Ala Ala Gly Ala Cys Cys Cys Ala Thr Ala 725 730 735 Cys Cys Thr Gly Cys Cys Cys Cys Cys Cys Cys Thr Gly Thr Cys Cys 740 745 750 Thr Gly Cys Thr Cys Cys Thr Gly Ala Ala Gly Cys Thr Gly Ala Gly 755 760 765 Gly Gly Cys Gly Cys Cys Cys Cys Thr Thr Cys Cys Gly Thr Cys Thr 770 775 780 Thr Cys Cys Thr Gly Thr Thr Cys Cys Cys Thr Cys Cys Thr Ala Ala 785 790 795 800 Gly Cys Cys Cys Ala Ala Gly Gly Ala Cys Ala Cys Cys Cys Thr Gly 805 810 815 Ala Thr Gly Ala Thr Cys Thr Cys Cys Ala Gly Gly Ala Cys Cys Cys 820 825 830 Cys Cys Gly Ala Gly Gly Thr Gly Ala Cys Cys Thr Gly Thr Gly Thr 835 840 845 Gly Gly Thr Gly Gly Thr Gly Gly Ala Thr Gly Thr Gly Thr Cys Cys 850 855 860 Cys Ala Cys Gly Ala Gly Gly Ala Cys Cys Cys Cys Gly Ala Gly Gly 865 870 875 880 Thr Gly Ala Ala Gly Thr Thr Cys Ala Ala Thr Thr Gly Gly Thr Ala 885 890 895 Cys Gly Thr Cys Gly Ala Cys Gly Gly Cys Gly Thr Gly Gly Ala Gly 900 905 910 Gly Thr Gly Cys Ala Cys Ala Ala Cys Gly Cys Cys Ala Ala Gly Ala 915 920 925 Cys Cys Ala Ala Ala Cys Cys Cys Cys Gly Gly Gly Ala Gly Gly Ala 930 935 940 Gly Cys Ala Gly Thr Ala Thr Ala Ala Cys Ala Gly Cys Ala Cys Cys 945 950 955 960 Thr Ala Cys Cys Gly Gly Gly Thr Gly Gly Thr Gly Thr Cys Cys Gly 965 970 975 Thr Gly Cys Thr Cys Ala Cys Cys Gly Thr Gly Cys Thr Gly Cys Ala 980 985 990 Cys Cys Ala Gly Gly Ala Cys Thr Gly Gly Cys Thr Gly Ala Ala Cys 995 1000 1005 Gly Gly Cys Ala Ala Gly Gly Ala Gly Thr Ala Cys Ala Ala Gly 1010 1015 1020 Thr Gly Thr Ala Ala Gly Gly Thr Cys Ala Gly Cys Ala Ala Thr 1025 1030 1035 Ala Ala Gly Gly Cys Cys Cys Thr Gly Cys Cys...

Claims

1. Use of an antagonistic anti-TREM-1 monoclonal antibody in the preparation of a medicament for treating a disease or disorder in a subject of need, wherein the subject exhibits increased expression levels of a TREM-1-related gene. The TREM-1-related genes mentioned above are: nicotinamide phosphoribosyltransferase (NAMPT); dehydrogenase / reductase 9 (DHRS9); cyclin-dependent kinase inhibitor 1A (CDKN1A); CD52 molecule (CD52); myotubule-associated protein 11 (MTMR11); EH domain-containing 1 (EHD1); solute carrier family 27 member 3 (SLC27A3); Pim-2 proto-oncogene serine / threonine kinase (PIM2); chitinase 3-like 1 (CHI3L1); polypeptide N-acetylgalactosyltransferase 6 (GALNT6); acyl-CoA thioesterase 7 (ACOT7); cytokine-inducible SH2-containing protein (CISH); sequence similarity family 129 member A (FAM129A); Pauli-like kinase 3 (PLK3); and major promoting factor superfamily 12 (MFSD12). C-type lectin domain family 12 member A (CLEC12A); interferon λ receptor 1 (IFNLR1) or a combination thereof, The disease or disorder mentioned above is ulcerative colitis (UC) or Crohn's disease (CD), and The subjects in question had previously been treated with infliximab and did not respond to the treatment.

2. The use as described in claim 1, wherein the TREM-1 related gene is chitinase 3-like 1 (CHI3L1).

3. The use as described in claim 1, wherein the antagonistic anti-TREM-1 monoclonal antibody is a fully human monoclonal antibody.

4. The use as described in claim 1, wherein the antagonistic anti-TREM-1 monoclonal antibody is a humanized monoclonal antibody.

5. Use of the reagent in the preparation of a kit for identifying subjects with a disease or disorder suitable for treatment with an antagonistic anti-TREM-1 monoclonal antibody, wherein said identification includes The expression levels of TREM-1-related genes in the samples of the subjects were measured, wherein subjects eligible for treatment with anti-TREM-1 monoclonal antibodies showed increased expression levels of TREM-1-related genes compared to the control. The TREM-1-related genes mentioned above are: nicotinamide phosphoribosyltransferase (NAMPT); dehydrogenase / reductase 9 (DHRS9); cyclin-dependent kinase inhibitor 1A (CDKN1A); CD52 molecule (CD52); myotubule-associated protein 11 (MTMR11); EH domain-containing 1 (EHD1); solute carrier family 27 member 3 (SLC27A3); Pim-2 proto-oncogene serine / threonine kinase (PIM2); and chitinase 3-like 1 (CH I3L1); N-acetylgalactosyltransferase 6 (GALNT6); acyl-CoA thioesterase 7 (ACOT7); cytokine-inducible SH2-containing protein (CISH); sequence similarity family 129 member A (FAM129A); Pauli-like kinase 3 (PLK3); major promoting factor superfamily 12 (MFSD12); C-type lectin domain family 12 member A (CLEC12A); interferon λ receptor 1 (IFNLR1) or combinations thereof, and The disease or disorder mentioned above is ulcerative colitis (UC) or Crohn's disease (CD).

6. The use as described in claim 5, wherein the TREM-1 related gene is chitinase 3-like 1 (CHI3L1).

7. The use as described in claim 5, wherein the antagonistic anti-TREM-1 monoclonal antibody is a fully human monoclonal antibody.

8. The use as described in claim 5, wherein the antagonistic anti-TREM-1 monoclonal antibody is a humanized monoclonal antibody.

9. Use of the reagent in the preparation of a kit for identifying non-responders to standards of care for a disease or disorder, wherein the identification includes The expression levels of TREM-1-related genes in samples from subjects who had received the stated standard of care were measured. The non-responders exhibited increased expression levels of the TREM-1-related genes, and The TREM-1-related genes mentioned above are: nicotinamide phosphoribosyltransferase (NAMPT); dehydrogenase / reductase 9 (DHRS9); cyclin-dependent kinase inhibitor 1A (CDKN1A); CD52 molecule (CD52); myotubule-associated protein 11 (MTMR11); EH domain-containing 1 (EHD1); solute carrier family 27 member 3 (SLC27A3); Pim-2 proto-oncogene serine / threonine kinase (PIM2); and chitinase 3-like 1 (C HI3L1); N-acetylgalactosyltransferase 6 (GALNT6); acyl-CoA thioesterase 7 (ACOT7); cytokine-inducible SH2-containing protein (CISH); sequence similarity family 129 member A (FAM129A); Pauli-like kinase 3 (PLK3); major facultative factor superfamily 12 (MFSD12); C-type lectin domain family 12 member A (CLEC12A); interferon λ receptor 1 (IFNLR1) or combinations thereof. The disease or disorder mentioned above is ulcerative colitis (UC) or Crohn's disease (CD), and The standard of care mentioned therein is infliximab treatment.

10. The use as claimed in claim 9, wherein the TREM-1 related gene is chitinase 3-like 1 (CHI3L1).

11. Use of an antagonistic anti-TREM-1 monoclonal antibody in the preparation of a kit for determining the efficacy of an antagonistic anti-TREM-1 monoclonal antibody in treating a disease or disorder in a subject of need, wherein the determination comprises administering the antagonistic anti-TREM-1 monoclonal antibody to the subject and measuring the expression level of a TREM-1-related gene in a sample of the subject, wherein the subject exhibits a reduced expression level of the TREM-1-related gene after the administration. The TREM-1-related genes mentioned above are: nicotinamide phosphoribosyltransferase (NAMPT); dehydrogenase / reductase 9 (DHRS9); cyclin-dependent kinase inhibitor 1A (CDKN1A); CD52 molecule (CD52); myotubule-associated protein 11 (MTMR11); EH domain-containing 1 (EHD1); solute carrier family 27 member 3 (SLC27A3); Pim-2 proto-oncogene serine / threonine kinase (PIM2); and chitinase 3-like 1 (CH I3L1); N-acetylgalactosyltransferase 6 (GALNT6); acyl-CoA thioesterase 7 (ACOT7); cytokine-inducible SH2-containing protein (CISH); sequence similarity family 129 member A (FAM129A); Pauli-like kinase 3 (PLK3); major promoting factor superfamily 12 (MFSD12); C-type lectin domain family 12 member A (CLEC12A); interferon λ receptor 1 (IFNLR1) or combinations thereof, and The disease or disorder mentioned above is ulcerative colitis (UC) or Crohn's disease (CD).

12. The use as claimed in claim 11, wherein the TREM-1 related gene is chitinase 3-like 1 (CHI3L1).

13. The use as described in claim 11, wherein the antagonistic anti-TREM-1 monoclonal antibody is a fully human monoclonal antibody.

14. The use as described in claim 11, wherein the antagonistic anti-TREM-1 monoclonal antibody is a humanized monoclonal antibody.

15. The use as described in any one of claims 1-2, 5-6, and 11-12, wherein the subject also exhibited one or more of the following prior to administration of the antagonistic anti-TREM-1 monoclonal antibody: increased baseline Mayo score, increased 2B grade lamina propria neutrophil infiltration score, and increased fecal calprotectin levels, wherein (a) The subjects described showed an increase of at least 5% in their baseline Mayo score compared to the reference; (b) The subject had a baseline Mayo score greater than 6 prior to administration; (c) The subjects exhibited an increase of at least 5% in the lamina propria neutrophil infiltration fraction of grade 2B compared to the control; (d) The subjects exhibited a grade 2B lamina propria neutrophil infiltration fraction greater than 0; (e) The subjects described exhibited an increase of at least 5% in fecal calprotectin levels compared to the control; and / or (f) The subjects showed fecal calprotectin levels greater than 1.5 log10.

16. The use as described in any one of claims 1 to 14, wherein one or more scores are further measured before, simultaneously with or after measuring the expression level of the TREM-1-related gene and / or administering an antagonistic anti-TREM-1 monoclonal antibody, said scores including baseline Mayo score, grade 2B lamina propria neutrophil infiltration score and fecal calprotectin level.

17. The use according to any one of claims 1-2, 5-6, and 11-12, wherein administration of the antagonistic anti-TREM-1 monoclonal antibody reduces the expression of said TREM-1-related genes, wherein (a) The baseline Mayo score decreased by at least 5%; (b) The neutrophil infiltration fraction in the lamina propria of grade 2B is reduced by at least 5%; and / or (c) The fecal calprotectin level is reduced by at least 5%.

18. The use as described in any one of claims 1-2, 5-6, and 11-12, wherein administration of the antagonistic anti-TREM-1 monoclonal antibody reduces the expression of the TREM-1-related gene, wherein administration of the antagonistic anti-TREM-1 monoclonal antibody also reduces the subject's baseline Mayo score, grade 2B lamina propria neutrophil infiltration fraction, and / or fecal calprotectin level, wherein (a) The baseline Mayo score decreased by at least 5%; (b) The neutrophil infiltration fraction in the lamina propria of grade 2B is reduced by at least 5%; and / or (c) The fecal calprotectin level is reduced by at least 5%.

19. The use as described in any one of claims 1 to 14, wherein the expression level of the TREM-1-related gene increases in the presence of a natural TREM-1 ligand, but does not increase in the presence of an agonistic anti-TREM-1 monoclonal antibody.

20. The use as claimed in any one of claims 5 to 14, wherein the sample comprises tissue, blood, serum, plasma, saliva, urine, or a combination thereof.

21. The use according to any one of claims 1 to 6 and 11 to 12, wherein the antagonistic anti-TREM-1 monoclonal antibody comprises heavy chain CDR1, CDR2, and CDR3 and light chain CDR1, CDR2, and CDR3, wherein (a) The light chain CDR1 consists of the sequence RASQSVDTFDYSFLH (SEQ ID NO: 24). (b) The light chain CDR2 consists of the sequence of RASNLES (SEQ ID NO: 21). (c) The light chain CDR3 consists of the sequence QQSNQDPYT (SEQ ID NO: 25). (d) The heavy chain CDR1 consists of the sequence of TYAMH (SEQ ID NO: 17). (e) The heavy chain CDR2 consists of the sequence RIRTKSSNYATYYAASVKG (SEQ ID NO: 18), and (f) wherein the heavy chain CDR3 is composed of the sequence of DMGIRRQFAY (SEQ ID NO: 19) or wherein the heavy chain CDR3 is composed of the sequence of DQGIRRQFAY (SEQ ID NO: 72).

22. The use as claimed in claim 21, wherein the antagonistic anti-TREM-1 monoclonal antibody comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein the VH comprises an amino acid sequence as shown in SEQ ID NO: 15 or 26-29, and the VL comprises an amino acid sequence as shown in SEQ ID NO:

23.

23. The use as claimed in claim 21, wherein the antagonistic anti-TREM-1 monoclonal antibody comprises a heavy chain (HC) and a light chain (LC), wherein the HC comprises an amino acid sequence as shown in SEQ ID NO: 30, 31, 32 or 33, and the LC comprises an amino acid sequence as shown in SEQ ID NO:

34.

24. The use according to any one of claims 1 to 6 and 11 to 12, wherein the antagonistic anti-TREM-1 monoclonal antibody comprises heavy chain CDR1, CDR2, and CDR3 and light chain CDR1, CDR2, and CDR3, wherein (a) The heavy chains CDR1, CDR2 and CDR3 are composed of the amino acid sequences shown in SEQ ID NO: 61, 62 and 63, respectively, and the light chains CDR1, CDR2 and CDR3 are composed of the amino acid sequences shown in SEQ ID NO: 64, 65 and 66, respectively; (b) The heavy chains CDR1, CDR2 and CDR3 are composed of the amino acid sequences shown in SEQ ID NO: 67, 68 and 69, respectively, and the light chains CDR1, CDR2 and CDR3 are composed of the amino acid sequences shown in SEQ ID NO: 70, 71 and 72, respectively; (c) The heavy chains CDR1, CDR2 and CDR3 are composed of the amino acid sequences shown in SEQ ID NO: 67, 68 and 69, respectively, and the light chains CDR1, CDR2 and CDR3 are composed of the amino acid sequences shown in SEQ ID NO: 64, 65 and 73, respectively; (d) The heavy chains CDR1, CDR2 and CDR3 are composed of the amino acid sequences shown in SEQ ID NO: 74, 75 and 76, respectively, and the light chains CDR1, CDR2 and CDR3 are composed of the amino acid sequences shown in SEQ ID NO: 70, 77 and 78, respectively; (e) The heavy chains CDR1, CDR2 and CDR3 are composed of the amino acid sequences shown in SEQ ID NO: 79, 80 and 81, respectively, and the light chains CDR1, CDR2 and CDR3 are composed of the amino acid sequences shown in SEQ ID NO: 70, 71 and 72, respectively; (f) The heavy chains CDR1, CDR2, and CDR3 are composed of the amino acid sequences shown in SEQ ID NO: 159, 160, and 161, respectively, and the light chains CDR1, CDR2, and CDR3 are composed of the amino acid sequences shown in SEQ ID NO: 70, 71, and 162, respectively; or (g) The heavy chains CDR1, CDR2 and CDR3 are composed of the amino acid sequences shown in SEQ ID NO: 159, 160 and 161, respectively, and the light chains CDR1, CDR2 and CDR3 are composed of the amino acid sequences shown in SEQ ID NO: 70, 71 and 133, respectively.

25. The use as claimed in claim 24, wherein the antagonistic anti-TREM-1 monoclonal antibody comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein the VH comprises an amino acid sequence as shown in SEQ ID NO: 53, 55, 58, 60 or 153, and wherein the VL comprises an amino acid sequence as shown in SEQ ID NO: 54, 56, 57, 59, 154 or 155.

26. The use as claimed in claim 24, wherein the antagonistic anti-TREM-1 monoclonal antibody further comprises a heavy chain (HC) constant region and a light chain (LC) constant region, wherein the HC constant region comprises an amino acid sequence as shown in SEQ ID NO: 48, SEQ ID NO: 47, SEQ ID NO: 11 or SEQ ID NO: 12, and the LC constant region comprises an amino acid sequence as shown in SEQ ID NO:

35.

27. The use according to any one of claims 1 to 6 and 11 to 12, wherein the antagonistic anti-TREM-1 monoclonal antibody comprises heavy chain CDR1, CDR2, and CDR3 and light chain CDR1, CDR2, and CDR3, wherein (a) The amino acid sequence of the heavy chain CDR1 is TYAMH; (b) The amino acid sequence of the heavy chain CDR2 is RIRTKSSNYATYYAASVKG; (c) The amino acid sequence of the heavy chain CDR3 is DMGQRRQFAY; (d) The amino acid sequence of the light chain CDR1 is RASESVDTFDYSFLH; (e) The amino acid sequence of the light chain CDR2 is RASNLES; and (f) The amino acid sequence of the light chain CDR3 is QQSNEDPYT.

28. The use as claimed in claim 27, wherein the antagonistic anti-TREM-1 monoclonal antibody comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein the VH comprises amino acids 1 to 121 of SEQ ID NO: 13, and wherein the VL comprises amino acids 1 to 111 of SEQ ID NO:

14.

29. The use as claimed in claim 27, wherein the antagonistic anti-TREM-1 monoclonal antibody comprises a heavy chain (HC) and a light chain (LC), wherein the HC comprises an amino acid sequence as shown in SEQ ID NO: 13, and wherein the LC comprises an amino acid sequence as shown in SEQ ID NO: 14.

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