Depletion and inhibition of plasmacytoid dendritic cells

Abstract

Disclosed are antibodies or fragments thereof, compositions and methods of use related to the depletion, inhibition, or depletion and inhibition of plasmacytoid dendritic cells (pDCs).

Description

[Technical Field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims priority to U.S. Provisional Patent Application No. 63 / 395,003, filed August 4, 2022, the contents of which are incorporated herein by reference in their entirety.

[0002] Sequence Listing Description This application incorporates by reference the material in the ST.26 XML file entitled NPBUS-2_ST.26.xml, which is 20 KB and was created on August 4, 2022.

[0003] The present disclosure provides novel antibodies and compositions and uses thereof. [Background technology]

[0004] Plasmacytoid dendritic cells (pDCs) have been implicated in several disorders. U.S. Patent No. 9,670,283 discloses compositions that may be useful for depleting pDCs in patients in need thereof. Summary of the Invention

[0005] The present disclosure provides a method for detecting a human leukemia virus (HV) comprising administering to a mammalian subject the present invention a method for detecting a human leukemia virus (HV) virus ... The present invention provides an antibody or fragment thereof comprising:

[0006] In embodiments, the antibody or fragment thereof has an affinity (K d ) binds the epitope having SEQ ID NO:11.

[0007] In embodiments, the antibody or fragment thereof is a monoclonal antibody or fragment thereof.

[0008] In embodiments, the antibody or fragment thereof is a humanized antibody or fragment thereof.

[0009] In embodiments, the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:1 or a sequence at least about 90% identical to SEQ ID NO:1, and the light chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, and a sequence at least about 90% identical to SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, or SEQ ID NO:10.

[0010] In embodiments, the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:2 or a sequence at least about 90% identical to SEQ ID NO:2, and the light chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, and a sequence at least about 90% identical to SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, or SEQ ID NO:10.

[0011] In embodiments, the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:3 or a sequence at least about 90% identical to SEQ ID NO:3, and the light chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, and a sequence at least about 90% identical to SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, or SEQ ID NO:10.

[0012] In embodiments, the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:4 or a sequence at least about 90% identical to SEQ ID NO:4, and the light chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, and a sequence at least about 90% identical to SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, or SEQ ID NO:10.

[0013] In embodiments, the heavy chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, and a sequence at least about 90% identical to SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, or SEQ ID NO:4; The light chain variable region comprises the amino acid sequence given by SEQ ID NO:5 or a sequence at least about 90% identical to SEQ ID NO:5.

[0014] In embodiments, the heavy chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, and a sequence at least about 90% identical to SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, or SEQ ID NO:4; The light chain variable region comprises the amino acid sequence given by SEQ ID NO:6, or a sequence at least about 90% identical to SEQ ID NO:6.

[0015] In embodiments, the heavy chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, and a sequence at least about 90% identical to SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, or SEQ ID NO:4; The light chain variable region comprises the amino acid sequence given by SEQ ID NO:7 or a sequence at least about 90% identical to SEQ ID NO:7.

[0016] In embodiments, the heavy chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, and a sequence at least about 90% identical to SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, or SEQ ID NO:4; The light chain variable region comprises the amino acid sequence given by SEQ ID NO:8 or a sequence at least about 90% identical to SEQ ID NO:8.

[0017] In embodiments, the heavy chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, and a sequence at least about 90% identical to SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, or SEQ ID NO:4; The light chain variable region comprises the amino acid sequence given by SEQ ID NO:9 or a sequence at least about 90% identical to SEQ ID NO:9.

[0018] In embodiments, the heavy chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, and a sequence at least about 90% identical to SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, or SEQ ID NO:4; The light chain variable region comprises the amino acid sequence given by SEQ ID NO:10, or a sequence at least about 90% identical to SEQ ID NO:10.

[0019] In embodiments, the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:1 or a sequence at least about 90% identical to SEQ ID NO:1, and the light chain variable region comprises the amino acid sequence given by SEQ ID NO:5 or a sequence at least about 90% identical to SEQ ID NO:5.

[0020] In embodiments, the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:2 or a sequence at least about 90% identical to SEQ ID NO:2, and the light chain variable region comprises the amino acid sequence given by SEQ ID NO:5 or a sequence at least about 90% identical to SEQ ID NO:5.

[0021] In embodiments, the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:3 or a sequence at least about 90% identical to SEQ ID NO:3, and the light chain variable region comprises the amino acid sequence given by SEQ ID NO:5 or a sequence at least about 90% identical to SEQ ID NO:5.

[0022] In embodiments, the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:4 or a sequence at least about 90% identical to SEQ ID NO:4, and the light chain variable region comprises the amino acid sequence given by SEQ ID NO:5 or a sequence at least about 90% identical to SEQ ID NO:5.

[0023] In embodiments, the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:1 or a sequence at least about 90% identical to SEQ ID NO:1, and the light chain variable region comprises the amino acid sequence given by SEQ ID NO:6 or a sequence at least about 90% identical to SEQ ID NO:6.

[0024] In embodiments, the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:1 or a sequence at least about 90% identical to SEQ ID NO:1, and the light chain variable region comprises the amino acid sequence given by SEQ ID NO:7 or a sequence at least about 90% identical to SEQ ID NO:7.

[0025] In embodiments, the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:1 or a sequence at least about 90% identical to SEQ ID NO:1, and the light chain variable region comprises the amino acid sequence given by SEQ ID NO:8 or a sequence at least about 90% identical to SEQ ID NO:8.

[0026] In embodiments, the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:1 or a sequence at least about 90% identical to SEQ ID NO:1, and the light chain variable region comprises the amino acid sequence given by SEQ ID NO:9 or a sequence at least about 90% identical to SEQ ID NO:9.

[0027] In embodiments, the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:1 or a sequence at least about 90% identical to SEQ ID NO:1, and the light chain variable region comprises the amino acid sequence given by SEQ ID NO:10 or a sequence at least about 90% identical to SEQ ID NO:10.

[0028] In embodiments, the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:2 or a sequence at least about 90% identical to SEQ ID NO:2, and the light chain variable region comprises the amino acid sequence given by SEQ ID NO:6 or a sequence at least about 90% identical to SEQ ID NO:6.

[0029] In embodiments, the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:3 or a sequence at least about 90% identical to SEQ ID NO:3, and the light chain variable region comprises the amino acid sequence given by SEQ ID NO:6 or a sequence at least about 90% identical to SEQ ID NO:6.

[0030] In embodiments, the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:4 or a sequence at least about 90% identical to SEQ ID NO:4, and the light chain variable region comprises the amino acid sequence given by SEQ ID NO:6 or a sequence at least about 90% identical to SEQ ID NO:6.

[0031] In embodiments, the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:2 or a sequence at least about 90% identical to SEQ ID NO:2, and the light chain variable region comprises the amino acid sequence given by SEQ ID NO:7 or a sequence at least about 90% identical to SEQ ID NO:7.

[0032] In embodiments, the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:3 or a sequence at least about 90% identical to SEQ ID NO:3, and the light chain variable region comprises the amino acid sequence given by SEQ ID NO:7 or a sequence at least about 90% identical to SEQ ID NO:7.

[0033] In embodiments, the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:4 or a sequence at least about 90% identical to SEQ ID NO:4, and the light chain variable region comprises the amino acid sequence given by SEQ ID NO:7 or a sequence at least about 90% identical to SEQ ID NO:7.

[0034] In embodiments, the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:2 or a sequence at least about 90% identical to SEQ ID NO:2, and the light chain variable region comprises the amino acid sequence given by SEQ ID NO:8 or a sequence at least about 90% identical to SEQ ID NO:8.

[0035] In embodiments, the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:3 or a sequence at least about 90% identical to SEQ ID NO:3, and the light chain variable region comprises the amino acid sequence given by SEQ ID NO:8 or a sequence at least about 90% identical to SEQ ID NO:8.

[0036] In embodiments, the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:4 or a sequence at least about 90% identical to SEQ ID NO:4, and the light chain variable region comprises the amino acid sequence given by SEQ ID NO:8 or a sequence at least about 90% identical to SEQ ID NO:8.

[0037] In embodiments, the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:2 or a sequence at least about 90% identical to SEQ ID NO:2, and the light chain variable region comprises the amino acid sequence given by SEQ ID NO:9 or a sequence at least about 90% identical to SEQ ID NO:9.

[0038] In embodiments, the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:3 or a sequence at least about 90% identical to SEQ ID NO:3, and the light chain variable region comprises the amino acid sequence given by SEQ ID NO:9 or a sequence at least about 90% identical to SEQ ID NO:9.

[0039] In embodiments, the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:4 or a sequence at least about 90% identical to SEQ ID NO:4, and the light chain variable region comprises the amino acid sequence given by SEQ ID NO:9 or a sequence at least about 90% identical to SEQ ID NO:9.

[0040] In embodiments, the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:2 or a sequence at least about 90% identical to SEQ ID NO:2, and the light chain variable region comprises the amino acid sequence given by SEQ ID NO:10 or a sequence at least about 90% identical to SEQ ID NO:10.

[0041] In embodiments, the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:3 or a sequence at least about 90% identical to SEQ ID NO:3, and the light chain variable region comprises the amino acid sequence given by SEQ ID NO:10 or a sequence at least about 90% identical to SEQ ID NO:10.

[0042] In embodiments, the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:4 or a sequence at least about 90% identical to SEQ ID NO:4, and the light chain variable region comprises the amino acid sequence given by SEQ ID NO:10 or a sequence at least about 90% identical to SEQ ID NO:10.

[0043] In several embodiments, a method for depleting, inhibiting, or depleting and inhibiting plasmacytoid dendritic cells (pDCs) in a patient in need thereof comprises administering to the patient an effective amount of an antibody or fragment thereof that specifically binds to an epitope comprising blood dendritic cell antigen-2 (BDCA2) and depletes, inhibits, or depletes and inhibits pDCs, thereby depleting, inhibiting, or depleting and inhibiting pDCs. The antibody or fragment thereof comprises a heavy chain variable region comprising an amino acid sequence selected from the group consisting of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, and a sequence at least about 90% identical to SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3 or SEQ ID NO:4, and a light chain variable region comprising an amino acid sequence selected from the group consisting of SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, and a sequence at least about 90% identical to SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9 or SEQ ID NO:10.

[0044] In embodiments of the method for depleting, inhibiting, or depleting and inhibiting pDCs, the epitope comprises the amino acid sequence given by SEQ ID NO: 11, and the antibody or fragment thereof has an affinity (K d ) binds the epitope.

[0045] In embodiments of the methods for depleting, inhibiting, or depleting and inhibiting pDCs, the antibody or fragment thereof is a monoclonal antibody or fragment thereof.

[0046] In embodiments of the methods for depleting, inhibiting, or depleting and inhibiting pDCs, the antibody or fragment thereof is a humanized antibody or fragment thereof.

[0047] In embodiments of the methods for depleting, inhibiting, or depleting and inhibiting pDC, the BDCA2 is human BDCA2.

[0048] In some embodiments, a method for treating a plasmacytoid dendritic cell (pDC)-associated disorder in a patient in need thereof comprises administering to the patient an effective amount of an antibody or fragment thereof that specifically binds to an epitope comprising blood dendritic cell antigen-2 (BDCA2) and depletes, inhibits, or both depletes and inhibits pDCs, thereby treating the disorder. The antibody or fragment thereof comprises a heavy chain variable region comprising an amino acid sequence selected from the group consisting of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, and sequences at least about 90% identical to SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, or SEQ ID NO:4, and a light chain variable region comprising an amino acid sequence selected from the group consisting of SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, and sequences at least about 90% identical to SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, or SEQ ID NO:10.

[0049] In embodiments of the method for treating a disorder associated with pDC, the disorder is selected from the group consisting of an infectious disease, an autoimmune disease, an inflammatory condition, a respiratory condition, a skin disorder, a cancer, and a combination of two or more thereof.

[0050] In an embodiment of the method for treating a disorder associated with pDC, the disorder is an infectious disease associated with infection by an infectious agent.

[0051] In embodiments of the methods for treating a disorder associated with pDC, the infectious agent is one or more viruses or one or more microorganisms.

[0052] In embodiments of methods for treating disorders associated with pDCs, the infectious agent is selected from the group consisting of Hepadnaviridae; Flaviviridae; Retroviridae; Herpesviridae; Papovaviridae; Rhabdoviridae; Paramyxoviridae; Reoviridae; Bunyaviridae; Filoviridae; Adenovirus Adenoviridae; Parvoviridae; Arenaviridae; Orthomyxoviridae; Poxviridae; Togaviridae; Coronaviridae; Picornaviridae; Rhinoviruses; Orbiviruses; Picodnaviruses; Encephalomyocarditis virus virus (EMV); Parainfluenza viruses; Adenoviruses; Coxsackieviruses; Echoviruses; Rubeola virus; Rubella virus; Human papillomaviruses; Canine distemper virus; Canine contagious hepatitis virus; Feline calicivirus; Feline rhinotracheitis virus; TGE virus (porcine); Foot and mouth disease virus; Simian virus 5;A virus selected from the group consisting of human parainfluenza virus type 2; human metapneumovirus; and enteroviruses;

[0053] In embodiments of methods for treating disorders associated with pDCs, the infectious agent is selected from the group consisting of Rickettsia, Chlamydia, Mycobacteria, Clostridia, Corynebacteria, Mycoplasma, Ureaplasma, Legionella, Shigella, Salmonella, pathogenic Escherichia coli species, Bordatella, Neisseria, Treponema, Bacillus, Haemophilus, Moraxella, Vibrio, Staphylococcus spp., Streptococcus spp., and / or Streptococcus spp. The microorganism is selected from the group consisting of Campylobacter spp., Borrelia spp., Leptospira spp., Erlichia spp., Klebsiella spp., Pseudomonas spp. and Helicobacter spp.

[0054] In embodiments of the method for treating a disorder associated with pDC, the infectious agent is selected from the group consisting of Helicobacter pylori, Chlamydia pneumoniae, Chlamydia trachomatis, Ureaplasma urealyticum, Mycoplasma pneumoniae, Staphylococcus aureus, Streptococcus pyogenes, Streptococcus pneumoniae, Streptococcus viridans, Enterococcus faecalis, Neisseria meningitidis, Neisseria gonorrhoeae, Treponema pallidum, and the like. pallidum, Bacillus anthracis, Salmonella typhi, Vibrio cholera, Pasteurella pestis (Yersinia pestis), Pseudomonas aeruginosa, Campylobacter jejuni, Clostridium difficile, Clostridium botulinum, Mycobacterium tuberculosis, Borrelia burgdorferi, Haemophilus ducreyi, Corynebacterium diphtheria, Bordetella pertussis, Bordetella parapertussis parapertussis, Bordetella bronchiseptica, Haemophilus influenzae, and enterotoxic Escherichia coliThe microorganism is selected from the group consisting of:

[0055] In embodiments of methods for treating a disorder associated with pDC, the disorder comprises an autoimmune disease selected from the group consisting of diabetes, psoriasis of the skin, hyperthyroidism, autoimmune adrenal insufficiency, hemolytic anemia, rheumatic carditis, systemic lupus erythematosus, cutaneous lupus erythematosus, psoriatic arthritis, Sjogren's syndrome polymyositis, myositis, multiple sclerosis, Crohn's disease, ulcerative colitis, lupus, inflammatory bowel syndrome, dermatomyositis, dermatitis, atopic dermatitis, scleroderma, polymyositis, psoriasis, alopecia areata, rheumatoid arthritis, graft versus host disease, interferonopathy, irritable bowel syndrome, and combinations of two or more thereof.

[0056] In embodiments of the methods for treating a disorder associated with pDC, the disorder comprises an inflammatory condition, and the inflammatory condition comprises acute kidney injury.

[0057] In embodiments of the methods for treating a disorder associated with pDC, the disorder comprises a respiratory condition, and the respiratory condition comprises asthma, pulmonary fibrosis, or a combination of asthma and pulmonary fibrosis.

[0058] In embodiments of the methods for treating a disorder associated with pDC, the disorder comprises a skin disorder selected from the group consisting of scleroderma, psoriasis, atopic dermatitis, dermatomyositis, and combinations of two or more thereof.

[0059] In embodiments of methods for treating a disorder associated with pDC, the disorder is selected from the group consisting of breast cancer, prostate cancer, lymphoma, skin cancer, pancreatic cancer, colon cancer, melanoma, malignant melanoma, ovarian cancer, brain cancer, primary brain carcinoma, head and neck cancer, glioma, glioblastoma, liver cancer, bladder cancer, non-small cell lung cancer, head or neck carcinoma, breast carcinoma, ovarian carcinoma, lung carcinoma, small cell lung carcinoma, Wilms' tumor, cervical carcinoma, testicular carcinoma, bladder carcinoma, pancreatic carcinoma, gastric carcinoma, colon carcinoma, prostate carcinoma, genitourinary carcinoma, thyroid carcinoma, esophageal carcinoma, myeloma, multiple myeloma, adrenal carcinoma, renal cell carcinoma, endometrial carcinoma, adrenocortical carcinoma, malignant pancreatic insulinoma, malignant carcinoid carcinoma, choriocarcinoma, mycosis fungoides, malignant hypercalcemia, cervical hyperplasia, leukemia, acute lymphoblastic leukemia, and the like. and cancers selected from the group consisting of hematologic malignancies, B-cell acute lymphoblastic leukemia, blastic plasmacytoid dendritic cell neoplasm ("BPDCN" or "pDC leukemia"), chronic lymphocytic leukemia, acute myeloid leukemia, chronic myelogenous leukemia ("AML"), chronic granulocytic leukemia, acute granulocytic leukemia, age-related hairy cell leukemia, chronic myelomonocytic leukemia ("CMML"), myelodysplastic syndromes, chronic myelofibrosis, multiple myeloma, neuroblastoma, rhabdomyosarcoma, Kaposi's sarcoma, Waldenstrom's macroglobulinemia, diffuse large B-cell lymphoma (DLBCL) (L265P), polycythemia vera, essential thrombocytosis, Hodgkin's disease, non-Hodgkin's lymphoma, soft tissue sarcoma, osteogenic sarcoma, primary macroglobulinemia, retinoblastoma, and combinations of two or more thereof.

[0060] In an embodiment of the method for treating a disorder associated with pDC, the epitope comprises the amino acid sequence given by SEQ ID NO: 11, and the antibody or fragment thereof has an affinity (K d ) binds the epitope.

[0061] In embodiments of the methods for treating a disorder associated with pDC, the antibody or fragment thereof is a monoclonal antibody or fragment thereof.

[0062] In embodiments of the methods for treating a disorder associated with pDC, the antibody or fragment thereof is a humanized antibody or fragment thereof.

[0063] In embodiments of methods for treating a disorder associated with pDC, the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:1 or a sequence at least about 90% identical to SEQ ID NO:1, and the light chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, and a sequence at least about 90% identical to SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, or SEQ ID NO:10.

[0064] In embodiments of methods for treating a disorder associated with pDC, the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:2 or a sequence at least about 90% identical to SEQ ID NO:2, and the light chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, and a sequence at least about 90% identical to SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, or SEQ ID NO:10.

[0065] In embodiments of methods for treating disorders associated with pDC, the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:3 or a sequence at least about 90% identical to SEQ ID NO:3, and the light chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, and a sequence at least about 90% identical to SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, or SEQ ID NO:10.

[0066] In embodiments of methods for treating a disorder associated with pDC, the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:4 or a sequence at least about 90% identical to SEQ ID NO:4, and the light chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, and a sequence at least about 90% identical to SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, or SEQ ID NO:10.

[0067] In embodiments of the method for treating a disorder associated with pDC, the heavy chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, and a sequence at least about 90% identical to SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, or SEQ ID NO:4; The light chain variable region comprises the amino acid sequence given by SEQ ID NO:5 or a sequence at least about 90% identical to SEQ ID NO:5.

[0068] In embodiments of the method for treating a disorder associated with pDC, the heavy chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, and a sequence at least about 90% identical to SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, or SEQ ID NO:4; The light chain variable region comprises the amino acid sequence given by SEQ ID NO:6, or a sequence at least about 90% identical to SEQ ID NO:6.

[0069] In embodiments of the method for treating a disorder associated with pDC, the heavy chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, and a sequence at least about 90% identical to SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, or SEQ ID NO:4; The light chain variable region comprises the amino acid sequence given by SEQ ID NO:7 or a sequence at least about 90% identical to SEQ ID NO:7.

[0070] In embodiments of the method for treating a disorder associated with pDC, the heavy chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, and a sequence at least about 90% identical to SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, or SEQ ID NO:4; The light chain variable region comprises the amino acid sequence given by SEQ ID NO:8 or a sequence at least about 90% identical to SEQ ID NO:8.

[0071] In embodiments of the method for treating a disorder associated with pDC, the heavy chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, and a sequence at least about 90% identical to SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, or SEQ ID NO:4; The light chain variable region comprises the amino acid sequence given by SEQ ID NO:9 or a sequence at least about 90% identical to SEQ ID NO:9.

[0072] In embodiments of the method for treating a disorder associated with pDC, the heavy chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, and a sequence at least about 90% identical to SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, or SEQ ID NO:4; The light chain variable region comprises the amino acid sequence given by SEQ ID NO:10, or a sequence at least about 90% identical to SEQ ID NO:10.

[0073] In an embodiment of the method for treating a disorder associated with pDC, the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:1 or a sequence at least about 90% identical to SEQ ID NO:1, and the light chain variable region comprises the amino acid sequence given by SEQ ID NO:5 or a sequence at least about 90% identical to SEQ ID NO:5.

[0074] In an embodiment of a method for treating a disorder associated with pDC, the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:2 or a sequence at least about 90% identical to SEQ ID NO:2, and the light chain variable region comprises the amino acid sequence given by SEQ ID NO:5 or a sequence at least about 90% identical to SEQ ID NO:5.

[0075] In an embodiment of a method for treating a disorder associated with pDC, the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:3 or a sequence at least about 90% identical to SEQ ID NO:3, and the light chain variable region comprises the amino acid sequence given by SEQ ID NO:5 or a sequence at least about 90% identical to SEQ ID NO:5.

[0076] In an embodiment of a method for treating a disorder associated with pDC, the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:4 or a sequence at least about 90% identical to SEQ ID NO:4, and the light chain variable region comprises the amino acid sequence given by SEQ ID NO:5 or a sequence at least about 90% identical to SEQ ID NO:5.

[0077] In an embodiment of a method for treating a disorder associated with pDC, the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:1 or a sequence at least about 90% identical to SEQ ID NO:1, and the light chain variable region comprises the amino acid sequence given by SEQ ID NO:6 or a sequence at least about 90% identical to SEQ ID NO:6.

[0078] In an embodiment of a method for treating a disorder associated with pDC, the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:1 or a sequence at least about 90% identical to SEQ ID NO:1, and the light chain variable region comprises the amino acid sequence given by SEQ ID NO:7 or a sequence at least about 90% identical to SEQ ID NO:7.

[0079] In an embodiment of the method for treating a disorder associated with pDC, the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:1 or a sequence at least about 90% identical to SEQ ID NO:1, and the light chain variable region comprises the amino acid sequence given by SEQ ID NO:8 or a sequence at least about 90% identical to SEQ ID NO:8.

[0080] In an embodiment of a method for treating a disorder associated with pDC, the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:1 or a sequence at least about 90% identical to SEQ ID NO:1, and the light chain variable region comprises the amino acid sequence given by SEQ ID NO:9 or a sequence at least about 90% identical to SEQ ID NO:9.

[0081] In an embodiment of a method for treating a disorder associated with pDC, the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:1 or a sequence at least about 90% identical to SEQ ID NO:1, and the light chain variable region comprises the amino acid sequence given by SEQ ID NO:10 or a sequence at least about 90% identical to SEQ ID NO:10.

[0082] In an embodiment of a method for treating a disorder associated with pDC, the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:2 or a sequence at least about 90% identical to SEQ ID NO:2, and the light chain variable region comprises the amino acid sequence given by SEQ ID NO:6 or a sequence at least about 90% identical to SEQ ID NO:6.

[0083] In an embodiment of a method for treating a disorder associated with pDC, the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:3 or a sequence at least about 90% identical to SEQ ID NO:3, and the light chain variable region comprises the amino acid sequence given by SEQ ID NO:6 or a sequence at least about 90% identical to SEQ ID NO:6.

[0084] In an embodiment of a method for treating a disorder associated with pDC, the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:4 or a sequence at least about 90% identical to SEQ ID NO:4, and the light chain variable region comprises the amino acid sequence given by SEQ ID NO:6 or a sequence at least about 90% identical to SEQ ID NO:6.

[0085] In an embodiment of a method for treating a disorder associated with pDC, the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:2 or a sequence at least about 90% identical to SEQ ID NO:2, and the light chain variable region comprises the amino acid sequence given by SEQ ID NO:7 or a sequence at least about 90% identical to SEQ ID NO:7.

[0086] In an embodiment of a method for treating a disorder associated with pDC, the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:3 or a sequence at least about 90% identical to SEQ ID NO:3, and the light chain variable region comprises the amino acid sequence given by SEQ ID NO:7 or a sequence at least about 90% identical to SEQ ID NO:7.

[0087] In an embodiment of a method for treating a disorder associated with pDC, the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:4 or a sequence at least about 90% identical to SEQ ID NO:4, and the light chain variable region comprises the amino acid sequence given by SEQ ID NO:7 or a sequence at least about 90% identical to SEQ ID NO:7.

[0088] In an embodiment of a method for treating a disorder associated with pDC, the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:2 or a sequence at least about 90% identical to SEQ ID NO:2, and the light chain variable region comprises the amino acid sequence given by SEQ ID NO:8 or a sequence at least about 90% identical to SEQ ID NO:8.

[0089] In an embodiment of a method for treating a disorder associated with pDC, the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:3 or a sequence at least about 90% identical to SEQ ID NO:3, and the light chain variable region comprises the amino acid sequence given by SEQ ID NO:8 or a sequence at least about 90% identical to SEQ ID NO:8.

[0090] In an embodiment of a method for treating a disorder associated with pDC, the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:4 or a sequence at least about 90% identical to SEQ ID NO:4, and the light chain variable region comprises the amino acid sequence given by SEQ ID NO:8 or a sequence at least about 90% identical to SEQ ID NO:8.

[0091] In an embodiment of a method for treating a disorder associated with pDC, the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:2 or a sequence at least about 90% identical to SEQ ID NO:2, and the light chain variable region comprises the amino acid sequence given by SEQ ID NO:9 or a sequence at least about 90% identical to SEQ ID NO:9.

[0092] In an embodiment of a method for treating a disorder associated with pDC, the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:3 or a sequence at least about 90% identical to SEQ ID NO:3, and the light chain variable region comprises the amino acid sequence given by SEQ ID NO:9 or a sequence at least about 90% identical to SEQ ID NO:9.

[0093] In an embodiment of a method for treating a disorder associated with pDC, the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:4 or a sequence at least about 90% identical to SEQ ID NO:4, and the light chain variable region comprises the amino acid sequence given by SEQ ID NO:9 or a sequence at least about 90% identical to SEQ ID NO:9.

[0094] In an embodiment of a method for treating a disorder associated with pDC, the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:2 or a sequence at least about 90% identical to SEQ ID NO:2, and the light chain variable region comprises the amino acid sequence given by SEQ ID NO:10 or a sequence at least about 90% identical to SEQ ID NO:10.

[0095] In an embodiment of a method for treating a disorder associated with pDC, the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:3 or a sequence at least about 90% identical to SEQ ID NO:3, and the light chain variable region comprises the amino acid sequence given by SEQ ID NO:10 or a sequence at least about 90% identical to SEQ ID NO:10.

[0096] In an embodiment of a method for treating a disorder associated with pDC, the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:4 or a sequence at least about 90% identical to SEQ ID NO:4, and the light chain variable region comprises the amino acid sequence given by SEQ ID NO:10 or a sequence at least about 90% identical to SEQ ID NO:10.

[0097] In some embodiments, a kit for depleting, inhibiting, or depleting and inhibiting plasmacytoid dendritic cells (pDCs) in a patient in need thereof comprises instructions and an antibody or fragment thereof, wherein the antibody or fragment thereof comprises a heavy chain variable region comprising an amino acid sequence selected from the group consisting of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, and a sequence at least about 90% identical to SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, or SEQ ID NO:4, and a light chain variable region comprising an amino acid sequence selected from the group consisting of SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, and a sequence at least about 90% identical to SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, or SEQ ID NO:10.

[0098] In some embodiments, the pharmaceutical composition comprises at least one excipient and an antibody or fragment thereof, wherein the antibody or fragment thereof comprises a heavy chain variable region comprising an amino acid sequence selected from the group consisting of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, and a sequence at least about 90% identical to SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, or SEQ ID NO:4, and a light chain variable region comprising an amino acid sequence selected from the group consisting of SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, and a sequence at least about 90% identical to SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, or SEQ ID NO:10.

[0099] In embodiments, antibodies or fragments thereof may be used in therapy.

[0100] In embodiments, the antibodies or fragments thereof may be used to deplete, inhibit, or deplete and inhibit plasmacytoid dendritic cells (pDCs).

[0101] In embodiments, the antibody or fragment thereof may be used for the manufacture of a medicament.

[0102] This summary is not intended to identify key features or essential features of the claimed subject matter, nor is it intended to be used as an aid in limiting the scope of the claimed subject matter. Further embodiments, forms, features, and aspects of the present disclosure will become apparent from the description and figures provided herewith.

[0103] The teachings of several embodiments of the present disclosure may be better understood by reference to the following description taken in conjunction with the accompanying drawings. [Brief explanation of the drawings]

[0104] [Figure 1] FIG. 1 shows a schematic diagram of the expression vectors used to produce chimeric and humanized anti-BDCA2 antibodies according to several embodiments. [Figure 2]FIG. 2 shows an SDS-PAGE analysis of NPB-303 according to several embodiments. [Figure 3] FIG. 3 shows protein analysis of NPB-303 on a Superose 6 column using an AKTA FPLC system according to multiple embodiments; FIG. 3A. BioRad gel filtration standards; B. NPB-303. [Figure 4] FIG. 4 shows the biophysical binding of 15b, 12b, and NPB-303 to the BDCA2 antigen from either monkey (Panel A) or human (Panel B) according to multiple embodiments. [Figure 5] FIG. 5 shows a structural model of the BDCA2 extracellular domain according to several embodiments; green: BDCA2 extracellular domain; yellow: NPB-303 epitope and orange: BDCA2 lectin (ligand) binding residues. [Figure 6] FIG. 6 shows a structural model of the NPB-303 variable (binding) domain according to multiple embodiments; green: VH framework; yellow: VH CDRs; blue: VL framework; brown: VL CDRs and red: aspartic acid isomerization motif. [Figure 7] FIG. 7 shows binding characterization of NPB-303 antibody by surface plasmon resonance (SPR) according to several embodiments. [Figure 8] FIG. 8 shows the binding characterization of NPB-303 mAb to human BDCA2 antigen in vitro according to several embodiments. [Figure 9] FIG. 9 shows the binding characterization of NPB-303 mAb to human pDCs in vitro according to multiple embodiments. [Figure 10] FIG. 10 shows a FACS plot showing that NPB-303 inhibits pDC IFNα responses to CpG-A stimulation in vitro, according to embodiments. [Figure 11] FIG. 11 shows a dose response curve of NPB-303 to IFNα according to several embodiments. [Figure 12] FIG. 12 shows the IC50 of NPB-303 in multiple human donor PBMC samples according to multiple embodiments. [Figure 13] FIG. 13 shows an in vitro efficacy analysis of NPB-303 according to embodiments. [Figure 14] FIG. 14 shows the pharmacokinetic profile of NPB-303 in genetically modified mice according to embodiments. [Figure 15] FIG. 15 shows FACS plots demonstrating pDC depletion and BDCA2 downregulation by NPB-303 in the blood, according to embodiments. [Figure 16] FIG. 16 shows summary data demonstrating pDC depletion (% CD4+CD123+ in Lin-) and BDCA2 downregulation on pDC (% CD4+BDCA2+ in Lin-) according to multiple embodiments. [Figure 17] FIG. 17 shows the pharmacodynamic profile of NPB-303 mAb in humanized mice in vivo according to embodiments. [Figure 18] FIG. 18 shows a summary of % pDC or % BDCA2+ cells from humanized mice treated with either isotype control or NPB-303 (normalized to isotype control antibody), according to multiple embodiments. [Figure 19] FIG. 19 shows a summarized analysis of human BDCA2+ pDCs among splenocytes of humanized mice according to embodiments. [Figure 20] FIG. 20 shows a summarized analysis of human CD123+ pDCs among splenocytes of humanized mice according to embodiments. [Figure 21] FIG. 21 shows a summary of other cell types (Lin-CD4-, CD4+CD123-, and CD4-CD123-) in the blood of humanized mice treated with isotype control mAb or NPB-303 according to embodiments. [Figure 22] FIG. 22 shows endotoxin in NPB-303 (1:1000 dilution) assay performed using kinetic turbidimetric and kinetic chromogenic assay (ENDOSAFE). [Figure 23]Figure 23 shows a summary of the anti-tumor activity of NPB-303 or KEYTRUDA® in mice with human immunity and tumor cells (“HIT mice”) according to embodiments. [Figure 24] FIG. 24 shows a summary of the body weights of tumor-bearing mice from various treatment groups according to embodiments. [Figure 25] FIG. 25 summarizes that pDCs were depleted in the spleen by NPB-303, but not by KEYTRUDA®, in tumor-bearing HIT mice according to multiple embodiments. [Figure 26] FIG. 26 summarizes that pDCs were depleted in tumors by NPB-303, but not by KEYTRUDA®, in tumor-bearing HIT mice, according to multiple embodiments. [Figure 27] FIG. 27 summarizes that, according to embodiments, BDCA2 was downregulated in the spleen by NPB-303, but not by KEYTRUDA®, in tumor-bearing HIT mice. [Figure 28] FIG. 28 summarizes that embodiments show that BDCA2 was downregulated in tumors by NPB-303, but not by KEYTRUDA®, in tumor-bearing HIT mice. [Figure 29] FIG. 29 shows a schematic diagram of a protocol for the establishment of a tumor-bearing humanized mouse model according to embodiments. [Figure 30] FIG. 30 shows a representative flow plot of human pDCs in tumors isolated from HIT-TNBC mice, according to embodiments. [Figure 31] FIG. 31 shows a summary of the percentage of human pDCs in tumors according to multiple embodiments. [Figure 32] FIG. 32 shows a representative flow plot of tumor versus blood human T cells isolated from HIT-TNBC mice, according to embodiments. [Figure 33] FIG. 33 shows mean fluorescence intensity (MFI) of Tim3 and PD1 on CD4+ T cells according to embodiments. [Figure 34] FIG. 34 shows the mean fluorescence intensity (MFI) of Tim3 and PD1 on CD8+ T cells according to embodiments. [Figure 35] FIG. 35 shows dose-dependent anti-tumor activity (by tumor volume measurement) by NPB-303 according to embodiments. [Figure 36] FIG. 36 shows stable body weight over time in HIT mice treated with NPB-303 according to embodiments. [Figure 37] FIG. 37 shows representative flow plots of Lin− and a summary of %CD4+CD123+ in the spleen according to embodiments. [Figure 38] FIG. 38 shows representative flow plots of Lin- and a summary of %CD4+CD123+ in tumors according to multiple embodiments. [Figure 39] FIG. 39 shows tumor volume over time for a second human TNBC cell line (MDA-MB-468) according to embodiments. [Figure 40] FIG. 40 shows dose-dependent anti-tumor activity (by tumor volume measurement) by NPB-303 according to embodiments. [Figure 41] FIG. 41 shows stable body weight over time in HIT mice treated with NPB-303 according to embodiments. [Figure 42] FIG. 42 shows a study protocol for combination therapy of NPB-303 and Abraxane® according to embodiments. [Figure 43] Figure 43 shows the anti-tumor activity (by tumor volume measurement) of NPB-303 alone, Abraxane® alone, and in combination therapy according to multiple embodiments. [Figure 44] FIG. 44 shows stable body weight over time in HIT mice treated with NPB-303 alone, Abraxane® alone, and combination therapy, according to embodiments. [Figure 45] Figure 45 shows that, according to embodiments, PD1 was downregulated in tumor-infiltrating CD8+ cells in both the NPB-303 and combination therapy groups compared to the isotype control group. [Figure 46] FIG. 46 shows that, according to embodiments, Tim3 was downregulated in tumor-infiltrating CD8+ cells in both the NPB-303 and combination therapy groups compared to the isotype control group. [Figure 47] Figure 47 shows representative liver images from each treatment group (isotype control group, NPB-303 alone, Abraxane® alone, and combination therapy) demonstrating a reduction in tumor nodules in groups of mice treated with NPB-303 alone or in combination with Abraxane®, according to embodiments. [Figure 48] Figure 48 shows a summary of liver infiltration scores for each treatment group (isotype control, NPB-303 alone, Abraxane® alone, and combination therapy) according to multiple embodiments. [Figure 49] Figure 49 shows representative H&E staining data of histological sections (100 microns) of liver tissue from each treatment group (isotype control, NPB-303 alone, Abraxane® alone, and combination therapy) according to multiple embodiments. [Figure 50] FIG. 50 shows representative H&E stained images (3x and 17x) of lungs according to several embodiments. [Figure 51] Figure 51 shows a summary of lung infiltration scores for each treatment group (isotype control, NPB-303 alone, Abraxane® alone, and combination therapy) according to multiple embodiments. [Figure 52] FIG. 52 shows the thermal denaturation (Tm) and aggregation onset (Tagg) of NPB-303 compared to therapeutic antibodies that have progressed to at least clinical trials according to multiple embodiments. [Figure 53] FIG. 53 shows the binding activity of NPB-303 under various storage temperature conditions according to several embodiments. [Figure 54] FIG. 54 shows detailed sequence modifications of the NPB-303 antibody according to multiple embodiments. [Figure 55] FIG. 55 shows sensorgrams of SPR-based affinity studies for 24 antibodies according to multiple embodiments. [Figure 56]FIG. 56 shows a series of graphs of an IFNα inhibition assay according to several embodiments. [Figure 57] Figure 57 shows a summary of antibody potency (IC50) according to several embodiments. [Figure 58] FIG. 58 shows sensorgrams of SPR-based affinity studies for three antibodies according to embodiments. [Figure 59] Figure 59 shows a panel of graphs demonstrating dose-dependent inhibition of TLR-9-induced IFNα by PBMCs from two healthy human donors using NPB-303 and three additional monoclonal antibodies according to embodiments. [Figure 60] Figure 60 shows a summary of antibody potency (IC50) for IFNα inhibition according to several embodiments. [Figure 61] Figure 61 shows a representative panel of FACS plots of human pDC (CD4+CD123+) and CD4+BDCA2+ cells (gated on human CD45+, then Lin− cells) in blood from humanized mice treated with an isotype control antibody, NPB-303, and three additional antibodies, according to multiple embodiments. [Figure 62] FIG. 62 shows a summary of % pDC of Lin − cells in blood from humanized mice treated with an isotype control antibody, NPB-303, and three additional antibodies according to embodiments. [Figure 63] FIG. 63 shows a summary of %BDCA2+ of Lin− cells in blood from humanized mice treated with an isotype control antibody, NPB-303, and three additional antibodies according to embodiments. [Figure 64] Figure 64 shows a representative panel of FACS plots of human pDC (CD4+CD123+) and CD4+BDCA2+ cells (gated on human CD45+, then Lin− cells) in spleens from humanized mice treated with an isotype control antibody, NPB-303, and three additional antibodies, according to multiple embodiments. [Figure 65]FIG. 65 shows a summary of % pDC of Lin- cells in spleens from humanized mice treated with an isotype control antibody, NPB-303, and three additional antibodies according to embodiments. [Figure 66] FIG. 66 shows a summary of %BDCA2+ of Lin- cells in spleens from humanized mice treated with an isotype control antibody, NPB-303, and three additional antibodies according to embodiments. [Figure 67] Figure 67 shows a representative panel of FACS plots of myeloid cells (Lin-CD4-), conventional DCs (CD4+CD123-), and other cell types (Lin-CD4-CD123-) in the blood from humanized mice treated with an isotype control antibody, NPB-303, and three additional antibodies, according to multiple embodiments. [Figure 68] FIG. 68 shows a summary of % Lin-CD4-, CD4+CD123-, and Lin-CD4-CD123- cells in blood from humanized mice treated with an isotype control antibody, NPB-303, and three additional antibodies according to embodiments. [Figure 69] FIG. 69 shows a summary of % Lin-CD4-, CD4+CD123-, and Lin-CD4-CD123- cells in spleens from humanized mice treated with an isotype control antibody, NPB-303, and three additional antibodies according to embodiments. DETAILED DESCRIPTION OF THE INVENTION

[0105] The embodiments described below are not intended to be exhaustive or to limit the invention to the precise forms disclosed in the following detailed description, but rather are chosen and described so that others skilled in the art can appreciate and understand the principles and practices of the present disclosure.

[0106] It is specifically contemplated that the various features of the invention described herein may be used in any combination, unless the context dictates otherwise. Furthermore, the present invention also contemplates that in some embodiments of the invention, any feature or combination of features described herein may be excluded or omitted. To illustrate, if the specification states that a composite comprises components A, B, and C, it is specifically contemplated that any or combination of A, B, or C, alone or in any combination, may be omitted and eliminated.

[0107] Amino acids are referred to herein in the style recommended by the IUPAC-IUB Biochemical Nomenclature Commission, or by either the one-letter or three-letter codes, in accordance with both 37 CFR § 1.822 and established usage.

[0108] The present disclosure provides a method for detecting a human leukemia virus (HV) comprising administering to a mammalian subject the present invention a method for detecting a human leukemia virus (HV) virus ... The present invention provides an antibody or fragment thereof comprising:

[0109] As used herein and in the appended claims, the singular forms "a," "an," and "the" are intended to include the plural forms unless the context clearly dictates otherwise. Also, as used herein, "and / or" refers to and includes any and all possible combinations of one or more of the associated listed items, as well as the lack of combinations when interpreted in the alternative ("or"). The term "about," as used herein when referring to a measurable value, such as a polypeptide amount, dosage, time, temperature, enzyme activity, or other biological activity, is meant to encompass variations of ±20%, ±10%, ±5%, ±1%, ±0.5%, or even ±0.1% of the specified amount.

[0110] The transitional phrase "consisting essentially of" means that the scope of a claim should be construed to include the specified materials or steps recited in the claim and "materials or steps that do not materially affect the basic and novel characteristic(s)" of the claimed subject matter. See In re Herz, 537 F.2d 549,551-52,190 USPQ 461,463 (CCPA 1976) (emphasis in original). See also MPEP § 2111.03 (9th ed., 10th rev.). The term "essentially consisting" (and grammatical variations), when applied to a polypeptide sequence, means a polypeptide consisting of both the recited sequence (e.g., SEQ ID NO:) and a total of 10 or fewer (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10) additional amino acids at the N-terminus and / or C-terminus of the recited sequence, such that the function of the polypeptide is not substantially altered. The additional amino acids include the total number of additional amino acids at both ends added together. The term "substantially altered" when applied to a polypeptide refers to an increase or decrease in epitope binding activity of at least about 50% or more compared to the activity of a polypeptide consisting of the recited sequence.

[0111] The term "isolated" can refer to a nucleic acid, nucleotide sequence, or polypeptide that is substantially free of cellular material, viral material, and / or culture medium (if produced by recombinant DNA techniques), or chemical precursors or other chemicals (if chemically synthesized). Furthermore, an "isolated fragment" is a fragment of a nucleic acid, nucleotide sequence, or polypeptide that is not naturally occurring as a fragment and would not be found in the natural state. "Isolated" does not imply that the preparation is technically pure (homogeneous), but rather that it is sufficiently pure to provide the polypeptide or nucleic acid in a form that can be used for its intended purpose.

[0112] When applied to a polypeptide, the term "fragment" may be understood to mean an amino acid sequence that is shortened in length relative to a reference polypeptide or amino acid sequence and that comprises, consists essentially of, and / or consists of an amino acid sequence of contiguous amino acids that is identical or nearly identical (e.g., about 90%, about 92%, about 95%, about 98%, about 99% identical) to the reference polypeptide or amino acid sequence. Such polypeptide fragments according to the present disclosure may, where appropriate, be included within a larger polypeptide of which they are a component. In some embodiments, such fragments may comprise, consist essentially of, and / or consist of peptides having a length of at least about 4, 6, 8, 10, 12, 15, 20, 25, 30, 35, 40, 45, 50, 75, 100, 150, 200, or more contiguous amino acids of the polypeptide or amino acid sequence.

[0113] As used herein, the terms "protein" and "polypeptide" are used interchangeably and include both peptides and proteins, unless otherwise indicated.

[0114] As used herein, a "functional polypeptide" or "functional fragment" substantially retains at least one biological activity normally associated with the polypeptide (e.g., target protein binding). In certain embodiments, a "functional polypeptide" or "functional fragment" substantially retains all of the activities possessed by the unmodified peptide. "Substantially retaining" biological activity means that the polypeptide retains at least about 20%, 30%, 40%, 50%, 60%, 75%, 85%, 90%, 95%, 97%, 98%, 99% or more of the biological activity of the native polypeptide (and may even have a higher level of activity than the native polypeptide). A "non-functional polypeptide" is one that exhibits little or essentially no detectable biological activity normally associated with the polypeptide (e.g., at most, only a minor amount, e.g., less than about 10% or even less than about 5%). Biological activity, such as protein binding, can be measured using assays known to those skilled in the art and described herein.

[0115] Disclosed herein are antibodies that specifically bind to an epitope comprising blood dendritic cell antigen-2 (BDCA2) and deplete, inhibit, or deplete and inhibit pDCs. Such antibodies can be used, for example, for research or therapeutic purposes, to deplete, inhibit, or deplete and inhibit pDCs in patients / animals. Such antibodies can be used to treat disorders associated with pDCs. Thus, one embodiment relates to an antibody or fragment thereof that specifically binds to BDCA2 and depletes, inhibits, or depletes and inhibits pDCs when administered to a patient / animal.

[0116] As used herein, the term "antibody" or "antibodies" refers to all types of immunoglobulins, including IgG, IgM, IgA, IgD, and IgE. Antibodies can be monoclonal or polyclonal and can be of any species of origin, including (for example) mouse, rat, rabbit, horse, goat, sheep, camel, or human, or can be chimeric. See, e.g., Walker et al., Molec. Immunol. 26:403 (1989). Antibodies can be recombinant monoclonal antibodies produced according to the methods disclosed in U.S. Pat. Nos. 4,474,893 or 4,816,567. Antibodies can also be chemically constructed according to the methods disclosed in U.S. Pat. No. 4,676,980.

[0117] Antibody fragments include, for example, Fab, Fab', F(ab')2, and Fv fragments; domain antibodies, diabodies; vaccibodies, linear antibodies; single-chain antibody molecules; and multispecific antibodies formed from antibody fragments. Such fragments can be produced by known techniques. For example, F(ab')2 fragments can be produced by pepsin digestion of antibody molecules, and Fab fragments can be generated by reducing the disulfide bridges of F(ab')2 fragments. Alternatively, Fab expression libraries can be constructed to allow rapid and easy identification of monoclonal Fab fragments with the desired specificity (Huse et al., Science 254:1275 (1989)).

[0118] Antibodies can be modified or mutated for compatibility with species other than the one in which they were produced. For example, antibodies can be humanized or camelized. If the patient is human, the antibody can be humanized. See, for example, Safdari, et al., Antibody humanization methods—a review and update. Biotechnol. Genet. Eng. Rev., 2013;29:175-86. Humanized forms of non-human (e.g., murine) antibodies are chimeric immunoglobulins, immunoglobulin chains, or fragments thereof (such as Fv, Fab, Fab', F(ab')2, or other antigen-binding subsequences of antibodies) that contain minimal sequence derived from non-human immunoglobulin. Humanized antibodies include human immunoglobulins (recipient antibodies) in which residues from the recipient's complementarity-determining regions (CDRs) are replaced by residues from the CDRs of a non-human species (donor antibody), such as mouse, rat, or rabbit, that have the desired specificity, affinity, and capacity. In some embodiments, Fv framework residues of the human immunoglobulin are replaced by corresponding non-human residues. Humanized antibodies may also comprise residues that are found neither in the recipient antibody nor in the imported CDR or framework sequences. Generally, 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 framework (FR) regions (i.e., the sequences between the CDR regions) are of human immunoglobulin consensus sequences. A humanized antibody may also comprise at least a portion of an immunoglobulin constant region (Fc), typically that of a human immunoglobulin (Jones et al., Nature 321:522 (1986); Riechmann et al., Nature, 332:323 (1988); and Presta, Curr. Op. Struct. Biol. 2:593 (1992)).

[0119] As mentioned above, methods for humanizing non-human antibodies are known. Generally, humanized antibodies have one or more amino acid residues introduced into them from a non-human source. These non-human amino acid residues are often referred to as "import" residues and are typically taken from an "import" variable domain. Humanization can essentially be performed by substituting rodent CDRs or CDR sequences for the corresponding sequences of a human antibody according to the method of Winter and coworkers (Jones et al., Nature 321:522 (1986); Riechmann et al., Nature 332:323 (1988); Verhoeyen et al., Science 239:1534 (1988)). Thus, such "humanized" antibodies are chimeric antibodies (U.S. Patent No. 4,816,567) in which substantially less than a complete human variable domain is substituted by the corresponding sequence from a non-human species. In practice, humanized antibodies may be human antibodies in which some CDR residues (e.g., all or part of the CDRs) and possibly some FR residues are substituted by residues from analogous sites in rodent (e.g., murine) antibodies.

[0120] Human antibodies can be produced using a variety of techniques known in the art, including phage display libraries (Hoogenboom and Winter, J. Mol. Biol. 227:381 (1991); Marks et al., J. Mol. Biol. 222:581 (1991)). The techniques of Cole et al. and Boerner et al. are also available for the preparation of human monoclonal antibodies (Cole et al., Monoclonal Antibodies and Cancer Therapy, Alan R. Liss, Inc.: New York, p. 77 (1985) and Boerner et al., J. Immunol. 147:86 (1991)). Similarly, human antibodies can be made by introducing human immunoglobulin loci into transgenic animals, e.g., mice, in which the endogenous immunoglobulin genes have been partially or completely inactivated. Upon antigenic challenge, human antibody production is observed, which closely resembles that seen in humans in all respects, including gene rearrangement, assembly, and antibody repertoire. This approach is described, for example, in U.S. Patent Nos. 5,545,807; 5,545,806; 5,569,825; 5,625,126; 5,633,425; 5,661,016, and the following scientific publications: Marks et al., Bio / Technology 10:779 (1992); Lonberg et al., Nature 368:856 (1994); Morrison, Nature 368:812 (1994); Fishwild et al., Nature Biotechnol. 14:845 (1996); Neuberger, Nature Biotechnol. 14:826 (1996); and Lonberg and Huszar, Intern. Rev. Immunol. 13:65 (1995).

[0121] Polyclonal antibodies used to practice the present disclosure can be produced, according to known procedures, by immunizing a suitable animal (e.g., rabbit, goat, etc.) with an antigen to which a monoclonal antibody against the target of interest binds, collecting immune serum from the animal, and isolating polyclonal antibodies from the immune serum. The polynucleotide and polypeptide sequences of BDCA2 are known and can be found in sequence databases such as GenBank. Exemplary sequences include the human BDCA2 polypeptide sequence (Accession No. Q8WTTO) and polynucleotide sequence (Accession No. AF293615), which are incorporated herein by reference in their entireties.

[0122] Monoclonal antibodies can be produced in hybridoma cell lines according to the technique of Kohler and Milstein, Nature 265:495 (1975). For example, a solution containing an appropriate antigen can be injected into a mouse, and after a sufficient time, the mouse can be sacrificed and spleen cells can be obtained. The spleen cells can then be immortalized by fusing them with myeloma cells or lymphoma cells, typically in the presence of polyethylene glycol, to produce hybridoma cells. The hybridoma cells are then grown in an appropriate medium, and the supernatant is screened for monoclonal antibodies with the desired specificity. Monoclonal Fab fragments can be produced in E. coli by recombinant techniques known to those skilled in the art. See, for example, Huse, Science 246:1275 (1989).

[0123] Antibodies specific to a target polypeptide can also be obtained by phage display techniques known in the art.

[0124] In some embodiments, the antibody or fragment thereof formed by the above method has an affinity (K d ) can bind the epitope having SEQ ID NO: 11. SEQ ID NO: 11 is a fragment of SEQ ID NO: 12, which provides the amino acid sequence of the epitope of the BDCA2 antigen. In some embodiments, the affinity (Kd ) is 0.1nM~90nM, 0.1nM~80nM, 0.1nM~70nM, 0.1nM~60nM, 0.1nM~50nM, 0.1nM~40nM, 0.1nM~30nM, 0.1nM~20nM, 0.1nM~10nM, 0.1nM~9nM, 0.1nM~8nM, 0.1nM~7nM, 0.1nM~6nM, 0.1n M~5nM, 0.1nM~4nM, 0.1nM~3nM, 0.1nM~2nM, 0.1nM~1nM, 0.1nM~0.9nM, 0.1nM~0.8nM, 0.1 nM~0.7nM, 0.1nM~0.6nM, 0.1nM~0.5nM, 0.1nM~0.4nM, 0.1nM~0.3nM, 0.1nM~0.2nM, 0.2nM ~100nM, 0.3nM~100nM, 0.4nM~100nM, 0.5nM~100nM, 0.6nM~100nM, 0.7nM~100nM, 0.8nM~ 100nM, 0.9nM~100nM, 1nM~100nM, 2nM~100nM, 3nM~100nM, 4nM~100nM, 5nM~100nM, 6nM~10 It may be 0 nM, 7 nM to 100 nM, 8 nM to 100 nM, 9 nM to 100 nM, 10 nM to 100 nM, 20 nM to 100 nM, 30 nM to 100 nM, 40 nM to 100 nM, 50 nM to 100 nM, 60 nM to 100 nM, 70 nM to 100 nM, 80 nM to 100 nM, or even 90 nM to 100 nM.

[0125] As used herein, the term "affinity" refers to the degree or strength of binding of an antibody to an epitope. Affinity is determined by the equilibrium dissociation constant (K D or K d ), apparent equilibrium dissociation constant (K D ' or K d ') and IC 50 Affinity can be measured and / or expressed in a number of ways known in the art, including, but not limited to, affinity (the amount required to produce 50% inhibition in a competitive assay). Affinity is understood to be the average affinity for a given population of antibodies that bind to an epitope. A variety of immunoassays can be used for screening to identify antibodies with the desired affinity for the peptide sequence given by SEQ ID NO: 11.

[0126] The antibody can be conjugated to a solid support (e.g., a bead, plate, slide, or well formed from a material such as latex or polystyrene) in accordance with known techniques. The antibody can also be radiolabeled (e.g., 35 S, 125 S, 131 I), enzyme labels (e.g., horseradish peroxidase, alkaline phosphatase), and fluorescent labels (e.g., fluorescein). Determination of the formation of antibody / antigen complexes in the methods disclosed herein can be by detection of, for example, precipitation, agglutination, flocculation, radioactivity, color development or change, fluorescence, luminescence, and the like.

[0127] In some embodiments, the CDRs of an antibody may comprise at least one heavy chain variable region and at least one light chain variable region.

[0128] In some embodiments, the antibody or fragment thereof comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 1, or a sequence at least about 90% identical thereto, e.g., a sequence at least about 95, 96, 97, 98, or 99% identical thereto. In some embodiments, the antibody or fragment thereof comprises a heavy chain variable region comprising at least 50 contiguous amino acids, e.g., at least about 100, 150, or 200 or more contiguous amino acids, of the amino acid sequence of SEQ ID NO: 1, or a sequence at least about 90% identical thereto.

[0129] In some embodiments, the antibody or fragment thereof comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 2, or a sequence at least about 90% identical thereto, e.g., a sequence at least about 95, 96, 97, 98, or 99% identical thereto. In some embodiments, the antibody or fragment thereof comprises a heavy chain variable region comprising at least about 50 contiguous amino acids, e.g., at least about 100, 150, or 200 or more contiguous amino acids, of the amino acid sequence of SEQ ID NO: 2, or a sequence at least about 90% identical thereto.

[0130] In some embodiments, the antibody or fragment thereof comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 3, or a sequence at least about 90% identical thereto, e.g., a sequence at least about 95, 96, 97, 98, or 99% identical thereto. In some embodiments, the antibody or fragment thereof comprises a heavy chain variable region comprising at least about 50 contiguous amino acids, e.g., at least about 100, 150, or 200 or more contiguous amino acids, of the amino acid sequence of SEQ ID NO: 3, or a sequence at least about 90% identical thereto.

[0131] In some embodiments, the antibody or fragment thereof comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 4, or a sequence at least about 90% identical thereto, e.g., a sequence at least about 95, 96, 97, 98, or 99% identical thereto. In some embodiments, the antibody or fragment thereof comprises a heavy chain variable region comprising at least about 50 contiguous amino acids, e.g., at least about 100, 150, or 200 or more contiguous amino acids, of the amino acid sequence of SEQ ID NO: 4, or a sequence at least about 90% identical thereto.

[0132] In some embodiments, the antibody or fragment thereof comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 5, or a sequence at least about 90% identical thereto, e.g., a sequence at least about 95, 96, 97, 98, or 99% identical thereto. In some embodiments, the antibody or fragment thereof comprises a light chain variable region comprising at least about 50 contiguous amino acids, e.g., at least about 100, 150, or 200 or more contiguous amino acids, of the amino acid sequence of SEQ ID NO: 5, or a sequence at least about 90% identical thereto.

[0133] In some embodiments, the antibody or fragment thereof comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 6, or a sequence at least about 90% identical thereto, e.g., a sequence at least about 95, 96, 97, 98, or 99% identical thereto. In some embodiments, the antibody or fragment thereof comprises a light chain variable region comprising at least about 50 contiguous amino acids, e.g., at least about 100, 150, or 200 or more contiguous amino acids, of the amino acid sequence of SEQ ID NO: 6, or a sequence at least about 90% identical thereto.

[0134] In some embodiments, the antibody or fragment thereof comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 7, or a sequence at least about 90% identical thereto, e.g., a sequence at least about 95, 96, 97, 98, or 99% identical thereto. In some embodiments, the antibody or fragment thereof comprises a light chain variable region comprising at least about 50 contiguous amino acids, e.g., at least about 100, 150, or 200 or more contiguous amino acids, of the amino acid sequence of SEQ ID NO: 7, or a sequence at least about 90% identical thereto.

[0135] In some embodiments, the antibody or fragment thereof comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 8, or a sequence at least about 90% identical thereto, e.g., a sequence at least about 95, 96, 97, 98, or 99% identical thereto. In some embodiments, the antibody or fragment thereof comprises a light chain variable region comprising at least about 50 contiguous amino acids, e.g., at least about 100, 150, or 200 or more contiguous amino acids, of the amino acid sequence of SEQ ID NO: 8, or a sequence at least about 90% identical thereto.

[0136] In some embodiments, the antibody or fragment thereof comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 9, or a sequence at least about 90% identical thereto, e.g., a sequence at least about 95, 96, 97, 98, or 99% identical thereto. In some embodiments, the antibody or fragment thereof comprises a light chain variable region comprising at least about 50 contiguous amino acids, e.g., at least about 100, 150, or 200 or more contiguous amino acids, of the amino acid sequence of SEQ ID NO: 9, or a sequence at least about 90% identical thereto.

[0137] In some embodiments, the antibody or fragment thereof comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 10, or a sequence at least about 90% identical thereto, e.g., a sequence at least about 95, 96, 97, 98, or 99% identical thereto. In some embodiments, the antibody or fragment thereof comprises a light chain variable region comprising at least about 50 contiguous amino acids, e.g., at least about 100, 150, or 200 or more contiguous amino acids, of the amino acid sequence of SEQ ID NO: 10, or a sequence at least about 90% identical thereto.

[0138] In some embodiments, the antibody or fragment thereof comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 1, or a sequence at least about 90% identical thereto, e.g., a sequence at least about 95, 96, 97, 98, or 99% identical thereto. Additionally, the antibody or fragment thereof comprises a light chain variable region comprising an amino acid sequence selected from the group consisting of SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, and a sequence at least about 90% identical to SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, or SEQ ID NO:10, e.g., a sequence at least about 95, 96, 97, 98, or 99% identical to SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, or SEQ ID NO:10.

[0139] In some embodiments, the antibody or fragment thereof comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:2, or a sequence at least about 90% identical thereto, e.g., a sequence at least about 95, 96, 97, 98, or 99% identical thereto. Additionally, the antibody or fragment thereof comprises a light chain variable region comprising an amino acid sequence selected from the group consisting of SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, and a sequence at least about 90% identical to SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, or SEQ ID NO:10, e.g., a sequence at least about 95, 96, 97, 98, or 99% identical to SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, or SEQ ID NO:10.

[0140] In some embodiments, the antibody or fragment thereof comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 3, or a sequence at least about 90% identical thereto, e.g., a sequence at least about 95, 96, 97, 98, or 99% identical thereto. Additionally, the antibody or fragment thereof comprises a light chain variable region comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, and a sequence at least about 90% identical to SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, or SEQ ID NO: 10, e.g., a sequence at least about 95, 96, 97, 98, or 99% identical to SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, or SEQ ID NO: 10.

[0141] In some embodiments, the antibody or fragment thereof comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:4, or a sequence at least about 90% identical thereto, e.g., a sequence at least about 95, 96, 97, 98, or 99% identical thereto. Additionally, the antibody or fragment thereof comprises a light chain variable region comprising an amino acid sequence selected from the group consisting of SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, and a sequence at least about 90% identical to SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, or SEQ ID NO:10, e.g., a sequence at least about 95, 96, 97, 98, or 99% identical to SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, or SEQ ID NO:10.

[0142] In some embodiments, the antibody or fragment thereof comprises a heavy chain variable region comprising an amino acid sequence selected from the group consisting of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, and a sequence at least about 90% identical to SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, or SEQ ID NO:4, e.g., a sequence at least about 95, 96, 97, 98, or 99% identical to SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, or SEQ ID NO:4. Additionally, the antibody or fragment thereof comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO:5, or a sequence at least about 90% identical thereto, e.g., a sequence at least about 95, 96, 97, 98, or 99% identical thereto.

[0143] In some embodiments, the antibody or fragment thereof comprises a heavy chain variable region comprising an amino acid sequence selected from the group consisting of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, and a sequence at least about 90% identical to SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, or SEQ ID NO:4, e.g., a sequence at least about 95, 96, 97, 98, or 99% identical to SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, or SEQ ID NO:4. Additionally, the antibody or fragment thereof comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO:6, or a sequence at least about 90% identical thereto, e.g., a sequence at least about 95, 96, 97, 98, or 99% identical thereto.

[0144] In some embodiments, the antibody or fragment thereof comprises a heavy chain variable region comprising an amino acid sequence selected from the group consisting of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, and a sequence at least about 90% identical to SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, or SEQ ID NO:4, e.g., a sequence at least about 95, 96, 97, 98, or 99% identical to SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, or SEQ ID NO:4. Additionally, the antibody or fragment thereof comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO:7, or a sequence at least about 90% identical thereto, e.g., a sequence at least about 95, 96, 97, 98, or 99% identical thereto.

[0145] In some embodiments, the antibody or fragment thereof comprises a heavy chain variable region comprising an amino acid sequence selected from the group consisting of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, and a sequence at least about 90% identical to SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, or SEQ ID NO:4, e.g., a sequence at least about 95, 96, 97, 98, or 99% identical to SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, or SEQ ID NO:4. Additionally, the antibody or fragment thereof comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO:8, or a sequence at least about 90% identical thereto, e.g., a sequence at least about 95, 96, 97, 98, or 99% identical thereto.

[0146] In some embodiments, the antibody or fragment thereof comprises a heavy chain variable region comprising an amino acid sequence selected from the group consisting of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, and a sequence at least about 90% identical to SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, or SEQ ID NO:4, e.g., a sequence at least about 95, 96, 97, 98, or 99% identical to SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, or SEQ ID NO:4. Additionally, the antibody or fragment thereof comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO:9, or a sequence at least about 90% identical thereto, e.g., a sequence at least about 95, 96, 97, 98, or 99% identical thereto.

[0147] In some embodiments, the antibody or fragment thereof comprises a heavy chain variable region comprising an amino acid sequence selected from the group consisting of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, and a sequence at least about 90% identical to SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, or SEQ ID NO:4, e.g., a sequence at least about 95, 96, 97, 98, or 99% identical to SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, or SEQ ID NO:4. Additionally, the antibody or fragment thereof comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO:10, or a sequence at least about 90% identical thereto, e.g., a sequence at least about 95, 96, 97, 98, or 99% identical thereto.

[0148] In some embodiments, the antibody or fragment thereof comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 1, or a sequence at least about 90% identical thereto, e.g., a sequence at least about 95, 96, 97, 98, or 99% identical thereto, and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 5, or a sequence at least about 90% identical thereto, e.g., a sequence at least about 95, 96, 97, 98, or 99% identical thereto.

[0149] In some embodiments, the antibody or fragment thereof comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 2, or a sequence at least about 90% identical thereto, e.g., a sequence at least about 95, 96, 97, 98, or 99% identical thereto, and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 5, or a sequence at least about 90% identical thereto, e.g., a sequence at least about 95, 96, 97, 98, or 99% identical thereto.

[0150] In some embodiments, the antibody or fragment thereof comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 3, or a sequence at least about 90% identical thereto, e.g., a sequence at least about 95, 96, 97, 98, or 99% identical thereto, and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 5, or a sequence at least about 90% identical thereto, e.g., a sequence at least about 95, 96, 97, 98, or 99% identical thereto.

[0151] In some embodiments, the antibody or fragment thereof comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 4, or a sequence at least about 90% identical thereto, e.g., a sequence at least about 95, 96, 97, 98, or 99% identical thereto, and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 5, or a sequence at least about 90% identical thereto, e.g., a sequence at least about 95, 96, 97, 98, or 99% identical thereto.

[0152] In some embodiments, the antibody or fragment thereof comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 1, or a sequence at least about 90% identical thereto, e.g., a sequence at least about 95, 96, 97, 98, or 99% identical thereto, and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 6, or a sequence at least about 90% identical thereto, e.g., a sequence at least about 95, 96, 97, 98, or 99% identical thereto.

[0153] In some embodiments, the antibody or fragment thereof comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 1, or a sequence at least about 90% identical thereto, e.g., a sequence at least about 95, 96, 97, 98, or 99% identical thereto, and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 7, or a sequence at least about 90% identical thereto, e.g., a sequence at least about 95, 96, 97, 98, or 99% identical thereto.

[0154] In some embodiments, the antibody or fragment thereof comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 1, or a sequence at least about 90% identical thereto, e.g., a sequence at least about 95, 96, 97, 98, or 99% identical thereto, and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 8, or a sequence at least about 90% identical thereto, e.g., a sequence at least about 95, 96, 97, 98, or 99% identical thereto.

[0155] In some embodiments, the antibody or fragment thereof comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 1, or a sequence at least about 90% identical thereto, e.g., a sequence at least about 95, 96, 97, 98, or 99% identical thereto, and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 9, or a sequence at least about 90% identical thereto, e.g., a sequence at least about 95, 96, 97, 98, or 99% identical thereto.

[0156] In some embodiments, the antibody or fragment thereof comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 1, or a sequence at least about 90% identical thereto, e.g., a sequence at least about 95, 96, 97, 98, or 99% identical thereto, and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 10, or a sequence at least about 90% identical thereto, e.g., a sequence at least about 95, 96, 97, 98, or 99% identical thereto.

[0157] In some embodiments, the antibody or fragment thereof comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 2, or a sequence at least about 90% identical thereto, e.g., a sequence at least about 95, 96, 97, 98, or 99% identical thereto, and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 6, or a sequence at least about 90% identical thereto, e.g., a sequence at least about 95, 96, 97, 98, or 99% identical thereto.

[0158] In some embodiments, the antibody or fragment thereof comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 2, or a sequence at least about 90% identical thereto, e.g., a sequence at least about 95, 96, 97, 98, or 99% identical thereto, and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 7, or a sequence at least about 90% identical thereto, e.g., a sequence at least about 95, 96, 97, 98, or 99% identical thereto.

[0159] In some embodiments, the antibody or fragment thereof comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 2, or a sequence at least about 90% identical thereto, e.g., a sequence at least about 95, 96, 97, 98, or 99% identical thereto, and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 8, or a sequence at least about 90% identical thereto, e.g., a sequence at least about 95, 96, 97, 98, or 99% identical thereto.

[0160] In some embodiments, the antibody or fragment thereof comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 2, or a sequence at least about 90% identical thereto, e.g., a sequence at least about 95, 96, 97, 98, or 99% identical thereto, and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 9, or a sequence at least about 90% identical thereto, e.g., a sequence at least about 95, 96, 97, 98, or 99% identical thereto.

[0161] In some embodiments, the antibody or fragment thereof comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 2, or a sequence at least about 90% identical thereto, e.g., a sequence at least about 95, 96, 97, 98, or 99% identical thereto, and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 10, or a sequence at least about 90% identical thereto, e.g., a sequence at least about 95, 96, 97, 98, or 99% identical thereto.

[0162] In some embodiments, the antibody or fragment thereof comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 3, or a sequence at least about 90% identical thereto, e.g., a sequence at least about 95, 96, 97, 98, or 99% identical thereto, and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 6, or a sequence at least about 90% identical thereto, e.g., a sequence at least about 95, 96, 97, 98, or 99% identical thereto.

[0163] In some embodiments, the antibody or fragment thereof comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 3, or a sequence at least about 90% identical thereto, e.g., a sequence at least about 95, 96, 97, 98, or 99% identical thereto, and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 7, or a sequence at least about 90% identical thereto, e.g., a sequence at least about 95, 96, 97, 98, or 99% identical thereto.

[0164] In some embodiments, the antibody or fragment thereof comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 3, or a sequence at least about 90% identical thereto, e.g., a sequence at least about 95, 96, 97, 98, or 99% identical thereto, and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 8, or a sequence at least about 90% identical thereto, e.g., a sequence at least about 95, 96, 97, 98, or 99% identical thereto.

[0165] In some embodiments, the antibody or fragment thereof comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 3, or a sequence at least about 90% identical thereto, e.g., a sequence at least about 95, 96, 97, 98, or 99% identical thereto, and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 9, or a sequence at least about 90% identical thereto, e.g., a sequence at least about 95, 96, 97, 98, or 99% identical thereto.

[0166] In some embodiments, the antibody or fragment thereof comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 3, or a sequence at least about 90% identical thereto, e.g., a sequence at least about 95, 96, 97, 98, or 99% identical thereto, and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 10, or a sequence at least about 90% identical thereto, e.g., a sequence at least about 95, 96, 97, 98, or 99% identical thereto.

[0167] In some embodiments, the antibody or fragment thereof comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 4, or a sequence at least about 90% identical thereto, e.g., a sequence at least about 95, 96, 97, 98, or 99% identical thereto, and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 6, or a sequence at least about 90% identical thereto, e.g., a sequence at least about 95, 96, 97, 98, or 99% identical thereto.

[0168] In some embodiments, the antibody or fragment thereof comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 4, or a sequence at least about 90% identical thereto, e.g., a sequence at least about 95, 96, 97, 98, or 99% identical thereto, and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 7, or a sequence at least about 90% identical thereto, e.g., a sequence at least about 95, 96, 97, 98, or 99% identical thereto.

[0169] In some embodiments, the antibody or fragment thereof comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 4, or a sequence at least about 90% identical thereto, e.g., a sequence at least about 95, 96, 97, 98, or 99% identical thereto, and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 8, or a sequence at least about 90% identical thereto, e.g., a sequence at least about 95, 96, 97, 98, or 99% identical thereto.

[0170] In some embodiments, the antibody or fragment thereof comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 4, or a sequence at least about 90% identical thereto, e.g., a sequence at least about 95, 96, 97, 98, or 99% identical thereto, and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 9, or a sequence at least about 90% identical thereto, e.g., a sequence at least about 95, 96, 97, 98, or 99% identical thereto.

[0171] In some embodiments, the antibody or fragment thereof comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 4, or a sequence at least about 90% identical thereto, e.g., a sequence at least about 95, 96, 97, 98, or 99% identical thereto, and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 10, or a sequence at least about 90% identical thereto, e.g., a sequence at least about 95, 96, 97, 98, or 99% identical thereto.

[0172] In some embodiments, the antibody or fragment thereof may be included in a composition. In some embodiments, the composition is a pharmaceutical composition comprising the antibody or fragment thereof in a pharmaceutically acceptable carrier, optionally containing one or more drugs, pharmaceutical agents, carriers, adjuvants, dispersants, diluents, etc. As used herein, the term "pharmaceutically acceptable" with respect to formulation components such as carriers, excipients, and diluents refers to a component that is not harmful to the patient and is compatible with other components, active ingredients, salts, or ingredients. "Pharmaceutically acceptable" includes "veterinarily acceptable" and thus independently includes both human and non-human mammalian use.

[0173] More specifically, as used herein, the term "pharmaceutical composition" refers to a pharmaceutical preparation containing an antibody or fragment thereof of the present disclosure and suitable for administration to a patient for therapeutic purposes. As used herein, the term "patient" refers to any organism to be treated with an antibody or fragment thereof of the present disclosure, including, but not limited to, any mammal, such as humans, other primates (e.g., monkeys, chimpanzees, etc.), companion animals (e.g., dogs, cats, horses, etc.), livestock (e.g., goats, sheep, pigs, cows, etc.), laboratory animals (e.g., mice, rats, etc.), and wild and zoo animals (e.g., wolves, bears, deer, etc.).

[0174] In some embodiments, pharmaceutical compositions may contain at least one pharmaceutically acceptable ingredient to provide improved formulations of the disclosed compounds, including, but not limited to, one or more pharmaceutically acceptable carriers, excipients, or diluents. The carriers, excipients, or diluents may take a wide variety of forms depending on the form of preparation desired for administration.

[0175] As used herein, the term "carrier" includes, but is not limited to, physiologically acceptable liquids for use as solvents or suspensions, including calcium carbonate, calcium phosphate, various sugars such as lactose, glucose, or sucrose, various starches, cellulose derivatives, gelatin, lipids, liposomes, nanoparticles, such as sterile solutions of water for injection (WFI), saline solution, dextrose solution, Hank's solution, Ringer's solution, vegetable oils, mineral oil, animal oils, polyethylene glycols, liquid paraffin, and the like.

[0176] As used herein, the term "excipient" generally includes, but is not limited to, fillers, binders, disintegrants, glidants, lubricants, complexing agents, solubilizers, stabilizers, preservatives, and surfactants, which may be selected to facilitate administration of the compound by a particular route. Suitable excipients include, for example, colloidal silicon dioxide, silica gel, talc, magnesium silicate, calcium silicate, sodium aluminosilicate, magnesium trisilicate, powdered cellulose, macrocrystalline cellulose, carboxymethylcellulose, crosslinked sodium carboxymethylcellulose, sodium benzoate, calcium carbonate, magnesium carbonate, stearic acid, aluminum stearate, calcium stearate, magnesium stearate, zinc stearate, sodium stearyl fumarate, Syloid®, stearowet C, magnesium oxide, starch, sodium starch glycolate, glyceryl monostearate, glyceryl dibehenate, glyceryl palmitostearate, hydrogenated vegetable oils, hydrogenated cottonseed oil, castor oil, mineral oil, polyethylene glycol (e.g., PEG-10 stearate), PEG-10 stearate, ...4000-8000), polyoxyethylene glycol, poloxamer, povidone, crospovidone, croscarmellose sodium, alginic acid, casein, methacrylate divinylbenzene copolymer, docusate sodium, cyclodextrin (e.g., 2-hydroxypropyl-δ-cyclodextrin), polysorbate (e.g., polysorbate 80), cetrimide, TPGS (d-α-tocopheryl polyethylene glycol 1000 succinate), magnesium lauryl sulfate, sodium lauryl sulfate, polyethylene glycol ether, difatty acid ester of polyethylene glycol, or polyoxyalkylene sorbitan fatty acid ester (e.g., polyoxyethylene sorbitan ester Tween®), polyoxyethylene Sorbitan fatty acid esters, sorbitan fatty acid esters, for example from fatty acids such as oleic acid, stearic acid or palmitic acid, mannitol, xylitol, sorbitol, maltose, lactose, lactose monohydrate or spray dried lactose, sucrose, fructose, calcium phosphate, dibasic calcium phosphate, tribasic calcium phosphate, calcium sulfate, dextrates, dextran, dextrin, dextrose, cellulose acetate, maltodextrin, simethicone, polydextrosem, chitosan, gelatin, HPMC (hydroxypropyl methylcellulose), HPC (hydroxypropyl cellulose), hydroxyethyl cellulose, and the like may also be included.

[0177] As will be understood by those skilled in the art, any diluent known in the art can be utilized in accordance with the present disclosure. In some embodiments of the present disclosure, the diluent is water-soluble. In some embodiments of the present disclosure, the diluent is water-insoluble. As used herein, the term "diluent" includes, but is not limited to, water, saline, phosphate-buffered saline (PBS), dextrose, glycerol, ethanol, buffered sodium acetate or ammonium acetate solution, and the like, and combinations thereof.

[0178] In some embodiments, the pharmaceutical composition of the present disclosure comprises at least one additional active ingredient. As used herein, the term "active ingredient" refers to a therapeutically active compound, as well as any prodrugs thereof, and pharmaceutically acceptable salts, hydrates, and solvates of the compound and prodrug. The additional active ingredient may be combined with the antibody or fragment thereof of the present disclosure, administered separately, or administered in the same pharmaceutical composition. The amount of the additional active ingredient to be administered can be determined by one skilled in the art based on the treatment with the antibody or fragment thereof of the present disclosure.

[0179] In some embodiments, the pharmaceutical composition is a human pharmaceutical composition. As used herein, the term "human pharmaceutical composition" refers to a pharmaceutical composition intended for administration to a human.

[0180] The pharmaceutical compositions of the present disclosure are suitable for administration to patients by any appropriate means, including, but not limited to, those used to administer conventional pharmaceuticals. The pharmaceutical compositions of the present disclosure may be administered using any applicable route envisioned by those skilled in the art, including, but not limited to, oral, intravenous ("IV") injection or infusion, intravesical, subcutaneous ("SC"), intramuscular ("IM"), intraperitoneal, intradermal, intraocular, inhalation (and intrapulmonary), intranasal, transdermal, epicutaneous, subcutaneous, topical, mucosal, nasal, ocular, skin impression, intravaginal, intrauterine, intracervical, and rectal. Such dosage forms should enable the antibody or fragment thereof of the present disclosure to reach target cells. Other factors are well known in the art and include considerations such as toxicity and dosage forms that delay the compound or composition from exerting its effect. Techniques and formulations generally may be found in Remington: The Science and Practice of Pharmacy, 21st edition, Lippincott, Williams and Wilkins, Philadelphia, Pa., 2005.

[0181] In some embodiments, the pharmaceutical compositions of the present disclosure are adapted for topical administration. As used herein, the term "topical administration" refers to administering an antibody or fragment thereof of the present disclosure to the skin surface of a patient / animal such that the antibody or fragment thereof passes through the skin layer. Transdermal administration and transmucosal administration are also encompassed by the term topical administration. As used herein, the term "transdermal" refers to the passage of an antibody or fragment thereof of the present disclosure across at least one skin layer of a patient / animal. As used herein, "transmucosal" refers to the passage of an antibody or fragment thereof of the present disclosure across the mucosa of a patient / animal. Unless otherwise indicated or suggested, the terms "topical administration," "transdermal administration," and "transmucosal administration" are used interchangeably herein.

[0182] Various topical delivery systems for delivering bioactive compounds to microorganisms within a patient are known in the art. Such systems include, but are not limited to, lotions, creams, gels, oils, ointments, solutions, suspensions, emulsions, etc., with the appropriate carrier selected in the art. In some embodiments, the pharmaceutical composition is administered in the form of a gel containing a polyhydric alcohol.

[0183] Suitable carriers include vegetable or mineral oils, white petrolatum (e.g., white soft paraffin), branched chain fats or oils, animal fats, and high molecular weight alcohols (e.g., C 12In some embodiments, the carrier is selected so that the antibody or fragment thereof of the present disclosure is soluble. In some embodiments, emulsifiers, stabilizers, humectants, and antioxidants may also be included, as well as agents that impart color or fragrance, if desired. In some embodiments, organic solvents or cosolvents, such as ethanol or propanol, may be used in the pharmaceutical compositions of the present disclosure. In some embodiments, evaporation of the solvent leaves a residue on the treated surface. In some embodiments, a penetrant appropriate for the barrier to be penetrated is used. Such penetrants are commonly known in the art and include, but are not limited to, bile salts and fusidic acid derivatives. In some embodiments, detergents may be used to facilitate penetration. In some embodiments, a cream for topical administration is formulated from a mixture of mineral oil, self-emulsifying beeswax, and water, into which is mixed the antibody or fragment thereof of the present disclosure dissolved in a small amount of solvent (e.g., oil). The particular topical delivery system used will depend on the intended site of administration.

[0184] In some embodiments, other materials can be added to the topical pharmaceutical composition of the present disclosure to have moisturizing effects and improve the consistency of the pharmaceutical composition.Examples of such compounds include, but are not limited to, cetyl ester wax, stearyl alcohol, cetyl alcohol, glycerin, methylparaben, propylparaben, quaternium-15, humectants, volatile methylsiloxane fluids, and polydiorganosiloxane-polyoxyalkylenes.See, for example, U.S. Patent Nos. 5,153,230 and 4,421,769.In some embodiments, if it is desired that the pharmaceutical composition has an additional cleansing effect, chemicals such as sodium lauryl sulfate or metal salts of carboxylic acids can be added.

[0185] In some embodiments, a wide variety of nonvolatile emollients are useful in the pharmaceutical compositions of the present disclosure. Non-limiting examples of such nonvolatile emollients are listed in McCutcheon's, Vol. 2 Functional Materials, North American Edition, (1992), pp. 137-168, and in the CTFA Cosmetic Ingredient Handbook, Second Edition (1992), which lists skin conditioning agents on pages 572-575 and skin protectants on page 580. In some embodiments, nonvolatile emollients include silicones, hydrocarbons, esters, and mixtures thereof. In some embodiments, esters include esters of monofunctional and difunctional fatty acids esterified with alcohols and polyols (i.e., alcohols having two or more hydroxyl groups). In some embodiments, long-chain esters of long-chain fatty acids are utilized in the pharmaceutical compositions of the present disclosure (i.e., C10-40 fatty acids esterified with C10-40 fatty alcohols). Non-limiting examples of esters useful in the pharmaceutical compositions of the present disclosure include, but are not limited to, those selected from the group consisting of diisopropyl adipate, isopropyl myristate, isopropyl palmitate, myristyl propionate, ethylene glycol distearate, 2-ethylhexyl palmitate, isodecyl neopentanoate, C12-15 alcohol benzoate, di-2-ethylhexyl maleate, ceryl palmitate, myristyl myristate, stearyl stearate, cetyl stearate, behenyl behenrate, and mixtures thereof.

[0186] Examples of silicone emollients useful in the pharmaceutical compositions of the present disclosure include, but are not limited to, polyalkylsiloxanes, cyclic polyalkylsiloxanes, and polyalkylarylsiloxanes. Suitable commercially available polyalkylsiloxanes include polydimethylsiloxanes, also known as dimethicones, non-limiting examples of which include the VICASIL™ series sold by General Electric Company and the DOW CORNING™ 200 series sold by Dow Corning Corporation. Commercially available polyalkylsiloxanes include, among others, cyclomethicones (DOW CORNING™ 244 fluid, DOW CORNING™ 344 fluid, DOW CORNING™ 245 fluid, and DOW CORNING™ 345). Suitable commercially available trimethylsiloxysilicates are sold in mixtures with dimethicones as DOW CORNING™ 593 fluid. Dimethiconol, a hydroxyl-terminated dimethyl silicone, is also useful in the pharmaceutical compositions of the present disclosure. Suitable commercially available dimethiconols are typically sold as mixtures with dimethicone or cyclomethicone (e.g., DOW CORNING™ 1401, 1402, and 1403 fluids). Suitable commercially available polyalkylaryl siloxanes include SF1075 methylphenyl fluid (sold by General Electric Company) and 556 cosmetic-grade phenyl trimethicone fluid (sold by Dow Corning Corporation).

[0187] Hydrocarbons suitable for use in the pharmaceutical compositions of the present disclosure include, but are not limited to, straight-chain and branched-chain hydrocarbons having from about 10 to about 30 carbon atoms. In some embodiments, the straight-chain and branched-chain hydrocarbons have from about 12 to about 24 carbon atoms. In some embodiments, the straight-chain and branched-chain hydrocarbons have from about 16 to about 22 carbon atoms. Non-limiting examples of such hydrocarbon materials include, among others, dodecane, squalane, cholesterol, pentahydrogenated polyisobutylene, docosane (i.e., a C22 hydrocarbon), hexadecane, and isohexadecane (a commercially available hydrocarbon sold as PERMETHYL™ 101A by Presperse, South Plainsfield, NJ).

[0188] In some embodiments, the topical pharmaceutical composition of the present disclosure comprises propylene glycol. In some embodiments, propylene glycol acts as a surfactant, aiding in the penetration, contact, and absorption of the antibody or fragment thereof of the present disclosure. In some embodiments, propylene glycol functions as a preservative. In some embodiments, the pharmaceutical composition of the present disclosure comprises a non-ionic surfactant, such as polysorbate. Such surfactants further reduce surface tension, thereby providing better surface contact of the pharmaceutical composition of the present disclosure with mucous membranes (such as the vaginal mucosa).

[0189] The topical pharmaceutical compositions of the present disclosure may optionally be formulated with a lipophilic phase, such as emulsions and liposomal dispersions. In some embodiments, liposomal formulations may prolong the circulation time of the antibody or fragment thereof of the present disclosure, increase the permeability of the antibody or fragment thereof, and improve the overall efficacy of the antibody or fragment thereof as a pharmaceutical. In some embodiments, the antibody or fragment thereof of the present disclosure may be combined with a lipid, a cationic lipid, or an anionic lipid. In some embodiments, the resulting emulsion or liposomal suspension may effectively increase the in vivo half-life of the activity of the pharmaceutical composition of the present disclosure. Examples of suitable anionic lipids for use with the pharmaceutical compositions of the present disclosure include, but are not limited to, anionic forms of cardiolipin, dimyristoyl, dipalmitoyl, dioleoylphosphatidylcholine, phosphatidylglycerol, palmitoyloleoylphosphatidylcholine, phosphatidylglycerol, phosphatidic acid, lysophosphatidic acid, phosphatidylserine, phosphatidylinositol, and cholesterol.

[0190] In some embodiments, the antibody or fragment thereof of the present disclosure is incorporated into a liposome. In some embodiments, a neutral lipid, cholesterol, and / or polyethylene glycol (PEG) is utilized in such a liposome. In some embodiments, the liposome composition is composed of partially hydrogenated soybean phosphatidylcholine (PHSC), cholesterol, methoxy-terminated PEG (mPEG), and / or distearoylphosphatidylethanolamine (DSPE). Liposomes can be prepared according to any suitable method known in the art.

[0191] In some embodiments, topical administration is via nasal spray or suppository (rectal or vaginal). Suppositories are prepared by mixing an antibody or fragment thereof of the present disclosure with a lipid vehicle such as theobroma oil, cocoa butter, glycerin, gelatin, or polyoxyethylene glycol. In some embodiments, topical administration involves a dressing, such as a transdermal patch or bandage, impregnated with an antibody or fragment thereof of the present disclosure and, optionally, one or more carriers, excipients, or diluents known in the art. In some embodiments, such dressings include, but are not limited to, semipermeable films, foams, hydrocolloids, and calcium alginate swabs. In some embodiments, dosage administration is continuous rather than intermittent throughout the dosing regimen.

[0192] In some embodiments, the pharmaceutical compositions of the present disclosure are adapted for oral administration. As used herein, the term "oral administration" refers to administration of an antibody or fragment thereof of the present disclosure to the mouth of a patient for ingestion into the digestive tract. In some embodiments, the pharmaceutical compositions of the present disclosure can be formulated into conventional oral dosage forms, including, but not limited to, capsules, tablets, powders, and liquid preparations such as suspensions, solutions, elixirs, syrups, concentrated drops, etc. In some embodiments, the antibody or fragment thereof of the present disclosure can be combined with solid excipients, optionally after adding appropriate excipients, to obtain, for example, tablets, coated tablets, hard capsules, soft capsules, solutions (e.g., aqueous, alcoholic, or oily solutions), etc., and the resulting mixture can be optionally milled and the resulting mixture can be optionally processed into granules. In some embodiments, excipients suitable for use in oral pharmaceutical compositions of the present disclosure include, but are not limited to, fillers such as sugars including lactose, glucose, sucrose, mannitol, or sorbitol; cellulose preparations such as corn starch, wheat starch, rice starch, potato starch, gelatin, tragacanth gum, methylcellulose, hydroxypropylmethylcellulose, sodium carboxymethylcellulose (CMC), and / or polyvinylpyrrolidone (PVP or povidone); and oily excipients including vegetable and animal oils such as sunflower oil, olive oil, or cod liver oil. In some embodiments, the oral pharmaceutical compositions of the present disclosure may also contain disintegrating agents such as cross-linked polyvinylpyrrolidone, agar, or alginic acid or a salt thereof such as sodium alginate; lubricants such as talc or magnesium stearate; plasticizers such as glycerol or sorbitol; sweeteners such as sucrose, fructose, lactose, or aspartame; natural or artificial flavors such as peppermint, oil of wintergreen, or cherry flavor; or dyes or pigments that can be used to identify or characterize different doses or combinations. In some embodiments, the oral pharmaceutical compositions of the present disclosure may also contain dragee cores with appropriate coatings.In some embodiments, for example, gum arabic, talc, polyvinylpyrrolidone, carbopol gel, polyethylene glycol, titanium dioxide, lacquer solutions, and concentrated sugar solutions that may optionally contain suitable organic solvents or solvent mixtures may be used.

[0193] In some embodiments, pharmaceutical compositions of the present disclosure that can be used orally include, but are not limited to, push-fit capsules made of gelatin ("gelcaps") and soft, sealed capsules made of gelatin and a plasticizer such as glycerol or sorbitol. In some embodiments, push-fit capsules can contain an antibody or fragment thereof of the present disclosure mixed with filler such as lactose, binders such as starches, and / or lubricants such as talc or magnesium stearate, and optionally stabilizers. In some embodiments involving soft capsules, the antibody or fragment thereof can be dissolved or suspended in a suitable liquid, such as fatty oils, liquid paraffin, or liquid polyethylene glycol.

[0194] In some embodiments, the pharmaceutical compositions of the present disclosure are adapted for inhalation administration. As used herein, the term "inhalation administration" refers to delivery of an antibody or fragment thereof of the present disclosure by passing through the patient's nose or mouth during inhalation, with the antibody or fragment passing through the wall of the patient's lungs. In some embodiments, pharmaceutical compositions of the present disclosure suitable for inhalation administration can be formulated as a dry powder or a suitable solution, suspension, or aerosol. In some embodiments, powders and solutions can be formulated using suitable additives known in the art. In some embodiments, powders can contain a suitable powder base, such as lactose or starch. In some embodiments, solutions can contain propylene glycol, sterile water, ethanol, sodium chloride, and other additives, such as acids, alkalis, and buffer salts. In some embodiments, such solutions or suspensions can be administered by inhalation via a spray, pump, atomizer, nebulizer, or the like. In some embodiments, pharmaceutical compositions of the present disclosure suitable for inhaled administration may also be used in combination with other inhaled therapies, including, but not limited to, corticosteroids such as, for example, fluticasone propionate, beclomethasone dipropionate, triamcinolone acetonide, budesonide, and mometasone furoate; beta-agonists such as, for example, albuterol, salmeterol, and formoterol; anticholinergics such as, for example, ipratroprium bromide or tiotropium; vasodilators such as, for example, treprostinal and iloprost; enzymes such as, for example, DNAase; therapeutic proteins; immunoglobulin antibodies; oligonucleotides such as, for example, single- or double-stranded DNA or RNA, siRNA; antibiotics such as, for example, tobramycin; muscarinic receptor antagonists; leukotriene antagonists; cytokine antagonists; protease inhibitors; cromolyn sodium; nedocril sodium; and sodium cromoglycate.

[0195] In some embodiments, the pharmaceutical composition of the present disclosure is adapted for intravesical administration. As used herein, the term "intravesical administration" refers to delivering an antibody or fragment thereof of the present disclosure directly into the bladder of a patient. In some embodiments, the pharmaceutical composition is administered via a catheter. In some embodiments, the catheter is a urethral catheter.

[0196] In some embodiments, pharmaceutical compositions of the present disclosure are adapted for parenteral administration. As used herein, the term "parenteral administration" refers to an antibody or fragment thereof of the present disclosure injected or infused into a patient, including, but not limited to, intravenous, intramuscular, intraarterial, intrathecal, intraventricular, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subcuticular, intraarticular, subcapsular, intrathecal, intraspinal, intracerebrospinal, and intrasternal injection and infusion. In some embodiments, pharmaceutical compositions of the present disclosure suitable for parenteral administration can be formulated in a sterile liquid solution, including, but not limited to, physiologically compatible buffers or solutions, such as saline solution, Hank's solution, or Ringer's solution. In some embodiments, pharmaceutical compositions of the present disclosure suitable for parenteral administration can be prepared as dispersions in non-aqueous solutions, such as, for example, glycerol, propylene glycol, ethanol, liquid polyethylene glycol, triacetin, vegetable oils, and the like. In some embodiments, the solution may also contain a preservative, such as methylparaben, propylparaben, chlorobutanol, phenol, sorbic acid, or thimerosal. Furthermore, pharmaceutical compositions of the present disclosure suitable for parenteral administration may be formulated in solid form, including, for example, lyophilized forms, and redissolved or suspended prior to use. In some embodiments, the pharmaceutical composition is administered via needle. For injection, the carrier may be a liquid, such as sterile pyrogen-free water, pyrogen-free phosphate-buffered saline solution, bacteriostatic water, or CREMOPHOR EL® (BASF, Parsippany, NJ). For other methods of administration, the carrier may be either a solid or a liquid.

[0197] The present disclosure also provides kits. In some embodiments, the kit comprises an antibody or fragment thereof according to the present disclosure. As used herein, the term "kit" refers to any manufactured article, such as a package, container, or the like, containing an antibody or fragment thereof according to the present disclosure. In some embodiments, the antibody or fragment thereof according to the present disclosure is packaged in a vial, bottle, tube, flask, or patch, which may be further packaged in a box, packet, bag, or the like. In some embodiments, the antibody or fragment thereof according to the present disclosure has been approved for administration to a patient by the U.S. Food and Drug Administration or a similar regulatory agency within the United States or a jurisdiction or territory outside the United States. In some embodiments, the kit includes instructions for use and / or other indication that the antibody or fragment thereof according to the present disclosure is suitable or approved for administration to a patient. In some embodiments, the antibody or fragment thereof according to the present disclosure is packaged in a unit dose or single-unit dose form, such as, for example, a single-unit dose pill, capsule, or the like. In some embodiments, the kit includes a dispenser.

[0198] The present disclosure also provides use of an antibody or fragment thereof of the present disclosure for the manufacture of a medicament. As used herein, the term "medicament" refers to a pharmaceutical composition according to the present disclosure. In some embodiments, the pharmaceutical composition is contained within any article of manufacture, such as, for example, a package, a container, etc.

[0199] In one embodiment, a method for depleting, inhibiting, or depleting and inhibiting pDCs in a patient / animal includes delivering to the patient / animal an effective amount of an antibody or fragment thereof that specifically binds to an epitope comprising BDCA2 and depletes, inhibits, or depletes and inhibits pDCs, thereby depleting, inhibiting, or depleting and inhibiting pDCs. The antibody or fragment thereof is as described above. Furthermore, the antibody or fragment thereof can be formulated into a pharmaceutical composition as described above.

[0200] The term "deplete," as used herein with respect to pDCs, refers to a measurable reduction in the number of pDCs in a patient or sample. The reduction can be at least about 10%, e.g., at least about 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, 99%, or more. In certain embodiments, the term refers to a reduction in the number of pDCs in a patient or sample to below detectable limits. In some embodiments, pDCs are depleted by at least about 50%, e.g., at least about 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, or 99% or more, compared to a patient not receiving the antibody or fragment thereof. In other embodiments, the present disclosure provides methods for reducing the number of pDCs in a sample, e.g., a mixed population of cells, and / or depleting pDCs in a sample, e.g., a mixed population of cells, ex vivo or in vitro, comprising delivering to the sample an effective amount of an antibody or fragment thereof that specifically binds to BDCA2 and reduces the number of pDCs and / or depletes pDCs, thereby reducing the number of pDCs and / or depleting pDCs. In some embodiments, the BDCA2 is human BDCA2.

[0201] The term "inhibit," as used herein with respect to pDCs, refers to slowing or eliminating some activity of pDCs, such as cell survival, pDC activation (e.g., type 1 interferon production), and any other activity of the cell. Various methods of quantifying and / or qualifying cell inhibition are within the purview of those skilled in the art.

[0202] In some embodiments, a method for treating a pDC-associated disorder in a patient in need thereof comprises delivering to the patient an effective amount of an antibody or fragment thereof that specifically binds to an epitope comprising blood dendritic cell antigen-2 (BDCA2) and depletes, inhibits, or both depletes and inhibits pDCs, thereby treating the disorder. The antibody or fragment thereof is as described above. Furthermore, the antibody or fragment thereof can be formulated into a pharmaceutical composition as described above.

[0203] The present disclosure provides methods of treatment. As used herein, the terms "treat," "treatment," "therapy," and similar terms refer to the administration of an antibody or fragment thereof or pharmaceutical composition of the present disclosure in an amount effective to prevent, alleviate, or ameliorate one or more symptoms of a disease or condition (i.e., an indication) and / or prolong the survival of the patient being treated. In some embodiments, "treat," "treatment," "therapy," and similar terms also include, but are not limited to, reducing or eliminating an infection in the patient.

[0204] In practicing the methods of the present disclosure, an effective amount of an antibody or fragment thereof of the present disclosure is administered to a patient / animal in need thereof. As used herein, the term "effective amount" in the context of administration refers to the amount of an antibody or fragment thereof or pharmaceutical composition of the present disclosure that, when administered to a patient / animal, is sufficient to prevent, alleviate, or ameliorate one or more symptoms of a disease or condition (i.e., an indication) and / or prolong the survival of the patient / animal being treated. Such an amount should result in no or few adverse events in the treated patient / animal. Similarly, such an amount should result in no or few toxic effects in the treated patient / animal. As one of ordinary skill in the art will understand, the amount of the antibody or fragment thereof or pharmaceutical composition of the present disclosure will vary depending on many factors, including, but not limited to, the activity of the antibody or fragment thereof of the present disclosure (in vitro, e.g., a compound of the present disclosure versus a target, or in vivo activity in an animal efficacy model), pharmacokinetic results in the animal model (e.g., biological half-life or bioavailability), the type of patient / animal being treated, the age, size, weight, and general physical condition of the patient / animal, the disorder associated with the patient / animal, and the dosing regimen being used in the treatment. One of ordinary skill in the art will understand that the therapeutic effect need not be complete or curative, as long as some benefit is provided to the patient / animal.

[0205] In some embodiments of the present disclosure, the effective amount of an antibody or fragment thereof of the present disclosure to be delivered to a patient / animal subject in need thereof can be quantified by determining micrograms of the antibody or fragment thereof of the present disclosure per kilogram of the patient / animal subject's body weight. In some embodiments, the amount of the antibody or fragment thereof administered to the patient / animal subject is about 0.1 to about 1000 milligrams (mg) of the antibody or fragment thereof of the present disclosure per kilogram (kg) of the patient's body weight. In some embodiments, the amount of the antibody or fragment thereof administered to the patient / animal subject is about 0.1 to about 500 mg of the antibody or fragment thereof of the present disclosure per kg of the patient's body weight. In some embodiments, the amount of the antibody or fragment thereof administered to the patient / animal subject is about 0.1 to about 300 mg of the antibody or fragment thereof of the present disclosure per kg of the patient's body weight. In some embodiments, the amount of the antibody or fragment thereof administered to the patient / animal subject is about 0.1 to about 200 mg of the antibody or fragment thereof of the present disclosure per kg of the patient's body weight. In some embodiments, the amount of an antibody or fragment thereof of the present disclosure administered to a patient is about 0.1 to about 100 mg of an antibody or fragment thereof of the present disclosure per kg of patient body weight. In some embodiments, the amount of an antibody or fragment thereof of the present disclosure administered to a patient is about 0.1 to about 80 mg of an antibody or fragment thereof of the present disclosure per kg of patient body weight. In some embodiments, the amount of an antibody or fragment thereof of the present disclosure administered to a patient is about 0.1 to about 50 mg of an antibody or fragment thereof of the present disclosure per kg of patient body weight. In some embodiments, the amount of an antibody or fragment thereof of the present disclosure administered to a patient is about 0.1 to about 20 mg of an antibody or fragment thereof of the present disclosure per kg of patient body weight. In some embodiments, the amount of an antibody or fragment thereof administered to a patient is about 0.1 to about 10 mg of an antibody or fragment thereof of the present disclosure per kg of patient body weight. As one of ordinary skill in the art will appreciate, multiple administrations may be used.

[0206] In some embodiments of the present disclosure, the antibody or fragment thereof of the present disclosure is administered as a multiple-dose regimen. As used herein, the term "multiple-dose regimen" refers to a treatment period of more than one day. In some embodiments of the present disclosure, the multiple-dose regimen is up to about two days in duration. In some embodiments of the present disclosure, the multiple-dose regimen is up to about three days in duration. In some embodiments of the present disclosure, the multiple-dose regimen is up to about four days in duration. In some embodiments of the present disclosure, the multiple-dose regimen is up to about five days in duration. In some embodiments of the present disclosure, the multiple-dose regimen is up to about six days in duration. In some embodiments of the present disclosure, the multiple-dose regimen is up to about seven days in duration. In some embodiments of the present disclosure, the multiple-dose regimen is up to about 14 days in duration. In some embodiments of the present disclosure, the multiple-dose regimen is up to about one month in duration. In some embodiments of the present disclosure, the multiple-dose regimen is up to about two months in duration. In some embodiments of the present disclosure, the multiple-dose regimen is up to about three months in duration. In some embodiments of the present disclosure, the multiple dose regimen is for a duration of up to about 4 months. In some embodiments of the present disclosure, the multiple dose regimen is for a duration of up to about 5 months. In some embodiments of the present disclosure, the multiple dose regimen is for a duration of up to about 6 months. Other durations may be used herein.

[0207] In some embodiments of the present disclosure, the antibody or fragment thereof of the present disclosure is administered as part of a chronic treatment regimen. As used herein, the term "chronic treatment regimen" refers to treatment with the antibody or fragment thereof of the present disclosure for an extended period of time, for the lifetime of the patient. In some embodiments, chronic treatment is lifelong treatment.

[0208] In some embodiments of the present disclosure, an antibody or fragment thereof of the present disclosure is administered as a single dose. In some embodiments of the present disclosure, an antibody or fragment thereof of the present disclosure is administered as a single unit dose. As used herein, the term "unit dose" refers to a predetermined amount of an antibody or fragment thereof of the present disclosure. The amount of an antibody or fragment thereof of the present disclosure is generally equal to the dosage of an antibody or fragment thereof of the present disclosure that would be administered to a patient, or a convenient fraction of such a dosage, such as, for example, half or one-third of such a dosage. According to the methods of the present disclosure, the terms "single dose" and "single unit dose" include embodiments in which an antibody or fragment thereof of the present disclosure can be administered as a single application and embodiments in which it can be administered as multiple applications.

[0209] In some embodiments, an antibody or fragment thereof of the present disclosure may also be used in combination with one or more additional active ingredients to treat the same disease or condition. In some embodiments, such combinations include administration of an antibody or fragment thereof of the present disclosure and one or more additional active ingredients at different times, or simultaneous administration of an antibody or fragment thereof of the present disclosure and one or more additional active ingredients. In some embodiments, the dosage may be modified for an antibody or fragment thereof of the present disclosure or one or more additional active ingredients used in combination by methods well known to those skilled in the art, for example, the amount administered may be reduced compared to an antibody or fragment thereof of the present disclosure or one or more additional active ingredients used alone. In some embodiments, simultaneous administration includes simultaneous administration of an antibody or fragment thereof of the present disclosure and an additional active ingredient in the same dosage form, simultaneous administration of an antibody or fragment thereof of the present disclosure and an additional active ingredient in separate dosage forms, and separate administration of an antibody or fragment thereof of the present disclosure and an additional active ingredient.

[0210] It is understood that combination includes use with one or more additional active ingredients or other medical procedures, where the one or more additional active ingredients or other medical procedures may be administered at a different time from the antibody or fragment thereof or pharmaceutical composition of the present disclosure (e.g., within a short period of time, such as within a few hours (e.g., about 1, 2, 3, 4 to 24 hours, etc.), or within a longer period of time (e.g., about 1 to 2 days, 2 to 4 days, 4 to 7 days, 1 to 4 weeks, etc.)), or simultaneously with the antibody or fragment thereof or pharmaceutical composition of the present disclosure. Combination also includes use with one or more additional active ingredients or other medical procedures, such as surgery, that are administered once or infrequently, in addition to the antibody or fragment thereof or pharmaceutical composition of the present disclosure, that are administered within a short period of time or within a longer period of time before or after the administration of the one or more additional active ingredients or the completion of the other medical procedure.

[0211] In some embodiments, the present disclosure provides for the delivery of an antibody or fragment thereof or pharmaceutical composition of the present disclosure and one or more additional active ingredients delivered by different administration routes or by the same administration route. In some embodiments, combinations for any administration route include the delivery of an antibody or fragment thereof or pharmaceutical composition of the present disclosure and one or more additional active ingredients delivered together by the same administration route in any pharmaceutical composition, including pharmaceutical compositions in which the two compounds are chemically linked so that the two compounds maintain their therapeutic activity when administered. In some embodiments, one or more additional active ingredients may be co-administered with an antibody or fragment thereof or pharmaceutical composition of the present disclosure. In some embodiments, combinations by co-administration include the administration of a co-formulation or formulation of chemically linked compounds, or the administration of two or more compounds in separate formulations administered by the same or different routes within a short time of each other (e.g., within about 1 hour, 2 hours, 3 hours, up to about 24 hours, etc.). In some embodiments, co-administration of separate formulations includes co-administration via a single device, e.g., the same inhalant device, the same syringe, etc., or administration from separate devices within a short time of each other. In some embodiments, co-formulation of an antibody or fragment thereof or pharmaceutical composition of the present disclosure with one or more additional active ingredients delivered by the same route involves preparing the materials together so that they can be administered by one device, including separate compounds combined in one formulation, or compounds that are chemically linked but modified so as to still retain their biological activity. In some embodiments, such chemically linked compounds may have a bond that is substantially maintained in vivo, or the bond may break down in vivo to separate the two active ingredients.

[0212] As used herein, the phrase "pDC-associated disorder" refers to any disease, disorder, or condition in which pDCs play a role in the cause, side effects, symptoms, or other aspects of the disease, disorder, or condition. Examples of such disorders include, but are not limited to, autoimmune diseases, inflammatory conditions, respiratory conditions, skin disorders, cancer, and combinations of two or more of these.

[0213] As used herein, the term "infectious disease" refers to any disease associated with infection by an infectious agent. Examples of infectious agents include, but are not limited to, viruses and microorganisms. Viruses include the Hepadnaviridae, which includes hepatitis A, B, C, D, E, F, and G; the Flaviviridae, which includes human hepatitis C virus (HCV), yellow fever virus, and dengue virus; the Retroviridae, which includes human immunodeficiency virus (HIV) and human T-lymphotropic viruses (HITLV1 and HTLV2); the Herpesviridae, which includes herpes simplex viruses (HSV-1 and HSV-2), Epstein-Barr virus (EBV), cytomegalovirus, varicella-zoster virus (VZV), human herpesvirus 6 (HHV-6), human herpesvirus 8 (HHV-8), and herpesvirus B; the Papovaviridae, which includes human papillomavirus; the Rhabdoviridae, which includes rabies virus; and the Paramyxovirus, which includes respiratory syncytial virus. Family Iridaceae; Family Reoviridae, including Rotavirus; Family Bunyaviridae, including Hantavirus; Family Filoviridae, including Ebolavirus; Family Adenoviridae; Family Parvoviridae, including Parvovirus B-19; Family Arenaviridae, including Lassavirus; Family Orthomyxoviridae, including Influenza virus; Family Poxviridae, including Orf virus, Molluscum contagiosum virus, Variola virus, and Monkeypox virus; Family Togaviridae, including Venezuelan Equine Encephalitis virus; Family Coronaviridae, including coronaviruses such as Severe Acute Respiratory Syndrome (SARS) virus (e.g., SARS-CoV-2); and Family Picornaviridae, including Poliovirus; Family Rhinovirus; Family Orbivirus; Family Picodnavirus; Family Encephalomyocarditis virus (EMV);These include, but are not limited to, parainfluenza virus, adenovirus, coxsackievirus, echovirus, measles virus, rubella virus, human papillomavirus, canine distemper virus, infectious canine hepatitis virus, feline calicivirus, feline rhinotracheitis virus, TGE virus (porcine), foot-and-mouth disease virus, simian virus 5, human parainfluenza virus type 2, human metapneumovirus, enterovirus, and any other pathogenic virus now known or later identified (see, e.g., Fundamental Virology, Fields et al., Eds., 3d ed Lippincott-Raven, New York, 1996, the entire contents of which are incorporated herein by reference for its teaching of pathogenic viruses);

[0214] Pathogenic microorganisms include Rickettsia, Chlamydia, Mycobacteria, Clostridia, Corynebacteria, Mycoplasma, Ureaplasma, Legionella, Shigella, Salmonella, pathogenic Escherichia coli species, Bordatella, Neisseria, Treponema, Bacillus, Haemophilus, Moraxella, Vibrio, Staphylococcus spp., and Streptococcus spp. spp., Campylobacter spp., Borrelia spp., Leptospira spp., Erlichia spp., Klebsiella spp., Pseudomonas spp., Helicobacter spp., and any other pathogenic microorganisms now known or later identified (see, e.g., Microbiology, Davis et al., Eds., 4th ed., Lippincott, New York, 1990, the entire contents of which are incorporated herein by reference for its teachings of pathogenic microorganisms). Specific examples of microorganisms include Helicobacter pylori, Chlamydia pneumoniae, Chlamydia trachomatis, Ureaplasma urealyticum, Mycoplasma pneumoniae, Staphylococcus aureus,aureus, Streptococcus pyogenes, Streptococcus pneumoniae, Streptococcus viridans, Enterococcus faecalis, Neisseria meningitidis, Neisseria gonorrhoeae, Treponema pallidum, Bacillus anthracis, Salmonella typhi, Vibrio cholera, Pasteurella pestis, Pseudomonas aeruginosa, Campylobacter jejuni, Clostridium difficile dificile, Clostridium botulinum, Mycobacterium tuberculosis, Borrelia burgdorferi, Haemophilus ducreyi, Corynebacterium diphtheria, Bordetella pertussis, Bordetella parapertussis, Bordetella bronchiseptica, Haemophilus influenza, and enterotoxic Escherichia coli.

[0215] The term " autoimmune disorder " used herein refers to any disorder associated with autoimmune reaction.Examples include but are not limited to diabetes, psoriasis of the skin, hyperthyroidism, autoimmune adrenal insufficiency, hemolytic anemia, rheumatic carditis, systemic lupus erythematosus, cutaneous lupus erythematosus, psoriatic arthritis, Sjogren's syndrome polymyositis, myositis, multiple sclerosis, Crohn's disease, ulcerative colitis, lupus, inflammatory bowel syndrome, dermatomyositis, dermatitis, atopic dermatitis, scleroderma, polymyositis, psoriasis, alopecia areata, rheumatoid arthritis, graft-versus-host disease, interferonism and irritable bowel syndrome.

[0216] As used herein, the term "inflammatory condition" refers to a condition in which the tissues of an organism undergo a response to a particular stimulus, including autoimmune disorders, commonly referred to as "inflammation." Symptoms of inflammation include swelling, pain, heat, redness, and loss of tissue function. Exemplary inflammatory conditions include neurodegenerative diseases; diabetes-related nephropathy and retinopathy; protein wasting, muscle fatigue or inflammation; infectious diseases; various cardiovascular diseases or disorders including atherosclerosis; neurodegenerative diseases such as Alzheimer's disease; infectious diseases such as myocarditis, cardiomyopathy, acute endocarditis, and pericarditis; systemic inflammatory response syndrome (SIRS) / sepsis; adult respiratory distress syndrome (ARDS); asthma; rheumatoid arthritis; osteoarthritis; juvenile arthritis; dry eye syndrome; systemic lupus erythematosus; airway hyperresponsiveness (AHR); bronchial hyperresponsiveness; chronic obstructive pulmonary disease (COPD); congestive heart failure (CHF); inflammatory complications of diabetes; metabolic syndrome; ankylosing spondylitis; end-stage renal disease (ESRD); hepatitis; liver fibrosis; pulmonary fibrosis; organ transplant rejection; acute kidney injury; and diseases of aging.

[0217] As used herein, the term "respiratory condition" refers to a pathological condition affecting organs and tissues that compromises gas exchange in air-breathing animals. These conditions include those of the respiratory tract, including the trachea, bronchi, bronchioles, alveoli, pleura, pleural cavity, nerves, and respiratory muscles. Respiratory diseases range from mild, self-limited conditions such as the common cold, influenza, and pharyngitis to life-threatening conditions such as bacterial pneumonia, pulmonary embolism, tuberculosis, acute or chronic asthma, pulmonary fibrosis, lung cancer, and severe acute respiratory syndromes, such as COVID-19. Respiratory diseases can be classified in many different ways, including by the organ or tissue involved, the type and pattern of accompanying signs and symptoms, or the cause of the disease.

[0218] As used herein, the term "skin disorder" refers to any medical condition that affects a patient's integumentary system, i.e., the organ system that surrounds the body and includes skin, hair, nails, and associated muscles and glands, whose primary function is to provide a barrier to the external environment. Exemplary skin disorders include, but are not limited to, scleroderma, psoriasis, dermatomyositis, and atopic dermatitis.

[0219] As used herein, the term "cancer" refers to any benign or malignant abnormal growth of cells. Examples include breast cancer, prostate cancer, lymphoma, skin cancer, pancreatic cancer, colon cancer, melanoma, malignant melanoma, ovarian cancer, brain cancer, primary brain carcinoma, head and neck cancer, glioma, glioblastoma, liver cancer, bladder cancer, non-small cell lung cancer, head or neck carcinoma, breast carcinoma, ovarian carcinoma, lung carcinoma, small cell lung carcinoma, Wilms' tumor, cervical carcinoma, testicular carcinoma, bladder carcinoma, pancreatic carcinoma, gastric carcinoma, colon carcinoma, prostate carcinoma, genitourinary carcinoma, thyroid carcinoma, esophageal carcinoma, myeloma, multiple myeloma, adrenal carcinoma, renal cell carcinoma, endometrial carcinoma, adrenocortical carcinoma, malignant pancreatic insulinoma, malignant carcinoid carcinoma, choriocarcinoma, mycosis fungoides, malignant hypercalcemia, cervical hyperplasia, leukemia, acute lymphoblastic leukemia, B In some embodiments, the cancer is selected from the group of tumor-forming cancers, including, but not limited to, acute lymphocytic leukemia, blastic plasmacytoid dendritic cell neoplasm ("BPDCN" or "pDC leukemia"), chronic lymphocytic leukemia, acute myeloid leukemia, chronic myelogenous leukemia ("AML"), chronic granulocytic leukemia, acute granulocytic leukemia, age-related hairy cell leukemia, chronic myelomonocytic leukemia ("CMML"), myelodysplastic syndrome, chronic myelofibrosis, multiple myeloma, neuroblastoma, rhabdomyosarcoma, Kaposi's sarcoma, Waldenstrom's macroglobulinemia, diffuse large B-cell lymphoma (DLBCL) (L265P), polycythemia vera, essential thrombocytosis, Hodgkin's disease, non-Hodgkin's lymphoma, soft tissue sarcoma, osteogenic sarcoma, primary macroglobulinemia, and retinoblastoma.

[0220] In addition to the aspects and embodiments described and provided elsewhere in this disclosure, the following non-limiting list of embodiments is also contemplated.

[0221] 1. a heavy chain variable region comprising an amino acid sequence selected from the group consisting of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, and a sequence at least about 90% identical to SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, or SEQ ID NO:4; a light chain variable region comprising an amino acid sequence selected from the group consisting of SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, and a sequence at least about 90% identical to SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, or SEQ ID NO:10; An antibody or fragment thereof comprising:

[0222] 2. The antibody or fragment thereof has an affinity (K d 11. The antibody or fragment thereof according to clause 1, which binds an epitope having SEQ ID NO: 11.

[0223] 3. The antibody or fragment thereof of clause 1 or clause 2, wherein said antibody or fragment thereof is a monoclonal antibody or fragment thereof.

[0224] 4. The antibody or fragment thereof according to clauses 1 to 3, wherein said antibody or fragment thereof is a humanized antibody or fragment thereof.

[0225] 5. the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:1, or a sequence at least about 90% identical to SEQ ID NO:1; the light chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, and a sequence at least about 90% identical to SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, or SEQ ID NO:10; 5. The antibody or fragment thereof according to any one of clauses 1 to 4.

[0226] 6. the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:2, or a sequence at least about 90% identical to SEQ ID NO:2; the light chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, and a sequence at least about 90% identical to SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, or SEQ ID NO:10; 5. The antibody or fragment thereof according to any one of clauses 1 to 4.

[0227] 7. the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:3, or a sequence at least about 90% identical to SEQ ID NO:3; the light chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, and a sequence at least about 90% identical to SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, or SEQ ID NO:10; 5. The antibody or fragment thereof according to any one of clauses 1 to 4.

[0228] 8. the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:4, or a sequence at least about 90% identical to SEQ ID NO:4; the light chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, and a sequence at least about 90% identical to SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, or SEQ ID NO:10; 5. The antibody or fragment thereof according to any one of clauses 1 to 4.

[0229] 9. the heavy chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, and a sequence at least about 90% identical to SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, or SEQ ID NO:4; the light chain variable region comprises the amino acid sequence given by SEQ ID NO:5 or a sequence at least about 90% identical to SEQ ID NO:5; 5. The antibody or fragment thereof according to any one of clauses 1 to 4.

[0230] 10. the heavy chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, and a sequence at least about 90% identical to SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, or SEQ ID NO:4; the light chain variable region comprises the amino acid sequence given by SEQ ID NO:6 or a sequence at least about 90% identical to SEQ ID NO:6; 5. The antibody or fragment thereof according to any one of clauses 1 to 4.

[0231] 11. the heavy chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, and a sequence at least about 90% identical to SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, or SEQ ID NO:4; the light chain variable region comprises the amino acid sequence given by SEQ ID NO:7 or a sequence at least about 90% identical to SEQ ID NO:7; 5. The antibody or fragment thereof according to any one of clauses 1 to 4.

[0232] 12. the heavy chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, and a sequence at least about 90% identical to SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, or SEQ ID NO:4; the light chain variable region comprises the amino acid sequence given by SEQ ID NO:8 or a sequence at least about 90% identical to SEQ ID NO:8. 5. The antibody or fragment thereof according to any one of clauses 1 to 4.

[0233] 13. The heavy chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, and a sequence at least about 90% identical to SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, or SEQ ID NO:4; the light chain variable region comprises the amino acid sequence given by SEQ ID NO:9 or a sequence at least about 90% identical to SEQ ID NO:9; 5. The antibody or fragment thereof according to any one of clauses 1 to 4.

[0234] 14. The heavy chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, and a sequence at least about 90% identical to SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, or SEQ ID NO:4; the light chain variable region comprises the amino acid sequence given by SEQ ID NO: 10, or a sequence at least about 90% identical to SEQ ID NO: 10; 5. The antibody or fragment thereof according to any one of clauses 1 to 4.

[0235] 15. The heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:1, or a sequence at least about 90% identical to SEQ ID NO:1; the light chain variable region comprises the amino acid sequence given by SEQ ID NO:5 or a sequence at least about 90% identical to SEQ ID NO:5; 10. The antibody or fragment thereof according to clause 1 to 5 or 9.

[0236] 16. The heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:2, or a sequence at least about 90% identical to SEQ ID NO:2; the light chain variable region comprises the amino acid sequence given by SEQ ID NO:5 or a sequence at least about 90% identical to SEQ ID NO:5; 10. The antibody or fragment thereof according to clause 1 to 4, 6 or 9.

[0237] 17. The heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:3, or a sequence at least about 90% identical to SEQ ID NO:3; the light chain variable region comprises the amino acid sequence given by SEQ ID NO:5 or a sequence at least about 90% identical to SEQ ID NO:5; The antibody or fragment thereof according to clause 1 to 4, 7 or 9.

[0238] 18. The heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:4, or a sequence at least about 90% identical to SEQ ID NO:4; the light chain variable region comprises the amino acid sequence given by SEQ ID NO:5 or a sequence at least about 90% identical to SEQ ID NO:5; 10. The antibody or fragment thereof according to clause 1 to 4, 8 or 9.

[0239] 19. The heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:1 or a sequence at least about 90% identical to SEQ ID NO:1; the light chain variable region comprises the amino acid sequence given by SEQ ID NO:6 or a sequence at least about 90% identical to SEQ ID NO:6; 11. The antibody or fragment thereof according to clause 1 to 5 or 10.

[0240] 20. The heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:1 or a sequence at least about 90% identical to SEQ ID NO:1; the light chain variable region comprises the amino acid sequence given by SEQ ID NO:7 or a sequence at least about 90% identical to SEQ ID NO:7; 12. The antibody or fragment thereof according to any one of clauses 1 to 5 or 11.

[0241] 21. The heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:1 or a sequence at least about 90% identical to SEQ ID NO:1; the light chain variable region comprises the amino acid sequence given by SEQ ID NO:8 or a sequence at least about 90% identical to SEQ ID NO:8. 13. The antibody or fragment thereof according to clause 1 to 5 or 12.

[0242] 22. The heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:1 or a sequence at least about 90% identical to SEQ ID NO:1; the light chain variable region comprises the amino acid sequence given by SEQ ID NO:9 or a sequence at least about 90% identical to SEQ ID NO:9; 14. The antibody or fragment thereof according to clause 1 to 5 or 13.

[0243] 23. The heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:1 or a sequence at least about 90% identical to SEQ ID NO:1; the light chain variable region comprises the amino acid sequence given by SEQ ID NO: 10, or a sequence at least about 90% identical to SEQ ID NO: 10; 15. The antibody or fragment thereof according to clause 1 to 5 or 14.

[0244] 24. The heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:2, or a sequence at least about 90% identical to SEQ ID NO:2; the light chain variable region comprises the amino acid sequence given by SEQ ID NO:6 or a sequence at least about 90% identical to SEQ ID NO:6; The antibody or fragment thereof according to clause 1 to 4, 6 or 10.

[0245] 25. The heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:3, or a sequence at least about 90% identical to SEQ ID NO:3; the light chain variable region comprises the amino acid sequence given by SEQ ID NO:6 or a sequence at least about 90% identical to SEQ ID NO:6; The antibody or fragment thereof according to clause 1 to 4, 7 or 10.

[0246] 26. The heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:4, or a sequence at least about 90% identical to SEQ ID NO:4; the light chain variable region comprises the amino acid sequence given by SEQ ID NO:6 or a sequence at least about 90% identical to SEQ ID NO:6; 11. The antibody or fragment thereof according to clause 1 to 4, 8 or 10.

[0247] 27. The heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:2 or a sequence at least about 90% identical to SEQ ID NO:2; the light chain variable region comprises the amino acid sequence given by SEQ ID NO:7 or a sequence at least about 90% identical to SEQ ID NO:7; 12. The antibody or fragment thereof according to clause 1 to 4, 6 or 11.

[0248] 28. The heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:3, or a sequence at least about 90% identical to SEQ ID NO:3; the light chain variable region comprises the amino acid sequence given by SEQ ID NO:7 or a sequence at least about 90% identical to SEQ ID NO:7; 12. The antibody or fragment thereof according to clause 1 to 4, 7 or 11.

[0249] 29. The heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:4, or a sequence at least about 90% identical to SEQ ID NO:4; the light chain variable region comprises the amino acid sequence given by SEQ ID NO:7 or a sequence at least about 90% identical to SEQ ID NO:7; 12. The antibody or fragment thereof according to clause 1 to 4, 8 or 11.

[0250] 30. The heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:2 or a sequence at least about 90% identical to SEQ ID NO:2; the light chain variable region comprises the amino acid sequence given by SEQ ID NO:8 or a sequence at least about 90% identical to SEQ ID NO:8. 13. The antibody or fragment thereof according to clause 1 to 4, 6 or 12.

[0251] 31. The heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:3 or a sequence at least about 90% identical to SEQ ID NO:3; the light chain variable region comprises the amino acid sequence given by SEQ ID NO:8 or a sequence at least about 90% identical to SEQ ID NO:8. 13. The antibody or fragment thereof according to clause 1 to 4, 7 or 12.

[0252] 32. The heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:4, or a sequence at least about 90% identical to SEQ ID NO:4; the light chain variable region comprises the amino acid sequence given by SEQ ID NO:8 or a sequence at least about 90% identical to SEQ ID NO:8. 13. The antibody or fragment thereof according to clause 1 to 4, 8 or 12.

[0253] 33. The heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:2 or a sequence at least about 90% identical to SEQ ID NO:2; the light chain variable region comprises the amino acid sequence given by SEQ ID NO:9 or a sequence at least about 90% identical to SEQ ID NO:9; 14. The antibody or fragment thereof according to clause 1 to 4, 6 or 13.

[0254] 34. The heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:3 or a sequence at least about 90% identical to SEQ ID NO:3; the light chain variable region comprises the amino acid sequence given by SEQ ID NO:9 or a sequence at least about 90% identical to SEQ ID NO:9; 14. The antibody or fragment thereof according to clause 1 to 4, 7 or 13.

[0255] 35. The heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:4, or a sequence at least about 90% identical to SEQ ID NO:4; the light chain variable region comprises the amino acid sequence given by SEQ ID NO:9 or a sequence at least about 90% identical to SEQ ID NO:9; 14. The antibody or fragment thereof according to clause 1 to 4, 8 or 13.

[0256] 36. The heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:2 or a sequence at least about 90% identical to SEQ ID NO:2; the light chain variable region comprises the amino acid sequence given by SEQ ID NO: 10, or a sequence at least about 90% identical to SEQ ID NO: 10; 15. The antibody or fragment thereof according to clause 1 to 4, 6 or 14.

[0257] 37. The heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:3, or a sequence at least about 90% identical to SEQ ID NO:3; the light chain variable region comprises the amino acid sequence given by SEQ ID NO: 10, or a sequence at least about 90% identical to SEQ ID NO: 10; 15. The antibody or fragment thereof according to clause 1 to 4, 7 or 14.

[0258] 38. The heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:4, or a sequence at least about 90% identical to SEQ ID NO:4; the light chain variable region comprises the amino acid sequence given by SEQ ID NO: 10, or a sequence at least about 90% identical to SEQ ID NO: 10; 15. The antibody or fragment thereof according to clause 1 to 4, 8 or 14.

[0259] 39. A method for depleting, inhibiting, or depleting and inhibiting plasmacytoid dendritic cells (pDCs) in a patient / animal in need thereof, comprising administering to said patient / animal an effective amount of an antibody or fragment thereof that specifically binds to an epitope comprising blood dendritic cell antigen-2 (BDCA2) and depletes, inhibits, or depletes and inhibits pDCs, thereby depleting, inhibiting, or depleting and inhibiting pDCs, wherein said antibody or fragment thereof is a heavy chain variable region comprising an amino acid sequence selected from the group consisting of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, and a sequence at least about 90% identical to SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, or SEQ ID NO:4; a light chain variable region comprising an amino acid sequence selected from the group consisting of SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, and a sequence at least about 90% identical to SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, or SEQ ID NO:10; A method comprising:

[0260] 40. The epitope comprises the amino acid sequence given by SEQ ID NO: 11, and the antibody or fragment thereof has an affinity (K d 39. The method of claim 39, wherein the epitope is bound by

[0261] 41. The method of clause 39 or clause 40, wherein the antibody or fragment thereof is a monoclonal antibody or fragment thereof.

[0262] 42. The method of clauses 39 to 41, wherein the antibody or fragment thereof is a humanized antibody or fragment thereof.

[0263] 43. The method of clauses 39 to 42, wherein said BDCA2 is human BDCA2.

[0264] 44. A method for treating a disorder associated with plasmacytoid dendritic cells (pDCs) in a patient in need thereof, comprising administering to said patient an effective amount of an antibody or fragment thereof that specifically binds to an epitope comprising blood dendritic cell antigen-2 (BDCA2) and depletes, inhibits, or both depletes and inhibits pDCs, thereby treating said disorder, wherein said antibody or fragment thereof: a heavy chain variable region comprising an amino acid sequence selected from the group consisting of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, and a sequence at least about 90% identical to SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, or SEQ ID NO:4; a light chain variable region comprising an amino acid sequence selected from the group consisting of SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, and a sequence at least about 90% identical to SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, or SEQ ID NO:10; A method comprising:

[0265] 45. The method of clause 44, wherein said disorder is selected from the group consisting of an infectious disease, an autoimmune disease, an inflammatory condition, a respiratory condition, a skin disorder, cancer, and combinations of two or more thereof.

[0266] 46. ​​The method of clause 45, wherein the disorder is an infectious disease associated with infection by an infectious agent.

[0267] 47. The method of clause 46, wherein the infectious agent is one or more viruses or one or more microorganisms.

[0268] 48. The infectious agent is selected from the group consisting of: Hepadnaviridae; Flaviviridae; Retroviridae; Herpesviridae; Papovaviridae; Rhabdoviridae; Paramyxoviridae; Reoviridae; Bunyaviridae; Filoviridae; Adenoviridae ae; Parvoviridae; Arenaviridae; Orthomyxoviridae; Poxviridae; Togaviridae; Coronaviridae; Picornaviridae; Rhinoviruses; Orbiviruses; Picodnaviruses; Encephalomyocarditis virus (EMV); Parainfluenza viruses; Adenoviruses; Coxsackieviruses; Echoviruses; Rubeola virus; Rubella virus; Human papillomaviruses; Canine distemper virus; Canine contagious hepatitis virus; Feline calicivirus; Feline rhinotracheitis virus; TGE virus (porcine); Foot and mouth disease virus; Simian virus 5;48. The method of clause 47, wherein the virus is selected from the group consisting of human parainfluenza virus type 2; human metapneumovirus; and enteroviruses.

[0269] 49. The infectious agent is selected from the group consisting of Rickettsia, Chlamydia, Mycobacteria, Clostridium, Corynebacteria, Mycoplasma, Ureaplasma, Legionella, Shigella, Salmonella, pathogenic Escherichia coli species, Bordatella, Neisseria, Treponema, Bacillus, Haemophilus, Moraxella, Vibrio, Staphylococcus spp., Streptococcus spp. 48. The method according to clause 47, wherein the microorganism is selected from the group consisting of Campylobacter spp., Borrelia spp., Leptospira spp., Erlichia spp., Klebsiella spp., Pseudomonas spp. and Helicobacter spp.

[0270] 50. The infectious agent is selected from the group consisting of Helicobacter pylori, Chlamydia pneumoniae, Chlamydia trachomatis, Ureaplasma urealyticum, Mycoplasma pneumoniae, Staphylococcus aureus, Streptococcus pyogenes, Streptococcus pneumoniae, Streptococcus viridans, Enterococcus faecalis, Neisseria meningitidis, Neisseria gonorrhoeae, Treponema pallidum, and Bacillus anthrax. anthracis, Salmonella typhi, Vibrio cholera, Pasteurella pestis (Yersinia pestis), Pseudomonas aeruginosa, Campylobacter jejuni, Clostridium difficile, Clostridium botulinum, Mycobacterium tuberculosis, Borrelia burgdorferi, Haemophilus ducreyi, Corynebacterium diphtheria, Bordetella pertussis, Bordetella parapertussis 48. The method according to clause 47, wherein the microorganism is selected from the group consisting of Bordetella parapertussis, Bordetella bronchiseptica, Haemophilus influenza and enterotoxic Escherichia coli.

[0271] 51. The method of clause 45, wherein said disorder comprises an autoimmune disease selected from the group consisting of diabetes, psoriasis of the skin, hyperthyroidism, autoimmune adrenal insufficiency, hemolytic anemia, rheumatic carditis, systemic lupus erythematosus, cutaneous lupus erythematosus, psoriatic arthritis, Sjogren's syndrome, polymyositis, myositis, multiple sclerosis, Crohn's disease, ulcerative colitis, lupus, inflammatory bowel syndrome, dermatomyositis, dermatitis, atopic dermatitis, scleroderma, polymyositis, psoriasis, alopecia areata, rheumatoid arthritis, graft versus host disease, interferonopathy, irritable bowel syndrome, and combinations of two or more thereof.

[0272] 52. The method of clause 45, wherein the disorder comprises an inflammatory condition, and the inflammatory condition comprises acute kidney injury.

[0273] 53. The method of clause 45, wherein said disorder comprises a respiratory condition, and said respiratory condition comprises asthma, pulmonary fibrosis, or a combination of asthma and pulmonary fibrosis.

[0274] 54. The method of clause 45, wherein said disorder comprises a skin disorder selected from the group consisting of scleroderma, psoriasis, dermatomyositis, atopic dermatitis, and combinations of two or more thereof.

[0275] 55. The disorder is selected from the group consisting of breast cancer, prostate cancer, lymphoma, skin cancer, pancreatic cancer, colon cancer, melanoma, malignant melanoma, ovarian cancer, brain cancer, primary brain carcinoma, head and neck cancer, glioma, glioblastoma, liver cancer, bladder cancer, non-small cell lung cancer, head or neck carcinoma, breast carcinoma, ovarian carcinoma, lung carcinoma, small cell lung carcinoma, Wilms' tumor, cervical carcinoma, testicular carcinoma, bladder carcinoma, pancreatic Carcinoma, gastric carcinoma, colon carcinoma, prostate carcinoma, genitourinary carcinoma, thyroid carcinoma, esophageal carcinoma, myeloma, multiple myeloma, adrenal carcinoma, renal cell carcinoma, endometrial carcinoma, adrenocortical carcinoma, malignant pancreatic insulinoma, malignant carcinoid carcinoma, choriocarcinoma, mycosis fungoides, malignant hypercalcemia, cervical hyperplasia, leukemia, acute lymphoblastic leukemia, B-cell acute lymphoblastic leukemia 46. ​​The method of clause 45, comprising a cancer selected from the group consisting of blastic plasmacytoid dendritic cell neoplasm ("BPDCN" or "pDC leukemia"), chronic lymphocytic leukemia, acute myeloid leukemia, chronic myelogenous leukemia ("AML"), chronic granulocytic leukemia, acute granulocytic leukemia, age-related hairy cell leukemia, chronic myelomonocytic leukemia ("CMML"), myelodysplastic syndrome, chronic myelofibrosis, multiple myeloma, neuroblastoma, rhabdomyosarcoma, Kaposi's sarcoma, Waldenstrom's macroglobulinemia, diffuse large B-cell lymphoma (DLBCL) (L265P), polycythemia vera, essential thrombocytosis, Hodgkin's disease, non-Hodgkin's lymphoma, soft tissue sarcoma, osteogenic sarcoma, primary macroglobulinemia, retinoblastoma, and combinations of two or more thereof.

[0276] 56. The epitope comprises the amino acid sequence given by SEQ ID NO: 11, and the antibody or fragment thereof has an affinity (K d 56. The method of clauses 44 to 55, wherein the epitope is bound by

[0277] 57. The method of clauses 44 to 56, wherein said antibody or fragment thereof is a monoclonal antibody or fragment thereof.

[0278] 58. The method of clauses 44 to 57, wherein said antibody or fragment thereof is a humanized antibody or fragment thereof.

[0279] 59. The heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:1 or a sequence at least about 90% identical to SEQ ID NO:1; the light chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, and a sequence at least about 90% identical to SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, or SEQ ID NO:10; The method described in clauses 44 to 58.

[0280] 60. The heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:2 or a sequence at least about 90% identical to SEQ ID NO:2; the light chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, and a sequence at least about 90% identical to SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, or SEQ ID NO:10; The method described in clauses 44 to 58.

[0281] 61. The heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:3 or a sequence at least about 90% identical to SEQ ID NO:3; the light chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, and a sequence at least about 90% identical to SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, or SEQ ID NO:10; The method described in clauses 44 to 58.

[0282] 62. The heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:4 or a sequence at least about 90% identical to SEQ ID NO:4; the light chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, and a sequence at least about 90% identical to SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, or SEQ ID NO:10; The method described in clauses 44 to 58.

[0283] 63. The heavy chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, and a sequence at least about 90% identical to SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, or SEQ ID NO:4; the light chain variable region comprises the amino acid sequence given by SEQ ID NO:5 or a sequence at least about 90% identical to SEQ ID NO:5; The method described in clauses 44 to 58.

[0284] 64. The heavy chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, and a sequence at least about 90% identical to SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, or SEQ ID NO:4; the light chain variable region comprises the amino acid sequence given by SEQ ID NO:6 or a sequence at least about 90% identical to SEQ ID NO:6; The method described in clauses 44 to 58.

[0285] 65. The heavy chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, and a sequence at least about 90% identical to SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, or SEQ ID NO:4; the light chain variable region comprises the amino acid sequence given by SEQ ID NO:7 or a sequence at least about 90% identical to SEQ ID NO:7; The method described in clauses 44 to 58.

[0286] 66. The heavy chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, and a sequence at least about 90% identical to SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, or SEQ ID NO:4; the light chain variable region comprises the amino acid sequence given by SEQ ID NO:8 or a sequence at least about 90% identical to SEQ ID NO:8. The method described in clauses 44 to 58.

[0287] 67. The heavy chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, and a sequence at least about 90% identical to SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, or SEQ ID NO:4; the light chain variable region comprises the amino acid sequence given by SEQ ID NO:9 or a sequence at least about 90% identical to SEQ ID NO:9; The method described in clauses 44 to 58.

[0288] 68. The heavy chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, and a sequence at least about 90% identical to SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, or SEQ ID NO:4; the light chain variable region comprises the amino acid sequence given by SEQ ID NO: 10, or a sequence at least about 90% identical to SEQ ID NO: 10; The method described in clauses 44 to 58.

[0289] 69. The heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:1 or a sequence at least about 90% identical to SEQ ID NO:1; the light chain variable region comprises the amino acid sequence given by SEQ ID NO:5 or a sequence at least about 90% identical to SEQ ID NO:5; The method according to clauses 44 to 59 or 63.

[0290] 70. The heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:2 or a sequence at least about 90% identical to SEQ ID NO:2; the light chain variable region comprises the amino acid sequence given by SEQ ID NO:5 or a sequence at least about 90% identical to SEQ ID NO:5; 44. The method according to any one of clauses 44 to 58, 60 or 63.

[0291] 71. The heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:3 or a sequence at least about 90% identical to SEQ ID NO:3; the light chain variable region comprises the amino acid sequence given by SEQ ID NO:5 or a sequence at least about 90% identical to SEQ ID NO:5; 44. The method according to clauses 44 to 58, 61 or 63.

[0292] 72. The heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:4 or a sequence at least about 90% identical to SEQ ID NO:4; the light chain variable region comprises the amino acid sequence given by SEQ ID NO:5 or a sequence at least about 90% identical to SEQ ID NO:5; 44. The method according to any one of clauses 44 to 58, 62 or 63.

[0293] 73. The heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:1 or a sequence at least about 90% identical to SEQ ID NO:1; the light chain variable region comprises the amino acid sequence given by SEQ ID NO:6 or a sequence at least about 90% identical to SEQ ID NO:6; The method according to clauses 44 to 59 or 64.

[0294] 74. The heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:1 or a sequence at least about 90% identical to SEQ ID NO:1; the light chain variable region comprises the amino acid sequence given by SEQ ID NO:7 or a sequence at least about 90% identical to SEQ ID NO:7; The method according to clauses 44 to 59 or 65.

[0295] 75. The heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:1 or a sequence at least about 90% identical to SEQ ID NO:1; the light chain variable region comprises the amino acid sequence given by SEQ ID NO:8 or a sequence at least about 90% identical to SEQ ID NO:8. The method according to clauses 44 to 59 or 66.

[0296] 76. The heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:1 or a sequence at least about 90% identical to SEQ ID NO:1; the light chain variable region comprises the amino acid sequence given by SEQ ID NO:9 or a sequence at least about 90% identical to SEQ ID NO:9; The method according to clauses 44 to 59 or 67.

[0297] 77. The heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:1 or a sequence at least about 90% identical to SEQ ID NO:1; the light chain variable region comprises the amino acid sequence given by SEQ ID NO: 10, or a sequence at least about 90% identical to SEQ ID NO: 10; The method according to clauses 44 to 59 or 68.

[0298] 78. The heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:2 or a sequence at least about 90% identical to SEQ ID NO:2; the light chain variable region comprises the amino acid sequence given by SEQ ID NO:6 or a sequence at least about 90% identical to SEQ ID NO:6; The method according to clauses 44 to 58, 60 or 64.

[0299] 79. The heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:3 or a sequence at least about 90% identical to SEQ ID NO:3; the light chain variable region comprises the amino acid sequence given by SEQ ID NO:6 or a sequence at least about 90% identical to SEQ ID NO:6; The method according to clauses 44 to 58, 61 or 64.

[0300] 80. The heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:4 or a sequence at least about 90% identical to SEQ ID NO:4; the light chain variable region comprises the amino acid sequence given by SEQ ID NO:6 or a sequence at least about 90% identical to SEQ ID NO:6; The method according to clauses 44 to 58, 62 or 64.

[0301] 81. The heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:2 or a sequence at least about 90% identical to SEQ ID NO:2; the light chain variable region comprises the amino acid sequence given by SEQ ID NO:7 or a sequence at least about 90% identical to SEQ ID NO:7; 44. The method according to any one of clauses 44 to 58, 60 or 65.

[0302] 82. The heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:3 or a sequence at least about 90% identical to SEQ ID NO:3; the light chain variable region comprises the amino acid sequence given by SEQ ID NO:7 or a sequence at least about 90% identical to SEQ ID NO:7; 44. The method according to clauses 44 to 58, 61 or 65.

[0303] 83. The heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:4 or a sequence at least about 90% identical to SEQ ID NO:4; the light chain variable region comprises the amino acid sequence given by SEQ ID NO:7 or a sequence at least about 90% identical to SEQ ID NO:7; 44. The method according to any one of clauses 44 to 58, 62 or 65.

[0304] 84. The heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:2 or a sequence at least about 90% identical to SEQ ID NO:2; the light chain variable region comprises the amino acid sequence given by SEQ ID NO:8 or a sequence at least about 90% identical to SEQ ID NO:8. 44. The method according to clauses 44 to 58, 60 or 66.

[0305] 85. The heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:3 or a sequence at least about 90% identical to SEQ ID NO:3; the light chain variable region comprises the amino acid sequence given by SEQ ID NO:8 or a sequence at least about 90% identical to SEQ ID NO:8. 44. The method according to clauses 44 to 58, 61 or 66.

[0306] 86. The heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:4 or a sequence at least about 90% identical to SEQ ID NO:4; the light chain variable region comprises the amino acid sequence given by SEQ ID NO:8 or a sequence at least about 90% identical to SEQ ID NO:8. The method according to clauses 44 to 58, 62 or 66.

[0307] 87. The heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:2 or a sequence at least about 90% identical to SEQ ID NO:2; the light chain variable region comprises the amino acid sequence given by SEQ ID NO:9 or a sequence at least about 90% identical to SEQ ID NO:9; The method according to clauses 44 to 58, 60 or 67.

[0308] 88. The heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:3 or a sequence at least about 90% identical to SEQ ID NO:3; the light chain variable region comprises the amino acid sequence given by SEQ ID NO:9 or a sequence at least about 90% identical to SEQ ID NO:9; The method according to clauses 44 to 58, 61 or 67.

[0309] 89. The heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:4 or a sequence at least about 90% identical to SEQ ID NO:4; the light chain variable region comprises the amino acid sequence given by SEQ ID NO:9 or a sequence at least about 90% identical to SEQ ID NO:9; The method according to clauses 44 to 58, 62 or 67.

[0310] 90. The heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:2 or a sequence at least about 90% identical to SEQ ID NO:2; the light chain variable region comprises the amino acid sequence given by SEQ ID NO: 10, or a sequence at least about 90% identical to SEQ ID NO: 10; The method according to clauses 44 to 58, 60 or 68.

[0311] 91. The heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:3 or a sequence at least about 90% identical to SEQ ID NO:3; the light chain variable region comprises the amino acid sequence given by SEQ ID NO: 10, or a sequence at least about 90% identical to SEQ ID NO: 10; The method according to clauses 44 to 58, 61 or 68.

[0312] 92. The heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:4 or a sequence at least about 90% identical to SEQ ID NO:4; the light chain variable region comprises the amino acid sequence given by SEQ ID NO: 10, or a sequence at least about 90% identical to SEQ ID NO: 10; The method according to clauses 44 to 58, 62 or 68.

[0313] 93. A kit for depleting, inhibiting, or depleting and inhibiting plasmacytoid dendritic cells (pDCs) in a patient / animal in need thereof, comprising: Instructions; and a heavy chain variable region comprising an amino acid sequence selected from the group consisting of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, and a sequence at least about 90% identical to SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, or SEQ ID NO:4; a light chain variable region comprising an amino acid sequence selected from the group consisting of SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, and a sequence at least about 90% identical to SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, or SEQ ID NO:10; an antibody or fragment thereof comprising A kit comprising:

[0314] 94. A pharmaceutical composition comprising: at least one excipient; and a heavy chain variable region comprising an amino acid sequence selected from the group consisting of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, and a sequence at least about 90% identical to SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, or SEQ ID NO:4; a light chain variable region comprising an amino acid sequence selected from the group consisting of SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, and a sequence at least about 90% identical to SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, or SEQ ID NO:10; A pharmaceutical composition comprising an antibody or fragment thereof comprising:

[0315] 95. An antibody or fragment thereof according to clause 1 for use in therapy.

[0316] 96. The antibody or fragment thereof according to clause 1 for use in depleting, inhibiting, or depleting and inhibiting plasmacytoid dendritic cells (pDCs).

[0317] 97. Use of an antibody or fragment thereof according to clause 1 for the manufacture of a medicament.

[0318] example Examples relevant to this disclosure are set forth below. In most cases, alternative techniques can be used. These examples are intended to be illustrative and are not intended to limit or restrict the scope of the invention as set forth in the claims.

[0319] To define the role of tumor-associated / TA-pDCs in the human breast cancer tumor microenvironment ("BC-TME"), which contributes to immunosuppression, and to elucidate their immune mechanisms, and to develop novel therapeutics (combining pDC-depleting antibodies with other immunotherapeutic agents), we developed a fully humanized antibody (NPB-303) that specifically binds human BDCA2 and inhibits / depletes human pDCs in vitro and in vivo. In a humanized mouse model harboring human immune cells and human BC tumor cells ("BC-HIT mice"), we demonstrated that TA-pDCs in the BC-TME play a role in the immunosuppressive TME, and that depletion of TA-pDCs in the BC-TME reverses immunosuppression and suppresses human BC growth in vivo. These findings can be used as the basis for clinical trials to test whether TA-pDC depletion, with or without other cancer immunotherapy ("IO") agents, exhibits enhanced antitumor activity in BC patients. The mechanism of TA-pDC in programming the BC-TME can be investigated and novel immunotherapeutics can be modeled in human BC-HIT mice.

[0320] Example 1 Humanization of mouse monoclonal antibody 12b The cDNA sequences of the heavy chain variable ("VH") and light chain variable ("VL") regions of mouse monoclonal antibody 12b were determined. These sequences were used to design optimized VH and VL region expression constructs, in which codons rarely used in mammals were replaced with frequently used codons. These gene constructs were synthesized and cloned into a human IgG1 antibody expression vector. The VH and VL region genes were cloned upstream of either the heavy chain constant region (CH1, hinge, CH2, and CH3) or the light chain constant region (VL / κ), as appropriate. The resulting chimeric antibody was designated NPB-301.

[0321] A three-dimensional molecular model of the mouse 12b variable region was constructed using JN Biosciences' proprietary algorithm. This was used to identify framework amino acid residues involved in the formation of the CDR structure. In parallel, the mouse VH and VL domains derived from 12b were searched in the GenBank® database to identify highly homologous human VH and VL domains, which were then selected as acceptors for CDR grafting. Finally, gene constructs were designed in which the 12b CDRs were grafted onto the human acceptor framework (i.e., replacing the original human CDRs), and human framework residues corresponding to various structural positions in 12b were backmutated to the mouse sequences to ensure functional CDR orientation.

[0322] Genes encoding the humanized VH and VL domains were cloned into an IgG1 expression vector as described above for chimeric antibodies. The resulting humanized antibody variant was designated NPB-30X, with different humanized variants designated by different final numbers (i.e., NPB-302, NPB-303, etc.).

[0323] Figure 1 provides a schematic diagram of the expression vectors used to produce chimeric and humanized anti-BDCA2 antibodies. The plasmid contains a heavy chain transcription unit starting with a human CMV promoter (CMV-P) to initiate transcription of the antibody heavy chain gene. The CMV promoter is followed by the VH, CH1, hinge, CH2, and CH3 exons. Following the heavy chain gene sequence, the light chain transcription unit starts with a CMV promoter, followed by the VL exon and CL (human kappa chain constant region) exon. The light chain gene is then followed by the SV40 early promoter (SV40-P), the puromycin N-acetyltransferase gene (puro) for resistance to puromycin, and a segment containing the SV40 polyadenylation site (SV40-A). Finally, the plasmid contains a portion of the pUC19 plasmid, including a bacterial origin of replication (pUC ori) and a beta-lactamase gene (β-lactamase). Arrows indicate the direction of transcription. Relevant restriction enzyme sites are also shown in Figure 1.

[0324] To obtain a cell line for stable antibody production, the expression vector in Figure 1 was stably transfected into CHO-K1 cells (ATCC). Briefly, CHO-K1 cells were cultured in SFM4CHO medium (HyClone) at 37°C and 7.5% CO2. For transfection, 20 μg of the expression vector was linearized using FspI restriction enzyme and transferred into approximately 107 cells by electroporation. The transfected cells were diluted in SFM4CHO medium and aliquoted into several 96-well plates. After 48 hours, 10 μg / ml puromycin was added to select for stable integrants. The cells were maintained under puromycin selection for 10 days.

[0325] Antibody expression was measured by ELISA. Briefly, plates were coated with a human IgG Fcγ chain-specific polyclonal antibody (Sigma-Aldrich). Then, wells were blocked with PBS containing 0.05% Tween 20 and 2% milk. Wells were washed, and 100 μL of supernatant from each stable cell line was added and incubated for 60 minutes. The supernatant was removed by washing, and HRP-conjugated goat anti-human kappa chain polyclonal antibody (Southern Biotech) was added and incubated for 30 minutes. After a final wash, ELISA signal was developed by adding ABTS substrate, and color development was stopped with 2% oxalic acid.

[0326] The assay signal was analyzed at 405 nM using a plate reader. All antibodies were diluted in PBS containing 0.1% skim milk and 0.025% Tween 20. A washing step using PBS-T (PBS containing 0.05% Tween 20) was applied between each incubation and signal development step.

[0327] Stable transfectants were grown in SFM4CHO medium using roller bottles until cell viability was less than 50%. The supernatant was collected and clarified by centrifugation and filtration. The sample was loaded onto a Protein A Sepharose column (HiTrap MabSelect Xtra, GE Healthcare). The column was washed with PBS, and the captured antibody was eluted with 0.1 M glycine-HCl (pH 3). The eluted antibody was neutralized with 1 M Tris-HCl (pH 8) and buffer-exchanged into PBS by dialysis. The antibody concentration was determined by measuring absorbance at 280 nm (1 mg / ml = 1.4 OD). Purified NPB-301, NPB-302, and NPB-303 were characterized by SDS-PAGE under reducing conditions according to standard procedures. Figure 2 provides an SDS-PAGE analysis of NPB-303. Figure 3 provides a protein analysis of NPB-303 on a Superose 6 column using an AKTA FPLC system. Panel A provides results with gel filtration standards made by BioRad. Panel B provides results with NPB-303. This preparation has greater than 98% antibody monomer, which has a molecular weight of 155 kD.

[0328] Production of recombinant human and rhesus BDCA2 proteins and measurement of their binding affinity to anti-BDCA2 antibodies by surface plasmon resonance (SPR) experiments were performed by Mass Biologics. The extracellular domains of either human BDCA2 (UniProt Q8WTT0, residues 45-213) or rhesus BDCA2 (UniProt A0A1D5QFY5, residues 45-212) were cloned into Mass Biologics' pYW7 vector or pcDNA3.4, respectively. Proteins were expressed using the ExpiCHO expression system (Thermo Fisher Scientific). Purification was performed by affinity chromatography using a Ni Sepharose Excel column (GE Life Sciences), followed by a standard size-exclusion chromatography step to remove aggregates and degradation products.

[0329] The BDCA2 antibody binding kinetics was measured using a Biacore surface plasmon resonance (SPR) instrument (GE Life Sciences). Briefly, anti-BDCA2 antibodies were captured on a Protein A sensor chip (GE Life Sciences). Recombinant BDCA2 protein was then flowed over the captured antibody over a range of concentrations at a rate of 50 μL / min. Data were analyzed to determine kinetic constants using the BIAevaluation software package (GE Life Sciences).

[0330] NPB-303 was selected for further study. The heavy and light chain sequences of NPB-303 are given by SEQ ID NO: 13 and SEQ ID NO: 14, respectively.

[0331] Figure 4 shows the biophysical binding of 15b, 12b, and NPB-303 to BDCA2 antigen from either monkey (panel A) or human (panel B). The binding data are also summarized in Table 1. In Table 1, monoclonal antibody (mAb) 15b is a mouse monoclonal antibody specific for human BDCA2, mAb 12b is a mouse monoclonal antibody that was humanized to produce NPB-303, which binds both human and monkey BDCA2, and NPB-303 is a humanized monoclonal antibody.

[0332] [Table 1]

[0333] Figure 5 shows a structural model of the BDCA2 extracellular domain (green). The yellow-shaded region is the NPB-303 epitope, and the orange-shaded region is the BDCA2 lectin (ligand) binding residues. Figure 6 shows a structural model of the NPB-303 variable (binding) domain. In the figure, the VH framework is shown in green, the VH CDRs are shown in yellow, the VL framework is shown in blue, the VL CDRs are shown in brown, and the aspartate isomerization motif is shown in red.

[0334] Example 2 In vitro and in vivo binding characterization of NPB-303 antibody An ELISA assay for NPB-303 binding was developed in-house. Briefly, Nunc MaxiSorp 96-well plates were coated with BDCA-2 antigen (Biorbyt CLEC4C orb358925) overnight at 4°C. After washing, the plates were blocked with blocking buffer (1% BSA in 1x PBS). Diluted standard NPB-303 antibody and other test lots were added to the plates. After 1 hour of incubation at room temperature, the plates were washed, and secondary antibody (HRP-conjugated goat anti-human IgG Fc antibody; ThermoFisher A18829) was added for 1 hour at room temperature. Signal was developed by adding TMB substrate reagent (ThermoFisher 34021). Signals at 450 nm and 590 nm were read on a plate reader.

[0335] NPB-303 and human IgG control antibodies were conjugated with APCCy7 labels using LIGHTNING-LINK™ Antibody Labeling Kits (Novus Biologicals, catalog no. 765-0005) according to the kit instructions. One million fresh or frozen human PBMCs were stained with Live / Dead™ Dye (Aqua Yellow) and either NPB-303-APCCy7 or human IgG-APCCy7 was added along with human CD45-BV786, CD3 / CD14 / CD19-FITC, CD4-PacBlue, CD123-PE, and BDCA2-APC. Cells were acquired on a BD FACSCelesta and analyzed using the FlowJo program.

[0336] Umbilical cord blood was obtained from the National Disease Research Interchange (NDRI, Philadelphia, PA), and its use was exempted by the New England Institutional Review Board (IRB). Human hematopoietic stem cells (HSCs) were enriched using ROSETTESEP™ Human CD3 Depletion Cocktail (Stem Cell Technologies). Cells were stained with Viability eF780 (BioLegend), FITC CD3 (BioLegend), PE CD34 (BioLegend), and APC CD133 / 2 (Miltenyi) and analyzed using a BD Accuri Flow Cytometer. Viable cells were counted using a ViCell (Beckman Coulter) system. All cells were isolated under sterile conditions. To construct humanized mice, 1–5 × 105 CD34+CD133+ cord blood cells supplemented with 1 μg / mL OKT3 mAb were injected intravenously into 4–6-week-old pre-irradiated mice. At 10-12 weeks post-engraftment, mice were analyzed for reconstitution by flow cytometry analysis of peripheral blood. Mice with 20% or more human leukocyte reconstitution were used for subsequent experiments.

[0337] All experiments involving animals were approved by the Institutional Animal Care and Use Committee (IACUC) of NPB. NSG mice (NOD.Cg-Prkdc) were from the Jackson Laboratory. scid Il2rg tm1Wjl / SzJ) and NBSGW (NOD.Cg-Kit W-41J Tyr + Prkdc scid Il2rg tm1Wjl Female immunodeficient mice, including NSG (Non-Dysplastic Syndrome) and NP1 (NOD-Rag1- / -IL2RgammaC-null) mice from NPB-USA, were used for the construction of humanized mice. NSG and NP1 mice were irradiated at least 4 hours prior to reconstitution (180 rads for NSG mice and 400 rads for NP1 mice using an Xrad IR225 X-ray irradiator). Some experiments were performed using humanized NSG mice (IHCM humanized CD34 cohort, >60% engraftment) purchased from the Jackson Laboratory.

[0338] Humanized mice were injected with NPB-303 (10 mg / kg IV) on days 0 and 2. Mice were sacrificed on day 3. Blood-derived cells were stained with Live / Dead™ Fixable Yellow (catalog no. L34959, Thermo Fisher), human CD45-BV786, mouse CD45.1-APCCy7, human CD3 / CD14 / CD19-FITC, human CD4-PacBlue, CD123-PE, and BDCA2-APC (all from BioLegend) and analyzed using the FlowJo program.

[0339] Human PBMCs were incubated with NPB-303 antibody or human IgG1 isotype control mAb (10 μg / mL) for 30 minutes at 37°C. PBMCs were then stimulated with CpG-ODN-2216 (Invivogen, catalog no. tlrl-2216, 2 μg / mL) for 16 hours. Cells were treated with GolgiPlug for 3 hours before flow cytometry analysis. After incubation, human PBMCs were stained with surface markers including Live-Dead Yellow Dye, human CD45-BV786, CD3-FITC, CD14-FITC, CD19-FITC, CD123-PECy7, BDCA2-APC, and CD4-Pacific Blue, followed by intracellular staining for IFNα-PE or IgG1-PE isotype control using Fixation / Permeabilization solution (BD Cytofix / Cytoperm, catalog no. 54714). Stained cells were analyzed using a BD Celesta Flow Cytometer, and data were analyzed using FlowJo software.

[0340] 1 × 10 cells in complete RPMI medium (10% FBS, 1 × non-essential amino acids, 1 × Pen-Strp, 1 mM sodium pyruvate, 10 mM HEPES, 50 μM 2-mercaptoethanol, and 2 mM L-glutamine) in a 96-well round-bottom plate. 6 Human PBMCs were seeded at 1 / well and treated with NPB-303 antibody at 10, 3.33, 1.11, 0.37, 0.124, 0.04, 0.014, 0.005, 0.0015, or 0.0005 μg / mL or an isotype control antibody at 10 μg / mL for 1 hour. All conditions were established in duplicate. Cells were then stimulated with CpG-ODN-2216 at a final concentration of 10 mg / mL and cultured at 37°C and 5% CO2 for 16 hours. Supernatants were collected, and IFNα was assessed using the PBL Human IFN Alpha Multi-Subtype ELISA Kit (serum, plasma, TCM). IC was calculated using the GraphPad Prism program. 50 was calculated.

[0341] Human PBMCs were incubated with NBP-303 antibody or human IgG1 isotype control mAb (10 μg / mL) for 30 minutes at 37°C. PBMCs were then stimulated with CpG-ODN-2216 (Invivogen, catalog no. tlrl-2216, 2 μg / mL) for 16 hours. Cells were treated with GolgiPlug for 3 hours before flow cytometry analysis. After incubation, human PBMCs were stained with surface markers including Live-Dead Yellow Dye, human CD45-BV786, CD3-FITC, CD14-FITC, CD19-FITC, CD123-PECy7, BDCA2-APC, and CD4-Pacific Blue, followed by intracellular staining for IFNα-PE or IgG1-PE isotype control using Fixation / Permeabilization solution (BD Cytofix / Cytoperm, catalog no. 54714). Stained cells were analyzed using a BD Celesta Flow Cytometer, and data were analyzed using FlowJo software.

[0342] NPB-303 binds BDCA2 on human pDCs (in vitro) and depletes / inhibits pDCs in vivo. To elucidate the role of the chronic pDC / IFN-I axis in suppressing antitumor immunity in the tumor microenvironment, we developed a novel humanized antibody to target human pDCs to treat human inflammatory diseases associated with TA-pDCs. We engineered a mouse monoclonal antibody that specifically binds to human BDCA2 on pDCs (see Figure 1) to replace the non-antigen-binding regions of the Fc and Fab domains with human antibody sequences. The NPB-303 antibody (huIgG1) showed a highly stable interaction with the BDCA2 protein (Figure 7) (Figure 8, K). d Approximately 1nM,k on (1 / ms) approx. 2.1×10 5 and k off (1 / s) approx. 3.3×10 -4). NPB-303 specifically binds to human pDCs in vitro (Figure 9). NPB-303 was examined for BDCA2-mediated inhibition of pDC activation in vitro. NPB303 effectively inhibited CpG-induced pDC activation, as demonstrated by intracellular IFNα detection (Figure 10) or by detection of IFNα in the supernatant (Figure 11). Similar studies in multiple human donor PBMC samples demonstrated that IC 50 was estimated to be approximately 3 ng / ml (FIGS. 12 and 13), significantly better than the recently reported Biogen BDCA2 Mab.

[0343] Example 3 Pharmacokinetic and pharmacodynamic characterization of NPB-303 antibody All animals used were NOD Rag Gamma (NRG) mice (catalog number: 007799) purchased from The Jackson Laboratory. All animals were male and 5-6 weeks old. Animals were injected with NPB-303 at a concentration of 10 mg / mL. The administration volume was 10 mL / kg, resulting in a 10 mg / kg antibody dose. Mice were administered a single dose at the beginning of the experiment via either IV (tail vein) or SC (flank). Blood was collected at various time points after NPB-303 injection via cheek bleed. Groups of three animals were treated per time point. In-life samples were limited to 50 μL per blood draw, with an overall limit of approximately 200 μL (10% blood volume) over the 2-week study period. Prior to the final sample collected from a given animal, mice were euthanized by CO2 asphyxiation, and a terminal blood draw was obtained by cardiac puncture. Blood was collected in K3EDTA tubes (Greiner Bio-One) and processed to plasma.

[0344] NPB-303 was detected in plasma samples using the IgG (Total) Human Uncoated ELISA Kit (ThermoFisher catalog number: 88-50550-22) according to the manufacturer's instructions. A pilot optimization was performed using 16 samples (n=1) at five different dilutions: 1:50, 1:2000, 1:500, 1:5000, and 1:10,000. The tested samples also included a standard curve constructed by preparing a solution of purified NPB-303 at 100 ng / mL. This standard curve was then used to generate two-fold serial dilutions (a total of seven concentrations, ranging from approximately 1.6 to 100 ng / mL). The NPB-303 concentration in the plasma samples was estimated by comparison with this curve. The data were used to calculate the half-life (T 1 / 2 ), maximum plasma concentration (C max ), time to reach maximum plasma concentration (T max) The mean residence time (MRT), area under the plasma concentration curve (AUC), apparent total body clearance (CL), and apparent terminal volume of distribution (VZ) were calculated. All data analyses were performed using the WinNonlin 7.0 software package (Certara).

[0345] NPB-303 was further characterized to quantify the endotoxin present.

[0346] Humanized female mice were injected intravenously with NPB-303 at 10 mg / kg. Mice were sacrificed at 0, 1, 6, 24, 48, 72, and 168 hours post-injection. Cells from mouse spleens were stained with Live / Dead Dye, mouse CD45.1 APCCy7, and human antibodies including CD3 FITC, CD14 FITC, CD19 FITC, CD123 PE, BDCA2 APC, CD4 Pacific Blue, and CD45 BV786 (all from BioLegend). Cells were analyzed using a BD Celesta Instrument, and data were analyzed using FlowJo software.

[0347] The pharmacokinetics of NPB-303 in NSG mice was investigated by intravenous or subcutaneous route (Figure 14 and Table 2). In Table 2, T 1 / 2 is the terminal half-life of the analyte in plasma, and T max is the time from administration to the maximum measured concentration, and C max is the maximum measured concentration of the analyte in plasma, MRT is the mean residence time, and AUC 0-∞ is the area under the concentration-time curve of the analyte in plasma over a time interval extrapolated from 0 to infinity, CL is the clearance, and V Z is the apparent volume of distribution during the terminal phase. In Figure 14, the crosses on the "Time Point (Hour)" axis indicate the plasma concentration of NPB-303 detected in samples taken 30 minutes before antibody injection. The estimated half-lives of NPB-303 were 6.2 and 4.2 days for the intravenous and subcutaneous routes, respectively. For the intravenous and subcutaneous routes, T max are 1 hour and 36 hours, respectively, and C max The values ​​calculated for these parameters and provided in Table 2 are highly dependent on the sampling protocol.

[0348] [Table 2]

[0349] When tested in mice transplanted with human CD34+ HSCs / HSPCs, NPB-303 efficiently and specifically depleted human pDCs in humanized mice in vivo (Figures 15, 16, 17, and 18). BDCA2 surface protein was efficiently internalized on remaining pDCs 10 days after mAb treatment (Figure 19). Human pDCs in lymphoid organs were efficiently depleted and / or inhibited by BDCA2 binding (Figures 19 and 20). Thus, humanized NPB-303 mAb specifically binds to BDCA2 on human pDCs and efficiently inhibits and depletes pDCs in vitro and in vivo. However, NPB-303 did not deplete other cell types in the blood (Lin-CD4-, CD4+CD123-, and CD4-CD123-), as shown in Figure 21.

[0350] Figure 22 provides endotoxin testing performed using a kinetic turbidimetric and kinetic chromogenic test method (ENDOSAFE). NPB-303 (1:1000 dilution) had an endotoxin level of less than 0.005 EU / mL with a reaction time of approximately 2520 seconds.

[0351] Example 4 Antitumor activity of NPB-303 antibody Two different human triple-negative breast cancer (TNBC) cell lines, MDA-MB-231 (ATCC HTB-26) and MDA-MB-468 (ATCC HTB-132), were purchased from the American Type Culture Collection (ATCC). Cells were cultured in Leibovitz's L-15 medium (Thermo Fisher Scientific, Waltham, MA, USA) supplemented with 10% heat-inactivated fetal bovine serum (GE Healthcare Life Sciences, HyClone Laboratories) and 1% penicillin-streptomycin (Thermo Fisher Scientific) at 37°C and 5% CO.

[0352] At least 12 weeks after human HSC transplantation, 5 × 10 cells suspended in serum-free medium mixed with Matrigel (Corning Life Sciences) were cultured. 6 TNBC MDA-MB-231 or MDA-MB-468 cells were injected into the thoracic mammary fat pad of female humanized mice. Tumor volume was monitored weekly using digital calipers.

[0353] In humanized mice bearing tumors (MDA-MB-231), the tumor size was approximately 100 mm 3 , about 300mm 3 or approximately 600 mm 3 Once tumor size reached approximately 300 mm, mice were sacrificed and tumor tissue was analyzed for human pDC infiltration using a flow cytometry pDC panel as previously described. 3 Upon reaching 100 μg / ml, blood, spleen, tumor, lung, and liver were isolated and analyzed for human pDC and T cell infiltration. For T cell flow cytometry analysis, cells were stained with Live / Dead Dye, mouse CD45.1 APCCy7, and human antibodies including CD3 PECy7, CD4 PE, CD8 BV515, PD1 BV421, and Tim3 APC (all from BioLegend). Cells were analyzed on a BD FACSCelesta Instrument, and data were analyzed using FlowJo software.

[0354] The tumor is approximately 80 to 100 mm 3 Once the tumor size reached 100 mg / kg, humanized NSG mice bearing TNBC (MDA-MB-231) were randomly assigned to four treatment groups. Mice were treated with NPB-303 or an isotype control antibody (30 mg / kg) at doses of 3, 10, or 30 mg / kg IV twice weekly for 4 weeks. Tumor volume, body weight, and general observations were recorded twice weekly. At the end of the study, spleens and tumors were analyzed for pDC infiltration as previously described.

[0355] For the development of a second TNBC (MDA-MB-468) model, HIT-TNBC mice were randomized and treated with NPB-303 at doses of 1 and 10 mg / kg and an isotype control antibody (10 mg / kg) intravenously twice weekly for 3 weeks. Tumor volume, body weight, and gross observations were recorded twice weekly.

[0356] For combination studies, humanized NBSGW (NOD.Cg-Kit) with TNBC (MDA-MB-231) W-41J Tyr + Prkdc scid Il2rg tm1Wjl Mice were used in the study. When tumor volumes reached approximately 200 mm3, they were randomized into four treatment groups: isotype control 30 mg / kg IV twice weekly for 3 weeks; Abraxane® alone 5 mg / kg ip daily for 5 days only; NPB-303 30 mg / kg iv twice weekly for 3 weeks; and a combination of Abraxane® (5 mg / kg) and NPB-303 (30 mg / kg) at the same treatment frequency as the single-agent treatment groups. Tumor volumes were recorded twice weekly. At the end of the study, mice were sacrificed, and spleen-, tumor-, and blood-derived cells were analyzed. Liver tissue was also isolated for histological analysis.

[0357] Liver tissues were removed from mice in the combination study and immediately fixed in 4% paraformaldehyde for up to 48 hours. The tissues were then transferred to 70% ethanol until embedding. Each tissue was sectioned at 100 μm intervals, typically yielding 10 slides per organ, and stained with H&E.

[0358] Samples were evaluated by a board-certified veterinary pathologist (HORUS Scientific & NOVA Preclinical Consultants, LLC) for morphological evidence and the extent of tumor infiltration into lung and liver tissue. Light microscopy was used to characterize tumor infiltration using the following semiquantitative scale: (0) no observable changes; (1) minimal—barely perceptible / inconspicuous features / changes in the tissue; (2) mild / slight features / presence in the tissue; (3) moderate / notable features / presence in the tissue; and (4) prominent / overwhelming features / presence in the tissue. Metastasis scores for all samples were analyzed in Microsoft EXCEL™, and p-values ​​between groups were calculated using a t-test.

[0359] Given the functional human immunity generated in NSG / NRG-hu HSC mice, these mice partially reject subcutaneously co-implanted human tumors. NSG-hu HSC mice can be enriched with human immune cells to support the growth of HLA-incompatible tumors and suppress tumor growth. Such humanized mice bearing human immune cells and tumors ("HIT mice") are described herein.

[0360] Furthermore, the immuno-oncology checkpoint inhibitor anti-PD1 mAb KEYTRUDA® can reactivate anti-tumor responses and suppress tumor growth. NPB-303 demonstrated comparable tumor volume reduction compared with KEYTRUDA® in tumor-bearing hu mice (Figure 23), and tumor-bearing hu mice appeared to tolerate NPB-303 treatment well (Figure 24). However, spleen and tumor pDCs were depleted by NPB-303 but not by KEYTRUDA® (Figures 25 and 26). Similarly, spleen and tumor BDCA-2 were downregulated by NPB-303 but not by KEYTRUDA® (Figures 27 and 28).

[0361] Using HIT mice bearing human triple-negative breast cancer cells ("TNBC-HIT"), we demonstrated that engrafted tumors were infiltrated with human CD45 leukocytes, including CD123+CD4+CD303+pDCs (Figures 30 and 31), as shown in Figure 29. Furthermore, human T cells were detected in tumors that displayed markers of immune exhaustion (PD1 and TIM3, Figures 32, 339, and 34).

[0362] NPB-303 was investigated to treat human TNBC in the TNBC-HIT model (Figure 35). NPB-303 at doses of 3 mg / kg, 10 mg / kg, and 30 mg / kg, or an isotype control mAb (30 mg / kg), resulted in approximately 100 mm 3 HIT mice bearing MDA-MB-231 TNBC were treated with NPB-303 and tumor growth was monitored for 31 days. All NPB-303-treated groups had significantly smaller tumor volumes than the isotype control mAb-treated group, and a dose-dependent response was observed (Figure 35). No obvious adverse effects were observed with the mAb (Figure 36). As expected, human pDCs were significantly depleted in both the tumors (Figure 37) and spleens (Figure 38) of NPB-303-treated mice. The remaining pDCs also lost BDCA2 / CD303 expression, indicating that NPB-303 binds to and inhibits all human pDCs.

[0363] To confirm the therapeutic activity of NPB-303 against human TNBC, we tested its efficacy in TNBC-HIT mice implanted with another human TNBC cell line (MDA-MB-468, Figure 39). Similar tumor-reducing activity was observed in MB-468-HIT mice (Figure 40), with no apparent adverse effects on HIT mice (Figure 41).

[0364] Patients with TNBC are currently treated with frontline chemotherapy, Abraxane®, followed by immune checkpoint inhibitors, such as anti-PD1 mAb (Keytruda®). Therefore, the combined activity of NPB-303 after Abraxane® treatment was tested in TNBC-HIT mice (Figure 42). When tumor size was tracked after treatment, Abraxane® or NPB-303 each inhibited tumor growth to a similar size (approximately 50%, Figure 43). Importantly, the Abx / NPB-303 combination further reduced tumor size by approximately 90% (Figure 43) without any apparent adverse effects on HIT mice (Figure 44). Compared with groups treated with ISO or Abraxane® alone, treatment with NPB-303 or Abraxane® in combination significantly restored tumor-infiltrating CD8 T cells through downregulation of PD1 and Tim3 expression (Figures 45 and 46).

[0365] Abraxane® treatment can lead to tumor metastasis. Therefore, metastatic tumors in TNBC-HIT mice were examined (Figure 43). Abraxane®-treated mice showed clear tumor metastasis to the liver, while NPB-303-treated mice showed no or less tumor metastasis. The combination of Abraxane® and NPB-303 showed significantly reduced tumor metastasis in the liver (Figures 47, 48, and 49) and lungs (Figures 50 and 51). These data indicate that tumor inhibition induced by NPB303 was not accompanied by enhanced tumor metastasis, and that the combination of NPB-303 with Abraxane® cooperatively suppressed tumor growth and reduced tumor metastasis.

[0366] NPB-303 also exhibits a thermal stability profile similar to that of other clinical-stage and commercially available monoclonal antibodies. As shown in Figure 52, the thermal denaturation (T m ) and aggregation onset (T agg) is at least comparable to that of therapeutic antibodies that have progressed into clinical trials. Furthermore, NPB-303 binding activity is stable under various heat stress conditions. Figure 53 shows the binding activity of NPB-303 under different conditions. There is no difference between NPB-303 immediately after thawing, NPB-303 incubated at 4°C for 2 weeks, and NPB-303 subjected to stress incubation at 42°C for 2 weeks.

[0367] pDC / IFN-I in the tumor microenvironment is associated with immunosuppression. NPB-303 inhibited TA-pDC in a dose-dependent manner in TNBC-HIT mice, a finding confirmed by using TNBC-HIT mice bearing two human TNBC cell lines (MB231 and MB468). This effect of NPB-303 was tested in combination with Abraxane®, a first-line chemotherapy agent. Either Abraxane® or NPB-303 reduced tumor growth, and their combination synergistically inhibited tumor growth. NPB-303 demonstrated significant activity in preventing tumor metastasis to the liver and lungs. Anti-PD1 antibodies, such as Keytruda®, have shown some efficacy in treating various types of human cancer, but the majority of cancer patients do not respond to Keytruda® due to other immunosuppressive mechanisms, including persistent inflammation. This example provides a new weapon for effective cancer treatment.

[0368] Example 5 NPB-303 Antibody Developability and Manufacturability Engineering As clinical trial participation and drug candidate development increase, safety and efficacy testing become more important. Compared to other molecular drugs, monoclonal antibodies are more susceptible to various post-translational modifications (PTMs) during production, transportation, and storage, which affect the stability of drug molecules and protein structure and function. Aspartate isomerization is a spontaneous, non-enzymatic PTM process. After aspartate isomerization, a methylene group is inserted into the protein backbone; that is, the aspartate side chain reduces the methylene group, thereby causing changes in protein structure and function. Many studies have demonstrated that aspartate isomerization can affect the efficacy of candidate drugs. For example, aspartate isomerization in the complementarity-determining region (CDR) of an antibody can reduce receptor binding efficiency and ultimately affect drug efficacy.

[0369] In the NPB-303 sequence, DG may be a problematic amino acid that causes isomerization problems. Feasibility engineering studies were performed on the antibody sequence by modifying it as follows: 1) changing DG in the heavy chain CDR3 to DA, EG, and SG without changing the light chain to determine whether these modifications affect affinity; 2) changing DGDG in the light chain CDR1 to EGEG, DADA, EGDA, DAEG, SGSG (possibly SGDG and DGSG) without changing the heavy chain to determine whether these modifications affect affinity; and 3) combining the best heavy and light chain modifications into the same Ab molecule. The detailed sequence modifications are shown in Figure 54 and include SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, and SEQ ID NO: 10.

[0370] Using transient antibody production, antibodies with different combinations of heavy and light chain combinations (Table 3) were produced at small scale (3 mL). After protein purification, antibodies were analyzed, including titer, A280, SEC-HPLC, and Caliper SDS (reduced and non-reduced). A summary of QC data for all antibodies is shown in Table 4. All antibodies were tested at a pH of 5.0 in a formulation containing 50 mM Na-AC, 33 mM Tris, and the indicated amount of antibody. [Table 3]

[0371] [Table 4]

[0372] BDCA2 antibody binding kinetics was measured using a Biacore surface plasmon resonance (SPR) instrument (GE Life Sciences). Briefly, anti-BDCA2 antibody was captured on an anti-human Fc chip. Recombinant BDCA2 protein was then flowed over the captured antibody over a range of concentrations at a rate of 50 μL / min. Data were analyzed to determine kinetic constants using the BIAevaluation software package (GE Life Sciences). Sensorgrams of SPR-based affinity studies for all 24 antibodies are shown in Figure 55, and biophysical characteristics of all 24 antibodies are summarized in Table 5. [Table 5]

[0373] The 24 antibodies were further tested for biological activity in inhibiting TLR-9-induced IFNα production in in vitro cultured PBMCs from a single healthy human donor. A set of graphs from the IFNα inhibition assay is shown in Figure 56, and the potency (IC) of all antibodies was 50 ) is summarized in Figure 57. IC 50 Although the inhibition rate differs between antibodies, all 24 antibodies show dose-dependent inhibition of TLR-9-induced IFNα production.

[0374] Based on the binding affinity, expression level, and potency of all 24 antibodies, the top three antibodies were selected (NPB-303_A.D; NPB-303_C.D, and NPB-303_C.G), and these antibodies were produced at a larger scale (100 mL) using transient transfection. A summary of the QC data for the top three antibodies is shown in Table 6. Binding affinity was analyzed using the SPR method, and sensorgrams for all three are shown in Figure 58. Biophysical activity is summarized in Table 7. The data indicate that the biophysical characterization of the large-scale production was similar to that of the small-scale production. [Table 6]

[0375] [Table 7]

[0376] NPB-303_A.D; NPB-303_C.D, and NPB-303_C.G obtained from large-scale production were also tested for inhibition of TLR-9-induced IFNα production. Dose-dependent IFNα inhibition was observed in two different healthy donors (Figure 59), with IC values ​​shown in Figure 60. 50 values ​​were obtained.

[0377] NPB-303_A.D, NPB-303_C.D, and NPB-303_C.G obtained from large-scale production were further tested in vivo in humanized mice for human pDC depletion. Briefly, 20 humanized mice were randomly assigned to five groups (four mice per group). Mice were treated with either isotype control antibody, NPB-303_WT, NPB-303_A.D, NPB-303_C.D, or NPB-303_C.G at 10 mg / kg iv. 72 hours later, the mice were sacrificed, and blood and spleens were analyzed for pDCs.

[0378] Representative flow plots of blood samples from one mouse per group are shown in Figure 61. Human pDCs were depleted in all mice treated with NPB-303 antibody compared with the isotype control (Figure 62), and BDCA2 expression was downregulated in all mice treated with NPB-303 antibody (Figure 63). Splenocytes were also analyzed for pDC depletion by flow cytometry. Representative flow plots of spleens from one mouse per group are shown in Figure 64. Human pDC depletion (Figure 65) and BDCA2 downregulation (Figure 66) were observed. Furthermore, other cell types, including Lin-CD4-, CD4+CD123-, and CD4-CD123- cells (Figure 67), were unaffected by NPB-303 antibody treatment in both the blood (Figure 68) and spleen (Figure 69).

[0379] While embodiments have been disclosed, the present invention is not limited to the disclosed embodiments. Instead, this application is intended to cover any variations, uses, or modifications of the invention using its general principles. Further, this application is intended to cover such departures from the present disclosure as come within known or customary practice in the art to which this invention pertains and which fall within the scope of the appended claims.

Claims

1. a heavy chain variable region comprising an amino acid sequence selected from the group consisting of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, and a sequence at least about 90% identical to SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, or SEQ ID NO:4; a light chain variable region comprising an amino acid sequence selected from the group consisting of SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, and a sequence at least about 90% identical to SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, or SEQ ID NO:10; An antibody or fragment thereof comprising:

2. The antibody or fragment thereof has an affinity (K ｄ 2. The antibody or fragment thereof according to claim 1, which binds an epitope having SEQ ID NO:

11.

3. The antibody or fragment thereof according to claim 1 or claim 2, wherein the antibody or fragment thereof is a monoclonal antibody or fragment thereof.

4. The antibody or fragment thereof according to claim 1 or claim 2, wherein the antibody or fragment thereof is a humanized antibody or fragment thereof.

5. the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:1, or a sequence at least about 90% identical to SEQ ID NO:1; the light chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, and a sequence at least about 90% identical to SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, or SEQ ID NO:10; An antibody or fragment thereof according to claim 1 or claim 2.

6. the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:2, or a sequence at least about 90% identical to SEQ ID NO:2; the light chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, and a sequence at least about 90% identical to SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, or SEQ ID NO:10; An antibody or fragment thereof according to claim 1 or claim 2.

7. the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:3, or a sequence at least about 90% identical to SEQ ID NO:3; the light chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, and a sequence at least about 90% identical to SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, or SEQ ID NO:10; An antibody or fragment thereof according to claim 1 or claim 2.

8. the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:4, or a sequence at least about 90% identical to SEQ ID NO:4; the light chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, and a sequence at least about 90% identical to SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, or SEQ ID NO:10; An antibody or fragment thereof according to claim 1 or claim 2.

9. the heavy chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, and a sequence at least about 90% identical to SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, or SEQ ID NO:4; the light chain variable region comprises the amino acid sequence given by SEQ ID NO:5 or a sequence at least about 90% identical to SEQ ID NO:5; An antibody or fragment thereof according to claim 1 or claim 2.

10. the heavy chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, and a sequence at least about 90% identical to SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, or SEQ ID NO:4; the light chain variable region comprises the amino acid sequence given by SEQ ID NO:6, or a sequence at least about 90% identical to SEQ ID NO:6; An antibody or fragment thereof according to claim 1 or claim 2.

11. the heavy chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, and a sequence at least about 90% identical to SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, or SEQ ID NO:4; the light chain variable region comprises the amino acid sequence given by SEQ ID NO:7, or a sequence at least about 90% identical to SEQ ID NO:7; An antibody or fragment thereof according to claim 1 or claim 2.

12. the heavy chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, and a sequence at least about 90% identical to SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, or SEQ ID NO:4; the light chain variable region comprises the amino acid sequence given by SEQ ID NO:8, or a sequence at least about 90% identical to SEQ ID NO:

8. An antibody or fragment thereof according to claim 1 or claim 2.

13. the heavy chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, and a sequence at least about 90% identical to SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, or SEQ ID NO:4; the light chain variable region comprises the amino acid sequence given by SEQ ID NO:9 or a sequence at least about 90% identical to SEQ ID NO:9; An antibody or fragment thereof according to claim 1 or claim 2.

14. the heavy chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, and a sequence at least about 90% identical to SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, or SEQ ID NO:4; the light chain variable region comprises the amino acid sequence given by SEQ ID NO: 10, or a sequence at least about 90% identical to SEQ ID NO: 10; An antibody or fragment thereof according to claim 1 or claim 2.

15. the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:1, or a sequence at least about 90% identical to SEQ ID NO:1; the light chain variable region comprises the amino acid sequence given by SEQ ID NO:5 or a sequence at least about 90% identical to SEQ ID NO:5; An antibody or fragment thereof according to claim 1 or claim 2.

16. the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:2, or a sequence at least about 90% identical to SEQ ID NO:2; the light chain variable region comprises the amino acid sequence given by SEQ ID NO:5 or a sequence at least about 90% identical to SEQ ID NO:5; An antibody or fragment thereof according to claim 1 or claim 2.

17. the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:3, or a sequence at least about 90% identical to SEQ ID NO:3; the light chain variable region comprises the amino acid sequence given by SEQ ID NO:5 or a sequence at least about 90% identical to SEQ ID NO:5; An antibody or fragment thereof according to claim 1 or claim 2.

18. the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:4, or a sequence at least about 90% identical to SEQ ID NO:4; the light chain variable region comprises the amino acid sequence given by SEQ ID NO:5 or a sequence at least about 90% identical to SEQ ID NO:5; An antibody or fragment thereof according to claim 1 or claim 2.

19. the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:1, or a sequence at least about 90% identical to SEQ ID NO:1; the light chain variable region comprises the amino acid sequence given by SEQ ID NO:6, or a sequence at least about 90% identical to SEQ ID NO:6; An antibody or fragment thereof according to claim 1 or claim 2.

20. the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:1, or a sequence at least about 90% identical to SEQ ID NO:1; the light chain variable region comprises the amino acid sequence given by SEQ ID NO:7, or a sequence at least about 90% identical to SEQ ID NO:7; An antibody or fragment thereof according to claim 1 or claim 2.

21. the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:1, or a sequence at least about 90% identical to SEQ ID NO:1; the light chain variable region comprises the amino acid sequence given by SEQ ID NO:8, or a sequence at least about 90% identical to SEQ ID NO:

8. An antibody or fragment thereof according to claim 1 or claim 2.

22. the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:1, or a sequence at least about 90% identical to SEQ ID NO:1; the light chain variable region comprises the amino acid sequence given by SEQ ID NO:9 or a sequence at least about 90% identical to SEQ ID NO:9; An antibody or fragment thereof according to claim 1 or claim 2.

23. the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:1, or a sequence at least about 90% identical to SEQ ID NO:1; the light chain variable region comprises the amino acid sequence given by SEQ ID NO: 10, or a sequence at least about 90% identical to SEQ ID NO: 10; An antibody or fragment thereof according to claim 1 or claim 2.

24. the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:2, or a sequence at least about 90% identical to SEQ ID NO:2; the light chain variable region comprises the amino acid sequence given by SEQ ID NO:6, or a sequence at least about 90% identical to SEQ ID NO:6; An antibody or fragment thereof according to claim 1 or claim 2.

25. the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:3, or a sequence at least about 90% identical to SEQ ID NO:3; the light chain variable region comprises the amino acid sequence given by SEQ ID NO:6, or a sequence at least about 90% identical to SEQ ID NO:6; An antibody or fragment thereof according to claim 1 or claim 2.

26. the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:4, or a sequence at least about 90% identical to SEQ ID NO:4; the light chain variable region comprises the amino acid sequence given by SEQ ID NO:6, or a sequence at least about 90% identical to SEQ ID NO:6; An antibody or fragment thereof according to claim 1 or claim 2.

27. the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:2, or a sequence at least about 90% identical to SEQ ID NO:2; the light chain variable region comprises the amino acid sequence given by SEQ ID NO:7, or a sequence at least about 90% identical to SEQ ID NO:7; An antibody or fragment thereof according to claim 1 or claim 2.

28. the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:3, or a sequence at least about 90% identical to SEQ ID NO:3; the light chain variable region comprises the amino acid sequence given by SEQ ID NO:7, or a sequence at least about 90% identical to SEQ ID NO:7; An antibody or fragment thereof according to claim 1 or claim 2.

29. the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:4, or a sequence at least about 90% identical to SEQ ID NO:4; the light chain variable region comprises the amino acid sequence given by SEQ ID NO:7, or a sequence at least about 90% identical to SEQ ID NO:7; An antibody or fragment thereof according to claim 1 or claim 2.

30. the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:2, or a sequence at least about 90% identical to SEQ ID NO:2; the light chain variable region comprises the amino acid sequence given by SEQ ID NO:8, or a sequence at least about 90% identical to SEQ ID NO:

8. An antibody or fragment thereof according to claim 1 or claim 2.

31. the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:3, or a sequence at least about 90% identical to SEQ ID NO:3; the light chain variable region comprises the amino acid sequence given by SEQ ID NO:8, or a sequence at least about 90% identical to SEQ ID NO:

8. An antibody or fragment thereof according to claim 1 or claim 2.

32. the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:4, or a sequence at least about 90% identical to SEQ ID NO:4; the light chain variable region comprises the amino acid sequence given by SEQ ID NO:8, or a sequence at least about 90% identical to SEQ ID NO:

8. An antibody or fragment thereof according to claim 1 or claim 2.

33. the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:2, or a sequence at least about 90% identical to SEQ ID NO:2; the light chain variable region comprises the amino acid sequence given by SEQ ID NO:9 or a sequence at least about 90% identical to SEQ ID NO:9; An antibody or fragment thereof according to claim 1 or claim 2.

34. the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:3, or a sequence at least about 90% identical to SEQ ID NO:3; the light chain variable region comprises the amino acid sequence given by SEQ ID NO:9 or a sequence at least about 90% identical to SEQ ID NO:9; An antibody or fragment thereof according to claim 1 or claim 2.

35. the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:4, or a sequence at least about 90% identical to SEQ ID NO:4; the light chain variable region comprises the amino acid sequence given by SEQ ID NO:9 or a sequence at least about 90% identical to SEQ ID NO:9; An antibody or fragment thereof according to claim 1 or claim 2.

36. the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:2, or a sequence at least about 90% identical to SEQ ID NO:2; the light chain variable region comprises the amino acid sequence given by SEQ ID NO: 10, or a sequence at least about 90% identical to SEQ ID NO: 10; An antibody or fragment thereof according to claim 1 or claim 2.

37. the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:3, or a sequence at least about 90% identical to SEQ ID NO:3; the light chain variable region comprises the amino acid sequence given by SEQ ID NO: 10, or a sequence at least about 90% identical to SEQ ID NO: 10; An antibody or fragment thereof according to claim 1 or claim 2.

38. the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:4, or a sequence at least about 90% identical to SEQ ID NO:4; the light chain variable region comprises the amino acid sequence given by SEQ ID NO: 10, or a sequence at least about 90% identical to SEQ ID NO: 10; An antibody or fragment thereof according to claim 1 or claim 2.

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