IL-8 antibodies and methods of use thereof

Abstract

Described herein are anti-IL-8 antibodies, compositions thereof, and methods of use thereof. Anti-IL-8 antibodies may be useful in methods of inhibiting tumor formation or growth, or a combination thereof. Also described herein are methods of using anti-IL-8 antibodies for treating disease, where the disease may include cancer, tumor, or viral infection.

Description

[Technical Field]

[0001] Sequence Listing Description

[0001] This application contains a Sequence Listing that has been submitted electronically in ASCII format and is incorporated herein by reference in its entirety. The ASCII copy, created on August 4, 2021, is named P-585889-PC_SL.txt and is 80,071 bytes in size.

[0002] Field of the Disclosure FIELD OF THE INVENTION

[0002] Disclosed herein are IL-8 antibodies and their use for treating diseases, such as, but not limited to, diseases associated with cancer or viral infection, or diseases associated with inflammation. [Background technology]

[0003] background IL-8 is a potent proinflammatory cytokine secreted by various cells, primarily macrophages. It binds to the seven-transmembrane G protein-coupled receptors (GPCRs), CXCR1 and CXCR2, on the cell surface of neutrophils and monocytes, affecting their activation and migration to sites of infection.

[0004]

[0004] Increasing evidence suggests a role for IL-8 signaling in cancer, both hematological malignancies and solid tumors. Studies have shown that IL-8 is important for cancer cell survival, invasion, and proliferation and may be a key regulator of cancer stem cell activity. Indeed, IL-8 is often overexpressed in many human cancers, and increased IL-8 expression has been correlated with poor prognosis in subjects with IL-8-expressing cancers. Studies have shown upregulation of the IL-8 gene in acute myeloid leukemia (AML) and myelodysplastic syndrome (MDS).

[0005] IL-8 has been observed to promote cancer progression by influencing the tumor microenvironment (TME), including recruiting neutrophils and bone marrow-derived immunosuppressive cells to the TME. Furthermore, IL-8 is known to function in both paracrine and autocrine modes within the TME. Interestingly, reducing the presence of the IL-8 receptor CXCR2 on the surface of tumor cells prolonged the survival of cancer patients and inhibited tumor angiogenesis in certain cancers.

[0006]

[0006] Therefore, targets for treating IL-8-expressing cancers or tumors include the cancer and tumor cells themselves, as well as the tumor microenvironment.

[0007]

[0007] Furthermore, IL-8 is known as a neutrophil chemotactic factor that induces chemotaxis in target cells, such as, but not limited to, neutrophils, and the presence of IL-8 is thought to be involved in the migration of these cells to the site of viral infection. There is increasing evidence that IL-8 is involved in viral infection, leading to increased cytokine release syndrome and cytokine storm during viral infection. Summary of the Invention [Problem to be solved by the invention]

[0008]

[0008] There remains a need to provide therapeutic molecules that target the deleterious effects of IL-8 in cancer and viral infections. The anti-IL-8 antibodies provided herein address this need by targeting cancer cell-expressed IL-8 and / or IL-8 receptors by modulating cell, e.g., neutrophil, activation and / or migration. [Means for solving the problem]

[0009] overview

[0009] In one aspect, disclosed herein is an isolated anti-IL-8 antibody comprising an antibody antigen-binding domain comprising a heavy chain variable region (VH) and a light chain variable region (VL), wherein the amino acid sequence of the VH-VL pair is selected from the pair sequences set forth in SEQ ID NOs: 2 and 3, SEQ ID NOs: 4 and 5, SEQ ID NOs: 6 and 7, SEQ ID NOs: 8 and 9, SEQ ID NOs: 10 and 11, SEQ ID NOs: 12 and 13, SEQ ID NOs: 14 and 15, SEQ ID NOs: 16 and 17, SEQ ID NOs: 18 and 19, SEQ ID NOs: 20 and 21, SEQ ID NOs: 22 and 23, SEQ ID NOs: 24 and 25, SEQ ID NOs: 26 and 27, SEQ ID NOs: 28 and 29, SEQ ID NOs: 30 and 31, SEQ ID NOs: 32 and 33, and SEQ ID NOs: 34 and 35.

[0010]

[0010] In one aspect, disclosed herein is a composition comprising an isolated anti-IL-8 antibody comprising an antibody antigen-binding domain comprising a heavy chain variable region (VH) and a light chain variable region (VL), wherein the amino acid sequence of the VH-VL pair is selected from the pair sequences set forth in SEQ ID NOs: 2 and 3, SEQ ID NOs: 4 and 5, SEQ ID NOs: 6 and 7, SEQ ID NOs: 8 and 9, SEQ ID NOs: 10 and 11, SEQ ID NOs: 12 and 13, SEQ ID NOs: 14 and 15, SEQ ID NOs: 16 and 17, SEQ ID NOs: 18 and 19, SEQ ID NOs: 20 and 21, SEQ ID NOs: 22 and 23, SEQ ID NOs: 24 and 25, SEQ ID NOs: 26 and 27, SEQ ID NOs: 28 and 29, SEQ ID NOs: 30 and 31, SEQ ID NOs: 32 and 33, and SEQ ID NOs: 34 and 35; and a pharmaceutically acceptable carrier.

[0011]

[0011] In one aspect, disclosed herein is an isolated polynucleotide sequence encoding the heavy chain variable region (VH) of an anti-IL-8 antibody and the light chain variable region (VL) of an anti-IL-8 antibody, wherein the amino acid sequence of the VH-VL pair is selected from the pair sequences set forth in SEQ ID NOs: 2 and 3, SEQ ID NOs: 4 and 5, SEQ ID NOs: 6 and 7, SEQ ID NOs: 8 and 9, SEQ ID NOs: 10 and 11, SEQ ID NOs: 12 and 13, SEQ ID NOs: 14 and 15, SEQ ID NOs: 16 and 17, SEQ ID NOs: 18 and 19, SEQ ID NOs: 20 and 21, SEQ ID NOs: 22 and 23, SEQ ID NOs: 24 and 25, SEQ ID NOs: 26 and 27, SEQ ID NOs: 28 and 29, SEQ ID NOs: 30 and 31, SEQ ID NOs: 32 and 33, and SEQ ID NOs: 34 and 35.

[0012]

[0012] In a related aspect, the polynucleotide sequence comprises two polynucleotide sequences, a first polynucleotide sequence encoding the VH of the anti-IL-8 antibody and a second polynucleotide sequence encoding the VL of the anti-IL-8 antibody.

[0013] In one aspect, disclosed herein is a vector comprising a polynucleotide sequence encoding an anti-IL-8 antibody.

[0014] In one aspect, disclosed herein is a host cell comprising a vector comprising a polynucleotide sequence encoding an anti-IL-8 antibody.

[0015]

[0015] In one aspect, the present specification discloses a method for producing an anti-IL-8 antibody comprising a heavy chain variable region (VH) and a light chain variable region (VL), the method comprising culturing a host cell described in claim 12 under conditions that promote expression of the vector in the host cell, and expressing the polynucleotide sequence contained in the vector, thereby producing an anti-IL-8 antibody comprising a VH and a VL.

[0016]

[0016] In one aspect, the present specification discloses isolated anti-IL-8 antibodies having the complementarity determining region (CDR) sequences set forth in Table 1F, wherein each antibody comprises a heavy chain variable region having heavy chain complementarity determining region (HCDR) 1, HCDR2 and HCDR3, and a light chain variable region having light chain complementarity determining region (LCDR) 1, LCDR2 and LCDR3, and wherein the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3 for each of the antibodies comprise the amino acid sequences set forth in Table 1F.

[0017]

[0017] In one aspect, disclosed herein is a composition comprising an isolated anti-IL-8 antibody having the complementarity determining region (CDR) sequences set forth in Table 1F, wherein each antibody comprises a heavy chain variable region having heavy chain complementarity determining region (HCDR) 1, HCDR2 and HCDR3, and a light chain variable region having light chain complementarity determining region (LCDR) 1, LCDR2 and LCDR3, and wherein the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3 for each of the antibodies comprise the amino acid sequences set forth in Table 1F; and a pharmaceutically acceptable carrier.

[0018]

[0018] In one aspect, disclosed herein is an isolated polynucleotide sequence encoding a heavy chain variable region (VH) of an anti-IL-8 antibody comprising a VH complementarity determining region (HCDR) as set forth in Table 1F and a light chain variable region (VL) of an anti-IL-8 antibody comprising a VL complementarity determining region (LCDR) as set forth in Table 1F, wherein the heavy chain variable region comprises heavy chain complementarity determining region (HCDR) 1, HCDR2 and HCDR3, and the light chain variable region comprises light chain complementarity determining region (LCDR) 1, LCDR2 and LCDR3, and the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3 for each of the VH and CL comprise the amino acid sequences set forth in Table 1F.

[0019]

[0019] In a related aspect, the polynucleotide sequence comprises two polynucleotide sequences, a first polynucleotide sequence encoding the VH of the anti-IL-8 antibody and a second polynucleotide sequence encoding the VL of the anti-IL-8 antibody.

[0020]

[0020] In one aspect, disclosed herein is an isolated polynucleotide sequence encoding the heavy chain variable region (VH) of an anti-IL-8 antibody and the light chain variable region (VL) of an anti-IL-8 antibody, wherein the polynucleotide sequence encoding the VH-VL pair is selected from the pair sequences set forth in SEQ ID NOs: 113 and 114, SEQ ID NOs: 115 and 116, SEQ ID NOs: 117 and 118, SEQ ID NOs: 119 and 120, SEQ ID NOs: 121 and 122, SEQ ID NOs: 123 and 124, SEQ ID NOs: 125 and 126, SEQ ID NOs: 127 and 128, SEQ ID NOs: 129 and 130, SEQ ID NOs: 131 and 132, SEQ ID NOs: 133 and 134, SEQ ID NOs: 135 and 136, SEQ ID NOs: 137 and 138, SEQ ID NOs: 139 and 140, SEQ ID NOs: 141 and 142, SEQ ID NOs: 36 and 37, and SEQ ID NOs: 38 and 39.

[0021] In a related aspect, the polynucleotide sequence comprises two polynucleotide sequences, a first polynucleotide sequence encoding the VH of an anti-IL-8 antibody and a second polynucleotide sequence encoding the VL of the anti-IL-8 antibody. In another related aspect, disclosed herein is a vector comprising polynucleotide sequences encoding the heavy chain variable region (VH) of an anti-IL-8 antibody and the light chain variable region (VL) of an anti-IL-8 antibody.

[0022] In related aspects, the antibodies disclosed herein comprise an IgG, Fv, scFv, Fab, F(ab')2, minibody, diabody, triabody, nanobody, single domain antibody, multispecific antibody, bispecific antibody, triabody, single chain antibody, heavy chain antibody, chimeric antibody, or humanized antibody. In further related aspects, the IgG comprises IgG1, IgG2, IgG3, or IgG4.

[0023] In one aspect, disclosed herein is a host cell comprising a vector comprising a polynucleotide sequence encoding an anti-IL-8 antibody VH or VL, or a combination thereof.

[0024]

[0024] In one aspect, disclosed herein is a method for producing an anti-IL-8 antibody comprising a heavy chain variable region (VH) and a light chain variable region (VL), the method comprising culturing a host cell under conditions that promote expression of the vector in the host cell, and expressing the polynucleotide sequence contained in the vector, thereby producing an anti-IL-8 antibody comprising a VH and a VL.

[0025]

[0025] In one aspect, disclosed herein is a vector comprising a polynucleotide sequence encoding a heavy chain variable region (VH) of an anti-IL-8 antibody comprising a VH complementarity determining region (HCDR) as described in Table 1F, and a light chain variable region (VL) of an anti-IL-8 antibody comprising a VL complementarity determining region (LCDR) as described in Table 1F.

[0026]

[0026] In one aspect, a host cell comprising a vector containing a polynucleotide sequence encoding a heavy chain variable region (VH) of an anti-IL-8 antibody comprising a VH complementarity determining region (HCDR) described in Table 1F, and a light chain variable region (VL) of an anti-IL-8 antibody comprising a VL complementarity determining region (LCDR) described in Table 1F is disclosed in the present specification.

[0027]

[0027] In one aspect, disclosed herein is a method for producing an anti-IL-8 antibody having a complementarity determining region (CDR) sequence set forth in Table 1F, comprising culturing a host cell described in claim 25 under conditions that promote expression of the vector in the host cell, and expressing the polynucleotide sequence contained in the vector, thereby producing an anti-IL-8 antibody having a complementarity determining region (CDR) sequence set forth in Table 1F.

[0028]

[0028] In one aspect, disclosed herein is a method for inhibiting tumor or cancer formation or growth, or a combination thereof, in a subject in need thereof, the method comprising administering to the subject an anti-IL-8 antibody comprising an antibody antigen-binding domain comprising the heavy chain variable region (VH) and light chain variable region (VL), thereby inhibiting tumor formation or growth, or a combination thereof, in the subject, wherein the amino acid sequences of the VH-VL pairs are selected from the pair sequences set forth in SEQ ID NOs: 2 and 3, SEQ ID NOs: 4 and 5, SEQ ID NOs: 6 and 7, SEQ ID NOs: 8 and 9, SEQ ID NOs: 10 and 11, SEQ ID NOs: 12 and 13, SEQ ID NOs: 14 and 15, SEQ ID NOs: 16 and 17, SEQ ID NOs: 18 and 19, SEQ ID NOs: 20 and 21, SEQ ID NOs: 22 and 23, SEQ ID NOs: 24 and 25, SEQ ID NOs: 26 and 27, SEQ ID NOs: 28 and 29, SEQ ID NOs: 30 and 31, SEQ ID NOs: 32 and 33, and SEQ ID NOs: 34 and 35.

[0029] In a related aspect, the inhibitory method inhibits neutrophil or monocyte activation within the tumor microenvironment, or a combination thereof. In a further related aspect, the inhibition reduces neutrophil or monocyte activation within the tumor microenvironment, or a combination thereof. In another further related aspect, the inhibition reduces the viability of pre-cancerous stem cells or tumor cells. In another related aspect, the pre-cancerous stem cells comprise pre-leukemic stem cells. In yet another related aspect, the cancer or tumor comprises a hematological cancer. In a further related aspect, the hematological cancer comprises leukemia, lymphoma, myeloma, acute myeloid leukemia (AML), acute promyelocytic leukemia, erythroleukemia, mixed B-myelomonocytic leukemia, or myelodysplastic syndrome (MDS). In yet another related aspect, the cancer or tumor comprises a solid cancer or tumor. In a further related aspect, the solid cancer or tumor comprises sarcoma, osteosarcoma, squamous cell carcinoma of the head and neck, non-small cell lung cancer, bladder cancer, pancreatic cancer, or pancreatic ductal adenocarcinoma.

[0030]

[0030] In one aspect, disclosed herein is a method for inhibiting tumor or cancer formation or growth, or a combination thereof, in a subject in need thereof, the method comprising administering to the subject anti-IL-8 antibodies having complementarity determining region (CDR) sequences set out in Table 1F, thereby inhibiting tumor formation or growth, or a combination thereof, in the subject, wherein each antibody comprises a heavy chain variable region having heavy chain complementarity determining region (HCDR) 1, HCDR2 and HCDR3, and a light chain variable region having light chain complementarity determining region (LCDR) 1, LCDR2 and LCDR3, and the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3 for each of the antibodies comprise the amino acid sequences set out in Table 1F.

[0031] In a related embodiment, the inhibition inhibits neutrophil or monocyte activation within the tumor microenvironment, or a combination thereof. In another related embodiment, the inhibition inhibits neutrophil or monocyte activation within the tumor microenvironment, or a combination thereof. In a further related embodiment, the inhibition reduces the viability of pre-cancerous stem cells or tumor cells. In yet another related embodiment, the pre-cancerous stem cells comprise pre-leukemic stem cells. In yet another related embodiment, the cancer or tumor comprises a hematological cancer. In a further related embodiment, the hematological cancer comprises leukemia, lymphoma, myeloma, acute myeloid leukemia (AML), acute promyelocytic leukemia, erythroleukemia, mixed B-myelomonocytic leukemia, or myelodysplastic syndrome (MDS). In a further related embodiment, the cancer or tumor comprises a solid cancer or tumor. In a further related embodiment, the solid cancer or tumor comprises sarcoma, osteosarcoma, squamous cell carcinoma of the head and neck, non-small cell lung cancer, bladder cancer, pancreatic cancer, or pancreatic ductal adenocarcinoma.

[0032] In another embodiment relating to tumor or cancer formation or growth, the subject is a human.

[0033]

[0033] In one aspect, disclosed herein is a method for treating a subject suffering from a disease, comprising administering to the subject an anti-IL-8 antibody comprising an antibody antigen-binding domain comprising a heavy chain variable region (VH) and a light chain variable region (VL), thereby treating the disease in the subject, wherein the disease comprises cancer or a tumor or a disease associated with viral infection or inflammation, or a combination thereof, and the amino acid sequence of the VH-VL pair is selected from the pair sequences set forth in SEQ ID NOs: 2 and 3, SEQ ID NOs: 4 and 5, SEQ ID NOs: 6 and 7, SEQ ID NOs: 8 and 9, SEQ ID NOs: 10 and 11, SEQ ID NOs: 12 and 13, SEQ ID NOs: 14 and 15, SEQ ID NOs: 16 and 17, SEQ ID NOs: 18 and 19, SEQ ID NOs: 20 and 21, SEQ ID NOs: 22 and 23, SEQ ID NOs: 24 and 25, SEQ ID NOs: 26 and 27, SEQ ID NOs: 28 and 29, SEQ ID NOs: 30 and 31, SEQ ID NOs: 32 and 33, and SEQ ID NOs: 34 and 35.

[0034] In a related embodiment, the cancer or tumor comprises a hematological cancer. In a further related embodiment, the hematological cancer comprises leukemia, lymphoma, myeloma, acute myeloid leukemia (AML), acute promyelocytic leukemia, erythroleukemia, mixed B-myelomonocytic leukemia, or myelodysplastic syndrome (MDS). In another related embodiment, the cancer or tumor comprises a solid cancer or tumor. In a further related embodiment, the solid cancer or tumor comprises a sarcoma, osteosarcoma, squamous cell carcinoma of the head and neck, non-small cell lung cancer, bladder cancer, pancreatic cancer, or pancreatic ductal adenocarcinoma.

[0035]

[0035] In one aspect, disclosed herein is a method for treating a subject suffering from a disease, comprising administering to the subject an anti-IL-8 antibody having a complementarity determining region (CDR) sequence as set forth in Table 1F, thereby treating the disease in the subject, wherein the disease comprises cancer or a tumor or a disease associated with a viral infection or inflammation, or a combination thereof, and each antibody comprises a heavy chain variable region having heavy chain complementarity determining region (HCDR) 1, HCDR2 and HCDR3, and a light chain variable region having light chain complementarity determining region (LCDR) 1, LCDR2 and LCDR3, and wherein the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3 for each of the antibodies comprise the amino acid sequences set forth in Table 1F.

[0036] In a related embodiment, the cancer or tumor comprises a hematological cancer. In a further related embodiment, the hematological cancer comprises leukemia, lymphoma, myeloma, acute myeloid leukemia (AML), acute promyelocytic leukemia, erythroleukemia, mixed B-myelomonocytic leukemia, or myelodysplastic syndrome (MDS). In another related embodiment, the cancer or tumor comprises a solid cancer or tumor. In a further related embodiment, the solid cancer or tumor comprises a sarcoma, osteosarcoma, squamous cell carcinoma of the head and neck, non-small cell lung cancer, bladder cancer, pancreatic cancer, or pancreatic ductal adenocarcinoma.

[0037] In embodiments relating to methods of treating a subject suffering from a disease, the subject is a human.

[0038] In one aspect, there is provided a method of inhibiting tumor or cancer formation or growth, or a combination thereof, in a subject in need thereof, said method comprising administering to said subject a polynucleotide encoding an anti-IL-8 antibody comprising an antibody antigen-binding domain comprising a heavy chain variable region (VH) and a light chain variable region (VL), thereby inhibiting tumor formation or growth, or a combination thereof, in said subject, wherein the polynucleotide sequences encoding the VH-VL pair are set forth in SEQ ID NOs: 113 and 114, SEQ ID NOs: 115 and 116, Disclosed herein are methods in which the paired sequences are selected from the paired sequences set forth in SEQ ID NOs: 6, 117 and 118, 119 and 120, 121 and 122, 123 and 124, 125 and 126, 127 and 128, 129 and 130, 131 and 132, 133 and 134, 135 and 136, 137 and 138, 139 and 140, 141 and 142, 36 and 37, and 38 and 39. In a related embodiment, the inhibition inhibits neutrophil or monocyte activation within the tumor microenvironment, or a combination thereof. In another related embodiment, the inhibition reduces neutrophil or monocyte activation within the tumor microenvironment, or a combination thereof. In yet another related embodiment, the inhibition reduces the viability of pre-cancerous stem cells or tumor cells. In another related embodiment, the pre-cancerous stem cells include pre-leukemic stem cells. In further related embodiments, the cancer or tumor comprises a hematological cancer. In yet other related embodiments, the hematological cancer comprises leukemia, lymphoma, myeloma, acute myeloid leukemia (AML), acute promyelocytic leukemia, erythroleukemia, mixed B-myelomonocytic leukemia, or myelodysplastic syndrome (MDS). In further related embodiments, the cancer or tumor comprises a solid cancer or tumor. In yet other related embodiments, the solid cancer or tumor comprises a sarcoma, osteosarcoma, squamous cell carcinoma of the head and neck, non-small cell lung cancer, bladder cancer, pancreatic cancer, or pancreatic ductal adenocarcinoma.

[0039] In one aspect, a method of treating a subject suffering from a disease comprises administering to the subject an anti-IL-8 antibody comprising an antibody antigen-binding domain comprising a heavy chain variable region (VH) and a light chain variable region (VL), thereby treating the disease in the subject, wherein the disease comprises cancer or tumor or a disease associated with viral infection or inflammation, or a combination thereof, and wherein the polynucleotide sequences encoding the VH-VL pair are selected from SEQ ID NOs: 113 and 114, SEQ ID NOs: 115 and 116, SEQ ID NO: 11 Disclosed herein is a method in which the paired sequences are selected from the paired sequences set forth in SEQ ID NOs: 7 and 118, SEQ ID NOs: 119 and 120, SEQ ID NOs: 121 and 122, SEQ ID NOs: 123 and 124, SEQ ID NOs: 125 and 126, SEQ ID NOs: 127 and 128, SEQ ID NOs: 129 and 130, SEQ ID NOs: 131 and 132, SEQ ID NOs: 133 and 134, SEQ ID NOs: 135 and 136, SEQ ID NOs: 137 and 138, SEQ ID NOs: 139 and 140, SEQ ID NOs: 141 and 142, SEQ ID NOs: 36 and 37, and SEQ ID NOs: 38 and 39.

[0040] In a related embodiment for a method of treating a subject afflicted with a disease, the cancer or tumor comprises a hematological cancer. In a further related embodiment, the hematological cancer comprises leukemia, lymphoma, myeloma, acute myeloid leukemia (AML), acute promyelocytic leukemia, erythroleukemia, mixed B-myelomonocytic leukemia, or myelodysplastic syndrome (MDS). In another related embodiment for a method of treating a subject afflicted with a disease, the cancer or tumor comprises a solid cancer or tumor. In a further related embodiment, the solid cancer or tumor comprises sarcoma, osteosarcoma, squamous cell carcinoma of the head and neck, non-small cell lung cancer, bladder cancer, pancreatic cancer, or pancreatic ductal adenocarcinoma.

[0041] In another related aspect of the methods disclosed herein, the subject is a human.

[0042] BRIEF DESCRIPTION OF THE DRAWINGS The subject matter which is regarded as the invention is particularly pointed out and distinctly claimed in the concluding portion of this specification. However, the invention, both as to organization and method of operation, together with its objects, features, and advantages, may best be understood by reference to the following detailed description when read in connection with the accompanying drawings. [Brief explanation of the drawings]

[0043] [Figure 1] mAb Expression Analysis: The expression of antibody molecules was examined under non-reducing and reducing conditions. [Figure 2]

[0044] Size exclusion analysis of mAbs. Size exclusion analysis was performed on HPLC. [Figure 3]

[0045] Binding of mAbs to human IL-8 by ELISA. The binding kinetics of mAbs to human IL-8 were measured by cell-free ELISA. [Figure 4]

[0046] Blocking of IL-8 binding to its cell surface receptor, CXCR2. Blocking of IL-8 binding to the CXCR2 receptor by mAbs was measured in a cell-based assay by flow cytometry. [Figure 5]

[0047] Inhibition of IL-8-induced NFκB activity by mAbs. Blocking of IL-8-induced NFκB activity by mAbs was measured in a HEK293-CXCR1 cell-based reporter assay. [Figure 6]

[0048] Cell surface expression of CXCR1 and CXCR2 in cell lines by IHC. Expression of CXCR1 and CXCR2 receptors was detected at various levels in osteosarcoma and pancreatic tumor cell lines by IHC. Photomicrograph images show that osteosarcoma cell lines (143B, SaOS-2, OS17) and pancreatic cancer cell line (PANC-1) expressed both CXCR2 and CXCR1 receptors. [Figure 7]

[0049] Cell line IL-8 secretion by ELISA. IL-8 secretion was monitored in the supernatants of MG-63, U2-OS, SAOS-2 and 143-B osteosarcoma cells by ELISA. [Figure 8]

[0050] Inhibition of IL-8-induced CXCR2 internalization by mAb. CXCR2 receptor internalization was monitored by flow cytometry. In THP-1 cells, CXCR2 receptor internalization was induced by IL-8 treatment, whereas CXCR2 internalization was dose-dependently blocked by co-treatment with mAb. [Figure 9]

[0051] Inhibition of IL-8 signaling in human neutrophils by STLX18 antibody. STLX18 monoclonal antibody inhibited IL-8-induced downstream signaling in human neutrophils as monitored by Western blot using phospho-specific antibodies against ERK and AKT. Actin was used as a loading control. [Figure 10]

[0052] Neutrophil migration toward IL8 was inhibited by STLX18 antibody. Human neutrophil migration (toward an IL8 source) across a porous membrane was inhibited by STLX18 antibody. [Figure 11A]

[0053] Monosodium urate (MSU)-induced knee swelling inhibited by STLX18 antibody. Treatment with STLX18 antibody reduced swelling in MSU-injected joints (FIG. 11B), while saline-injected control joints did not change size (FIG. 11A), indicating that STLX18 had no effect on control knee diameter. [Figure 11B] Monosodium urate (MSU)-induced knee swelling inhibited by STLX18 antibody. Treatment with STLX18 antibody reduced swelling in MSU-injected joints (FIG. 11B), while saline-injected control joints did not change size (FIG. 11A), demonstrating that STLX18 had no effect on control knee diameter. DETAILED DESCRIPTION OF THE INVENTION

[0044] Detailed Description

[0054] In the following detailed description, numerous specific details are set forth to provide a thorough understanding of the anti-IL-8 monoclonal antibodies (mAbs) described and exemplified herein, and their therapeutic uses. However, it will be understood by those skilled in the art that the production and use of IL-8-binding anti-IL-8 antibodies may, in some instances, be practiced without these specific details. In other instances, well-known methods, procedures, and components have not been described in detail so as not to obscure the disclosure provided herein.

[0045]

[0055] IL-8 is a potent proinflammatory cytokine secreted by various cells, primarily macrophages. It binds to the seven-transmembrane GPCRs, CXCR1 and CXCR2, on the cell surface of neutrophils and monocytes, influencing their activation and migration to sites of infection. Accumulating evidence suggests a role for IL-8 signaling in cancer, both hematological malignancies and solid tumors.

[0046]

[0056] Disclosed herein are humanized IL-8 binding mAbs or fragments thereof. In certain embodiments, these anti-IL-8 mAbs or fragments thereof can be used in therapeutic methods for inhibiting tumorigenesis or tumor growth, or a combination thereof. In certain embodiments, these anti-IL-8 mAbs or fragments thereof can be used in therapeutic methods for treating subjects suffering from diseases such as cancer or tumors, or viral infections.

[0047]

[0057] Throughout this application, various references or publications are cited. The disclosures of these references and publications in their entireties are hereby incorporated by reference into this application in order to more fully describe the state of the art to which this invention pertains.

[0048]

[0058] As used herein, the term "antibody" may be used synonymously with the term "immunoglobulin" or "IgG," all of which have the same quality and meaning. An antibody binding domain or antigen-binding site may be a fragment of an antibody or a genetically engineered product of one or more fragments of an antibody, which fragment is responsible for specific binding to a target antigen. "Specific binding" means that the binding is selective for the antigen of interest and can be distinguished from undesired or non-specific interactions. For example, an antibody may have an equilibrium dissociation constant of ≦10 -5 , 10 -6 , or 10 -7 M. In one embodiment, the equilibrium dissociation constant is ≦10 -8 M or 10 -9 In certain further embodiments, the equilibrium dissociation constant may be ≦10 -10 M, 10 -11 M or 10 -12 In some embodiments, the equilibrium dissociation constant is ≦10 -5 M~10 -12 M may be in the range.

[0049]

[0059] As used herein, the term "antibody" encompasses one or more antibody fragments that retain binding specificity, including, but not limited to, IgG, heavy chain variable region (VH), light chain variable region (VL), Fab fragment, F(ab')2 fragment, scFv fragment, Fv fragment, multispecific antibody, bispecific antibody, triabody, single-chain antibody, heavy chain antibody, nanobody, minibody, diabody, triabody, tetrabody, and single-domain antibody (see, e.g., Hudson and Souriau, Nature Med. 9:129-134 (2003)). Humanized, primatized, and chimeric antibodies are also encompassed, as these terms are commonly understood in the art. In certain embodiments, the antibodies disclosed herein are engineered, i.e., non-naturally occurring antibodies that are designed to bind IL-8 and provide a particular functional activity.

[0050]

[0060] In certain embodiments, the antibody comprises a heavy chain constant region such as an IgG1, IgG2, IgG3, IgG4, IgA, IgE, IgM, or IgD constant region. In some embodiments, the heavy chain constant region is an IgG1 heavy chain constant region. In some embodiments, the antibody comprises a light chain constant region, either a kappa light chain constant region or a lambda light chain constant region. In some embodiments, the antibody lacks a constant region, e.g., a Fab fragment or a single-chain Fv fragment.

[0051]

[0061] As used herein, the term "heavy chain variable region" may be used interchangeably with the terms "VH domain" or "VH region" or "VH", all of which have the same meaning and qualities. As used herein, the term "light chain variable region" may be used interchangeably with the terms "VL domain" or "VL region" or "VL", all of which have the same meaning and qualities.

[0052]

[0062] Those skilled in the art will recognize that a "heavy chain variable region" or "VH" in the context of an antibody encompasses a fragment of a heavy chain containing three complementarity-determining regions (CDRs) interposed between adjacent stretches known as framework regions. Framework regions are more highly conserved than the CDRs and form a scaffold to support the CDRs. Similarly, those skilled in the art will also recognize that a "light chain variable region" or "VL" in the context of an antibody encompasses a fragment of a light chain containing three CDRs interposed between framework regions.

[0053]

[0063] As used herein, the term "complementarity-determining region" or "CDR" refers to the hypervariable region of a heavy or light chain variable region. Proceeding from the N-terminus, each heavy or light chain polypeptide has three CDRs, designated "CDR1," "CDR2," and "CDR3." Crystallographic analysis of several antigen-antibody complexes has demonstrated that the amino acid residues of the CDRs form extensive contacts with the bound antigen, with the most extensive antigen contact being with the heavy chain CDR3. Thus, the CDR regions are primarily responsible for the specificity of the antigen-binding site. In one embodiment, the antigen-binding site contains six CDRs, including the CDRs from each of the heavy and light chain variable regions. An example of a set of six CDRs contained within an IL-8 antigen-binding site is shown in Table 1F (see Example 2).

[0054]

[0064] As used herein, the term "framework region" or "FR" refers to the four adjacent amino acid sequences that constitute the CDRs of a heavy or light chain variable region. Although some FR residues may contact the bound antigen, FR residues are primarily involved in folding the variable region into the antigen-binding site. In certain embodiments, the FR residues involved in folding the variable region include residues directly adjacent to the CDR. Within the FRs, certain amino acid residues and certain structural features are highly conserved. In this regard, all variable region sequences contain an internal disulfide loop of approximately 90 amino acid residues. When the variable region folds into the antigen-binding site, the CDRs are presented as protruding loop motifs that form the antigen-binding surface. It is generally recognized that there are conserved structural regions of the FRs that influence the shape of the folded CDR loops into specific "canonical" structures, regardless of the exact CDR amino acid sequence. Furthermore, certain FR residues are known to participate in contacts between non-covalent domains that stabilize the interaction of antibody heavy and light chains.

[0055]

[0065] Wu and Kabat (Tai Te Wu, Elvin A. Kabat. An analysis of the sequences of the variable regions of Benedict Jones proteins and myeloma light chains and their implications for antibody complementarity. Journal of Experimental Medicine, 132, 2, 8 (1970); Kabat EA, Wu TT, Bilofsky H, Reid-Miller M, Perry H. Sequence of proteins of immunological interest. Bethesda: National Institutes of Health; 1983. 323 (1983)) developed the alignment of antibody peptide sequences, and their contribution to this was severalfold: First, by studying sequence similarities between variable domains, they identified corresponding residues that are more or less homologous among all antibodies in all vertebrate species, so long as they adopt a similar three-dimensional structure, play a similar functional role, interact similarly with neighboring residues, and exist in a similar chemical environment. Second, they devised a peptide sequence numbering system in which homologous immunoglobulin residues are assigned the same position number. Those skilled in the art can unambiguously assign what is generally referred to herein as Kabat numbering to any variable domain sequence without relying on experimental data beyond the sequence itself. Third, Kabat and Wu calculated the variability for each Kabat-numbered sequence position, which represents the discovery of several or many possible amino acids when variable domain sequences are aligned. They identified three contiguous regions of high variability embedded within four contiguous regions of low variability. Kabat and Wu formally defined the residues that make up these variable pathways and designated these "complementarity-determining regions" (CDRs), which display chemical complementarity between antibody and antigen. The role in the three-dimensional folding of the variable domain, rather than in antigen recognition, was attributed to the remaining low-variability regions, referred to herein as "framework regions."Fourth, Kabat and Wu established a public database of antibody peptide and nucleic acid sequences that continues to be maintained and is well known to those skilled in the art.

[0056]

[0066] Chothia and colleagues (Cyrus Chothia, Arthur M. Lesk. Canonical structures for the hypervariable regions of immunoglobulins. Journal of Molecular Biology, 196, 4, 8 (1987)) found that certain subportions within the Kabat CDRs adopt nearly identical peptide backbone structures despite high diversity at the amino acid sequence level. These subportions are designated L1, L2, and L3 or H1, H2, and H3, where "L" and "H" represent the light chain and heavy chain regions, respectively. These regions are sometimes referred to as Chothia CDRs, which have overlapping boundaries with the Kabat CDRs.

[0057]

[0067] More recent studies have shown that nearly all antibody-binding residues fall within regions of structural consensus (Kunik, V. et al., PloS Computational Biology 8(2):el002388 (February 2012)). In certain embodiments, these regions are referred to as antibody-binding regions. It has been shown that these regions can also be identified from antibody sequences.

[0058]

[0068] IMGT® is the international ImMunoGeneTics information system® (see Nucleic Acids Res. 2015 Jan;43(Database issue):D413-22. doi:10.1093 / nar / gku1056. Epub 2014 Nov 5 Free article. PMID:25378316 LIGM:441 and Dev Comp Immunol. 2003 Jan;27(1):55-77). IMGT is a proprietary numbering system for immunoglobulin and T-cell receptor variable domains and Ig superfamily V-like domains (Lefranc et al., Dev Comp Immunol. 27:55-77(2003)). IMGT® is a unified numbering system for IG and TcR variable domain sequences based on the alignment of five or more IG and TcR variable region sequences, taking into account and combining the Kabat definitions of FRs and CDRs, structural data, and Chothia characterization of hypervariable loops. IMGT is well known in the art to be considered a universal numbering scheme for antibodies.

[0059]

[0069] In one embodiment, the identification of the CDR regions uses the IMGT analysis system. In one embodiment, the identification of the CDR regions uses a Kabat-based analysis system.

[0060]

[0070] Antigen-binding sequences are typically located within the heavy and light chain variable regions of antibodies. Antibodies can exist in various forms or with various domains, including but not limited to complementarity-determining regions (CDRs), variable regions (Fv), VH domains, VL domains, single-chain variable regions (scFv), and Fab fragments.

[0061]

[0071] Those skilled in the art will appreciate that an scFv is a fusion polypeptide comprising the variable heavy (VH) and variable light (VL) regions of an immunoglobulin linked by a short linker peptide, which linker can have, for example, from 10 to about 25 amino acids.

[0062]

[0072] Those skilled in the art will also understand that the term "Fab" in reference to an antibody generally encompasses the portion of an antibody consisting of a single light chain (both variable and constant regions) linked by disulfide bonds to the variable region and first constant region of a single heavy chain, while F(ab')2 includes a fragment of a heavy chain containing a VH domain and a light chain containing a VL domain.

[0063]

[0073] In some embodiments, antibodies include whole antibody molecules, including monoclonal (mAb) and polyclonal antibodies. In some embodiments, antibodies include one or more antibody fragments that retain binding specificity, including, but not limited to, variable heavy chain (VH) fragments, variable light chain (VL) fragments, Fab fragments, F(ab')2 fragments, scFv fragments, Fv fragments, minibodies, diabodies, triabodies, and tetrabodies.

[0064] Anti-IL-8 antibody

[0074] Described herein in one embodiment is an isolated anti-IL-8 antibody comprising an antibody antigen-binding domain comprising a heavy chain variable region (VH) and a light chain variable region (VL), wherein the amino acid sequence of the VH-VL pair is selected from the pair sequences set forth in SEQ ID NOs: 2 and 3, 4 and 5, 6 and 7, 8 and 9, 10 and 11, 12 and 13, 14 and 15, 16 and 17, 18 and 19, 20 and 21, 22 and 23, 24 and 25, 26 and 27, 28 and 29, 30 and 31, 32 and 33, and 34 and 35. In one embodiment, the amino acid sequence of the VH-VL pair is set forth in SEQ ID NOs: 2 and 3. In one embodiment, the amino acid sequence of the VH-VL pair is set forth in SEQ ID NOs: 4 and 5. In one embodiment, the amino acid sequence of the VH-VL pair is set forth in SEQ ID NOs: 6 and 7. In some embodiments, the amino acid sequences of the VH-VL pair are set forth in SEQ ID NOs: 8 and 9. In some embodiments, the amino acid sequences of the VH-VL pair are set forth in SEQ ID NOs: 10 and 11. In some embodiments, the amino acid sequences of the VH-VL pair are set forth in SEQ ID NOs: 12 and 13. In some embodiments, the amino acid sequences of the VH-VL pair are set forth in SEQ ID NOs: 14 and 15. In some embodiments, the amino acid sequences of the VH-VL pair are set forth in SEQ ID NOs: 16 and 17. In some embodiments, the amino acid sequences of the VH-VL pair are set forth in SEQ ID NOs: 18 and 19. In some embodiments, the amino acid sequences of the VH-VL pair are set forth in SEQ ID NOs: 20 and 21. In some embodiments, the amino acid sequences of the VH-VL pair are set forth in SEQ ID NOs: 22 and 23. In some embodiments, the amino acid sequences of the VH-VL pair are set forth in SEQ ID NOs: 24 and 25. In some embodiments, the amino acid sequences of the VH-VL pair are set forth in SEQ ID NOs: 26 and 27. In one embodiment, the amino acid sequences of the VH-VL pair are set forth in SEQ ID NOs: 28 and 29. In one embodiment, the amino acid sequences of the VH-VL pair are set forth in SEQ ID NOs: 30 and 31. In one embodiment, the amino acid sequences of the VH-VL pair are set forth in SEQ ID NOs: 32 and 33.In one embodiment, the amino acid sequences of the VH-VL pair are set forth in SEQ ID NOs:34 and 35.

[0065]

[0075] In certain embodiments, the VH and / or VL amino acid sequence is selected from homologs having at least 80% identity to any of the VH or VL sequences set forth in SEQ ID NOs: 2 and 3, SEQ ID NOs: 4 and 5, SEQ ID NOs: 6 and 7, SEQ ID NOs: 8 and 9, SEQ ID NOs: 10 and 11, SEQ ID NOs: 12 and 13, SEQ ID NOs: 14 and 15, SEQ ID NOs: 16 and 17, SEQ ID NOs: 18 and 19, SEQ ID NOs: 20 and 21, SEQ ID NOs: 22 and 23, SEQ ID NOs: 24 and 25, SEQ ID NOs: 26 and 27, SEQ ID NOs: 28 and 29, SEQ ID NOs: 30 and 31, SEQ ID NOs: 32 and 33, and SEQ ID NOs: 34 and 35. In certain embodiments, the VH or VL amino acid sequence is selected from homologs having at least 80% identity to the VH and / or VL sequences set forth in SEQ ID NOs: 2 and 3. In certain embodiments, the VH or VL amino acid sequence is selected from homologs having at least 80% identity to the VH and / or VL sequences set forth in SEQ ID NOs: 4 and 5. In some embodiments, the VH or VL amino acid sequence is selected from homologs having at least 80% identity to the VH and / or VL sequences set forth in SEQ ID NOs: 6 and 7. In some embodiments, the VH or VL amino acid sequence is selected from homologs having at least 80% identity to the VH and / or VL sequences set forth in SEQ ID NOs: 8 and 9. In some embodiments, the VH or VL amino acid sequence is selected from homologs having at least 80% identity to the VH and / or VL sequences set forth in SEQ ID NOs: 10 and 11. In some embodiments, the VH or VL amino acid sequence is selected from homologs having at least 80% identity to the VH and / or VL sequences set forth in SEQ ID NOs: 12 and 13. In some embodiments, the VH or VL amino acid sequence is selected from homologs having at least 80% identity to the VH and / or VL sequences set forth in SEQ ID NOs: 14 and 15. In one embodiment, the VH or VL amino acid sequence is selected from homologs having at least 80% identity to the VH and / or VL sequences set forth in SEQ ID NOs: 16 and 17. In one embodiment, the VH or VL amino acid sequence is selected from homologs having at least 80% identity to the VH and / or VL sequences set forth in SEQ ID NOs: 18 and 19.In some embodiments, the VH or VL amino acid sequence is selected from homologs having at least 80% identity to the VH and / or VL sequences set forth in SEQ ID NOs: 20 and 21. In some embodiments, the VH or VL amino acid sequence is selected from homologs having at least 80% identity to the VH and / or VL sequences set forth in SEQ ID NOs: 22 and 23. In some embodiments, the VH or VL amino acid sequence is selected from homologs having at least 80% identity to the VH and / or VL sequences set forth in SEQ ID NOs: 24 and 25. In some embodiments, the VH or VL amino acid sequence is selected from homologs having at least 80% identity to the VH and / or VL sequences set forth in SEQ ID NOs: 26 and 27. In some embodiments, the VH or VL amino acid sequence is selected from homologs having at least 80% identity to the VH and / or VL sequences set forth in SEQ ID NOs: 28 and 29. In one embodiment, the VH or VL amino acid sequence is selected from homologs having at least 80% identity to the VH and / or VL sequences set forth in SEQ ID NOs: 30 and 31. In one embodiment, the VH or VL amino acid sequence is selected from homologs having at least 80% identity to the VH and / or VL sequences set forth in SEQ ID NOs: 32 and 33. In one embodiment, the VH or VL amino acid sequence is selected from homologs having at least 80% identity to the VH and / or VL sequences set forth in SEQ ID NOs: 34 and 35.

[0066]

[0076] In some embodiments, the homologs share 100% sequence identity within the three VH CDRs (HCDR1, HCDR2, HCDR3). In some embodiments, the homologs share 100% sequence identity within the three VL CDRs LCDR1, LCDR2, and LCDR3. In some embodiments, the VH amino acid sequence is selected from homologs having at least 80% identity to any of the sequences set forth in SEQ ID NOs: 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, and 34. In some embodiments, the VH amino acid sequence comprises a homolog having at least 80% identity to the sequence set forth in SEQ ID NO: 2. In some embodiments, the VH amino acid sequence comprises a homolog having at least 80% identity to the sequence set forth in SEQ ID NO: 4. In some embodiments, the VH amino acid sequence comprises a homolog having at least 80% identity to the sequence set forth in SEQ ID NO: 6. In one embodiment, the amino acid sequence of the VH comprises a homologue having at least 80% identity to the sequence set forth in SEQ ID NO:8. In one embodiment, the amino acid sequence of the VH comprises a homologue having at least 80% identity to the sequence set forth in SEQ ID NO:10. In one embodiment, the amino acid sequence of the VH comprises a homologue having at least 80% identity to the sequence set forth in SEQ ID NO:12. In one embodiment, the amino acid sequence of the VH comprises a homologue having at least 80% identity to the sequence set forth in SEQ ID NO:14. In one embodiment, the amino acid sequence of the VH comprises a homologue having at least 80% identity to the sequence set forth in SEQ ID NO:16. In one embodiment, the amino acid sequence of the VH comprises a homologue having at least 80% identity to the sequence set forth in SEQ ID NO:18. In one embodiment, the amino acid sequence of the VH comprises a homologue having at least 80% identity to the sequence set forth in SEQ ID NO:20. In one embodiment, the amino acid sequence of the VH comprises a homologue having at least 80% identity to the sequence set forth in SEQ ID NO:22. In one embodiment, the amino acid sequence of VH comprises a homolog having at least 80% identity to the sequence set forth in SEQ ID NO:24.In some embodiments, the amino acid sequence of the VH comprises a homolog having at least 80% identity to the sequence set forth in SEQ ID NO: 26. In some embodiments, the amino acid sequence of the VH comprises a homolog having at least 80% identity to the sequence set forth in SEQ ID NO: 28. In some embodiments, the amino acid sequence of the VH comprises a homolog having at least 80% identity to the sequence set forth in SEQ ID NO: 30. In some embodiments, the amino acid sequence of the VH comprises a homolog having at least 80% identity to the sequence set forth in SEQ ID NO: 32. In some embodiments, the amino acid sequence of the VH comprises a homolog having at least 80% identity to the sequence set forth in SEQ ID NO: 34. In some embodiments, the amino acid sequence of the VL is selected from a homolog having at least 80% identity to any of the sequences set forth in SEQ ID NOs: 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25 27, 29, 31, 33, and 35. In some embodiments, the amino acid sequence of the VL comprises a homologue having at least 80% identity to the sequence set forth in SEQ ID NO:3. In some embodiments, the amino acid sequence of the VL comprises a homologue having at least 80% identity to the sequence set forth in SEQ ID NO:5. In some embodiments, the amino acid sequence of the VL comprises a homologue having at least 80% identity to the sequence set forth in SEQ ID NO:7. In some embodiments, the amino acid sequence of the VL comprises a homologue having at least 80% identity to the sequence set forth in SEQ ID NO:9. In some embodiments, the amino acid sequence of the VL comprises a homologue having at least 80% identity to the sequence set forth in SEQ ID NO:11. In some embodiments, the amino acid sequence of the VL comprises a homologue having at least 80% identity to the sequence set forth in SEQ ID NO:13. In some embodiments, the amino acid sequence of the VL comprises a homologue having at least 80% identity to the sequence set forth in SEQ ID NO:15. In some embodiments, the amino acid sequence of the VL comprises a homologue having at least 80% identity to the sequence set forth in SEQ ID NO:17. In one embodiment, the amino acid sequence of the VL comprises a homolog having at least 80% identity to the sequence set forth in SEQ ID NO:19.In some embodiments, the amino acid sequence of the VL comprises a homologue having at least 80% identity to the sequence set forth in SEQ ID NO:21. In some embodiments, the amino acid sequence of the VL comprises a homologue having at least 80% identity to the sequence set forth in SEQ ID NO:23. In some embodiments, the amino acid sequence of the VL comprises a homologue having at least 80% identity to the sequence set forth in SEQ ID NO:25. In some embodiments, the amino acid sequence of the VL comprises a homologue having at least 80% identity to the sequence set forth in SEQ ID NO:27. In some embodiments, the amino acid sequence of the VL comprises a homologue having at least 80% identity to the sequence set forth in SEQ ID NO:29. In some embodiments, the amino acid sequence of the VL comprises a homologue having at least 80% identity to the sequence set forth in SEQ ID NO:31. In some embodiments, the amino acid sequence of the VL comprises a homologue having at least 80% identity to the sequence set forth in SEQ ID NO:33. In some embodiments, the amino acid sequence of the VL comprises a homologue having at least 80% identity to the sequence set forth in SEQ ID NO:35. The variable chain region (VH or VL) and each CDR within each of these VH or VL regions can be determined based on Tables 1A-1F as provided in Example 2.

[0067]

[0077] In certain embodiments, a VH homolog has at least 85% identity, at least 87%, at least 89%, at least 91%, at least 93%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to a VH polypeptide described herein when determined using BlastP software from the National Center of Biotechnology Information (NCBI) with default parameters. In certain embodiments, a VL homolog has at least 85% identity, at least 87%, at least 89%, at least 91%, at least 93%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to a VL polypeptide described herein when determined using BlastP software from the National Center of Biotechnology Information (NCBI) with default parameters. In certain embodiments, a VH homolog has at least 85% identity, at least 87%, at least 89%, at least 91%, at least 93%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to a VH polypeptide described herein, and a VL homolog has at least 85% identity, at least 87%, at least 89%, at least 91%, at least 93%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to a VL polypeptide described herein as determined using BlastP software from the National Center of Biotechnology Information (NCBI) using default parameters.

[0068]

[0078] Those skilled in the art will understand that the term "homology" and its grammatical forms encompass the degree of similarity between two or more structures. The term "homologous sequence" refers to regions having similar order of monomers in a macromolecule.

[0069]

[0079] The term "homology," when used in reference to polypeptide (or protein) sequences, refers to the degree of similarity between two or more polypeptide (or protein) sequences (e.g., genes) or fragments thereof. Typically, the degree of similarity between two or more polypeptide (or protein) sequences refers to the similarity in the composition, order, or arrangement of two or more amino acids of the two or more polypeptides (or proteins). The two or more polypeptides (or proteins) can be from the same or different species or groups. The term "percent homology," when used in reference to polypeptide (or protein) sequences, generally refers to the percent similarity between the amino acid sequences of two or more polypeptide (or protein) sequences. The terms "homologous polypeptide" and "homologous protein," respectively, generally refer to polypeptides or proteins having similar amino acid sequences and functions. Such homologous polypeptides or proteins may be related by having similar amino acid sequences and functions, but may be derived from or originate from different or the same species, or may be synthesized using recombinant genetic techniques well known to those skilled in the art.

[0070]

[0080]

[0009] Described herein are isolated anti-IL-8 antibodies comprising an antibody antigen-binding domain comprising a heavy chain variable region (VH) and a light chain variable region (VL), wherein the heavy chain variable region comprises complementarity determining region 1 (HCDR1), HCDR2, and HCDR3, and the light chain variable region has complementarity determining region 1 (LCDR1), LCDR2, and LCDR3. In one embodiment, the isolated anti-IL-8 antibodies comprise the complementarity determining region (CDR) sequences set forth in Table 1F, wherein each antibody comprises a heavy chain variable region having heavy chain complementarity determining region (HCDR)1, HCDR2, and HCDR3, and a light chain variable region having light chain complementarity determining region (LCDR)1, LCDR2, and LCDR3, and the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 for each of the antibodies comprise the amino acid sequences set forth in Table 1F. In one embodiment, the isolated anti-IL-8 antibodies comprise the CDR sequences set forth in Table 1F, each antibody comprising a heavy chain variable region having heavy chain complementarity determining region (HCDR) 1, HCDR2, and HCDR3, and a light chain variable region having light chain complementarity determining region (LCDR) 1, LCDR2, and LCDR3, wherein the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 for the antibodies are (a) SEQ ID NO:41, SEQ ID NO:44, SEQ ID NO:53, SEQ ID NO:60, SEQ ID NO:67, and SEQ ID NO:76; (b) SEQ ID NO:81, SEQ ID NO:84, SEQ ID NO:53, SEQ ID NO:93, SEQ ID NO:100, and SEQ ID NO:109; (c) SEQ ID NO:40, SEQ ID NO:43, SEQ ID NO:50, SEQ ID NO:57, SEQ ID NO:64, and SEQ ID NO:73; (d) SEQ ID NO:40, SEQ ID NO:43, SEQ ID NO:51, SEQ ID NO:58, SEQ ID NO:65, and SEQ ID NO:74; (e) SEQ ID NO:41, SEQ ID NO:44, SEQ ID NO:52, SEQ ID NO:59, SEQ ID NO:66, and SEQ ID NO:75; (f) SEQ ID NO:40, SEQ ID NO:43, SEQ ID NO:50, SEQ ID NO:57, SEQ ID NO:68, and SEQ ID NO:73; (g) SEQ ID NO:40, SEQ ID NO:43, SEQ ID NO:50, SEQ ID NO:57, SEQ ID NO:69, and SEQ ID NO:73; (h) SEQ ID NO:41, SEQ ID NO:45, SEQ ID NO:53, SEQ ID NO:61, SEQ ID NO:70, and SEQ ID NO:77; (i) SEQ ID NO:40, SEQ ID NO:46, SEQ ID NO:54, SEQ ID NO:62, SEQ ID NO:71, and SEQ ID NO:78; (j) SEQ ID NO:42, SEQ ID NO:47, SEQ ID NO:53, SEQ ID NO:63, SEQ ID NO:72, and SEQ ID NO:79; (k) SEQ ID NO:41, SEQ ID NO:48, SEQ ID NO:53, SEQ ID NO:60, SEQ ID NO:67, and SEQ ID NO:76; (l) SEQ ID NO:41, SEQ ID NO:49, SEQ ID NO:53, SEQ ID NO:60, SEQ ID NO:67, and SEQ ID NO:76; (m) SEQ ID NO:41, SEQ ID NO:44, SEQ ID NO:55, SEQ ID NO:60, SEQ ID NO:67, and SEQ ID NO:76; (n) SEQ ID NO:41, SEQ ID NO:44, SEQ ID NO:56, SEQ ID NO:60, SEQ ID NO:67, and SEQ ID NO:76; (o) SEQ ID NO:41, SEQ ID NO:48, SEQ ID NO:55, SEQ ID NO:60, SEQ ID NO:67, and SEQ ID NO:76; (p) SEQ ID NO:41, SEQ ID NO:48, SEQ ID NO:56, SEQ ID NO:60, SEQ ID NO:67, and SEQ ID NO:76; (q) SEQ ID NO:41, SEQ ID NO:49, SEQ ID NO:55, SEQ ID NO:60, SEQ ID NO:67, and SEQ ID NO:76; (r) SEQ ID NO:41, SEQ ID NO:49, SEQ ID NO:56, SEQ ID NO:60, SEQ ID NO:67, and SEQ ID NO:76; (s) SEQ ID NO: 80, SEQ ID NO: 83, SEQ ID NO: 50, SEQ ID NO: 90, SEQ ID NO: 97, and SEQ ID NO: 106; (t) SEQ ID NO:80, SEQ ID NO:83, SEQ ID NO:51, SEQ ID NO:91, SEQ ID NO:98, and SEQ ID NO:107; (u) SEQ ID NO:81, SEQ ID NO:84, SEQ ID NO:52, SEQ ID NO:92, SEQ ID NO:99, and SEQ ID NO:108; (v) SEQ ID NO:80, SEQ ID NO:83, SEQ ID NO:50, SEQ ID NO:90, SEQ ID NO:101, and SEQ ID NO:106; (w) SEQ ID NO:80, SEQ ID NO:83, SEQ ID NO:50, SEQ ID NO:90, SEQ ID NO:102, and SEQ ID NO:106; (x) SEQ ID NO:81, SEQ ID NO:85, SEQ ID NO:53, SEQ ID NO:94, SEQ ID NO:103, and SEQ ID NO:110; (y) SEQ ID NO:80, SEQ ID NO:86, SEQ ID NO:54, SEQ ID NO:95, SEQ ID NO:104, and SEQ ID NO:111; (z) SEQ ID NO:82, SEQ ID NO:87, SEQ ID NO:53, SEQ ID NO:93, SEQ ID NO:105, and SEQ ID NO:112; (aa) SEQ ID NO: 81, SEQ ID NO: 88, SEQ ID NO: 53, SEQ ID NO: 93, SEQ ID NO: 100, and SEQ ID NO: 109; (bb) SEQ ID NO:81, SEQ ID NO:89, SEQ ID NO:53, SEQ ID NO:93, SEQ ID NO:100, and SEQ ID NO:109; (cc) SEQ ID NO:81, SEQ ID NO:84, SEQ ID NO:55, SEQ ID NO:93, SEQ ID NO:100, and SEQ ID NO:109; (dd) SEQ ID NO:81, SEQ ID NO:84, SEQ ID NO:56, SEQ ID NO:93, SEQ ID NO:100, and SEQ ID NO:109; (ee) SEQ ID NO:81, SEQ ID NO:88, SEQ ID NO:55, SEQ ID NO:93, SEQ ID NO:100, and SEQ ID NO:109; (ff) SEQ ID NO:81, SEQ ID NO:88, SEQ ID NO:56, SEQ ID NO:93, SEQ ID NO:100, and SEQ ID NO:109; (gg) SEQ ID NO:81, SEQ ID NO:89, SEQ ID NO:55, SEQ ID NO:93, SEQ ID NO:100, and SEQ ID NO:109; or (hh) SEQ ID NO: 81, SEQ ID NO: 89, SEQ ID NO: 56, SEQ ID NO: 93, SEQ ID NO: 100, and SEQ ID NO: 109 The amino acid sequence includes the amino acid sequence set forth in

[0071]

[0081] In one embodiment, an isolated anti-IL-8 antibody comprising CDR sequences comprises a heavy chain variable region having heavy chain complementarity determining region (HCDR)1, HCDR2, and HCDR3, and a light chain variable region having light chain complementarity determining region (LCDR)1, LCDR2, and LCDR3, wherein the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 for the antibody comprise the amino acid sequences set forth in SEQ ID NO:40, SEQ ID NO:43, SEQ ID NO:50, SEQ ID NO:57, SEQ ID NO:64, and SEQ ID NO:73, respectively. In one embodiment, the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 for the antibody comprise the amino acid sequences set forth in SEQ ID NO:40, SEQ ID NO:43, SEQ ID NO:51, SEQ ID NO:58, SEQ ID NO:65, and SEQ ID NO:74, respectively. In some embodiments, the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 for the antibody comprise the amino acid sequences set forth in SEQ ID NO: 41, SEQ ID NO: 44, SEQ ID NO: 52, SEQ ID NO: 59, SEQ ID NO: 66, and SEQ ID NO: 75, respectively. In some embodiments, the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 for the antibody comprise the amino acid sequences set forth in SEQ ID NO: 41, SEQ ID NO: 44, SEQ ID NO: 53, SEQ ID NO: 60, SEQ ID NO: 67, and SEQ ID NO: 76, respectively. In some embodiments, the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 for the antibody comprise the amino acid sequences set forth in SEQ ID NO: 40, SEQ ID NO: 43, SEQ ID NO: 50, SEQ ID NO: 57, SEQ ID NO: 68, and SEQ ID NO: 73, respectively. In some embodiments, the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 for the antibody comprise the amino acid sequences set forth in SEQ ID NO: 40, SEQ ID NO: 43, SEQ ID NO: 50, SEQ ID NO: 57, SEQ ID NO: 69, and SEQ ID NO: 73, respectively. In some embodiments, the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 for the antibody comprise the amino acid sequences set forth in SEQ ID NO: 41, SEQ ID NO: 45, SEQ ID NO: 53, SEQ ID NO: 61, SEQ ID NO: 70, and SEQ ID NO: 77, respectively.In some embodiments, the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 for the antibody comprise the amino acid sequences set forth in SEQ ID NO: 40, SEQ ID NO: 46, SEQ ID NO: 54, SEQ ID NO: 62, SEQ ID NO: 71, and SEQ ID NO: 78, respectively. In some embodiments, the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 for the antibody comprise the amino acid sequences set forth in SEQ ID NO: 42, SEQ ID NO: 47, SEQ ID NO: 53, SEQ ID NO: 63, SEQ ID NO: 72, and SEQ ID NO: 79, respectively. In some embodiments, the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 for the antibody comprise the amino acid sequences set forth in SEQ ID NO: 41, SEQ ID NO: 48, SEQ ID NO: 53, SEQ ID NO: 60, SEQ ID NO: 67, and SEQ ID NO: 76, respectively. In some embodiments, the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 for the antibody comprise the amino acid sequences set forth in SEQ ID NO: 41, SEQ ID NO: 49, SEQ ID NO: 53, SEQ ID NO: 60, SEQ ID NO: 67, and SEQ ID NO: 76, respectively. In some embodiments, the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 for the antibody comprise the amino acid sequences set forth in SEQ ID NO: 41, SEQ ID NO: 44, SEQ ID NO: 55, SEQ ID NO: 60, SEQ ID NO: 67, and SEQ ID NO: 76, respectively. In some embodiments, the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 for the antibody comprise the amino acid sequences set forth in SEQ ID NO: 41, SEQ ID NO: 44, SEQ ID NO: 56, SEQ ID NO: 60, SEQ ID NO: 67, and SEQ ID NO: 76, respectively. In some embodiments, the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 for the antibody comprise the amino acid sequences set forth in SEQ ID NO: 41, SEQ ID NO: 48, SEQ ID NO: 55, SEQ ID NO: 60, SEQ ID NO: 67, and SEQ ID NO: 76, respectively. In some embodiments, the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 for the antibody comprise the amino acid sequences set forth in SEQ ID NO: 41, SEQ ID NO: 48, SEQ ID NO: 56, SEQ ID NO: 60, SEQ ID NO: 67, and SEQ ID NO: 76, respectively.In some embodiments, the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 for the antibody comprise the amino acid sequences set forth in SEQ ID NO: 41, SEQ ID NO: 49, SEQ ID NO: 55, SEQ ID NO: 60, SEQ ID NO: 67, and SEQ ID NO: 76, respectively. In some embodiments, the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 for the antibody comprise the amino acid sequences set forth in SEQ ID NO: 41, SEQ ID NO: 49, SEQ ID NO: 56, SEQ ID NO: 60, SEQ ID NO: 67, and SEQ ID NO: 76, respectively. In some embodiments, the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 for the antibody comprise the amino acid sequences set forth in SEQ ID NO: 80, SEQ ID NO: 83, SEQ ID NO: 50, SEQ ID NO: 90, SEQ ID NO: 97, and SEQ ID NO: 106, respectively. In some embodiments, the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 for the antibody comprise the amino acid sequences set forth in SEQ ID NO: 80, SEQ ID NO: 83, SEQ ID NO: 51, SEQ ID NO: 91, SEQ ID NO: 98, and SEQ ID NO: 107, respectively. In some embodiments, the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 for the antibody comprise the amino acid sequences set forth in SEQ ID NO: 81, SEQ ID NO: 84, SEQ ID NO: 52, SEQ ID NO: 92, SEQ ID NO: 99, and SEQ ID NO: 108, respectively. In some embodiments, the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 for the antibody comprise the amino acid sequences set forth in SEQ ID NO: 81, SEQ ID NO: 84, SEQ ID NO: 53, SEQ ID NO: 93, SEQ ID NO: 100, and SEQ ID NO: 109, respectively. In some embodiments, the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 for the antibody comprise the amino acid sequences set forth in SEQ ID NO: 80, SEQ ID NO: 83, SEQ ID NO: 50, SEQ ID NO: 90, SEQ ID NO: 101, and SEQ ID NO: 106, respectively. In some embodiments, the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 for the antibody comprise the amino acid sequences set forth in SEQ ID NO: 80, SEQ ID NO: 83, SEQ ID NO: 50, SEQ ID NO: 90, SEQ ID NO: 102, and SEQ ID NO: 106, respectively.In some embodiments, the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 for the antibody comprise the amino acid sequences set forth in SEQ ID NO: 81, SEQ ID NO: 85, SEQ ID NO: 53, SEQ ID NO: 94, SEQ ID NO: 103, and SEQ ID NO: 110, respectively. In some embodiments, the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 for the antibody comprise the amino acid sequences set forth in SEQ ID NO: 80, SEQ ID NO: 86, SEQ ID NO: 54, SEQ ID NO: 95, SEQ ID NO: 104, and SEQ ID NO: 111, respectively. In some embodiments, the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 for the antibody comprise the amino acid sequences set forth in SEQ ID NO: 82, SEQ ID NO: 87, SEQ ID NO: 53, SEQ ID NO: 93, SEQ ID NO: 105, and SEQ ID NO: 112, respectively. In some embodiments, the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 for the antibody comprise the amino acid sequences set forth in SEQ ID NO: 81, SEQ ID NO: 88, SEQ ID NO: 53, SEQ ID NO: 93, SEQ ID NO: 100, and SEQ ID NO: 109, respectively. In some embodiments, the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 for the antibody comprise the amino acid sequences set forth in SEQ ID NO: 81, SEQ ID NO: 89, SEQ ID NO: 53, SEQ ID NO: 93, SEQ ID NO: 100, and SEQ ID NO: 109, respectively. In some embodiments, the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 for the antibody comprise the amino acid sequences set forth in SEQ ID NO: 81, SEQ ID NO: 84, SEQ ID NO: 55, SEQ ID NO: 93, SEQ ID NO: 100, and SEQ ID NO: 109, respectively. In some embodiments, the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 for the antibody comprise the amino acid sequences set forth in SEQ ID NO: 81, SEQ ID NO: 84, SEQ ID NO: 56, SEQ ID NO: 93, SEQ ID NO: 100, and SEQ ID NO: 109, respectively. In some embodiments, the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 for the antibody comprise the amino acid sequences set forth in SEQ ID NO: 81, SEQ ID NO: 88, SEQ ID NO: 55, SEQ ID NO: 93, SEQ ID NO: 100, and SEQ ID NO: 109, respectively.In some embodiments, the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 for the antibody comprise the amino acid sequences set forth in SEQ ID NO: 81, SEQ ID NO: 88, SEQ ID NO: 56, SEQ ID NO: 93, SEQ ID NO: 100, and SEQ ID NO: 109, respectively. In some embodiments, the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 for the antibody comprise the amino acid sequences set forth in SEQ ID NO: 81, SEQ ID NO: 89, SEQ ID NO: 55, SEQ ID NO: 93, SEQ ID NO: 100, and SEQ ID NO: 109, respectively. In some embodiments, the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 for the antibody comprise the amino acid sequences set forth in SEQ ID NO: 81, SEQ ID NO: 89, SEQ ID NO: 56, SEQ ID NO: 93, SEQ ID NO: 100, and SEQ ID NO: 109, respectively.

[0072]

[0082] Those skilled in the art will understand that "IL-8-binding antibody," in its broadest sense, encompasses an antibody that specifically binds to an antigenic determinant of an IL-8 polypeptide. Those skilled in the art will understand that specificity for binding to IL-8 reflects that the binding is selective for the IL-8 antigen and can be distinguished from undesired or nonspecific interactions. In certain embodiments, the IL-8-binding antibody comprises one or more antibody fragments.

[0073]

[0083] In certain embodiments, the antigenic determinant comprises an IL-8 epitope. The term "epitope" includes any determinant, in certain embodiments, a polypeptide determinant capable of specific binding to an anti-IL-8 binding domain. An epitope is a region of an antigen bound by an antibody or antigen-binding fragment thereof. In certain embodiments, the IL-8 antigen-binding fragment of an antibody comprises a heavy chain variable region, a light chain variable region, or a combination thereof, as described herein.

[0074]

[0084] In certain embodiments, epitopic determinants include chemically active surface groupings of molecules such as amino acids, sugar side chains, phosphoryl or sulfonyl groups, and in certain embodiments, may have specific three-dimensional structural characteristics, and / or specific charge characteristics. In certain embodiments, an IL-8 binding antibody is said to specifically bind an IL-8 epitope when it preferentially recognizes IL-8 in a complex mixture of proteins and / or macromolecules.

[0075]

[0085] In one embodiment, the IL-8 binding antibody has an equilibrium dissociation constant of ≦10 -5 , 10 -6 , or 10 -7 M. In one embodiment, the equilibrium dissociation constant is ≦10 -8 M or 10 -9 In certain further embodiments, the equilibrium dissociation constant may be ≦10 -10 M, 10 -11 M or 10 -12 In some embodiments, the equilibrium dissociation constant is ≦10 -5 M~10 -12 M may be in the range.

[0076]

[0086] An antibody-binding domain can be a fragment of an antibody or a genetically engineered product of one or more fragments of an antibody, which fragment is responsible for specific binding to an antigen. "Specific binding" means that the binding is selective for the antigen of interest, e.g., IL-8 in the embodiments described herein, and can be distinguished from undesired or non-specific interactions. As used herein, the term "IL-8-binding antibody" can, in certain embodiments, encompass an entire immunoglobulin structure, a fragment thereof, or a domain thereof.

[0077]

[0087] In certain embodiments, the IL-8 antibodies described herein bind to human IL-8. In certain embodiments, the IL-8 antibodies described herein bind to cynomolgus IL-8. In certain embodiments, the IL-8 antibodies described herein bind to both human and cynomolgus IL-8. In certain embodiments, the IL-8 binding antibodies described herein inhibit the binding of IL-8 to the CXCR1 receptor. In certain embodiments, the IL-8 binding antibodies described herein reduce the binding of IL-8 to the CXCR1 receptor. In certain embodiments, the IL-8 binding antibodies described herein inhibit the binding of IL-8 to the CXCR2 receptor. In certain embodiments, the IL-8 binding antibodies described herein reduce the binding of IL-8 to the CXCR2 receptor. In certain embodiments, the IL-8 binding antibodies described herein inhibit the binding of IL-8 to both the CXCR1 and CXCR2 receptors. In certain embodiments, the IL-8 binding antibodies described herein reduce the binding of IL-8 to the CXCR1 and CXCR2 receptors.

[0078]

[0088] In certain embodiments, the IL-8-binding antibodies described herein inhibit IL-8-induced internalization of the CXCR1 receptor. In certain embodiments, the IL-8-binding antibodies described herein reduce IL-8-induced internalization of the CXCR1 receptor. In certain embodiments, the IL-8-binding antibodies described herein inhibit IL-8-induced internalization of the CXCR2 receptor. In certain embodiments, the IL-8-binding antibodies described herein reduce IL-8-induced internalization of the CXCR2 receptor. In certain embodiments, the IL-8-binding antibodies described herein inhibit IL-8-induced internalization of the CXCR1 and CXCR2 receptors. In certain embodiments, the IL-8-binding antibodies described herein reduce IL-8-induced internalization of the CXCR1 and CXCR2 receptors.

[0079]

[0089] In certain embodiments, the IL-8 binding antibodies described herein inhibit IL-8-induced NFκB activity. In certain embodiments, the IL-8 binding antibodies described herein reduce IL-8-induced NFκB activity. In certain embodiments, the IL-8 binding antibodies described herein inhibit IL-8-activated neutrophil migration. In certain embodiments, the IL-8 binding antibodies described herein reduce IL-8-induced neutrophil migration. In certain embodiments, the IL-8 binding antibodies described herein inhibit IL-8-activated monocyte migration. In certain embodiments, the IL-8 binding antibodies described herein reduce IL-8-activated monocyte migration.

[0080]

[0090] In certain embodiments, the IL-8 binding antibodies described herein inhibit IL-8-induced AKT and ERK phosphorylation. In certain embodiments, the IL-8 binding antibodies described herein reduce IL-8-induced AKT and ERK phosphorylation in neutrophils.

[0081]

[0091] In some embodiments, the IL-8 binding antibodies described herein reduce inflammation. In some embodiments, the IL-8 binding antibodies described herein eliminate inflammation. In some embodiments, the IL-8 binding antibodies described herein reduce swelling in a joint. In some embodiments, the IL-8 binding antibodies described herein reduce swelling in a diarthrodial joint selected from a ball and socket joint, a saddle joint, a hinge joint, a condylar joint, a pivot joint, and a gliding joint. In some embodiments, the IL-8 binding antibodies described herein reduce swelling in a joint selected from a knee, a finger, a shoulder, an elbow, a wrist, an ankle, a toe, or a hip, or any combination thereof. In some embodiments, the IL-8 binding antibodies described herein reduce inflammation. In some embodiments, the IL-8 binding antibodies described herein reduce inflammation in a joint. In some embodiments, the IL-8 binding antibodies described herein reduce inflammation in a diarthrodial joint selected from a ball and socket joint, a saddle joint, a hinge joint, a condylar joint, a pivot joint, and a gliding joint. In certain embodiments, the IL-8 binding antibodies described herein reduce swelling in a joint selected from the knee, finger, shoulder, elbow, wrist, ankle, toe, or hip, or any combination thereof.

[0082]

[0092] Examples of antibody binding domains include, but are not limited to, complementarity determining regions (CDRs), variable regions (Fv), VH domains, light chain variable regions (VL), heavy chains, light chains, single-chain variable regions (scFv), and Fab fragments. Those skilled in the art will appreciate that scFvs are not actually fragments of antibodies, but instead are fusion polypeptides comprising the variable heavy (VH) and variable light (VL) chain regions of immunoglobulins linked by a short linker peptide, for example, but not limited to, 10 to about 25 amino acids. Those skilled in the art will also appreciate that the term "Fab," in reference to antibodies, generally encompasses the portion of an antibody consisting of a single light chain (both variable and constant regions) connected by disulfide bonds to the variable region and first constant region of a single heavy chain.

[0083]

[0093] In some embodiments, antibodies include whole antibody molecules, including monoclonal, polyclonal, and multispecific (e.g., bispecific) antibodies. In some embodiments, antibodies include one or more antibody fragments that retain binding specificity, including, but not limited to, variable heavy chain (VH) fragments, variable light chain (VL) fragments, Fab fragments, F(ab')2 fragments, scFv fragments, Fv fragments, minibodies, diabodies, triabodies, and tetrabodies (see, e.g., Hudson and Souriau, Nature Med. 9:129-134 (2003) (incorporated herein by reference in its entirety)). Humanized, primatized, and chimeric antibodies are also included.

[0084]

[0094] Those skilled in the art will understand that an "isolated IL-8-binding antibody" encompasses, in certain embodiments, (1) an antibody that is free from at least some other proteins with which it is not typically found in nature or with which it is typically found during its synthesis; (2) an antibody that is essentially free from other, non-identical IL-8-binding antibodies from the same source; (3) an antibody that can be recombinantly expressed by a cell; (4) an antibody that has been separated from at least about 50 percent of the polynucleotides, lipids, carbohydrates, or other materials with which it is associated during synthesis; or (5) an antibody that is not naturally occurring, or a combination thereof. Such an isolated antibody may be encoded by genomic DNA, cDNA, mRNA, or other RNA, or may be of synthetic origin, or any combination thereof. In certain embodiments, an isolated antibody is substantially free of proteins or polypeptides or other contaminants that may interfere with its use (therapeutic, diagnostic, prophylactic, research, or otherwise). As used throughout, the terms "IL-8 antibody," "IL-8-binding antibody," and the like all have the same meaning and quality and may be used interchangeably.

[0085]

[0095] In certain embodiments, the IL-8 antibody comprises a recombinant antibody. In certain embodiments, the IL-8 antibody comprises a humanized antibody. In certain embodiments, the IL-8 antibody comprises an engineered antibody. In certain embodiments, the engineered antibody comprises improved binding compared to available antibodies. In certain embodiments, the engineered antibody comprises improved binding and dissociation constants (K) compared to other available IL-8 binding antibodies. on and K. off In certain embodiments, the engineered antibody comprises improved stability compared to available IL-8 binding antibodies.

[0086]

[0096] In certain embodiments, the present disclosure provides polypeptides comprising VH and VL domains that can be dimerized under suitable conditions. For example, the VH and VL domains can be combined in a suitable buffer and dimerized through appropriate interactions, such as hydrophobic interactions. In another embodiment, the VH and VL domains can be combined in a suitable buffer containing an enzyme and / or cofactor that can promote dimerization of the VH and VL domains. In another embodiment, the VH and VL domains can be combined in a suitable vehicle that allows them to react with each other in the presence of suitable reagents and / or catalysts.

[0087]

[0097] In certain embodiments, VH and VL domains may be combined into a longer polypeptide sequence that may include, for example, but not limited to, a constant region, a hinge region, a linker region, an Fc region, or a disulfide bond region, or any combination thereof. A constant domain is an immunoglobulin folding unit of the constant part of an immunoglobulin molecule, also called a domain of the constant region (e.g., CH1, CH2, CH3, CH4, Ck, Cl).

[0088]

[0098] In some embodiments, the anti-IL-8 antibody comprises a mutant immunoglobulin. Examples of mutant immunoglobulins include immunoglobulins with engineered Fc portions. Cellular immune responses are primarily driven by interactions between antibodies and Fc gamma receptors (FcγRs). Non-limiting examples of immunoglobulins with engineered Fc portions are provided in Wang et al. (2018) Protein Cell, 9(1):63-73 (see Table 1 in Wang et al.), which is incorporated herein in its entirety. Examples of mutant immunoglobulins with altered IgG binding with cytotoxicity (ADCC) components are found, for example, in Xu D, Alegre ML, Varga SS, Rothermel AL, Collins AM, Pulito VL, et al. In vitro characterization of five humanized OKT3 effector function variant antibodies. Cell Immunol. (2000) 200:16-26, which is incorporated herein in its entirety. In certain embodiments, the anti-IL8 immunoglobulin comprises an engineered Fc portion such that the interaction between the antibody and an Fc gamma receptor is increased, decreased, or eliminated.

[0089]

[0099] In some embodiments, the mutant anti-IL-8 IgG comprises an Fc region-engineered IgG1. In some embodiments, the mutant anti-IL-8 IgG comprises an Fc region-engineered IgG2. In some embodiments, the mutant anti-IL-8 IgG comprises an Fc region-engineered IgG4. In some embodiments, Fc variants comprising mutations in the Fc region exhibit reduced effector function. In certain embodiments, mutations in the Fc region of an antibody abolish immune effector function of the antibody. In some embodiments, the Fc portion of an antibody described herein is engineered, and the engineered antibody comprises improved efficacy or safety, or both, compared to a human IgG isotype.

[0090]

[0100] In certain embodiments, the mutation comprises a missense mutation (substitution of at least one amino acid for another), a nonsense mutation (substitution of a stop codon such that translation of the antibody polypeptide is prematurely terminated), an insertion mutation (insertion of at least one amino acid), or a deletion mutation (deletion of at least one amino acid).

[0091]

[0101] In certain embodiments, altering the glycosylation of IgG affects antibody activity. In certain embodiments, a mutant anti-IL-8 IgG comprises an altered glycosylation pattern in the Fc portion of the antibody. In certain embodiments, a mutant anti-IL-8 IgG comprising an altered glycosylation pattern has enhanced effector function. In certain embodiments, a mutant anti-IL-8 antibody comprises a defucosylated antibody. In certain embodiments, a mutant anti-IL-8 IgG comprising an altered glycosylation pattern has reduced effector function.

[0092]

[0102] In one embodiment, modulating the effector function of an anti-IL-8 antibody involves mutating the Fc region of the antibody so that the antibody-dependent cellular cytotoxicity (ADCC) response is enhanced. In one embodiment, modulating the effector function of an anti-IL-8 antibody involves mutating the Fc region of the antibody so that the antibody no longer binds to ADCC components, thereby reducing or eliminating the ADCC response. For example, but not limited to, an IgG containing the L234A / L235A (LALA) mutations cannot bind to Fc receptors.

[0093]

[0103] In one embodiment, the anti-IL-8 antibody comprises a mutated Fc region of the antibody such that the antibody-dependent cellular phagocytosis (ADCP) response is enhanced. In one embodiment, the anti-IL-8 antibody comprises a mutated Fc region of the antibody such that the antibody-complement-dependent cytotoxicity (CDC) response is enhanced.

[0094]

[0104] In certain embodiments, mutations in the Fc region of an IL-8 antibody increase binding to Fc gamma receptors. In certain embodiments, mutations in the Fc region of an IL-8 antibody decrease binding to Fc gamma receptors. In certain embodiments, mutations in the Fc region of an IL-8 antibody abolish binding to Fc gamma receptors.

[0095]

[0105] In certain embodiments, mutations in the Fc region of an IL-8 antibody increase the half-life of the IgG. In certain embodiments, mutations in the Fc region of an IL-8 antibody increase co-engagement of the antibody with a target antigen and an Fc gamma receptor. In certain embodiments, mutations in the Fc region of an IL-8 antibody decrease co-engagement of the antibody with a target antigen and an Fc gamma receptor.

[0096]

[0106] In certain embodiments, the isolated anti-IL-8 antibodies described herein include IgG, Fv, scFv, Fab, F(ab')2, minibody, diabody, triabody, nanobody, single domain antibody, multispecific antibody, bispecific antibody, trispecific antibody, single chain antibody, heavy chain antibody, chimeric antibody, or humanized antibody, or a combination thereof. In certain embodiments, the IgG can be of the subclass IgG1, IgG2, IgG3, or IgG4. In certain embodiments, the IgG comprises IgG1. In certain embodiments, the IgG comprises IgG2. In certain embodiments, the IgG comprises IgG3. In certain embodiments, the IgG comprises IgG4.

[0097]

[0107] Disclosed herein in one embodiment is an isolated polynucleotide sequence encoding a heavy chain variable region (VH) of an anti-IL-8 antibody and a light chain variable region (VL) of an anti-IL-8 antibody, wherein the amino acid sequence of the VH-VL pair is selected from the paired sequences set forth in SEQ ID NOs: 2 and 3, 4 and 5, 6 and 7, 8 and 9, 10 and 11, 12 and 13, 14 and 15, 16 and 17, 18 and 19, 20 and 21, 22 and 23, 24 and 25, 26 and 27, 28 and 29, 30 and 31, 32 and 33, and 34 and 35. Disclosed herein in one embodiment is an isolated polynucleotide sequence encoding an anti-IL-8 antibody VH and an anti-IL-8 antibody VL, wherein the amino acid sequence of the VH-VL pair comprises the paired sequence set forth in SEQ ID NOs: 2 and 3. Disclosed herein in one embodiment is an isolated polynucleotide sequence encoding an anti-IL-8 antibody VH and an anti-IL-8 antibody VL, wherein the amino acid sequence of the VH-VL pair comprises the pairing sequences set forth in SEQ ID NOs: 4 and 5. Disclosed herein in one embodiment is an isolated polynucleotide sequence encoding an anti-IL-8 antibody VH and an anti-IL-8 antibody VL, wherein the amino acid sequence of the VH-VL pair comprises the pairing sequences set forth in SEQ ID NOs: 6 and 7. Disclosed herein in one embodiment is an isolated polynucleotide sequence encoding an anti-IL-8 antibody VH and an anti-IL-8 antibody VL, wherein the amino acid sequence of the VH-VL pair comprises the pairing sequences set forth in SEQ ID NOs: 8 and 9. In one embodiment, disclosed herein is an isolated polynucleotide sequence encoding a VH of an anti-IL-8 antibody and a VL of an anti-IL-8 antibody, wherein the amino acid sequence of the VH-VL pair comprises the paired sequences set forth in SEQ ID NOs: 10 and 11.In one embodiment, disclosed herein is an isolated polynucleotide sequence encoding an anti-IL-8 antibody VH and an anti-IL-8 antibody VL, wherein the amino acid sequence of the VH-VL pair comprises the pairing sequences set forth in SEQ ID NOs: 12 and 13. In one embodiment, disclosed herein is an isolated polynucleotide sequence encoding an anti-IL-8 antibody VH and an anti-IL-8 antibody VL, wherein the amino acid sequence of the VH-VL pair comprises the pairing sequences set forth in SEQ ID NOs: 14 and 15. In one embodiment, disclosed herein is an isolated polynucleotide sequence encoding an anti-IL-8 antibody VH and an anti-IL-8 antibody VL, wherein the amino acid sequence of the VH-VL pair comprises the pairing sequences set forth in SEQ ID NOs: 16 and 17. In one embodiment, disclosed herein is an isolated polynucleotide sequence encoding an anti-IL-8 antibody VH and an anti-IL-8 antibody VL, wherein the amino acid sequence of the VH-VL pair comprises the pairing sequences set forth in SEQ ID NOs: 18 and 19. In one embodiment, disclosed herein is an isolated polynucleotide sequence encoding an anti-IL-8 antibody VH and an anti-IL-8 antibody VL, wherein the amino acid sequence of the VH-VL pair comprises the pairing sequences set forth in SEQ ID NOs: 20 and 21. In one embodiment, disclosed herein is an isolated polynucleotide sequence encoding an anti-IL-8 antibody VH and an anti-IL-8 antibody VL, wherein the amino acid sequence of the VH-VL pair comprises the pairing sequences set forth in SEQ ID NOs: 22 and 23. In one embodiment, disclosed herein is an isolated polynucleotide sequence encoding the VH of an anti-IL-8 antibody and the VL of an anti-IL-8 antibody, wherein the amino acid sequence of the VH-VL pair comprises the paired sequences set forth in SEQ ID NOs: 24 and 25. In one embodiment, disclosed herein is an isolated polynucleotide sequence encoding the VH of an anti-IL-8 antibody and the VL of an anti-IL-8 antibody, wherein the amino acid sequence of the VH-VL pair comprises the paired sequences set forth in SEQ ID NOs: 26 and 27.In one embodiment, disclosed herein is an isolated polynucleotide sequence encoding an anti-IL-8 antibody VH and an anti-IL-8 antibody VL, wherein the amino acid sequence of the VH-VL pair comprises the pairing sequences set forth in SEQ ID NOs: 28 and 29. In one embodiment, disclosed herein is an isolated polynucleotide sequence encoding an anti-IL-8 antibody VH and an anti-IL-8 antibody VL, wherein the amino acid sequence of the VH-VL pair comprises the pairing sequences set forth in SEQ ID NOs: 30 and 31. In one embodiment, disclosed herein is an isolated polynucleotide sequence encoding an anti-IL-8 antibody VH and an anti-IL-8 antibody VL, wherein the amino acid sequence of the VH-VL pair comprises the pairing sequences set forth in SEQ ID NOs: 32 and 33. In one embodiment, disclosed herein is an isolated polynucleotide sequence encoding a VH of an anti-IL-8 antibody and a VL of an anti-IL-8 antibody, wherein the amino acid sequence of the VH-VL pair comprises the paired sequences set forth in SEQ ID NOs: 34 and 35.

[0098]

[0108] Described herein in one embodiment is an isolated anti-IL-8 antibody comprising an antibody antigen-binding domain comprising a heavy chain variable region (VH) and a light chain variable region (VL), wherein the polynucleotide sequence encoding the VH-VL pair is selected from the pair sequences set forth in SEQ ID NOs: 113 and 114, 115 and 116, 117 and 118, 119 and 120, 121 and 122, 123 and 124, 125 and 126, 127 and 128, 129 and 130, 131 and 132, 133 and 134, 135 and 136, 137 and 138, 139 and 140, 141 and 142, 36 and 37, and 38 and 39. In one embodiment, the nucleotide sequence encoding the VH-VL pair is set forth in SEQ ID NOs: 113 and 114. In certain embodiments, the nucleotide sequences encoding the VH-VL pair are set forth in SEQ ID NOs: 115 and 116. In certain embodiments, the nucleotide sequences encoding the VH-VL pair are set forth in SEQ ID NOs: 117 and 118. In certain embodiments, the nucleotide sequences encoding the VH-VL pair are set forth in SEQ ID NOs: 119 and 120. In certain embodiments, the nucleotide sequences encoding the VH-VL pair are set forth in SEQ ID NOs: 121 and 122. In certain embodiments, the nucleotide sequences encoding the VH-VL pair are set forth in SEQ ID NOs: 123 and 124. In certain embodiments, the nucleotide sequences encoding the VH-VL pair are set forth in SEQ ID NOs: 125 and 126. In certain embodiments, the nucleotide sequences encoding the VH-VL pair are set forth in SEQ ID NOs: 127 and 128. In certain embodiments, the nucleotide sequences encoding the VH-VL pair are set forth in SEQ ID NOs: 129 and 130. In certain embodiments, the nucleotide sequences encoding the VH-VL pair are set forth in SEQ ID NOs: 131 and 132. In one embodiment, the nucleotide sequences encoding the VH-VL pair are set forth in SEQ ID NOs: 133 and 134. In one embodiment, the nucleotide sequences encoding the VH-VL pair are set forth in SEQ ID NOs: 135 and 136.In some embodiments, the nucleotide sequence encoding the VH-VL pair is set forth in SEQ ID NOs: 137 and 138. In some embodiments, the nucleotide sequence encoding the VH-VL pair is set forth in SEQ ID NOs: 139 and 140. In some embodiments, the nucleotide sequence encoding the VH-VL pair is set forth in SEQ ID NOs: 141 and 142. In some embodiments, the nucleotide sequence encoding the VH-VL pair is set forth in SEQ ID NOs: 36 and 37. In some embodiments, the nucleotide sequence encoding the VH-VL pair is set forth in SEQ ID NOs: 38 and 39.

[0099]

[0109] Those skilled in the art will understand that as used herein, the terms "polynucleotide sequence" and "nucleotide sequence" all have the same meaning and quality and can be used interchangeably in certain embodiments.

[0100]

[0110] In a specific embodiment, disclosed herein is an isolated polynucleotide sequence encoding a heavy chain variable region (VH) of an anti-IL-8 antibody comprising a VH complementarity determining region (HCDR) set forth in Table 1F and a light chain variable region (VL) of an anti-IL-8 antibody comprising a VL complementarity determining region (LCDR) set forth in Table 1F, wherein the heavy chain variable region comprises heavy chain complementarity determining region (HCDR)1, HCDR2, and HCDR3, and the light chain variable region comprises light chain complementarity determining region (LCDR)1, LCDR2, and LCDR3, and the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 for the VH and VL, respectively, comprise the amino acid sequences set forth in Table 1F. In one embodiment, an isolated polynucleotide sequence encoding a heavy chain variable region (VH) of an anti-IL-8 antibody comprising a VH complementarity determining region (HCDR) set forth in Table 1F and a light chain variable region (VL) of an anti-IL-8 antibody comprising a VL complementarity determining region (LCDR) set forth in Table 1F, wherein the heavy chain variable region comprises heavy chain complementarity determining region (HCDR) 1, HCDR2, and HCDR3, and the light chain variable region comprises light chain complementarity determining region (LCDR) 1, LCDR2, and LCDR3, and wherein the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 for each of the antibodies are (a) SEQ ID NO:41, SEQ ID NO:44, SEQ ID NO:53, SEQ ID NO:60, SEQ ID NO:67, and SEQ ID NO:76; (b) SEQ ID NO:81, SEQ ID NO:84, SEQ ID NO:53, SEQ ID NO:93, SEQ ID NO:100, and SEQ ID NO:109; (c) SEQ ID NO:40, SEQ ID NO:43, SEQ ID NO:50, SEQ ID NO:57, SEQ ID NO:64, and SEQ ID NO:73; (d) SEQ ID NO:40, SEQ ID NO:43, SEQ ID NO:51, SEQ ID NO:58, SEQ ID NO:65, and SEQ ID NO:74; (e) SEQ ID NO:41, SEQ ID NO:44, SEQ ID NO:52, SEQ ID NO:59, SEQ ID NO:66, and SEQ ID NO:75; (f) SEQ ID NO:40, SEQ ID NO:43, SEQ ID NO:50, SEQ ID NO:57, SEQ ID NO:68, and SEQ ID NO:73; (g) SEQ ID NO:40, SEQ ID NO:43, SEQ ID NO:50, SEQ ID NO:57, SEQ ID NO:69, and SEQ ID NO:73; (h) SEQ ID NO:41, SEQ ID NO:45, SEQ ID NO:53, SEQ ID NO:61, SEQ ID NO:70, and SEQ ID NO:77; (i) SEQ ID NO:40, SEQ ID NO:46, SEQ ID NO:54, SEQ ID NO:62, SEQ ID NO:71, and SEQ ID NO:78; (j) SEQ ID NO:42, SEQ ID NO:47, SEQ ID NO:53, SEQ ID NO:63, SEQ ID NO:72, and SEQ ID NO:79; (k) SEQ ID NO:41, SEQ ID NO:48, SEQ ID NO:53, SEQ ID NO:60, SEQ ID NO:67, and SEQ ID NO:76; (l) SEQ ID NO:41, SEQ ID NO:49, SEQ ID NO:53, SEQ ID NO:60, SEQ ID NO:67, and SEQ ID NO:76; (m) SEQ ID NO:41, SEQ ID NO:44, SEQ ID NO:55, SEQ ID NO:60, SEQ ID NO:67, and SEQ ID NO:76; (n) SEQ ID NO:41, SEQ ID NO:44, SEQ ID NO:56, SEQ ID NO:60, SEQ ID NO:67, and SEQ ID NO:76; (o) SEQ ID NO:41, SEQ ID NO:48, SEQ ID NO:55, SEQ ID NO:60, SEQ ID NO:67, and SEQ ID NO:76; (p) SEQ ID NO:41, SEQ ID NO:48, SEQ ID NO:56, SEQ ID NO:60, SEQ ID NO:67, and SEQ ID NO:76; (q) SEQ ID NO:41, SEQ ID NO:49, SEQ ID NO:55, SEQ ID NO:60, SEQ ID NO:67, and SEQ ID NO:76; (r) SEQ ID NO:41, SEQ ID NO:49, SEQ ID NO:56, SEQ ID NO:60, SEQ ID NO:67, and SEQ ID NO:76; (s) SEQ ID NO: 80, SEQ ID NO: 83, SEQ ID NO: 50, SEQ ID NO: 90, SEQ ID NO: 97, and SEQ ID NO: 106; (t) SEQ ID NO:80, SEQ ID NO:83, SEQ ID NO:51, SEQ ID NO:91, SEQ ID NO:98, and SEQ ID NO:107; (u) SEQ ID NO:81, SEQ ID NO:84, SEQ ID NO:52, SEQ ID NO:92, SEQ ID NO:99, and SEQ ID NO:108; (v) SEQ ID NO:80, SEQ ID NO:83, SEQ ID NO:50, SEQ ID NO:90, SEQ ID NO:101, and SEQ ID NO:106; (w) SEQ ID NO:80, SEQ ID NO:83, SEQ ID NO:50, SEQ ID NO:90, SEQ ID NO:102, and SEQ ID NO:106; (x) SEQ ID NO:81, SEQ ID NO:85, SEQ ID NO:53, SEQ ID NO:94, SEQ ID NO:103, and SEQ ID NO:110; (y) SEQ ID NO:80, SEQ ID NO:86, SEQ ID NO:54, SEQ ID NO:95, SEQ ID NO:104, and SEQ ID NO:111; (z) SEQ ID NO:82, SEQ ID NO:87, SEQ ID NO:53, SEQ ID NO:93, SEQ ID NO:105, and SEQ ID NO:112; (aa) SEQ ID NO: 81, SEQ ID NO: 88, SEQ ID NO: 53, SEQ ID NO: 93, SEQ ID NO: 100, and SEQ ID NO: 109; (bb) SEQ ID NO:81, SEQ ID NO:89, SEQ ID NO:53, SEQ ID NO:93, SEQ ID NO:100, and SEQ ID NO:109; (cc) SEQ ID NO:81, SEQ ID NO:84, SEQ ID NO:55, SEQ ID NO:93, SEQ ID NO:100, and SEQ ID NO:109; (dd) SEQ ID NO:81, SEQ ID NO:84, SEQ ID NO:56, SEQ ID NO:93, SEQ ID NO:100, and SEQ ID NO:109; (ee) SEQ ID NO:81, SEQ ID NO:88, SEQ ID NO:55, SEQ ID NO:93, SEQ ID NO:100, and SEQ ID NO:109; (ff) SEQ ID NO:81, SEQ ID NO:88, SEQ ID NO:56, SEQ ID NO:93, SEQ ID NO:100, and SEQ ID NO:109; (gg) SEQ ID NO:81, SEQ ID NO:89, SEQ ID NO:55, SEQ ID NO:93, SEQ ID NO:100, and SEQ ID NO:109; or (hh) SEQ ID NO: 81, SEQ ID NO: 89, SEQ ID NO: 56, SEQ ID NO: 93, SEQ ID NO: 100, and SEQ ID NO: 109 Disclosed herein is an isolated polynucleotide sequence comprising the amino acid sequence set forth in

[0101]

[0111] In a specific embodiment, disclosed herein is an isolated polynucleotide sequence encoding a heavy chain variable region (VH) of an anti-IL-8 antibody comprising a VH complementarity-determining region (HCDR), wherein the polynucleotide sequence is as set forth in Table 1G with the CDR regions represented therein, and an isolated polynucleotide sequence encoding a light chain variable region (VL) of an anti-IL-8 antibody comprising a VL complementarity-determining region (LCDR), wherein the polynucleotide sequence is as set forth in Table 1G with the CDR regions represented therein, wherein the heavy chain variable region comprises heavy chain complementarity-determining region (HCDR)1, HCDR2, and HCDR3, and the light chain variable region comprises light chain complementarity-determining region (LCDR)1, LCDR2, and LCDR3, and the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 for the VH and VL, respectively, comprise the nucleotide sequences set forth in Table 1G.

[0102]

[0112] In some embodiments, the polynucleotide sequences disclosed herein comprise two polynucleotide sequences: a first polynucleotide sequence encoding the VH of an anti-IL-8 antibody and a second polynucleotide sequence encoding the VL of the anti-IL-8 antibody. In some embodiments, the polynucleotide sequences encode the VH and VL regions of an anti-IL-8 antibody comprising an IgG, Fv, scFv, Fab, F(ab')2, minibody, diabody, triabody, nanobody, single-domain antibody, multispecific antibody, bispecific antibody, triabody, single-chain antibody, heavy-chain antibody, chimeric antibody, or humanized antibody, or a combination thereof. In some embodiments, the polynucleotide sequences encode the VH and VL regions of an anti-IL-8 antibody comprising an IgG, wherein the IgG can be of the subclass IgG1, IgG2, IgG3, or IgG4.

[0103]

[0113] In some embodiments, the present disclosure also provides a vector comprising the above-described polynucleotide sequence. In some embodiments, the vector comprises two vectors, one comprising a polynucleotide encoding a VH and one comprising a polynucleotide encoding a VL. Given the amino acid sequences disclosed herein, one skilled in the art would readily construct a vector or plasmid to encode the amino acid sequence.

[0104]

[0114] Those skilled in the art will understand that the polynucleotides described herein, or fragments thereof, can be combined with other DNA sequences, such as promoters, polyadenylation signals, additional restriction enzyme sites, multiple cloning sites, other coding segments, etc., so that their total length can vary considerably, regardless of the length of the coding sequence itself. Thus, it is contemplated that nucleic acid fragments of almost any length can be used, with the total length preferably being limited by the ease of preparation and use in the intended recombinant DNA protocol. For example, exemplary polynucleotide segments having total lengths of about 10,000, 5000, 3000, 2000, 1000, 500, 200, 100, 50, and the like (including all intermediate lengths) are contemplated as useful.

[0105]

[0115] In certain embodiments, the isolated polynucleotide is inserted into a vector. As used herein, the term "vector" encompasses a vehicle into which a polynucleotide encoding a protein can be covalently inserted to result in expression of the protein and / or cloning of the polynucleotide. The isolated polynucleotide can be inserted into a vector using any suitable method known in the art, for example, but not limited to, a vector can be digested with an appropriate restriction enzyme and then ligated with an isolated polynucleotide having matching restriction ends.

[0106]

[0116] Examples of suitable vectors include, but are not limited to, plasmids, phagemids, cosmids, artificial chromosomes, such as yeast artificial chromosomes (YACs), bacterial artificial chromosomes (BACs), or P1-derived artificial chromosomes (PACs), bacteriophages, such as lambda phage or M13 phage, and animal viruses. Examples of animal virus categories useful as vectors include, but are not limited to, retroviruses (including lentiviruses), adenoviruses, adeno-associated viruses, herpesviruses (e.g., herpes simplex viruses), poxviruses, baculoviruses, papillomaviruses, and papovaviruses (e.g., SV40). In some embodiments, the vector comprises an expression vector.

[0107]

[0117] In some embodiments, an expression vector comprises a nucleic acid construct described herein. A suitable vector can be selected or constructed, containing appropriate regulatory sequences, including promoter sequences, transcription termination sequences, polyadenylation sequences, enhancer sequences, marker genes, and other sequences, as needed. The regulatory sequences can be operably linked to the nucleic acid sequence contained within the nucleic acid construct. The vector can be a plasmid, virus, e.g., a phage, or phagemid, as needed. For further details, see, for example, Molecular Cloning: a Laboratory Manual: 3rd edition, Sambrook and Russell, 2001, Cold Spring Harbor Laboratory Press. Many known techniques and protocols for the manipulation of nucleic acids, for example, in preparing nucleic acid constructs, mutagenesis, sequencing, introducing DNA into cells and gene expression, and analyzing proteins, are described in detail in Current Protocols in Molecular Biology, Second Edition, Ausubel et al. eds., John Wiley & Sons, 1988, and Short Protocols in Molecular Biology: A Compendium of Methods from Current Protocols in Molecular Biology, Ausubel et al. eds., John Wiley & Sons, 4th edition 1999. The disclosures of Sambrook et al. and Ausubel et al. (both) are incorporated herein by reference.

[0108]

[0118] Vectors can be introduced into host cells using any suitable method known in the art, including, but not limited to, DEAE-dextran mediated delivery, calcium phosphate precipitation methods, cationic lipid-mediated delivery, liposome-mediated transfection, electroporation, biolistics, receptor-mediated gene delivery, polylysine, histones, chitosan, and peptide-mediated delivery. Standard methods for transfection and transformation of cells for expression of a vector of interest are well known in the art.

[0109]

[0119] For expression of an IL-8 antibody or a component thereof, a vector can be introduced into a host cell to allow expression of the polypeptide in the host cell. The expression vector can contain various elements for controlling expression, including, but not limited to, a promoter sequence, a transcription initiation sequence, an enhancer sequence, a selectable marker, and a signal sequence. These elements can be selected as needed by those skilled in the art. In certain embodiments, these elements can be considered "control" elements.

[0110]

[0120] Those skilled in the art will understand that the term "control sequence" can encompass polynucleotide sequences that can affect the expression, processing, or subcellular localization of coding sequences to which they are ligated or operably linked. The nature of such control sequences can depend on the host organism. In certain embodiments, transcription control sequences for prokaryotes can include a promoter, a ribosomal binding site, and a transcription termination sequence. In other particular embodiments, transcription control sequences for eukaryotes can include a promoter containing one or more recognition sites for transcription factors, transcription enhancer sequences, transcription termination sequences, and polyadenylation sequences. In certain embodiments, "control sequences" can include leader sequences and / or fusion partner sequences.

[0111]

[0121] In certain embodiments, for example and without limitation, a promoter sequence may be selected to promote transcription of a polynucleotide in a vector. Suitable promoter sequences include, but are not limited to, T7 promoter, T3 promoter, SP6 promoter, β-actin promoter, EF1a promoter, CMV promoter, and SV40 promoter. An enhancer sequence may be selected to enhance transcription of a polynucleotide. A selectable marker may be selected to allow selection of host cells inserted with the vector from those not; for example, a selectable marker may be a gene that confers antibiotic resistance. A signal sequence may be selected to allow the expressed polypeptide to be transported out of the host cell.

[0112]

[0122] The vector may also include materials that aid in its entry into a cell, including, but not limited to, a viral particle, liposome, or protein coating.

[0113]

[0123] In some embodiments, the expression vector comprises an isolated polynucleotide sequence encoding an IL-8 antibody or a component thereof, for example, but not limited to, a VH domain, a VL domain, or a combined VH-VL domain, such as may be present in a Fab element, a F(ab')2 element, an IgG, an Fv, or an scFv. In some embodiments, the expression vector comprises a polynucleotide sequence encoding an IL-8 VH domain or a VL domain, or a combination thereof, wherein the polynucleotide sequence is selected from those set forth in Table 1G. In some embodiments, the expression vector comprises a polynucleotide sequence encoding an IL-8 VH domain or a VL domain, or a combination thereof, wherein the polynucleotide sequence is selected from the sequences set forth in SEQ ID NOs: 113-142 and 36-39. In some embodiments, the isolated polynucleotide sequence encodes an anti-IL-8 antibody component, such as a minibody, diabody, triabody, nanobody, single-domain antibody, multispecific antibody, bispecific antibody, triabody, single-chain antibody, heavy-chain antibody, chimeric antibody, or humanized antibody, or a combination thereof, as described above. The IL-8 binding domain and its components are described in detail above.

[0114]

[0124] In some embodiments, the expression vector comprises an isolated polynucleotide sequence encoding a VH domain. In some embodiments, the expression vector comprises an isolated nucleic acid sequence encoding a VL domain. In some embodiments, the expression vector comprises an isolated nucleic acid sequence encoding a VH and VL domain. In some embodiments, the expression vector comprises an isolated nucleic acid sequence encoding a set of CDRs of a VH region. In some embodiments, the expression vector comprises an isolated nucleic acid sequence encoding a set of CDRs of a VL region. In some embodiments, the expression vector comprises an isolated nucleic acid sequence encoding a set of CDRs of a VH region and a VL region. In some embodiments, the CDR regions of the VH and VL regions are those shown in Table 1G.

[0115]

[0125] In another embodiment, the present disclosure also provides a host cell comprising the vector provided herein. Depending on the use and experimental conditions, one skilled in the art will readily use a suitable host cell to harbor and / or express the above-described polynucleotide sequences.

[0116]

[0126] For cloning a polynucleotide, a vector can be introduced into a host cell (isolated host cell) to allow the vector to replicate itself, thereby amplifying copies of the polynucleotide contained therein. Cloning vectors can contain sequence components that generally include, but are not limited to, an origin of replication, a promoter sequence, a transcription initiation sequence, an enhancer sequence, and a selectable marker. These elements can be selected as needed by those skilled in the art. For example, an origin of replication can be selected to promote autonomous replication of the vector within the host cell.

[0117]

[0127] In certain embodiments, the present disclosure provides an isolated host cell containing a vector provided herein. The host cell containing the vector may be useful for expressing or cloning a polynucleotide contained in the vector.

[0118]

[0128] In some embodiments, the recombinant host cell comprises one or more of the constructs described above. A polynucleotide encoding any CDR or set of CDRs, or a VH domain or VL domain, or an antibody antigen-binding site or antibody molecule, such as, but not limited to, an IgG, Fv, scFv, Fab, F(ab')2, minibody, diabody, triabody, nanobody, single-domain antibody, multispecific antibody, bispecific antibody, triabody, single-chain antibody, heavy-chain antibody, chimeric antibody, or humanized antibody, or a combination thereof. In some embodiments, the host cell comprises one or more of the constructs described above encoding an IgG subclass selected from IgG1, IgG2, IgG3, and IgG4.

[0119]

[0129] Disclosed herein, in certain embodiments, are methods for producing an encoded product, the method comprising expression from a polynucleotide construct. In certain embodiments, the polynucleotide construct comprises a polynucleotide sequence selected from SEQ ID NOs: 113-142 and 36-39, or a CDR portion thereof. Expression may, in certain embodiments, be achieved by culturing recombinant host cells containing the nucleic acid construct under appropriate conditions. Following production by expression, the antibody or IL-8 antigen-binding fragment thereof may be isolated and / or purified using any suitable technique and then used, as desired, for example, in the therapeutic methods described herein.

[0120]

[0130] In certain embodiments, systems for cloning and expression of polypeptides in a variety of different host cells are well known. Suitable host cells may include, but are not limited to, prokaryotic cells, fungal cells, yeast cells, or higher eukaryotic cells, such as insect cells or mammalian cells.

[0121]

[0131] Suitable prokaryotic cells for this purpose include, but are not limited to, eubacteria, such as gram-negative or gram-positive bacteria, for example, Enterobacteriaceae, such as Escherichia, for example, E. coli, Enterobacter, Erwinia, Klebsiella, Proteus, Salmonella, for example, Salmonella typhimurium, Serratia, for example, Serratia marcescans, and Shigella, as well as Bacilli, such as B. subtilis and B. licheniformis, Pseudomonas, such as P. aeruginosa, and Streptomyces.

[0122]

[0132] Expression of antibodies and antigen-binding fragments in prokaryotic cells such as E. coli is well established in the art. For a review, see, e.g., Pluckthun, A. Bio / Technology 9:545-551 (1991). Expression in eukaryotic cells in culture is also available to those skilled in the art as an option for producing antibodies or antigen-binding fragments thereof; see recent reviews, e.g., Ref, ME (1993) Curr. Opinion Biotech. 4:573-576; Trill JJ et al. (1995) Curr. Opinion Biotech 6:553-560.

[0123]

[0133] Suitable fungal cells for this purpose include, but are not limited to, filamentous fungi and yeast. Illustrative examples of fungal cells include Saccharomyces cerevisiae, common baker's yeast, Schizosaccharomyces pombe, Kluyveromyces hosts, such as K. lactis, K. fragilis (ATCC 12,424), K. bulgaricus (ATCC 16,045), K. wickeramii (ATCC 24,178), K. waltii (ATCC 56,500), K. drosophilarum (ATCC 16,045), and the like. 36,906, K. thermotolerans, and K. marxianus; Yarrowia (EP 402,226); Pichia pastoris (EP 183,070); Candida; Trichoderma reesia (EP 244,234); Neurospora crassa; Schwanniomyces, e.g., Schwanniomyces occidentalis; occidentalis); and filamentous fungi such as Neurospora, Penicillium, Tolypocladium, and Aspergillus hosts such as A. nidulans and A. niger.

[0124]

[0134] Higher eukaryotic cells, particularly those derived from multicellular organisms, can be used for expressing glycosylated VH and VL domains as provided herein. Suitable higher eukaryotic cells include, but are not limited to, invertebrate cells and insect cells, and vertebrate cells. Examples of invertebrate cells include plant and insect cells. Many baculovirus strains and variants have been identified, as well as corresponding permissive insect host cells from hosts such as Spodoptera frugiperda (caterpillar), Aedes aegypti (mosquito), Aedes albopictus (mosquito), Drosophila melanogaster (fruit fly), and Bombyx mori (silkworm). Various viral strains for transfection are publicly available, such as the K-1 variant of Autographa californica NPV and the Bm-5 strain of Bombyx mori NPV, and such viruses may be used as viruses herein, particularly for transfection of Spodoptera frugiperda cells, as described herein. Plant cell cultures of cotton, corn, potato, soybean, petunia, tomato, and tobacco may also be used as hosts. Mammalian cell lines available in the art for expression of heterologous polypeptides include Chinese hamster ovary (CHO) cells, HeLa cells, baby hamster kidney cells, NS0 mouse melanoma cells, YB2 / 0 rat myeloma cells, human embryonic kidney cells, human embryonic retina cells, and many others.Non-limiting examples of vertebrate cells include mammalian host cell lines, such as the SV40-transformed monkey kidney CV1 line (COS-7, ATCC CRL 1651); human embryonic kidney line (293 or 293 cells subcloned for growth in suspension culture, Graham et al., J. Gen Virol. 36:59 (1977)); baby hamster kidney cells (BHK, ATCC CCL 10); Chinese hamster ovary cells / -DHFR (CHO, Urlaub et al., Proc. Natl. Acad. Sci. USA 77:4216 (1980)); ExpiCHO-S™ cells (ThermoFisher Scientific cat. #A29133); mouse Sertoli cells (TM4, Mather, Biol. Reprod. 23:243-251 (1980)); monkey kidney cells (CV1 ATCC CCL 70); African green monkey kidney cells (VERO-76, ATCC CRK-1587); human cervical carcinoma cells (HELA, ATCC CCL 2); canine kidney cells (MDCK, ATCC CCL 34); buffalo rat hepatocytes (BRL 3A, ATCC CRL 1442); human lung cells (W138, ATCC CCL 75); human hepatocytes (Hep G2, HB 8065); mouse mammary tumor (MMT 060562, ATCC CCL51); TRI cells (Mather et al., Annals NY Acad. Sci. 383:44-68 (1982)); MRC 5 cells; FS4 cells; and a human hepatocellular carcinoma line (Hep G2).

[0125]

[0135] A non-limiting example of an expression system well known in the art is the Lonza (USA) GS Gene Expression System®. In certain embodiments, vectors encoding the polypeptides described herein include Lonza (USA) GS® vectors, such as, but not limited to, pXC-IgG1zaDK (based on pXC-18.4) and pXC-κ (based on pXC-17.4). These GS® vectors and other similar vectors known in the art encompass a range of vector options, including universal base vectors, IgG constant region vectors, IgG site-specific conjugation vectors, pXC Multigene vectors, and GS piggyBac™ vectors (+ transposase). In certain embodiments, host cells in which the encoded polypeptides described herein can be expressed include the GS Xceed® CHOK1SV GS-KO® cell line or other similar cells known in the art or generated to optimize protein expression. In certain embodiments, the vector and host cell combination optimizes expression of an IL-8 antibody polypeptide or an IL-8 binding fragment thereof.

[0126]

[0136] In certain embodiments, provided herein are host cells containing the nucleic acids disclosed herein. Such host cells may be in vitro, or in culture. Such host cells may be in vivo. The in vivo presence of the host cells may allow for intracellular expression of the IL-8 binding antibodies described herein as "intrabodies" or intracellular antibodies. Intrabodies may be used for gene therapy.

[0127]

[0137] In certain embodiments, a host cell comprises a first vector encoding a first polypeptide, e.g., a VH domain, and a second vector encoding a second polypeptide, e.g., a VL domain. In certain embodiments, a host cell comprises a vector encoding a first polypeptide, e.g., a VH domain, and a second polypeptide, e.g., a VL domain.

[0128]

[0138] In certain embodiments, the host cell comprises a first vector encoding a VH domain and a second vector encoding a VL domain, hi certain embodiments, the host cell comprises a single vector encoding a VH domain and a VL domain.

[0129]

[0139] In some embodiments, the isolated cell comprises an isolated nucleic acid sequence disclosed herein. In some embodiments, the isolated cell comprises two isolated nucleic acid sequences disclosed herein, where one nucleic acid encodes a VH domain and the other nucleic acid encodes a VL domain. In some embodiments, the isolated cell comprises a single isolated nucleic acid sequence disclosed herein encoding a VH domain and a VL domain.

[0130]

[0140] In certain embodiments, the first vector and the second vector may or may not be introduced simultaneously. In certain embodiments, the first vector and the second vector may be introduced into the host cell together. In certain embodiments, the first vector may be introduced into the host cell first, and then the second vector may be introduced. In certain embodiments, the first vector may be introduced into the host cell, which may then be established into a stable cell line expressing the first polypeptide, and then the second vector may be introduced into the stable cell line.

[0131]

[0141] The introduction can be followed by causing or allowing expression from the nucleic acid, e.g., culturing host cells under conditions for expression of the gene. In certain embodiments, the present disclosure provides a method of expressing a polypeptide provided herein, comprising culturing a host cell containing a vector under conditions whereby an inserted polynucleotide in the vector is expressed.

[0132]

[0142] In some embodiments, the nucleic acid is integrated into the genome (e.g., chromosome) of the host cell. Integration can be facilitated by the inclusion of sequences that facilitate recombination with the genome, according to standard techniques. In some embodiments, the nucleic acid construct is not integrated into the genome, and the vector is episomal.

[0133]

[0143] In one embodiment, disclosed herein is a method comprising using the construct described above in an expression system to express an IL-8 binding antibody or fragment thereof, as described herein above.

[0134]

[0144] Suitable conditions for expression of a polynucleotide may include, but are not limited to, a suitable culture medium, a suitable density of host cells in the culture medium, the presence of necessary nutrients, the presence of supplementary factors, suitable temperature and humidity, and the absence of microbial contaminants. Those skilled in the art can select suitable conditions for expression as needed.

[0135]

[0145] In certain embodiments, the IL-8 binding antibodies described herein can be prepared, isolated, and / or purified in a substantially pure or homogeneous form. Disclosed herein is a method for producing an anti-IL-8 antibody comprising a heavy chain variable region (VH) and a light chain variable region (VH), the method comprising culturing host cells under conditions conducive to expression of a vector in the host cells, thereby expressing a polynucleotide sequence contained in the vector, and thereby producing an anti-IL-8 antibody or an IL-8 antigen-binding domain thereof. In certain embodiments, the method for producing an anti-IL-8 antibody comprising a heavy chain variable region (VH) and a light chain variable region (VH), the method comprises culturing host cells comprising a vector comprising an isolated polynucleotide sequence encoding the heavy chain variable region (VH) of the anti-IL-8 antibody and the light chain variable region (VL) of the anti-IL-8 antibody under conditions conducive to expression of the vector in the host cells, thereby expressing the polynucleotide sequence contained in the vector, and thereby producing an anti-IL-8 antibody or an IL-8 antigen-binding domain thereof comprising a VH and a VL. Disclosed herein is a method for producing an anti-IL-8 antibody, comprising culturing a host cell comprising a vector comprising an isolated polynucleotide sequence encoding the VH and VL of the anti-IL-8 antibody, wherein the amino acid sequence of the VH-VL pair comprises the pair sequence set forth in SEQ ID NOs: 2 and 3, 4 and 5, 6 and 7, 8 and 9, 10 and 11, 12 and 13, 14 and 15, 16 and 17, 18 and 19, 20 and 21, 22 and 23, 24 and 25, 26 and 27, 28 and 29, 30 and 31, 32 and 33, and 34 and 35. In one embodiment, the method for producing an anti-IL-8 antibody comprises culturing a host cell comprising a vector comprising an isolated polynucleotide sequence encoding the VH and VL of the anti-IL-8 antibody, wherein the amino acid sequence of the VH-VL pair comprises the pair sequence set forth in SEQ ID NOs: 2 and 3. In one embodiment, a method for producing an anti-IL-8 antibody comprises culturing a host cell containing a vector comprising an isolated polynucleotide sequence encoding the VH and VL of the anti-IL-8 antibody, wherein the amino acid sequence of the VH-VL pair comprises the paired sequences set forth in SEQ ID NOs: 4 and 5.In certain embodiments, a method for producing an anti-IL-8 antibody comprises culturing a host cell comprising a vector comprising an isolated polynucleotide sequence encoding the VH and VL of the anti-IL-8 antibody, wherein the amino acid sequence of the VH-VL pair comprises the pairing sequences set forth in SEQ ID NOs: 6 and 7. In certain embodiments, a method for producing an anti-IL-8 antibody comprises culturing a host cell comprising a vector comprising an isolated polynucleotide sequence encoding the VH and VL of the anti-IL-8 antibody, wherein the amino acid sequence of the VH-VL pair comprises the pairing sequences set forth in SEQ ID NOs: 8 and 9. In certain embodiments, a method for producing an anti-IL-8 antibody comprises culturing a host cell comprising a vector comprising an isolated polynucleotide sequence encoding the VH and VL of the anti-IL-8 antibody, wherein the amino acid sequence of the VH-VL pair comprises the pairing sequences set forth in SEQ ID NOs: 10 and 11. In certain embodiments, a method for producing an anti-IL-8 antibody comprises culturing a host cell comprising a vector comprising an isolated polynucleotide sequence encoding the VH and VL of the anti-IL-8 antibody, wherein the amino acid sequence of the VH-VL pair comprises the pairing sequences set forth in SEQ ID NOs: 12 and 13. In certain embodiments, a method for producing an anti-IL-8 antibody comprises culturing a host cell comprising a vector comprising an isolated polynucleotide sequence encoding the VH and VL of the anti-IL-8 antibody, wherein the amino acid sequence of the VH-VL pair comprises the pairing sequences set forth in SEQ ID NOs: 14 and 15. In certain embodiments, a method for producing an anti-IL-8 antibody comprises culturing a host cell comprising a vector comprising an isolated polynucleotide sequence encoding the VH and VL of the anti-IL-8 antibody, wherein the amino acid sequence of the VH-VL pair comprises the pairing sequences set forth in SEQ ID NOs: 16 and 17. In one embodiment, a method for producing an anti-IL-8 antibody comprises culturing a host cell comprising a vector comprising an isolated polynucleotide sequence encoding the VH and VL of the anti-IL-8 antibody, wherein the amino acid sequence of the VH-VL pair comprises the paired sequences set forth in SEQ ID NOs: 18 and 19. In one embodiment, a method for producing an anti-IL-8 antibody comprises culturing a host cell comprising a vector comprising an isolated polynucleotide sequence encoding the VH and VL of the anti-IL-8 antibody, wherein the amino acid sequence of the VH-VL pair comprises the paired sequences set forth in SEQ ID NOs: 20 and 21.In certain embodiments, a method for producing an anti-IL-8 antibody comprises culturing a host cell comprising a vector comprising an isolated polynucleotide sequence encoding the VH and VL of the anti-IL-8 antibody, wherein the amino acid sequence of the VH-VL pair comprises the pairing sequences set forth in SEQ ID NOs: 22 and 23. In certain embodiments, a method for producing an anti-IL-8 antibody comprises culturing a host cell comprising a vector comprising an isolated polynucleotide sequence encoding the VH and VL of the anti-IL-8 antibody, wherein the amino acid sequence of the VH-VL pair comprises the pairing sequences set forth in SEQ ID NOs: 24 and 25. In certain embodiments, a method for producing an anti-IL-8 antibody comprises culturing a host cell comprising a vector comprising an isolated polynucleotide sequence encoding the VH and VL of the anti-IL-8 antibody, wherein the amino acid sequence of the VH-VL pair comprises the pairing sequences set forth in SEQ ID NOs: 26 and 27. In certain embodiments, a method for producing an anti-IL-8 antibody comprises culturing a host cell comprising a vector comprising an isolated polynucleotide sequence encoding the VH and VL of the anti-IL-8 antibody, wherein the amino acid sequence of the VH-VL pair comprises the pairing sequences set forth in SEQ ID NOs: 28 and 29. In certain embodiments, a method for producing an anti-IL-8 antibody comprises culturing a host cell comprising a vector comprising an isolated polynucleotide sequence encoding the VH and VL of the anti-IL-8 antibody, wherein the amino acid sequence of the VH-VL pair comprises the pairing sequences set forth in SEQ ID NOs: 30 and 31. In certain embodiments, a method for producing an anti-IL-8 antibody comprises culturing a host cell comprising a vector comprising an isolated polynucleotide sequence encoding the VH and VL of the anti-IL-8 antibody, wherein the amino acid sequence of the VH-VL pair comprises the pairing sequences set forth in SEQ ID NOs: 32 and 33. In one embodiment, a method for producing an anti-IL-8 antibody comprises culturing a host cell containing a vector comprising an isolated polynucleotide sequence encoding the VH and VL of the anti-IL-8 antibody, wherein the amino acid sequence of the VH-VL pair comprises the paired sequences set forth in SEQ ID NOs: 34 and 35.

[0136]

[0146] In one embodiment, a method for producing an anti-IL-8 antibody comprising a heavy chain variable region (VH) and a light chain variable region (VL) comprises culturing a host cell comprising a vector comprising an isolated polynucleotide sequence encoding the heavy chain variable region (VH) of the anti-IL-8 antibody and the light chain variable region (VL) of the anti-IL-8 antibody under conditions conducive to expression of the vector in the host cell, thereby expressing the polynucleotide sequence contained in the vector and thereby producing an anti-IL-8 antibody comprising the VH and VL or an IL-8 antigen-binding domain thereof; Disclosed herein are methods wherein the polypeptide sequence is selected from the pair sequences set forth in SEQ ID NOs: 113 and 114, SEQ ID NOs: 115 and 116, SEQ ID NOs: 117 and 118, SEQ ID NOs: 119 and 120, SEQ ID NOs: 121 and 122, SEQ ID NOs: 123 and 124, SEQ ID NOs: 125 and 126, SEQ ID NOs: 127 and 128, SEQ ID NOs: 129 and 130, SEQ ID NOs: 131 and 132, SEQ ID NOs: 133 and 134, SEQ ID NOs: 135 and 136, SEQ ID NOs: 137 and 138, SEQ ID NOs: 139 and 140, SEQ ID NOs: 141 and 142, SEQ ID NOs: 36 and 37, and SEQ ID NOs: 38 and 39. In certain embodiments, a method of producing an anti-IL-8 antibody comprises culturing a host cell comprising a vector comprising isolated polynucleotide sequences encoding the VH and VL of the anti-IL-8 antibody, wherein the polynucleotide sequence encoding the VH-VL comprises the pair sequences set forth in SEQ ID NOs: 113 and 114. In certain embodiments, a method for producing an anti-IL-8 antibody comprises culturing a host cell comprising a vector comprising isolated polynucleotide sequences encoding the VH and VL of the anti-IL-8 antibody, wherein the polynucleotide sequence encoding the VH-VL comprises the paired sequences set forth in SEQ ID NOs: 115 and 116. In certain embodiments, a method for producing an anti-IL-8 antibody comprises culturing a host cell comprising a vector comprising isolated polynucleotide sequences encoding the VH and VL of the anti-IL-8 antibody, wherein the polynucleotide sequence encoding the VH-VL comprises the paired sequences set forth in SEQ ID NOs: 117 and 118.In certain embodiments, a method for producing an anti-IL-8 antibody comprises culturing a host cell comprising a vector comprising an isolated polynucleotide sequence encoding the VH and VL of the anti-IL-8 antibody, wherein the polynucleotide sequence encoding the VH-VL comprises the paired sequences set forth in SEQ ID NOs: 119 and 120. In certain embodiments, a method for producing an anti-IL-8 antibody comprises culturing a host cell comprising a vector comprising an isolated polynucleotide sequence encoding the VH and VL of the anti-IL-8 antibody, wherein the polynucleotide sequence encoding the VH-VL comprises the paired sequences set forth in SEQ ID NOs: 121 and 122. In certain embodiments, a method for producing an anti-IL-8 antibody comprises culturing a host cell comprising a vector comprising an isolated polynucleotide sequence encoding the VH and VL of the anti-IL-8 antibody, wherein the polynucleotide sequence encoding the VH-VL comprises the paired sequences set forth in SEQ ID NOs: 123 and 124. In certain embodiments, a method for producing an anti-IL-8 antibody comprises culturing a host cell comprising a vector comprising an isolated polynucleotide sequence encoding the VH and VL of the anti-IL-8 antibody, wherein the polynucleotide sequence encoding the VH-VL comprises the paired sequences set forth in SEQ ID NOs: 125 and 126. In certain embodiments, a method for producing an anti-IL-8 antibody comprises culturing a host cell comprising a vector comprising an isolated polynucleotide sequence encoding the VH and VL of the anti-IL-8 antibody, wherein the polynucleotide sequence encoding the VH-VL comprises the paired sequences set forth in SEQ ID NOs: 127 and 128. In certain embodiments, a method for producing an anti-IL-8 antibody comprises culturing a host cell comprising a vector comprising an isolated polynucleotide sequence encoding the VH and VL of the anti-IL-8 antibody, wherein the polynucleotide sequence encoding the VH-VL comprises the paired sequences set forth in SEQ ID NOs: 129 and 130. In one embodiment, a method for producing an anti-IL-8 antibody comprises culturing a host cell containing a vector comprising an isolated polynucleotide sequence encoding the VH and VL of the anti-IL-8 antibody, wherein the polynucleotide sequence encoding the VH-VL comprises the paired sequences set forth in SEQ ID NOs: 131 and 132.In certain embodiments, a method for producing an anti-IL-8 antibody comprises culturing a host cell comprising a vector comprising an isolated polynucleotide sequence encoding the VH and VL of the anti-IL-8 antibody, wherein the polynucleotide sequence encoding the VH-VL comprises the paired sequences set forth in SEQ ID NOs: 133 and 134. In certain embodiments, a method for producing an anti-IL-8 antibody comprises culturing a host cell comprising a vector comprising an isolated polynucleotide sequence encoding the VH and VL of the anti-IL-8 antibody, wherein the polynucleotide sequence encoding the VH-VL comprises the paired sequences set forth in SEQ ID NOs: 135 and 136. In certain embodiments, a method for producing an anti-IL-8 antibody comprises culturing a host cell comprising a vector comprising an isolated polynucleotide sequence encoding the VH and VL of the anti-IL-8 antibody, wherein the polynucleotide sequence encoding the VH-VL comprises the paired sequences set forth in SEQ ID NOs: 137 and 138. In certain embodiments, a method for producing an anti-IL-8 antibody comprises culturing a host cell comprising a vector comprising an isolated polynucleotide sequence encoding the VH and VL of the anti-IL-8 antibody, wherein the polynucleotide sequence encoding the VH-VL comprises the paired sequences set forth in SEQ ID NOs: 139 and 140. In certain embodiments, a method for producing an anti-IL-8 antibody comprises culturing a host cell comprising a vector comprising an isolated polynucleotide sequence encoding the VH and VL of the anti-IL-8 antibody, wherein the polynucleotide sequence encoding the VH-VL comprises the paired sequences set forth in SEQ ID NOs: 141 and 142. In certain embodiments, a method for producing an anti-IL-8 antibody comprises culturing a host cell comprising a vector comprising an isolated polynucleotide sequence encoding the VH and VL of the anti-IL-8 antibody, wherein the polynucleotide sequence encoding the VH-VL comprises the paired sequences set forth in SEQ ID NOs: 36 and 37. In one embodiment, a method for producing an anti-IL-8 antibody comprises culturing a host cell containing a vector comprising an isolated polynucleotide sequence encoding the VH and VL of the anti-IL-8 antibody, wherein the polynucleotide sequence encoding the VH-VL comprises the paired sequences set forth in SEQ ID NOs: 38 and 39.

[0137]

[0147] In one embodiment, a method for producing an anti-IL-8 antibody comprising a complementarity determining region (CDR) sequence set forth in Table 1F comprises culturing a host cell containing a vector comprising an isolated polynucleotide sequence encoding a heavy chain variable region (VH) of an anti-IL-8 antibody comprising a VH complementarity determining region (HCDR) set forth in Table 1F and a light chain variable region (VL) of an anti-IL-8 antibody comprising a VL complementarity determining region (LCDR) set forth in Table 1F, wherein the heavy chain variable region has heavy chain complementarity determining region (HCDR)1, HCDR2, and HCDR3 under conditions conducive to expression of the vector in the host cell, wherein the light chain variable region has light chain complementarity determining region (LCDR)1, LCDR2, and LCDR3, and wherein the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 for the antibody are (a) SEQ ID NO:41, SEQ ID NO:44, SEQ ID NO:53, SEQ ID NO:60, SEQ ID NO:67, and SEQ ID NO:76; (b) SEQ ID NO:81, SEQ ID NO:84, SEQ ID NO:53, SEQ ID NO:93, SEQ ID NO:100, and SEQ ID NO:109; (c) SEQ ID NO:40, SEQ ID NO:43, SEQ ID NO:50, SEQ ID NO:57, SEQ ID NO:64, and SEQ ID NO:73; (d) SEQ ID NO:40, SEQ ID NO:43, SEQ ID NO:51, SEQ ID NO:58, SEQ ID NO:65, and SEQ ID NO:74; (e) SEQ ID NO:41, SEQ ID NO:44, SEQ ID NO:52, SEQ ID NO:59, SEQ ID NO:66, and SEQ ID NO:75; (f) SEQ ID NO:40, SEQ ID NO:43, SEQ ID NO:50, SEQ ID NO:57, SEQ ID NO:68, and SEQ ID NO:73; (g) SEQ ID NO:40, SEQ ID NO:43, SEQ ID NO:50, SEQ ID NO:57, SEQ ID NO:69, and SEQ ID NO:73; (h) SEQ ID NO:41, SEQ ID NO:45, SEQ ID NO:53, SEQ ID NO:61, SEQ ID NO:70, and SEQ ID NO:77; (i) SEQ ID NO:40, SEQ ID NO:46, SEQ ID NO:54, SEQ ID NO:62, SEQ ID NO:71, and SEQ ID NO:78; (j) SEQ ID NO:42, SEQ ID NO:47, SEQ ID NO:53, SEQ ID NO:63, SEQ ID NO:72, and SEQ ID NO:79; (k) SEQ ID NO:41, SEQ ID NO:48, SEQ ID NO:53, SEQ ID NO:60, SEQ ID NO:67, and SEQ ID NO:76; (l) SEQ ID NO:41, SEQ ID NO:49, SEQ ID NO:53, SEQ ID NO:60, SEQ ID NO:67, and SEQ ID NO:76; (m) SEQ ID NO:41, SEQ ID NO:44, SEQ ID NO:55, SEQ ID NO:60, SEQ ID NO:67, and SEQ ID NO:76; (n) SEQ ID NO:41, SEQ ID NO:44, SEQ ID NO:56, SEQ ID NO:60, SEQ ID NO:67, and SEQ ID NO:76; (o) SEQ ID NO:41, SEQ ID NO:48, SEQ ID NO:55, SEQ ID NO:60, SEQ ID NO:67, and SEQ ID NO:76; (p) SEQ ID NO:41, SEQ ID NO:48, SEQ ID NO:56, SEQ ID NO:60, SEQ ID NO:67, and SEQ ID NO:76; (q) SEQ ID NO:41, SEQ ID NO:49, SEQ ID NO:55, SEQ ID NO:60, SEQ ID NO:67, and SEQ ID NO:76; (r) SEQ ID NO:41, SEQ ID NO:49, SEQ ID NO:56, SEQ ID NO:60, SEQ ID NO:67, and SEQ ID NO:76; (s) SEQ ID NO: 80, SEQ ID NO: 83, SEQ ID NO: 50, SEQ ID NO: 90, SEQ ID NO: 97, and SEQ ID NO: 106; (t) SEQ ID NO:80, SEQ ID NO:83, SEQ ID NO:51, SEQ ID NO:91, SEQ ID NO:98, and SEQ ID NO:107; (u) SEQ ID NO:81, SEQ ID NO:84, SEQ ID NO:52, SEQ ID NO:92, SEQ ID NO:99, and SEQ ID NO:108; (v) SEQ ID NO:80, SEQ ID NO:83, SEQ ID NO:50, SEQ ID NO:90, SEQ ID NO:101, and SEQ ID NO:106; (w) SEQ ID NO:80, SEQ ID NO:83, SEQ ID NO:50, SEQ ID NO:90, SEQ ID NO:102, and SEQ ID NO:106; (x) SEQ ID NO:81, SEQ ID NO:85, SEQ ID NO:53, SEQ ID NO:94, SEQ ID NO:103, and SEQ ID NO:110; (y) SEQ ID NO:80, SEQ ID NO:86, SEQ ID NO:54, SEQ ID NO:95, SEQ ID NO:104, and SEQ ID NO:111; (z) SEQ ID NO:82, SEQ ID NO:87, SEQ ID NO:53, SEQ ID NO:93, SEQ ID NO:105, and SEQ ID NO:112; (aa) SEQ ID NO: 81, SEQ ID NO: 88, SEQ ID NO: 53, SEQ ID NO: 93, SEQ ID NO: 100, and SEQ ID NO: 109; (bb) SEQ ID NO:81, SEQ ID NO:89, SEQ ID NO:53, SEQ ID NO:93, SEQ ID NO:100, and SEQ ID NO:109; (cc) SEQ ID NO:81, SEQ ID NO:84, SEQ ID NO:55, SEQ ID NO:93, SEQ ID NO:100, and SEQ ID NO:109; (dd) SEQ ID NO:81, SEQ ID NO:84, SEQ ID NO:56, SEQ ID NO:93, SEQ ID NO:100, and SEQ ID NO:109; (ee) SEQ ID NO:81, SEQ ID NO:88, SEQ ID NO:55, SEQ ID NO:93, SEQ ID NO:100, and SEQ ID NO:109; (ff) SEQ ID NO:81, SEQ ID NO:88, SEQ ID NO:56, SEQ ID NO:93, SEQ ID NO:100, and SEQ ID NO:109; (gg) SEQ ID NO:81, SEQ ID NO:89, SEQ ID NO:55, SEQ ID NO:93, SEQ ID NO:100, and SEQ ID NO:109; or (hh) SEQ ID NO: 81, SEQ ID NO: 89, SEQ ID NO: 56, SEQ ID NO: 93, SEQ ID NO: 100, and SEQ ID NO: 109 2. The method of claim 1, wherein the amino acid sequence is and expressing the polynucleotide sequence contained in the vector, thereby producing an anti-IL-8 antibody having a complementarity-determining region (CDR) sequence set forth in Table 1F.

[0138]

[0148] In some embodiments of the method for producing an IL-8 antibody, the antibody is produced in vivo. In some embodiments of the method for producing an IL-8 antibody, the antibody is produced in vitro. In some embodiments of the method for producing an IL-8 antibody, if the antibody is produced in vitro, it may be isolated in a further step.

[0139] Composition of Use

[0149] The anti-IL-8 antibodies disclosed herein can be administered to a subject (e.g., a human or animal) alone or in combination with a carrier, i.e., a pharmaceutically acceptable carrier. Pharmaceutically acceptable means a material that is not biologically or otherwise harmful; i.e., the material can be administered to a subject without causing any undesired biological effects or interacting in a deleterious manner with any of the other components of the pharmaceutical composition in which it is contained. As will be known to those of skill in the art, a carrier will be selected to minimize degradation of the polypeptides disclosed herein and to minimize any adverse side effects in the subject. Those of skill in the art will understand that the term "physiologically acceptable carrier, diluent, or excipient" can, in certain embodiments, be used interchangeably with the term "pharmaceutically acceptable carrier," all of which have the same meaning and qualities.

[0140]

[0150] Pharmaceutical compositions may be prepared by methodology well known in the pharmaceutical art.

[0141]

[0151] Disclosed herein in one embodiment is a composition comprising an isolated anti-IL-8 antibody comprising an antibody antigen-binding domain comprising a heavy chain variable region (VH) and a light chain variable region (VL), wherein the amino acid sequence of the VH-VL pair is selected from the pair sequences set forth in SEQ ID NOs: 2 and 3, SEQ ID NOs: 4 and 5, SEQ ID NOs: 6 and 7, SEQ ID NOs: 8 and 9, SEQ ID NOs: 10 and 11, SEQ ID NOs: 12 and 13, SEQ ID NOs: 14 and 15, SEQ ID NOs: 16 and 17, SEQ ID NOs: 18 and 19, SEQ ID NOs: 20 and 21, SEQ ID NOs: 22 and 23, SEQ ID NOs: 24 and 25, SEQ ID NOs: 26 and 27, SEQ ID NOs: 28 and 29, SEQ ID NOs: 30 and 31, SEQ ID NOs: 32 and 33, and SEQ ID NOs: 34 and 35; and a pharmaceutically acceptable carrier. In one embodiment, disclosed herein is a composition comprising isolated anti-IL-8 antibodies having the complementarity determining region (CDR) sequences set forth in Table 1F, wherein each antibody comprises a heavy chain variable region having heavy chain complementarity determining region (HCDR)1, HCDR2, and HCDR3, and a light chain variable region having light chain complementarity determining region (LCDR)1, LCDR2, and LCDR3, and wherein the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 for each of the antibodies comprise the amino acid sequences set forth in Table 1F; and a pharmaceutically acceptable carrier. In certain embodiments, a composition comprises isolated anti-IL-8 antibodies comprising the CDR sequences set forth in Table 1F, each antibody comprising a heavy chain variable region having heavy chain complementarity determining region (HCDR) 1, HCDR2, and HCDR3, and a light chain variable region having light chain complementarity determining region (LCDR) 1, LCDR2, and LCDR3, wherein the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 for each antibody are (a) SEQ ID NO:41, SEQ ID NO:44, SEQ ID NO:53, SEQ ID NO:60, SEQ ID NO:67, and SEQ ID NO:76; (b) SEQ ID NO:81, SEQ ID NO:84, SEQ ID NO:53, SEQ ID NO:93, SEQ ID NO:100, and SEQ ID NO:109; (c) SEQ ID NO:40, SEQ ID NO:43, SEQ ID NO:50, SEQ ID NO:57, SEQ ID NO:64, and SEQ ID NO:73; (d) SEQ ID NO:40, SEQ ID NO:43, SEQ ID NO:51, SEQ ID NO:58, SEQ ID NO:65, and SEQ ID NO:74; (e) SEQ ID NO:41, SEQ ID NO:44, SEQ ID NO:52, SEQ ID NO:59, SEQ ID NO:66, and SEQ ID NO:75; (f) SEQ ID NO:40, SEQ ID NO:43, SEQ ID NO:50, SEQ ID NO:57, SEQ ID NO:68, and SEQ ID NO:73; (g) SEQ ID NO:40, SEQ ID NO:43, SEQ ID NO:50, SEQ ID NO:57, SEQ ID NO:69, and SEQ ID NO:73; (h) SEQ ID NO:41, SEQ ID NO:45, SEQ ID NO:53, SEQ ID NO:61, SEQ ID NO:70, and SEQ ID NO:77; (i) SEQ ID NO:40, SEQ ID NO:46, SEQ ID NO:54, SEQ ID NO:62, SEQ ID NO:71, and SEQ ID NO:78; (j) SEQ ID NO:42, SEQ ID NO:47, SEQ ID NO:53, SEQ ID NO:63, SEQ ID NO:72, and SEQ ID NO:79; (k) SEQ ID NO:41, SEQ ID NO:48, SEQ ID NO:53, SEQ ID NO:60, SEQ ID NO:67, and SEQ ID NO:76; (l) SEQ ID NO:41, SEQ ID NO:49, SEQ ID NO:53, SEQ ID NO:60, SEQ ID NO:67, and SEQ ID NO:76; (m) SEQ ID NO:41, SEQ ID NO:44, SEQ ID NO:55, SEQ ID NO:60, SEQ ID NO:67, and SEQ ID NO:76; (n) SEQ ID NO:41, SEQ ID NO:44, SEQ ID NO:56, SEQ ID NO:60, SEQ ID NO:67, and SEQ ID NO:76; (o) SEQ ID NO:41, SEQ ID NO:48, SEQ ID NO:55, SEQ ID NO:60, SEQ ID NO:67, and SEQ ID NO:76; (p) SEQ ID NO:41, SEQ ID NO:48, SEQ ID NO:56, SEQ ID NO:60, SEQ ID NO:67, and SEQ ID NO:76; (q) SEQ ID NO:41, SEQ ID NO:49, SEQ ID NO:55, SEQ ID NO:60, SEQ ID NO:67, and SEQ ID NO:76; (r) SEQ ID NO:41, SEQ ID NO:49, SEQ ID NO:56, SEQ ID NO:60, SEQ ID NO:67, and SEQ ID NO:76; (s) SEQ ID NO: 80, SEQ ID NO: 83, SEQ ID NO: 50, SEQ ID NO: 90, SEQ ID NO: 97, and SEQ ID NO: 106; (t) SEQ ID NO:80, SEQ ID NO:83, SEQ ID NO:51, SEQ ID NO:91, SEQ ID NO:98, and SEQ ID NO:107; (u) SEQ ID NO:81, SEQ ID NO:84, SEQ ID NO:52, SEQ ID NO:92, SEQ ID NO:99, and SEQ ID NO:108; (v) SEQ ID NO:80, SEQ ID NO:83, SEQ ID NO:50, SEQ ID NO:90, SEQ ID NO:101, and SEQ ID NO:106; (w) SEQ ID NO:80, SEQ ID NO:83, SEQ ID NO:50, SEQ ID NO:90, SEQ ID NO:102, and SEQ ID NO:106; (x) SEQ ID NO:81, SEQ ID NO:85, SEQ ID NO:53, SEQ ID NO:94, SEQ ID NO:103, and SEQ ID NO:110; (y) SEQ ID NO:80, SEQ ID NO:86, SEQ ID NO:54, SEQ ID NO:95, SEQ ID NO:104, and SEQ ID NO:111; (z) SEQ ID NO:82, SEQ ID NO:87, SEQ ID NO:53, SEQ ID NO:93, SEQ ID NO:105, and SEQ ID NO:112; (aa) SEQ ID NO: 81, SEQ ID NO: 88, SEQ ID NO: 53, SEQ ID NO: 93, SEQ ID NO: 100, and SEQ ID NO: 109; (bb) SEQ ID NO:81, SEQ ID NO:89, SEQ ID NO:53, SEQ ID NO:93, SEQ ID NO:100, and SEQ ID NO:109; (cc) SEQ ID NO:81, SEQ ID NO:84, SEQ ID NO:55, SEQ ID NO:93, SEQ ID NO:100, and SEQ ID NO:109; (dd) SEQ ID NO:81, SEQ ID NO:84, SEQ ID NO:56, SEQ ID NO:93, SEQ ID NO:100, and SEQ ID NO:109; (ee) SEQ ID NO:81, SEQ ID NO:88, SEQ ID NO:55, SEQ ID NO:93, SEQ ID NO:100, and SEQ ID NO:109; (ff) SEQ ID NO:81, SEQ ID NO:88, SEQ ID NO:56, SEQ ID NO:93, SEQ ID NO:100, and SEQ ID NO:109; (gg) SEQ ID NO:81, SEQ ID NO:89, SEQ ID NO:55, SEQ ID NO:93, SEQ ID NO:100, and SEQ ID NO:109; or (hh) SEQ ID NO: 81, SEQ ID NO: 89, SEQ ID NO: 56, SEQ ID NO: 93, SEQ ID NO: 100, and SEQ ID NO: 109 The amino acid sequence includes the amino acid sequence set forth in

[0142]

[0152] In certain embodiments of the compositions described herein, the antibody comprises an IgG, Fv, scFv, Fab, F(ab')2, minibody, diabody, triabody, nanobody, single domain antibody, multispecific antibody, bispecific antibody, triabody, single chain antibody, heavy chain antibody, chimeric antibody, or humanized antibody, or a combination thereof. In certain embodiments, the composition comprises an anti-IL-8 antibody comprising an IgG subclass selected from IgG1, IgG2, IgG3, or IgG4.

[0143]

[0153] Those skilled in the art will recognize that, in certain embodiments, the term "IL-8 binding antibody" can be used interchangeably with the term "drug" or "medicament," all of which have the same meaning and quality. In certain embodiments, a medicament comprising an IL-8 binding antibody includes a pharmaceutical composition.

[0144]

[0154] In certain embodiments, the anti-IL-8 antibodies disclosed herein can be administered to a subject (e.g., a human or animal) alone or as part of a combination therapy with an additional therapeutic agent. In certain embodiments, the compositions described herein comprise an anti-IL-8 antibody in combination with an additional therapeutic agent. In certain embodiments, the additional therapeutic agent comprises a chemotherapeutic agent, an antiviral agent, an immunomodulatory agent, or an additional therapeutic antibody, or any combination thereof.

[0145]

[0155] In certain embodiments, the anti-IL-8 antibody described herein and the additional therapeutic agent are comprised in the same composition. In certain embodiments, the anti-IL-8 antibody described herein and the additional therapeutic agent are comprised in different compositions. In certain embodiments, the administration of a combination of an anti-IL-8 antibody described herein and an additional therapeutic agent, or composition thereof, is simultaneous. In certain embodiments, the administration of a combination of an anti-IL-8 antibody described herein and an additional therapeutic agent, or composition thereof, comprises administration of the anti-IL-8 antibody or composition thereof before administration of the additional therapeutic agent. In certain embodiments, the administration of a combination of an anti-IL-8 antibody described herein and an additional therapeutic agent, or composition thereof, comprises administration of the anti-IL-8 antibody or composition thereof after administration of the additional therapeutic agent.

[0146]

[0156] The pharmaceutical compositions comprising the anti-IL-8 antibodies disclosed herein can be administered in any suitable manner (e.g., to a mammal, cell, or tissue) depending on whether local or systemic treatment is required. For example, the compositions can be administered topically (e.g., intraocularly, vaginally, rectally, intranasally, transdermally, etc.), orally, by inhalation, or parenterally (including by intravenous infusion or subcutaneous, intracavitary, intraperitoneal, intradermal, or intramuscular injection). Local intranasal administration refers to delivery of the composition to the nose and nasal cavity through one or both nostrils. The compositions can be delivered by a spray or infusion mechanism or by aerosolization. Delivery can also be directed to any site in the respiratory system (e.g., lungs) via intubation. Alternatively, administration can be intratumoral, e.g., by topical or intravenous injection.

[0147]

[0157] When the composition is to be administered parenterally, administration is generally by injection. Injectables can be prepared in conventional forms, either as liquid solutions or suspensions, solid forms suitable for suspension in liquid prior to injection, or as emulsions. Furthermore, parenteral administration can include formulations of slow-release or sustained-release systems to maintain a constant dosage.

[0148] How to use

[0158] The anti-IL-8 antibodies disclosed herein can be used in therapeutic methods. In certain embodiments, the anti-IL-8 antibodies can be used, for example, as immunotherapeutic agents to treat a subject suffering from a disease, wherein the disease includes cancer, tumor, or viral infection, as described herein. The anti-IL-8 antibodies disclosed herein can be used in therapeutic methods. In certain embodiments, the anti-IL-8 antibodies can be used, for example, as immunotherapeutic agents to treat a subject suffering from a disease, wherein the disease includes cancer, tumor, viral infection, inflammation, or a combination thereof, as described herein. In certain embodiments, the use of the anti-IL-8 antibodies described herein includes use as immunotherapeutic agents. In certain embodiments, the anti-IL-8 antibodies can be used, for example, as immunotherapeutic agents to address cytokine release syndrome or cytokine storm. In certain embodiments, the use of the anti-IL-8 antibodies reduces the severity of cytokine release syndrome or cytokine storm. In certain embodiments, the use of the anti-IL-8 antibodies reduces the severity of the effects of cytokine release syndrome or cytokine storm on a subject. In some embodiments, the use of anti-IL-8 antibodies reduces the harmful effects of cytokine release syndrome or cytokine storm on organs or tissues affected by viral infection. In some embodiments, the anti-IL-8 antibodies described herein are used to treat a subject suffering from cytokine release syndrome or cytokine storm. In some embodiments, the use of the anti-IL-8 antibodies described herein includes use as an anti-inflammatory agent. In some embodiments, methods of using IL-8 antibodies include treating inflammation in a subject in need thereof, for example, but not limited to, to reduce swelling in one or more joints of the subject.

[0149]

[0159] In certain embodiments, anti-IL-8 antibodies may be used as immunotherapeutic agents, for example, to inhibit tumor formation or growth, or a combination thereof. In certain embodiments, anti-IL-8 antibodies may be used as immunotherapeutic agents for specific inhibition of immune cell activation, for example, as described herein. In certain embodiments, anti-IL-8 antibodies may be used as immunotherapeutic agents to inhibit neutrophil or monocyte activation, or a combination thereof. In certain embodiments, anti-IL-8 antibodies may be used as immunotherapeutic agents to inhibit neutrophil or monocyte migration into a tumor microenvironment, or a combination thereof. In certain embodiments, anti-IL-8 antibodies may be used as immunotherapeutic agents to inhibit neutrophil or monocyte migration into a virally infected microenvironment, or a combination thereof. In certain embodiments, anti-IL-8 antibodies inhibit neutrophil and monocyte activation and migration to tissues and organs affected by viral infection. In certain embodiments, these tissues and / or organs are not infected by the virus but are adversely affected by indirect effects.

[0150]

[0160] In certain embodiments, anti-IL-8 antibodies block the ability of IL-8 to bind to its cell surface receptor, thereby preventing IL-8's ability to transduce cell signals. In certain embodiments, anti-IL-8 antibodies can be used as immunotherapeutic agents, where the anti-IL-8 antibody inhibits cell proliferation, for example, but not limited to, inhibiting tumor cell proliferation. In certain embodiments, anti-IL-8 antibodies can be used as immunotherapeutic agents, where the anti-IL-8 antibody reduces proliferation, for example, but not limited to, reducing tumor cell proliferation. In certain embodiments, anti-IL-8 antibodies can be used as immunotherapeutic agents, where the anti-IL-8 antibody inhibits cancer or tumor cell metastasis. In certain embodiments, anti-IL-8 antibodies can be used as immunotherapeutic agents, where the anti-IL-8 antibody reduces cancer or tumor cell metastasis.

[0151]

[0161]

[0014] In one embodiment, disclosed herein is a method of inhibiting tumor formation or growth, or a combination thereof, in a subject in need thereof, the method comprising the step of administering to the subject an anti-IL-8 antibody disclosed herein comprising an antibody antigen-binding domain comprising a heavy chain variable region (VH) and a light chain variable region (VL), thereby inhibiting tumor formation or growth, or a combination thereof, in the subject, wherein the amino acid sequences of the VH-VL pairs are selected from the paired sequences set forth in SEQ ID NOs: 2 and 3, SEQ ID NOs: 4 and 5, SEQ ID NOs: 6 and 7, SEQ ID NOs: 8 and 9, SEQ ID NOs: 10 and 11, SEQ ID NOs: 12 and 13, SEQ ID NOs: 14 and 15, SEQ ID NOs: 16 and 17, SEQ ID NOs: 18 and 19, SEQ ID NOs: 20 and 21, SEQ ID NOs: 22 and 23, SEQ ID NOs: 24 and 25, SEQ ID NOs: 26 and 27, SEQ ID NOs: 28 and 29, SEQ ID NOs: 30 and 31, SEQ ID NOs: 32 and 33, and SEQ ID NOs: 34 and 35. In one embodiment, a method for inhibiting tumor formation or growth, or a combination thereof, in a subject in need thereof comprises administering to the subject an anti-IL-8 antibody comprising an antibody antigen-binding domain comprising a heavy chain variable region (VH) and a light chain variable region (VL), wherein the amino acid sequence of the VH-VL pair comprises the pairing sequences set forth in SEQ ID NOs: 2 and 3. In one embodiment, a method for inhibiting tumor formation or growth, or a combination thereof, in a subject in need thereof comprises administering to the subject an anti-IL-8 antibody comprising an antibody antigen-binding domain comprising a heavy chain variable region (VH) and a light chain variable region (VL), wherein the amino acid sequence of the VH-VL pair comprises the pairing sequences set forth in SEQ ID NOs: 4 and 5. In one embodiment, a method for inhibiting tumor formation or growth, or a combination thereof, in a subject in need thereof comprises administering to the subject an anti-IL-8 antibody comprising an antibody antigen-binding domain comprising a heavy chain variable region (VH) and a light chain variable region (VL), wherein the amino acid sequence of the VH-VL pair comprises the pairing sequences set forth in SEQ ID NOs: 6 and 7. In one embodiment, a method for inhibiting tumor formation or growth, or a combination thereof, in a subject in need thereof comprises administering to the subject an anti-IL-8 antibody comprising an antibody antigen-binding domain comprising a heavy chain variable region (VH) and a light chain variable region (VL), wherein the amino acid sequences of the VH-VL pair comprise the paired sequences set forth in SEQ ID NOs: 8 and 9.In one embodiment, a method for inhibiting tumor formation or growth, or a combination thereof, in a subject in need thereof comprises administering to the subject an anti-IL-8 antibody comprising an antibody antigen-binding domain comprising a heavy chain variable region (VH) and a light chain variable region (VL), wherein the amino acid sequence of the VH-VL pair comprises the pairing sequences set forth in SEQ ID NOs: 10 and 11. In one embodiment, a method for inhibiting tumor formation or growth, or a combination thereof, in a subject in need thereof comprises administering to the subject an anti-IL-8 antibody comprising an antibody antigen-binding domain comprising a heavy chain variable region (VH) and a light chain variable region (VL), wherein the amino acid sequence of the VH-VL pair comprises the pairing sequences set forth in SEQ ID NOs: 12 and 13. In one embodiment, a method for inhibiting tumor formation or growth, or a combination thereof, in a subject in need thereof comprises administering to the subject an anti-IL-8 antibody comprising an antibody antigen-binding domain comprising a heavy chain variable region (VH) and a light chain variable region (VL), wherein the amino acid sequence of the VH-VL pair comprises the pairing sequences set forth in SEQ ID NOs: 14 and 15. In one embodiment, a method for inhibiting tumor formation or growth, or a combination thereof, in a subject in need thereof comprises administering to the subject an anti-IL-8 antibody comprising an antibody antigen-binding domain comprising a heavy chain variable region (VH) and a light chain variable region (VL), wherein the amino acid sequence of the VH-VL pair comprises the pairing sequences set forth in SEQ ID NOs: 16 and 17. In one embodiment, a method for inhibiting tumor formation or growth, or a combination thereof, in a subject in need thereof comprises administering to the subject an anti-IL-8 antibody comprising an antibody antigen-binding domain comprising a heavy chain variable region (VH) and a light chain variable region (VL), wherein the amino acid sequence of the VH-VL pair comprises the pairing sequences set forth in SEQ ID NOs: 18 and 19. In one embodiment, a method for inhibiting tumor formation or growth, or a combination thereof, in a subject in need thereof comprises administering to the subject an anti-IL-8 antibody comprising an antibody antigen-binding domain comprising a heavy chain variable region (VH) and a light chain variable region (VL), wherein the amino acid sequence of the VH-VL pair comprises the pairing sequences set forth in SEQ ID NOs: 20 and 21.In one embodiment, a method for inhibiting tumor formation or growth, or a combination thereof, in a subject in need thereof comprises administering to the subject an anti-IL-8 antibody comprising an antibody antigen-binding domain comprising a heavy chain variable region (VH) and a light chain variable region (VL), wherein the amino acid sequence of the VH-VL pair comprises the pairing sequences set forth in SEQ ID NOs: 22 and 23. In one embodiment, a method for inhibiting tumor formation or growth, or a combination thereof, in a subject in need thereof comprises administering to the subject an anti-IL-8 antibody comprising an antibody antigen-binding domain comprising a heavy chain variable region (VH) and a light chain variable region (VL), wherein the amino acid sequence of the VH-VL pair comprises the pairing sequences set forth in SEQ ID NOs: 24 and 25. In one embodiment, a method for inhibiting tumor formation or growth, or a combination thereof, in a subject in need thereof comprises administering to the subject an anti-IL-8 antibody comprising an antibody antigen-binding domain comprising a heavy chain variable region (VH) and a light chain variable region (VL), wherein the amino acid sequence of the VH-VL pair comprises the pairing sequences set forth in SEQ ID NOs: 26 and 27. In one embodiment, a method for inhibiting tumor formation or growth, or a combination thereof, in a subject in need thereof comprises administering to the subject an anti-IL-8 antibody comprising an antibody antigen-binding domain comprising a heavy chain variable region (VH) and a light chain variable region (VL), wherein the amino acid sequence of the VH-VL pair comprises the pairing sequences set forth in SEQ ID NOs: 28 and 29. In one embodiment, a method for inhibiting tumor formation or growth, or a combination thereof, in a subject in need thereof comprises administering to the subject an anti-IL-8 antibody comprising an antibody antigen-binding domain comprising a heavy chain variable region (VH) and a light chain variable region (VL), wherein the amino acid sequence of the VH-VL pair comprises the pairing sequences set forth in SEQ ID NOs: 30 and 31. In one embodiment, a method for inhibiting tumor formation or growth, or a combination thereof, in a subject in need thereof comprises administering to the subject an anti-IL-8 antibody comprising an antibody antigen-binding domain comprising a heavy chain variable region (VH) and a light chain variable region (VL), wherein the amino acid sequence of the VH-VL pair comprises the pairing sequences set forth in SEQ ID NOs: 32 and 33.In one embodiment, a method for inhibiting tumor formation or growth, or a combination thereof, in a subject in need thereof comprises administering to the subject an anti-IL-8 antibody comprising an antibody antigen-binding domain comprising a heavy chain variable region (VH) and a light chain variable region (VL), wherein the amino acid sequences of the VH-VL pair comprise the paired sequences set forth in SEQ ID NOs: 34 and 35.

[0152]

[0162] In one embodiment, disclosed herein is a method of inhibiting tumor formation or growth, or a combination thereof, in a subject in need thereof, comprising the step of administering to the subject anti-IL-8 antibodies having complementarity determining region (CDR) sequences set forth in Table 1F, thereby inhibiting tumor formation or growth, or a combination thereof, in the subject, wherein each antibody comprises a heavy chain variable region having heavy chain complementarity determining region (HCDR) 1, HCDR2, and HCDR3, and a light chain variable region having light chain complementarity determining region (LCDR) 1, LCDR2, and LCDR3, and wherein the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 for each of the antibodies comprise the amino acid sequences set forth in Table 1F. In one embodiment, a method of inhibiting tumor formation or growth, or a combination thereof, in a subject in need thereof, comprising the step of administering to the subject anti-IL-8 antibodies having complementarity determining region (CDR) sequences as set forth in Table 1F, thereby inhibiting tumor formation or growth, or a combination thereof, in the subject, wherein each antibody comprises a heavy chain variable region having heavy chain complementarity determining region (HCDR) 1, HCDR2, and HCDR3, and a light chain variable region having light chain complementarity determining region (LCDR) 1, LCDR2, and LCDR3, and wherein the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 for each of the antibodies are (a) SEQ ID NO:41, SEQ ID NO:44, SEQ ID NO:53, SEQ ID NO:60, SEQ ID NO:67, and SEQ ID NO:76; (b) SEQ ID NO:81, SEQ ID NO:84, SEQ ID NO:53, SEQ ID NO:93, SEQ ID NO:100, and SEQ ID NO:109; (c) SEQ ID NO:40, SEQ ID NO:43, SEQ ID NO:50, SEQ ID NO:57, SEQ ID NO:64, and SEQ ID NO:73; (d) SEQ ID NO:40, SEQ ID NO:43, SEQ ID NO:51, SEQ ID NO:58, SEQ ID NO:65, and SEQ ID NO:74; (e) SEQ ID NO:41, SEQ ID NO:44, SEQ ID NO:52, SEQ ID NO:59, SEQ ID NO:66, and SEQ ID NO:75; (f) SEQ ID NO:40, SEQ ID NO:43, SEQ ID NO:50, SEQ ID NO:57, SEQ ID NO:68, and SEQ ID NO:73; (g) SEQ ID NO:40, SEQ ID NO:43, SEQ ID NO:50, SEQ ID NO:57, SEQ ID NO:69, and SEQ ID NO:73; (h) SEQ ID NO:41, SEQ ID NO:45, SEQ ID NO:53, SEQ ID NO:61, SEQ ID NO:70, and SEQ ID NO:77; (i) SEQ ID NO:40, SEQ ID NO:46, SEQ ID NO:54, SEQ ID NO:62, SEQ ID NO:71, and SEQ ID NO:78; (j) SEQ ID NO:42, SEQ ID NO:47, SEQ ID NO:53, SEQ ID NO:63, SEQ ID NO:72, and SEQ ID NO:79; (k) SEQ ID NO:41, SEQ ID NO:48, SEQ ID NO:53, SEQ ID NO:60, SEQ ID NO:67, and SEQ ID NO:76; (l) SEQ ID NO:41, SEQ ID NO:49, SEQ ID NO:53, SEQ ID NO:60, SEQ ID NO:67, and SEQ ID NO:76; (m) SEQ ID NO:41, SEQ ID NO:44, SEQ ID NO:55, SEQ ID NO:60, SEQ ID NO:67, and SEQ ID NO:76; (n) SEQ ID NO:41, SEQ ID NO:44, SEQ ID NO:56, SEQ ID NO:60, SEQ ID NO:67, and SEQ ID NO:76; (o) SEQ ID NO:41, SEQ ID NO:48, SEQ ID NO:55, SEQ ID NO:60, SEQ ID NO:67, and SEQ ID NO:76; (p) SEQ ID NO:41, SEQ ID NO:48, SEQ ID NO:56, SEQ ID NO:60, SEQ ID NO:67, and SEQ ID NO:76; (q) SEQ ID NO:41, SEQ ID NO:49, SEQ ID NO:55, SEQ ID NO:60, SEQ ID NO:67, and SEQ ID NO:76; (r) SEQ ID NO:41, SEQ ID NO:49, SEQ ID NO:56, SEQ ID NO:60, SEQ ID NO:67, and SEQ ID NO:76; (s) SEQ ID NO: 80, SEQ ID NO: 83, SEQ ID NO: 50, SEQ ID NO: 90, SEQ ID NO: 97, and SEQ ID NO: 106; (t) SEQ ID NO:80, SEQ ID NO:83, SEQ ID NO:51, SEQ ID NO:91, SEQ ID NO:98, and SEQ ID NO:107; (u) SEQ ID NO:81, SEQ ID NO:84, SEQ ID NO:52, SEQ ID NO:92, SEQ ID NO:99, and SEQ ID NO:108; (v) SEQ ID NO:80, SEQ ID NO:83, SEQ ID NO:50, SEQ ID NO:90, SEQ ID NO:101, and SEQ ID NO:106; (w) SEQ ID NO:80, SEQ ID NO:83, SEQ ID NO:50, SEQ ID NO:90, SEQ ID NO:102, and SEQ ID NO:106; (x) SEQ ID NO:81, SEQ ID NO:85, SEQ ID NO:53, SEQ ID NO:94, SEQ ID NO:103, and SEQ ID NO:110; (y) SEQ ID NO:80, SEQ ID NO:86, SEQ ID NO:54, SEQ ID NO:95, SEQ ID NO:104, and SEQ ID NO:111; (z) SEQ ID NO:82, SEQ ID NO:87, SEQ ID NO:53, SEQ ID NO:93, SEQ ID NO:105, and SEQ ID NO:112; (aa) SEQ ID NO: 81, SEQ ID NO: 88, SEQ ID NO: 53, SEQ ID NO: 93, SEQ ID NO: 100, and SEQ ID NO: 109; (bb) SEQ ID NO:81, SEQ ID NO:89, SEQ ID NO:53, SEQ ID NO:93, SEQ ID NO:100, and SEQ ID NO:109; (cc) SEQ ID NO:81, SEQ ID NO:84, SEQ ID NO:55, SEQ ID NO:93, SEQ ID NO:100, and SEQ ID NO:109; (dd) SEQ ID NO:81, SEQ ID NO:84, SEQ ID NO:56, SEQ ID NO:93, SEQ ID NO:100, and SEQ ID NO:109; (ee) SEQ ID NO:81, SEQ ID NO:88, SEQ ID NO:55, SEQ ID NO:93, SEQ ID NO:100, and SEQ ID NO:109; (ff) SEQ ID NO:81, SEQ ID NO:88, SEQ ID NO:56, SEQ ID NO:93, SEQ ID NO:100, and SEQ ID NO:109; (gg) SEQ ID NO:81, SEQ ID NO:89, SEQ ID NO:55, SEQ ID NO:93, SEQ ID NO:100, and SEQ ID NO:109; or (hh) SEQ ID NO: 81, SEQ ID NO: 89, SEQ ID NO: 56, SEQ ID NO: 93, SEQ ID NO: 100, and SEQ ID NO: 109 Disclosed herein is a method comprising the amino acid sequence set forth in

[0153]

[0163] In certain embodiments, the method of inhibiting tumor formation or growth, or both, inhibits tumor formation. In certain embodiments, the method of inhibiting tumor formation or growth, or both, reduces the rate of tumor formation. In certain embodiments, the method of inhibiting tumor formation or growth, or both, inhibits tumor growth. In certain embodiments, the method of inhibiting tumor formation or growth, or both, reduces the rate of tumor growth. In certain embodiments, the method of inhibiting tumor formation or growth, or both, halts tumor growth. In certain embodiments, the method of inhibiting tumor formation or growth, or both, inhibits de novo tumor formation and reduces tumor growth. In certain embodiments, the method of inhibiting tumor formation or growth, or both, inhibits de novo tumor formation, inhibits tumor growth, and inhibits metastasis. In certain embodiments, the method of inhibiting tumor formation or growth, or both, reduces the rate of de novo tumor formation, reduces tumor growth, and reduces the rate of tumor metastasis. In certain embodiments, the method of inhibiting tumor formation or growth, or both, inhibits tumor metastasis. In certain embodiments, the method of inhibiting tumor formation or growth, or both, reduces the rate of tumor metastasis.

[0154]

[0164] In certain embodiments, the method of inhibiting tumor formation or growth, or both, inhibits neutrophil or monocyte activation, or a combination thereof, within the tumor microenvironment. In certain embodiments, the method of inhibiting tumor formation or growth, or both, reduces neutrophil or monocyte activation, or a combination thereof, within the tumor microenvironment. Those skilled in the art will understand that the terms "tumor microenvironment," "cancer microenvironment," "TME," and "tumor environment" have the same qualities and meanings, can be used interchangeably, and encompass the microenvironment for tumor development. While a normal cell microenvironment may inhibit malignant cell growth, modulations occurring within the tumor microenvironment may synergistically support cell growth.

[0155]

[0165] Tumors shape their microenvironment, supporting the development of both tumor and non-malignant cells. The tumor microenvironment influences angiogenesis by disrupting signaling pathways required for cell recruitment and blood vessel assembly. Endothelial progenitor cells (EPCs), which are recruited under hypoxic conditions for angiogenesis, have also been associated with metastasis. Furthermore, proteins such as IL-8 can be secreted by tumors or cancers, where the presence of secreted proteins can modulate the microenvironment by contributing growth factors and proteases that degrade the extracellular matrix and affect cell motility and adhesion.

[0156]

[0166] In certain embodiments, the method of inhibiting tumor formation or growth, or both, reduces the viability of pre-cancerous stem cells or tumor cells. Cell viability can be assessed by known techniques, such as trypan blue dye exclusion assays. Viability or, conversely, toxicity can also be measured based on cell survival, e.g., comparing the viability of normal and cancerous cell cultures exposed to anti-IL-8 antibodies. Toxicity can also be measured based on cell lysis, e.g., comparing the lysis of normal and cancerous cell cultures exposed to anti-IL-8 antibodies. Cell lysis can be assessed by known techniques, such as chromium (Cr) release assays or dead cell indicator dyes (propidium iodide, TO-PRO-3 iodide).

[0157]

[0167] In some embodiments of the methods for inhibiting tumor formation or growth, or both, the pre-cancerous stem cells comprise pre-hematopoietic cancer stem cells. In some embodiments of the methods for inhibiting tumor formation or growth, or both, the tumor cells comprise hematopoietic cancer cells. In some embodiments, hematopoietic tumors are types of cancer that affect the blood, bone marrow, and lymph nodes. Hematopoietic tumors can originate from either of two major blood cell lineages: myeloid and lymphoid cell lines. Myeloid cell lines typically produce granulocytes, erythrocytes, platelets, macrophages, and mast cells, while lymphoid cell lines produce B, T, and plasma cells. Lymphomas (e.g., Hodgkin's lymphoma), lymphocytic leukemias, and myelomas originate from the lymphatic system, while acute and chronic myeloid leukemias (AML, CML), myelodysplastic syndromes, and myeloproliferative disorders are myeloid in origin. In some embodiments of the methods for inhibiting tumor formation or growth, or both, the pre-cancerous stem cells comprise pre-leukemic cancer stem cells. In certain embodiments, the hematological cancer comprises leukemia, lymphoma, myeloma, acute myeloid leukemia (AML), acute promyelocytic leukemia, erythroleukemia, mixed B-myelomonocytic leukemia, or myelodysplastic syndrome (MDS).

[0158]

[0168] In some embodiments of the method of inhibiting tumor formation or growth, or both, the pre-cancerous stem cells comprise a solid cancer or tumor, hi some embodiments, the solid cancer or tumor comprises a sarcoma, osteosarcoma, squamous cell carcinoma of the head and neck, non-small cell lung cancer, bladder cancer, pancreatic cancer, or pancreatic ductal adenocarcinoma.

[0159]

[0169] In certain embodiments of the method of inhibiting tumor formation or growth, or both, the subject is a human.

[0160]

[0170] Disclosed herein in one embodiment is a method of treating a subject suffering from a disease, the method comprising administering to the subject an anti-IL-8 antibody comprising an antibody antigen-binding domain comprising a heavy chain variable region (VH) and a light chain variable region (VL), thereby treating the disease in the subject, wherein the disease comprises cancer or tumor or viral infection or inflammation-related disease, or a combination thereof, and the amino acid sequences of the VH-VL pairs are selected from the paired sequences set forth in SEQ ID NOs: 2 and 3, SEQ ID NOs: 4 and 5, SEQ ID NOs: 6 and 7, SEQ ID NOs: 8 and 9, SEQ ID NOs: 10 and 11, SEQ ID NOs: 12 and 13, SEQ ID NOs: 14 and 15, SEQ ID NOs: 16 and 17, SEQ ID NOs: 18 and 19, SEQ ID NOs: 20 and 21, SEQ ID NOs: 22 and 23, SEQ ID NOs: 24 and 25, SEQ ID NOs: 26 and 27, SEQ ID NOs: 28 and 29, SEQ ID NOs: 30 and 31, SEQ ID NOs: 32 and 33, and SEQ ID NOs: 34 and 35. In one embodiment, a method for treating a subject suffering from a disease comprises administering to the subject an anti-IL-8 antibody comprising an antibody antigen-binding domain comprising a heavy chain variable region (VH) and a light chain variable region (VL), wherein the disease comprises cancer, tumor, viral infection, inflammation-associated disease, or a combination thereof, and the amino acid sequence of the VH-VL pair comprises the paired sequences set forth in SEQ ID NOs: 2 and 3. In one embodiment, a method for treating a subject suffering from a disease comprises administering to the subject an anti-IL-8 antibody comprising an antibody antigen-binding domain comprising a heavy chain variable region (VH) and a light chain variable region (VL), wherein the disease comprises cancer, tumor, viral infection, inflammation-associated disease, or a combination thereof, and the amino acid sequence of the VH-VL pair comprises the paired sequences set forth in SEQ ID NOs: 4 and 5. In one embodiment, a method for treating a subject suffering from a disease comprises administering to the subject an anti-IL-8 antibody comprising an antibody antigen-binding domain comprising a heavy chain variable region (VH) and a light chain variable region (VL), wherein the disease comprises cancer, tumor, viral infection, inflammation-associated disease, or a combination thereof, and the amino acid sequence of the VH-VL pair comprises the paired sequences set forth in SEQ ID NOs: 6 and 7.In one embodiment, a method for treating a subject suffering from a disease comprises administering to the subject an anti-IL-8 antibody comprising an antibody antigen-binding domain comprising a heavy chain variable region (VH) and a light chain variable region (VL), wherein the disease comprises cancer, or a tumor, or a disease associated with viral infection or inflammation, or a combination thereof, and the amino acid sequence of the VH-VL pair comprises the paired sequences set forth in SEQ ID NOs: 8 and 9. In one embodiment, a method for treating a subject suffering from a disease comprises administering to the subject an anti-IL-8 antibody comprising an antibody antigen-binding domain comprising a heavy chain variable region (VH) and a light chain variable region (VL), wherein the disease comprises cancer, or a tumor, or a disease associated with viral infection or inflammation, or a combination thereof, and the amino acid sequence of the VH-VL pair comprises the paired sequences set forth in SEQ ID NOs: 10 and 11. In one embodiment, a method for treating a subject suffering from a disease comprises administering to the subject an anti-IL-8 antibody comprising an antibody antigen-binding domain comprising a heavy chain variable region (VH) and a light chain variable region (VL), wherein the disease comprises cancer, tumor, viral infection, inflammation-associated disease, or a combination thereof, and the amino acid sequence of the VH-VL pair comprises the paired sequences set forth in SEQ ID NOs: 12 and 13. In one embodiment, a method for treating a subject suffering from a disease comprises administering to the subject an anti-IL-8 antibody comprising an antibody antigen-binding domain comprising a heavy chain variable region (VH) and a light chain variable region (VL), wherein the disease comprises cancer, tumor, viral infection, inflammation-associated disease, or a combination thereof, and the amino acid sequence of the VH-VL pair comprises the paired sequences set forth in SEQ ID NOs: 14 and 15. In one embodiment, a method for treating a subject suffering from a disease comprises administering to the subject an anti-IL-8 antibody comprising an antibody antigen-binding domain comprising a heavy chain variable region (VH) and a light chain variable region (VL), wherein the disease comprises cancer, tumor, viral infection, inflammation-associated disease, or a combination thereof, and the amino acid sequence of the VH-VL pair comprises the paired sequences set forth in SEQ ID NOs: 16 and 17.In one embodiment, a method for treating a subject suffering from a disease comprises administering to the subject an anti-IL-8 antibody comprising an antibody antigen-binding domain comprising a heavy chain variable region (VH) and a light chain variable region (VL), wherein the disease comprises cancer, tumor, viral infection, inflammation-associated disease, or a combination thereof, and wherein the amino acid sequence of the VH-VL pair comprises the paired sequences set forth in SEQ ID NOs: 18 and 19. In one embodiment, a method for treating a subject suffering from a disease comprises administering to the subject an anti-IL-8 antibody comprising an antibody antigen-binding domain comprising a heavy chain variable region (VH) and a light chain variable region (VL), wherein the disease comprises cancer, tumor, viral infection, inflammation-associated disease, or a combination thereof, and wherein the amino acid sequence of the VH-VL pair comprises the paired sequences set forth in SEQ ID NOs: 20 and 21. In one embodiment, a method for treating a subject suffering from a disease comprises administering to the subject an anti-IL-8 antibody comprising an antibody antigen-binding domain comprising a heavy chain variable region (VH) and a light chain variable region (VL), wherein the disease comprises cancer, tumor, viral infection, inflammation-associated disease, or a combination thereof, and the amino acid sequence of the VH-VL pair comprises the paired sequences set forth in SEQ ID NOs: 22 and 23. In one embodiment, a method for treating a subject suffering from a disease comprises administering to the subject an anti-IL-8 antibody comprising an antibody antigen-binding domain comprising a heavy chain variable region (VH) and a light chain variable region (VL), wherein the disease comprises cancer, tumor, viral infection, inflammation-associated disease, or a combination thereof, and the amino acid sequence of the VH-VL pair comprises the paired sequences set forth in SEQ ID NOs: 24 and 25. In one embodiment, a method for treating a subject suffering from a disease comprises administering to the subject an anti-IL-8 antibody comprising an antibody antigen-binding domain comprising a heavy chain variable region (VH) and a light chain variable region (VL), wherein the disease comprises cancer, tumor, viral infection, inflammation-associated disease, or a combination thereof, and the amino acid sequence of the VH-VL pair comprises the paired sequences set forth in SEQ ID NOs: 26 and 27.In one embodiment, a method for treating a subject suffering from a disease comprises administering to the subject an anti-IL-8 antibody comprising an antibody antigen-binding domain comprising a heavy chain variable region (VH) and a light chain variable region (VL), wherein the disease comprises cancer, tumor, viral infection, inflammation-associated disease, or a combination thereof, and the amino acid sequence of the VH-VL pair comprises the paired sequences set forth in SEQ ID NOs: 28 and 29. In one embodiment, a method for treating a subject suffering from a disease comprises administering to the subject an anti-IL-8 antibody comprising an antibody antigen-binding domain comprising a heavy chain variable region (VH) and a light chain variable region (VL), wherein the disease comprises cancer, tumor, viral infection, inflammation-associated disease, or a combination thereof, and the amino acid sequence of the VH-VL pair comprises the paired sequences set forth in SEQ ID NOs: 30 and 31. In one embodiment, a method for treating a subject suffering from a disease comprises administering to the subject an anti-IL-8 antibody comprising an antibody antigen-binding domain comprising a heavy chain variable region (VH) and a light chain variable region (VL), wherein the disease comprises cancer, tumor, viral infection, inflammation-associated disease, or a combination thereof, and wherein the amino acid sequences of the VH-VL pair comprise the paired sequences set forth in SEQ ID NOs: 32 and 33. In one embodiment, a method for treating a subject suffering from a disease comprises administering to the subject an anti-IL-8 antibody comprising an antibody antigen-binding domain comprising a heavy chain variable region (VH) and a light chain variable region (VL), wherein the disease comprises cancer, tumor, viral infection, inflammation-associated disease, or a combination thereof, and wherein the amino acid sequences of the VH-VL pair comprise the paired sequences set forth in SEQ ID NOs: 34 and 35. In one embodiment, the method for treating a subject suffering from a disease comprises treating cancer or tumor. In one embodiment, the method for treating a subject suffering from a disease comprises treating viral infection. In certain embodiments, the method of treating a subject suffering from a disease comprises treating a disease associated with inflammation, or a combination thereof.

[0161]

[0171] In certain embodiments, disclosed herein are methods of treating a subject suffering from a disease, the method comprising administering to the subject anti-IL-8 antibodies having complementarity determining region (CDR) sequences set forth in Table 1F, thereby treating the disease in the subject, wherein the disease comprises cancer or a tumor or a viral infection or a disease associated with inflammation, or a combination thereof, and wherein each antibody comprises a heavy chain variable region having heavy chain complementarity determining region (HCDR) 1, HCDR2, and HCDR3, and a light chain variable region having light chain complementarity determining region (LCDR) 1, LCDR2, and LCDR3, and wherein the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 for each of the antibodies comprise the amino acid sequences set forth in Table 1F. In certain embodiments, there is provided a method of treating a subject suffering from a disease, the method comprising administering to the subject anti-IL-8 antibodies having complementarity determining region (CDR) sequences set forth in Table 1F, thereby treating the disease in the subject, wherein the disease comprises cancer or a tumor or a disease associated with a viral infection or inflammation, or a combination thereof, and wherein each antibody comprises a heavy chain variable region having heavy chain complementarity determining region (HCDR) 1, HCDR2, and HCDR3, and a light chain variable region having light chain complementarity determining region (LCDR) 1, LCDR2, and LCDR3, and wherein the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 for each of the antibodies are (a) SEQ ID NO:41, SEQ ID NO:44, SEQ ID NO:53, SEQ ID NO:60, SEQ ID NO:67, and SEQ ID NO:76; (b) SEQ ID NO:81, SEQ ID NO:84, SEQ ID NO:53, SEQ ID NO:93, SEQ ID NO:100, and SEQ ID NO:109; (c) SEQ ID NO:40, SEQ ID NO:43, SEQ ID NO:50, SEQ ID NO:57, SEQ ID NO:64, and SEQ ID NO:73; (d) SEQ ID NO:40, SEQ ID NO:43, SEQ ID NO:51, SEQ ID NO:58, SEQ ID NO:65, and SEQ ID NO:74; (e) SEQ ID NO:41, SEQ ID NO:44, SEQ ID NO:52, SEQ ID NO:59, SEQ ID NO:66, and SEQ ID NO:75; (f) SEQ ID NO:40, SEQ ID NO:43, SEQ ID NO:50, SEQ ID NO:57, SEQ ID NO:68, and SEQ ID NO:73; (g) SEQ ID NO:40, SEQ ID NO:43, SEQ ID NO:50, SEQ ID NO:57, SEQ ID NO:69, and SEQ ID NO:73; (h) SEQ ID NO:41, SEQ ID NO:45, SEQ ID NO:53, SEQ ID NO:61, SEQ ID NO:70, and SEQ ID NO:77; (i) SEQ ID NO:40, SEQ ID NO:46, SEQ ID NO:54, SEQ ID NO:62, SEQ ID NO:71, and SEQ ID NO:78; (j) SEQ ID NO:42, SEQ ID NO:47, SEQ ID NO:53, SEQ ID NO:63, SEQ ID NO:72, and SEQ ID NO:79; (k) SEQ ID NO:41, SEQ ID NO:48, SEQ ID NO:53, SEQ ID NO:60, SEQ ID NO:67, and SEQ ID NO:76; (l) SEQ ID NO:41, SEQ ID NO:49, SEQ ID NO:53, SEQ ID NO:60, SEQ ID NO:67, and SEQ ID NO:76; (m) SEQ ID NO:41, SEQ ID NO:44, SEQ ID NO:55, SEQ ID NO:60, SEQ ID NO:67, and SEQ ID NO:76; (n) SEQ ID NO:41, SEQ ID NO:44, SEQ ID NO:56, SEQ ID NO:60, SEQ ID NO:67, and SEQ ID NO:76; (o) SEQ ID NO:41, SEQ ID NO:48, SEQ ID NO:55, SEQ ID NO:60, SEQ ID NO:67, and SEQ ID NO:76; (p) SEQ ID NO:41, SEQ ID NO:48, SEQ ID NO:56, SEQ ID NO:60, SEQ ID NO:67, and SEQ ID NO:76; (q) SEQ ID NO:41, SEQ ID NO:49, SEQ ID NO:55, SEQ ID NO:60, SEQ ID NO:67, and SEQ ID NO:76; (r) SEQ ID NO:41, SEQ ID NO:49, SEQ ID NO:56, SEQ ID NO:60, SEQ ID NO:67, and SEQ ID NO:76; (s) SEQ ID NO: 80, SEQ ID NO: 83, SEQ ID NO: 50, SEQ ID NO: 90, SEQ ID NO: 97, and SEQ ID NO: 106; (t) SEQ ID NO:80, SEQ ID NO:83, SEQ ID NO:51, SEQ ID NO:91, SEQ ID NO:98, and SEQ ID NO:107; (u) SEQ ID NO:81, SEQ ID NO:84, SEQ ID NO:52, SEQ ID NO:92, SEQ ID NO:99, and SEQ ID NO:108; (v) SEQ ID NO:80, SEQ ID NO:83, SEQ ID NO:50, SEQ ID NO:90, SEQ ID NO:101, and SEQ ID NO:106; (w) SEQ ID NO:80, SEQ ID NO:83, SEQ ID NO:50, SEQ ID NO:90, SEQ ID NO:102, and SEQ ID NO:106; (x) SEQ ID NO:81, SEQ ID NO:85, SEQ ID NO:53, SEQ ID NO:94, SEQ ID NO:103, and SEQ ID NO:110; (y) SEQ ID NO:80, SEQ ID NO:86, SEQ ID NO:54, SEQ ID NO:95, SEQ ID NO:104, and SEQ ID NO:111; (z) SEQ ID NO:82, SEQ ID NO:87, SEQ ID NO:53, SEQ ID NO:93, SEQ ID NO:105, and SEQ ID NO:112; (aa) SEQ ID NO: 81, SEQ ID NO: 88, SEQ ID NO: 53, SEQ ID NO: 93, SEQ ID NO: 100, and SEQ ID NO: 109; (bb) SEQ ID NO:81, SEQ ID NO:89, SEQ ID NO:53, SEQ ID NO:93, SEQ ID NO:100, and SEQ ID NO:109; (cc) SEQ ID NO:81, SEQ ID NO:84, SEQ ID NO:55, SEQ ID NO:93, SEQ ID NO:100, and SEQ ID NO:109; (dd) SEQ ID NO:81, SEQ ID NO:84, SEQ ID NO:56, SEQ ID NO:93, SEQ ID NO:100, and SEQ ID NO:109; (ee) SEQ ID NO:81, SEQ ID NO:88, SEQ ID NO:55, SEQ ID NO:93, SEQ ID NO:100, and SEQ ID NO:109; (ff) SEQ ID NO:81, SEQ ID NO:88, SEQ ID NO:56, SEQ ID NO:93, SEQ ID NO:100, and SEQ ID NO:109; (gg) SEQ ID NO:81, SEQ ID NO:89, SEQ ID NO:55, SEQ ID NO:93, SEQ ID NO:100, and SEQ ID NO:109; or (hh) SEQ ID NO: 81, SEQ ID NO: 89, SEQ ID NO: 56, SEQ ID NO: 93, SEQ ID NO: 100, and SEQ ID NO: 109 Disclosed herein is a method comprising the amino acid sequence set forth in

[0162]

[0172] In certain embodiments, the therapeutic methods disclosed herein reduce minimal residual disease, increase remission, increase the duration of remission, reduce the rate of tumor recurrence, prevent tumor or cancer metastasis or reduce the rate of tumor or cancer metastasis, reduce the severity of a viral infection, improve the immune response to a viral infection, reduce inflammation, or reduce swelling, or any combination thereof, in a treated subject compared to a subject not administered an anti-IL-8 antibody or pharmaceutical composition thereof.

[0163]

[0173] Those skilled in the art will understand that the term "treatment" and its grammatical forms can, in certain embodiments, encompass both therapeutic treatment and prophylactic or preventative measures related to a tumor or cancer or viral infection described herein, where the goal is to prevent or alleviate the targeted tumor or cancer or viral infection described herein. Thus, in certain embodiments of the methods disclosed herein, treatment can include directly affecting or curing, suppressing, inhibiting, preventing, reducing the severity of, delaying the onset of, or alleviating the symptoms associated with a disease, disorder, or condition, or combination thereof, for example, where the disease or disorder includes a cancer or tumor, or a viral infection, or inflammation-related disease, or combination thereof. Thus, in certain embodiments, "treatment" includes preventing, slowing progression, inhibiting growth, slowing disease progression, reducing tumor burden, reducing incidence, promoting remission, inducing remission, increasing remission, hastening recovery, increasing the effectiveness of or reducing resistance to alternative treatments, reducing the impact of infection, improving the immune response to infection, or a combination thereof. In certain embodiments, "prevention" includes delaying the onset of symptoms, preventing disease recurrence, reducing the number or frequency of recurrent episodes, increasing the latency period between symptomatic episodes, reducing the impact of cytokine release syndrome or cytokine storm, attenuating cytokine release syndrome or cytokine storm, or a combination thereof. In certain embodiments, "suppression" or "inhibition" includes reducing the severity of symptoms, reducing the severity of acute episodes, reducing the number of symptoms, reducing the incidence of disease-related symptoms, reducing the latency period of symptoms, ameliorating symptoms, reducing secondary symptoms, reducing secondary infections, prolonging patient survival, or a combination thereof.

[0164]

[0174] In some embodiments of the methods of treatment disclosed herein, the size of a cancer or tumor is reduced. In some embodiments, the growth rate of a cancer or tumor is reduced. In some embodiments, the size or growth rate of a cancer or tumor, or a combination thereof, is reduced. In some embodiments, the adverse effects of a viral infection are reduced, for example, but not limited to, a reduction in cytokine release syndrome or cytokine storm in an affected tissue or organ. In some embodiments of the methods of treatment, administration of an anti-IL-8 antibody reduces the duration of a viral infection. In some embodiments of the methods of treatment, administration of an anti-IL-8 antibody increases clearance of a viral infection. In some embodiments, the survival of a subject in need is increased.

[0165]

[0175] Many diseases and cancers are known to be caused by viruses. Examples of disease-causing viruses include, but are not limited to, norovirus, rotavirus, hepatitis A, B, C, D, or E virus, rabies virus, West Nile virus, enterovirus, echovirus, coxsackievirus, herpes simplex virus (HSV), HSV-2, varicella-zoster virus, mosquito-borne virus, arbovirus, St. Louis encephalitis virus, California encephalitis virus, lymphocytic choriomeningitis virus, human immunodeficiency virus (HIV), poliovirus, Zika virus, rubella virus, cytomegalovirus, human papillomavirus (HPV), enterovirus D68, severe acute respiratory syndrome (SARS) coronavirus, Middle East respiratory syndrome coronavirus, SARS coronavirus 2, Epstein-Barr virus, influenza virus, respiratory syncytial virus, polyomavirus (e.g., JC virus, BK virus), Ebola virus, dengue virus, or any combination thereof. In certain embodiments of the methods of treating a disease, the disease comprises a viral infection.

[0166]

[0176] In some embodiments of the methods of treating a disease, the disease comprises a cancer or tumor. In some embodiments of the methods of treating a cancer or tumor, the cancer or tumor comprises a hematological cancer. In some embodiments, the hematological cancer comprises leukemia, lymphoma, myeloma, acute myeloid leukemia (AML), acute promyelocytic leukemia, erythroleukemia, mixed B-myelomonocytic leukemia, or myelodysplastic syndrome (MDS).

[0167]

[0177] In some embodiments of the methods for treating cancer or tumor, the cancer or tumor comprises a solid cancer or tumor. In some embodiments of the methods for treating cancer or tumor, the solid cancer or tumor comprises a sarcoma, osteosarcoma, squamous cell carcinoma of the head and neck, non-small cell lung cancer, bladder cancer, pancreatic cancer, or pancreatic ductal adenocarcinoma.

[0168]

[0178] Many diseases are known to be associated with inflammation, including, but not limited to, asthma, cancer, chronic inflammatory diseases, atherosclerosis, diabetes, and autoimmune and degenerative diseases, arthritis, and rheumatoid arthritis.

[0169]

[0179] In certain embodiments, the method of treating a disease associated with inflammation includes treating any of a number of diseases known to be associated with inflammation, including, but not limited to, asthma, cancer, chronic inflammatory diseases, atherosclerosis, diabetes, and autoimmune and degenerative diseases, arthritis, or rheumatoid arthritis, or any combination thereof.

[0170]

[0180] In some embodiments of treating inflammation-associated diseases, the treatment reduces or eliminates joint swelling. Disclosed herein are methods of treating inflammation-associated diseases, comprising administering an IL-8 antibody described herein, wherein the treatment reduces or eliminates joint swelling. In some embodiments, the method of treating inflammation-associated diseases reduces swelling in a diarthrodial joint selected from a ball and socket joint, a saddle joint, a hinge joint, a condylar joint, a pivot joint, and a gliding joint. In some embodiments, the method of treating inflammation-associated diseases reduces swelling in a joint selected from a knee, a finger, a shoulder, an elbow, a wrist, an ankle, a toe, or a hip, or any combination thereof. In some embodiments, the method of treating inflammation-associated diseases reduces inflammation in a subject in need thereof. In some embodiments, the method of treating inflammation-associated diseases reduces inflammation in a diarthrodial joint selected from a ball and socket joint, a saddle joint, a hinge joint, a condylar joint, a pivot joint, and a gliding joint.

[0171]

[0181] In certain embodiments of the methods of treating a disease including a cancer, a tumor, or a disease associated with a viral infection, or inflammation, or a combination thereof, the subject is a human.

[0172]

[0182] In some embodiments of the methods for treating a disease comprising cancer, a tumor, or a viral infection, or a disease associated with inflammation, or a combination thereof, administering comprises administering an anti-IL-8 antibody. In some embodiments of the methods for treating a disease comprising cancer, a tumor, or a viral infection, or a disease associated with inflammation, or a combination thereof, administering comprises administering a pharmaceutical composition comprising an anti-IL-8 antibody. In some embodiments of the methods for treating a disease comprising cancer, a tumor, or a viral infection, or a disease associated with inflammation, or a combination thereof, administering comprises administering a polynucleotide encoding the anti-IL-8 antibody. In some embodiments of the methods for treating a disease comprising cancer, a tumor, or a viral infection, or a disease associated with inflammation, or a combination thereof, administering comprises administering a pharmaceutical composition comprising a polynucleotide encoding the anti-IL-8 antibody.

[0173]

[0183] In some embodiments of the method for treating a disease comprising cancer, a tumor, or a disease associated with a viral infection, or inflammation, or a combination thereof, administering comprises administering a polynucleotide encoding an anti-IL-8 antibody comprising an antibody antigen-binding domain comprising a heavy chain variable region (VH) and a light chain variable region (VL), wherein the polynucleotide sequence encoding the VH-VL pair is selected from the pair sequences set forth in SEQ ID NOs: 113 and 114, SEQ ID NOs: 115 and 116, SEQ ID NOs: 117 and 118, SEQ ID NOs: 119 and 120, SEQ ID NOs: 121 and 122, SEQ ID NOs: 123 and 124, SEQ ID NOs: 125 and 126, SEQ ID NOs: 127 and 128, SEQ ID NOs: 129 and 130, SEQ ID NOs: 131 and 132, SEQ ID NOs: 133 and 134, SEQ ID NOs: 135 and 136, SEQ ID NOs: 137 and 138, SEQ ID NOs: 139 and 140, SEQ ID NOs: 141 and 142, SEQ ID NOs: 36 and 37, and SEQ ID NOs: 38 and 39. In some embodiments of the method for treating a disease associated with cancer, tumor, or viral infection, or inflammation, or a combination thereof, the administering comprises administering a polynucleotide encoding an anti-IL-8 antibody comprising an antibody antigen-binding domain comprising a VH and a VL, wherein the polynucleotide sequence encoding the VH-VL pair comprises the pairing sequences set forth in SEQ ID NOs: 113 and 114. In some embodiments of the method for treating a disease associated with cancer, tumor, or viral infection, or inflammation, or a combination thereof, the administering comprises administering a polynucleotide encoding an anti-IL-8 antibody comprising an antibody antigen-binding domain comprising a VH and a VL, wherein the polynucleotide sequence encoding the VH-VL pair comprises the pairing sequences set forth in SEQ ID NOs: 115 and 116. In some embodiments of the method for treating a disease associated with cancer, tumor, or viral infection, or inflammation, or a combination thereof, the administering comprises administering a polynucleotide encoding an anti-IL-8 antibody comprising an antibody antigen-binding domain comprising a VH and a VL, wherein the polynucleotide sequence encoding the VH-VL pair comprises the pairing sequences set forth in SEQ ID NOs: 117 and 118.In some embodiments of the method for treating a disease associated with cancer, tumor, or viral infection, or inflammation, or a combination thereof, the administering comprises administering a polynucleotide encoding an anti-IL-8 antibody comprising an antibody antigen-binding domain comprising a VH and a VL, wherein the polynucleotide sequence encoding the VH-VL pair comprises the pairing sequences set forth in SEQ ID NOs: 119 and 120. In some embodiments of the method for treating a disease associated with cancer, tumor, or viral infection, or inflammation, or a combination thereof, the administering comprises administering a polynucleotide encoding an anti-IL-8 antibody comprising an antibody antigen-binding domain comprising a VH and a VL, wherein the polynucleotide sequence encoding the VH-VL pair comprises the pairing sequences set forth in SEQ ID NOs: 121 and 122. In some embodiments of the method for treating a disease associated with cancer, tumor, or viral infection, or inflammation, or a combination thereof, the administering comprises administering a polynucleotide encoding an anti-IL-8 antibody comprising an antibody antigen-binding domain comprising a VH and a VL, wherein the polynucleotide sequence encoding the VH-VL pair comprises the pairing sequences set forth in SEQ ID NOs: 123 and 124. In certain embodiments of the method for treating a disease associated with cancer, tumor, or viral infection, or inflammation, or a combination thereof, the administering comprises administering a polynucleotide encoding an anti-IL-8 antibody comprising an antibody antigen-binding domain comprising a VH and a VL, wherein the polynucleotide sequence encoding the VH-VL pair comprises the pairing sequences set forth in SEQ ID NOs: 125 and 126. In certain embodiments of the method for treating a disease associated with cancer, tumor, or viral infection, or inflammation, or a combination thereof, the administering comprises administering a polynucleotide encoding an anti-IL-8 antibody comprising an antibody antigen-binding domain comprising a VH and a VL, wherein the polynucleotide sequence encoding the VH-VL pair comprises the pairing sequences set forth in SEQ ID NOs: 127 and 128.In some embodiments of the method for treating a disease associated with cancer, tumor, or viral infection, or inflammation, or a combination thereof, the administering comprises administering a polynucleotide encoding an anti-IL-8 antibody comprising an antibody antigen-binding domain comprising a VH and a VL, wherein the polynucleotide sequence encoding the VH-VL pair comprises the pairing sequences set forth in SEQ ID NOs: 129 and 130. In some embodiments of the method for treating a disease associated with cancer, tumor, or viral infection, or inflammation, or a combination thereof, the administering comprises administering a polynucleotide encoding an anti-IL-8 antibody comprising an antibody antigen-binding domain comprising a VH and a VL, wherein the polynucleotide sequence encoding the VH-VL pair comprises the pairing sequences set forth in SEQ ID NOs: 131 and 132. In some embodiments of the method for treating a disease associated with cancer, tumor, or viral infection, or inflammation, or a combination thereof, the administering comprises administering a polynucleotide encoding an anti-IL-8 antibody comprising an antibody antigen-binding domain comprising a VH and a VL, wherein the polynucleotide sequence encoding the VH-VL pair comprises the pairing sequences set forth in SEQ ID NOs: 133 and 134. In certain embodiments of the method for treating a disease associated with cancer, tumor, or viral infection, or inflammation, or a combination thereof, the administering comprises administering a polynucleotide encoding an anti-IL-8 antibody comprising an antibody antigen-binding domain comprising a VH and a VL, wherein the polynucleotide sequence encoding the VH-VL pair comprises the pairing sequences set forth in SEQ ID NOs: 135 and 136. In certain embodiments of the method for treating a disease associated with cancer, tumor, or viral infection, or inflammation, or a combination thereof, the administering comprises administering a polynucleotide encoding an anti-IL-8 antibody comprising an antibody antigen-binding domain comprising a VH and a VL, wherein the polynucleotide sequence encoding the VH-VL pair comprises the pairing sequences set forth in SEQ ID NOs: 137 and 138.In some embodiments of the method for treating a disease associated with cancer, tumor, or viral infection, or inflammation, or a combination thereof, the administering comprises administering a polynucleotide encoding an anti-IL-8 antibody comprising an antibody antigen-binding domain comprising a VH and a VL, wherein the polynucleotide sequence encoding the VH-VL pair comprises the pairing sequences set forth in SEQ ID NOs: 139 and 140. In some embodiments of the method for treating a disease associated with cancer, tumor, or viral infection, or inflammation, or a combination thereof, the administering comprises administering a polynucleotide encoding an anti-IL-8 antibody comprising an antibody antigen-binding domain comprising a VH and a VL, wherein the polynucleotide sequence encoding the VH-VL pair comprises the pairing sequences set forth in SEQ ID NOs: 141 and 142. In some embodiments of the method for treating a disease associated with cancer, tumor, or viral infection, or inflammation, or a combination thereof, the administering comprises administering a polynucleotide encoding an anti-IL-8 antibody comprising an antibody antigen-binding domain comprising a VH and a VL, wherein the polynucleotide sequence encoding the VH-VL pair comprises the pairing sequences set forth in SEQ ID NOs: 36 and 37. In one embodiment of the method for treating a disease including cancer, a tumor, or a disease associated with a viral infection, or inflammation, or a combination thereof, the administering comprises administering a polynucleotide encoding an anti-IL-8 antibody comprising an antibody antigen-binding domain comprising a VH and a VL, wherein the polynucleotide sequence encoding the VH-VL pair comprises the paired sequences set forth in SEQ ID NOs: 38 and 39.

[0174]

[0184] In certain embodiments, anti-IL-8 antibodies can be administered to a subject directly or by administering to a subject a polynucleotide sequence encoding the polypeptide, such that such nucleic acid sequence can be carried by a vector.

[0175]

[0185] The exact amount of the anti-IL-8 antibody of the present invention or a composition thereof required to produce the desired effect will vary from subject to subject, depending on the species, age, sex, weight, and general condition of the subject, the particular polypeptide, the route of administration, and whether other drugs are included in the regimen. Therefore, it is impossible to specify an exact amount for every composition. However, an appropriate amount can be determined by one of ordinary skill in the art using routine experimentation. Dosages can vary, and polypeptides can be administered in one or more doses (e.g., two or more, three or more, four or more, or five or more) per day for one or more days. Guidance for selecting the appropriate dose for an antibody can be readily found in the literature.

[0176]

[0186] In some embodiments of the methods described herein, the tumor or cancer cells secrete IL-8. In some embodiments of the methods described herein, the tumor or cancer cells have increased secretion of IL-8 compared to non-tumor or non-cancerous cells of the same cell type. In some embodiments of the methods described herein, the tumor or cancer cells express CXCR1 and / or CXCR2 receptors on their cell surface. In some embodiments of the methods described herein, the tumor or cancer cells express CXCR1 and / or CXCR2 receptors on their cell surface compared to non-tumor or non-cancerous cells of the same cell type.

[0177]

[0187] In certain embodiments of the methods of inhibiting tumor formation or growth, or a combination thereof, the anti-IL-8 antibody comprises an IgG, Fv, scFv, Fab, F(ab')2, minibody, diabody, triabody, nanobody, single domain antibody, multispecific antibody, bispecific antibody, triabody, single chain antibody, heavy chain antibody, chimeric antibody, or humanized antibody, or a combination thereof. In certain embodiments of the methods of inhibiting tumor formation or growth, or a combination thereof, the anti-IL-8 antibody comprising an IgG subclass is selected from IgG1, IgG2, IgG3, or IgG4.

[0178]

[0188] In certain embodiments of the methods of treating a subject suffering from a disease including cancer or a tumor or a viral infection, the anti-IL-8 antibody comprises an IgG, Fv, scFv, Fab, F(ab')2, minibody, diabody, triabody, nanobody, single domain antibody, multispecific antibody, bispecific antibody, triabody, single chain antibody, heavy chain antibody, chimeric antibody, or humanized antibody, or a combination thereof. In certain embodiments of the methods of inhibiting tumor formation or growth, or a combination thereof, the anti-IL-8 antibody comprising an IgG subclass is selected from IgG1, IgG2, IgG3, or IgG4.

[0179]

[0189] In certain embodiments of the methods disclosed herein, the administering comprises administering a pharmaceutical composition comprising an anti-IL-8 antibody.

[0180]

[0190] In certain embodiments, polynucleotide sequences encoding the anti-IL-8 antibodies described herein are used in methods of inhibiting tumor formation or growth, or a combination thereof, wherein the polynucleotide sequences encode antibodies comprising an anti-IL-8 antibody heavy chain variable region (VH) and an anti-IL-8 antibody light chain variable region (VL), and the amino acid sequences of the VH-VL pairs are selected from the paired sequences set forth in SEQ ID NOs: 2 and 3, SEQ ID NOs: 4 and 5, SEQ ID NOs: 6 and 7, SEQ ID NOs: 8 and 9, SEQ ID NOs: 10 and 11, SEQ ID NOs: 12 and 13, SEQ ID NOs: 14 and 15, SEQ ID NOs: 16 and 17, SEQ ID NOs: 18 and 19, SEQ ID NOs: 20 and 21, SEQ ID NOs: 22 and 23, SEQ ID NOs: 24 and 25, SEQ ID NOs: 26 and 27, SEQ ID NOs: 28 and 29, SEQ ID NOs: 30 and 31, SEQ ID NOs: 32 and 33, and SEQ ID NOs: 34 and 35. In certain embodiments, polynucleotide sequences encoding the anti-IL-8 antibodies described herein are used in a method of inhibiting tumor formation or growth, or a combination thereof, wherein the polynucleotide encodes a heavy chain variable region (VH) of the anti-IL-8 antibody comprising the VH complementarity determining regions (HCDRs) set forth in Table 1F and a light chain variable region (VL) of the anti-IL-8 antibody comprising the VL complementarity determining regions (LCDRs) set forth in Table 1F, wherein the heavy chain variable region has heavy chain complementarity determining regions (HCDR) 1, HCDR2, and HCDR3, and the light chain variable region has light chain complementarity determining regions (LCDR) 1, LCDR2, and LCDR3, and wherein the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 for each of the antibodies are (a) SEQ ID NO:41, SEQ ID NO:44, SEQ ID NO:53, SEQ ID NO:60, SEQ ID NO:67, and SEQ ID NO:76; (b) SEQ ID NO:81, SEQ ID NO:84, SEQ ID NO:53, SEQ ID NO:93, SEQ ID NO:100, and SEQ ID NO:109; (c) SEQ ID NO:40, SEQ ID NO:43, SEQ ID NO:50, SEQ ID NO:57, SEQ ID NO:64, and SEQ ID NO:73; (d) SEQ ID NO:40, SEQ ID NO:43, SEQ ID NO:51, SEQ ID NO:58, SEQ ID NO:65, and SEQ ID NO:74; (e) SEQ ID NO:41, SEQ ID NO:44, SEQ ID NO:52, SEQ ID NO:59, SEQ ID NO:66, and SEQ ID NO:75; (f) SEQ ID NO:40, SEQ ID NO:43, SEQ ID NO:50, SEQ ID NO:57, SEQ ID NO:68, and SEQ ID NO:73; (g) SEQ ID NO:40, SEQ ID NO:43, SEQ ID NO:50, SEQ ID NO:57, SEQ ID NO:69, and SEQ ID NO:73; (h) SEQ ID NO:41, SEQ ID NO:45, SEQ ID NO:53, SEQ ID NO:61, SEQ ID NO:70, and SEQ ID NO:77; (i) SEQ ID NO:40, SEQ ID NO:46, SEQ ID NO:54, SEQ ID NO:62, SEQ ID NO:71, and SEQ ID NO:78; (j) SEQ ID NO:42, SEQ ID NO:47, SEQ ID NO:53, SEQ ID NO:63, SEQ ID NO:72, and SEQ ID NO:79; (k) SEQ ID NO:41, SEQ ID NO:48, SEQ ID NO:53, SEQ ID NO:60, SEQ ID NO:67, and SEQ ID NO:76; (l) SEQ ID NO:41, SEQ ID NO:49, SEQ ID NO:53, SEQ ID NO:60, SEQ ID NO:67, and SEQ ID NO:76; (m) SEQ ID NO:41, SEQ ID NO:44, SEQ ID NO:55, SEQ ID NO:60, SEQ ID NO:67, and SEQ ID NO:76; (n) SEQ ID NO:41, SEQ ID NO:44, SEQ ID NO:56, SEQ ID NO:60, SEQ ID NO:67, and SEQ ID NO:76; (o) SEQ ID NO:41, SEQ ID NO:48, SEQ ID NO:55, SEQ ID NO:60, SEQ ID NO:67, and SEQ ID NO:76; (p) SEQ ID NO:41, SEQ ID NO:48, SEQ ID NO:56, SEQ ID NO:60, SEQ ID NO:67, and SEQ ID NO:76; (q) SEQ ID NO:41, SEQ ID NO:49, SEQ ID NO:55, SEQ ID NO:60, SEQ ID NO:67, and SEQ ID NO:76; (r) SEQ ID NO:41, SEQ ID NO:49, SEQ ID NO:56, SEQ ID NO:60, SEQ ID NO:67, and SEQ ID NO:76; (s) SEQ ID NO: 80, SEQ ID NO: 83, SEQ ID NO: 50, SEQ ID NO: 90, SEQ ID NO: 97, and SEQ ID NO: 106; (t) SEQ ID NO:80, SEQ ID NO:83, SEQ ID NO:51, SEQ ID NO:91, SEQ ID NO:98, and SEQ ID NO:107; (u) SEQ ID NO:81, SEQ ID NO:84, SEQ ID NO:52, SEQ ID NO:92, SEQ ID NO:99, and SEQ ID NO:108; (v) SEQ ID NO:80, SEQ ID NO:83, SEQ ID NO:50, SEQ ID NO:90, SEQ ID NO:101, and SEQ ID NO:106; (w) SEQ ID NO:80, SEQ ID NO:83, SEQ ID NO:50, SEQ ID NO:90, SEQ ID NO:102, and SEQ ID NO:106; (x) SEQ ID NO:81, SEQ ID NO:85, SEQ ID NO:53, SEQ ID NO:94, SEQ ID NO:103, and SEQ ID NO:110; (y) SEQ ID NO:80, SEQ ID NO:86, SEQ ID NO:54, SEQ ID NO:95, SEQ ID NO:104, and SEQ ID NO:111; (z) SEQ ID NO:82, SEQ ID NO:87, SEQ ID NO:53, SEQ ID NO:93, SEQ ID NO:105, and SEQ ID NO:112; (aa) SEQ ID NO: 81, SEQ ID NO: 88, SEQ ID NO: 53, SEQ ID NO: 93, SEQ ID NO: 100, and SEQ ID NO: 109; (bb) SEQ ID NO:81, SEQ ID NO:89, SEQ ID NO:53, SEQ ID NO:93, SEQ ID NO:100, and SEQ ID NO:109; (cc) SEQ ID NO:81, SEQ ID NO:84, SEQ ID NO:55, SEQ ID NO:93, SEQ ID NO:100, and SEQ ID NO:109; (dd) SEQ ID NO:81, SEQ ID NO:84, SEQ ID NO:56, SEQ ID NO:93, SEQ ID NO:100, and SEQ ID NO:109; (ee) SEQ ID NO:81, SEQ ID NO:88, SEQ ID NO:55, SEQ ID NO:93, SEQ ID NO:100, and SEQ ID NO:109; (ff) SEQ ID NO:81, SEQ ID NO:88, SEQ ID NO:56, SEQ ID NO:93, SEQ ID NO:100, and SEQ ID NO:109; (gg) SEQ ID NO:81, SEQ ID NO:89, SEQ ID NO:55, SEQ ID NO:93, SEQ ID NO:100, and SEQ ID NO:109; or (hh) SEQ ID NO: 81, SEQ ID NO: 89, SEQ ID NO: 56, SEQ ID NO: 93, SEQ ID NO: 100, and SEQ ID NO: 109 The amino acid sequence includes the amino acid sequence set forth in

[0181]

[0191] In certain embodiments, polynucleotide sequences encoding the anti-IL-8 antibodies described herein are used in methods of treating a disease including cancer or tumor or viral infection, wherein the polynucleotide sequences encode an antibody comprising an anti-IL-8 antibody heavy chain variable region (VH) and an anti-IL-8 antibody light chain variable region (VL), and the amino acid sequences of the VH-VL pair are selected from the paired sequences set forth in SEQ ID NOs: 2 and 3, SEQ ID NOs: 4 and 5, SEQ ID NOs: 6 and 7, SEQ ID NOs: 8 and 9, SEQ ID NOs: 10 and 11, SEQ ID NOs: 12 and 13, SEQ ID NOs: 14 and 15, SEQ ID NOs: 16 and 17, SEQ ID NOs: 18 and 19, SEQ ID NOs: 20 and 21, SEQ ID NOs: 22 and 23, SEQ ID NOs: 24 and 25, SEQ ID NOs: 26 and 27, SEQ ID NOs: 28 and 29, SEQ ID NOs: 30 and 31, SEQ ID NOs: 32 and 33, and SEQ ID NOs: 34 and 35. In certain embodiments, polynucleotide sequences encoding the anti-IL-8 antibodies described herein are used in methods of treating a disease, including cancer or tumor or viral infection, wherein the polynucleotide sequences encode a heavy chain variable region (VH) of the anti-IL-8 antibody comprising the VH complementarity determining region (HCDR) set forth in Table 1F and a light chain variable region (VL) of the anti-IL-8 antibody comprising the VL complementarity determining region (LCDR) set forth in Table 1F, wherein the heavy chain variable region has heavy chain complementarity determining region (HCDR)1, HCDR2, and HCDR3, and the light chain variable region has light chain complementarity determining region (LCDR)1, LCDR2, and LCDR3, and wherein the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 for each of the antibodies are (a) SEQ ID NO:41, SEQ ID NO:44, SEQ ID NO:53, SEQ ID NO:60, SEQ ID NO:67, and SEQ ID NO:76; (b) SEQ ID NO:81, SEQ ID NO:84, SEQ ID NO:53, SEQ ID NO:93, SEQ ID NO:100, and SEQ ID NO:109; (c) SEQ ID NO:40, SEQ ID NO:43, SEQ ID NO:50, SEQ ID NO:57, SEQ ID NO:64, and SEQ ID NO:73; (d) SEQ ID NO:40, SEQ ID NO:43, SEQ ID NO:51, SEQ ID NO:58, SEQ ID NO:65, and SEQ ID NO:74; (e) SEQ ID NO:41, SEQ ID NO:44, SEQ ID NO:52, SEQ ID NO:59, SEQ ID NO:66, and SEQ ID NO:75; (f) SEQ ID NO:40, SEQ ID NO:43, SEQ ID NO:50, SEQ ID NO:57, SEQ ID NO:68, and SEQ ID NO:73; (g) SEQ ID NO:40, SEQ ID NO:43, SEQ ID NO:50, SEQ ID NO:57, SEQ ID NO:69, and SEQ ID NO:73; (h) SEQ ID NO:41, SEQ ID NO:45, SEQ ID NO:53, SEQ ID NO:61, SEQ ID NO:70, and SEQ ID NO:77; (i) SEQ ID NO:40, SEQ ID NO:46, SEQ ID NO:54, SEQ ID NO:62, SEQ ID NO:71, and SEQ ID NO:78; (j) SEQ ID NO:42, SEQ ID NO:47, SEQ ID NO:53, SEQ ID NO:63, SEQ ID NO:72, and SEQ ID NO:79; (k) SEQ ID NO:41, SEQ ID NO:48, SEQ ID NO:53, SEQ ID NO:60, SEQ ID NO:67, and SEQ ID NO:76; (l) SEQ ID NO:41, SEQ ID NO:49, SEQ ID NO:53, SEQ ID NO:60, SEQ ID NO:67, and SEQ ID NO:76; (m) SEQ ID NO:41, SEQ ID NO:44, SEQ ID NO:55, SEQ ID NO:60, SEQ ID NO:67, and SEQ ID NO:76; (n) SEQ ID NO:41, SEQ ID NO:44, SEQ ID NO:56, SEQ ID NO:60, SEQ ID NO:67, and SEQ ID NO:76; (o) SEQ ID NO:41, SEQ ID NO:48, SEQ ID NO:55, SEQ ID NO:60, SEQ ID NO:67, and SEQ ID NO:76; (p) SEQ ID NO:41, SEQ ID NO:48, SEQ ID NO:56, SEQ ID NO:60, SEQ ID NO:67, and SEQ ID NO:76; (q) SEQ ID NO:41, SEQ ID NO:49, SEQ ID NO:55, SEQ ID NO:60, SEQ ID NO:67, and SEQ ID NO:76; (r) SEQ ID NO:41, SEQ ID NO:49, SEQ ID NO:56, SEQ ID NO:60, SEQ ID NO:67, and SEQ ID NO:76; (s) SEQ ID NO: 80, SEQ ID NO: 83, SEQ ID NO: 50, SEQ ID NO: 90, SEQ ID NO: 97, and SEQ ID NO: 106; (t) SEQ ID NO:80, SEQ ID NO:83, SEQ ID NO:51, SEQ ID NO:91, SEQ ID NO:98, and SEQ ID NO:107; (u) SEQ ID NO:81, SEQ ID NO:84, SEQ ID NO:52, SEQ ID NO:92, SEQ ID NO:99, and SEQ ID NO:108; (v) SEQ ID NO:80, SEQ ID NO:83, SEQ ID NO:50, SEQ ID NO:90, SEQ ID NO:101, and SEQ ID NO:106; (w) SEQ ID NO:80, SEQ ID NO:83, SEQ ID NO:50, SEQ ID NO:90, SEQ ID NO:102, and SEQ ID NO:106; (x) SEQ ID NO:81, SEQ ID NO:85, SEQ ID NO:53, SEQ ID NO:94, SEQ ID NO:103, and SEQ ID NO:110; (y) SEQ ID NO:80, SEQ ID NO:86, SEQ ID NO:54, SEQ ID NO:95, SEQ ID NO:104, and SEQ ID NO:111; (z) SEQ ID NO:82, SEQ ID NO:87, SEQ ID NO:53, SEQ ID NO:93, SEQ ID NO:105, and SEQ ID NO:112; (aa) SEQ ID NO: 81, SEQ ID NO: 88, SEQ ID NO: 53, SEQ ID NO: 93, SEQ ID NO: 100, and SEQ ID NO: 109; (bb) SEQ ID NO:81, SEQ ID NO:89, SEQ ID NO:53, SEQ ID NO:93, SEQ ID NO:100, and SEQ ID NO:109; (cc) SEQ ID NO:81, SEQ ID NO:84, SEQ ID NO:55, SEQ ID NO:93, SEQ ID NO:100, and SEQ ID NO:109; (dd) SEQ ID NO:81, SEQ ID NO:84, SEQ ID NO:56, SEQ ID NO:93, SEQ ID NO:100, and SEQ ID NO:109; (ee) SEQ ID NO:81, SEQ ID NO:88, SEQ ID NO:55, SEQ ID NO:93, SEQ ID NO:100, and SEQ ID NO:109; (ff) SEQ ID NO:81, SEQ ID NO:88, SEQ ID NO:56, SEQ ID NO:93, SEQ ID NO:100, and SEQ ID NO:109; (gg) SEQ ID NO:81, SEQ ID NO:89, SEQ ID NO:55, SEQ ID NO:93, SEQ ID NO:100, and SEQ ID NO:109; or (hh) SEQ ID NO: 81, SEQ ID NO: 89, SEQ ID NO: 56, SEQ ID NO: 93, SEQ ID NO: 100, and SEQ ID NO: 109 The amino acid sequence includes the amino acid sequence set forth in

[0182]

[0192] In certain embodiments of the methods disclosed herein, the polynucleotide encoding the anti-IL-8 antibody may encode an IgG, Fv, scFv, Fab, or F(ab')2. The IgG may be of the IgG1, IgG2, IgG3, or IgG4 subclass. In certain embodiments of the methods disclosed herein, the polynucleotide encoding the anti-IL-8 antibody may encode a portion of a minibody, diabody, triabody, nanobody, or single-domain antibody.

[0183]

[0193] In one embodiment, the subject is a mammal, eg, a human, suffering from one or more IL-18-related disorders, including cancer or viral infection or inflammation-related disorders, or a combination thereof.

[0184]

[0194] In certain embodiments, the method of inhibiting tumor formation or growth, or a combination thereof, comprises the use of an anti-IL-8 antibody disclosed herein as part of a combination therapy with an additional therapeutic agent. In certain embodiments, the method of inhibiting tumor formation or growth, or a combination thereof, comprises the use of a composition described herein comprising an anti-IL-8 antibody described herein in combination with an additional therapeutic agent. In certain embodiments, in the method of inhibiting tumor formation or growth, or a combination thereof, the additional therapeutic agent comprises a chemotherapeutic agent, an antiviral agent, an immunomodulatory agent, or an additional therapeutic antibody, or any combination thereof.

[0185]

[0195] In certain embodiments, a method for inhibiting tumor formation or growth, or a combination thereof, comprises the use of a composition comprising an anti-IL-8 antibody described herein and an additional therapeutic agent contained in the same composition. In certain embodiments, a method for inhibiting tumor formation or growth, or a combination thereof, comprises the use of an anti-IL-8 antibody described herein and an additional therapeutic agent contained in different compositions. In certain embodiments, a method for inhibiting tumor formation or growth, or a combination thereof, comprises the use of one or more compositions comprising a combination of an anti-IL-8 antibody described herein and an additional therapeutic agent, wherein administration is simultaneous. In certain embodiments, a method for inhibiting tumor formation or growth, or a combination thereof, comprises the use of an anti-IL-8 antibody described herein and an additional therapeutic agent, or a composition thereof, wherein administration of the anti-IL-8 antibody or composition thereof occurs before administration of the additional therapeutic agent. In certain embodiments, a method for inhibiting tumor formation or growth, or a combination thereof, comprises the use of an anti-IL-8 antibody described herein and an additional therapeutic agent, or a composition thereof, wherein administration of the anti-IL-8 antibody or composition thereof occurs after administration of the additional therapeutic agent.

[0186]

[0196] In certain embodiments, a method of treating a subject suffering from a disease comprises the use of an anti-IL-8 antibody disclosed herein as part of a combination therapy with an additional therapeutic agent. In certain embodiments, a method of treating a subject suffering from a disease comprises the use of a composition described herein comprising an anti-IL-8 antibody described herein in combination with an additional therapeutic agent. In certain embodiments, in the method of treating a subject suffering from a disease, the additional therapeutic agent comprises a chemotherapeutic agent, an antiviral agent, an immunomodulatory agent, or an additional therapeutic antibody, or any combination thereof.

[0187]

[0197] In certain embodiments, a method for treating a subject suffering from a disease comprises the use of a composition comprising an anti-IL-8 antibody described herein and an additional therapeutic agent contained in the same composition. In certain embodiments, a method for treating a subject suffering from a disease comprises the use of an anti-IL-8 antibody described herein and an additional therapeutic agent contained in different compositions. In certain embodiments, a method for treating a subject suffering from a disease comprises the use of one or more compositions comprising a combination of an anti-IL-8 antibody described herein and an additional therapeutic agent, wherein administration is simultaneous. In certain embodiments, a method for treating a subject suffering from a disease comprises the use of an anti-IL-8 antibody described herein and an additional therapeutic agent, or a composition thereof, wherein administration of the anti-IL-8 antibody or composition thereof occurs before administration of the additional therapeutic agent. In certain embodiments, a method for treating a subject suffering from a disease comprises the use of an anti-IL-8 antibody described herein and an additional therapeutic agent, or a composition thereof, wherein administration of the anti-IL-8 antibody or composition thereof occurs after administration of the additional therapeutic agent.

[0188]

[0198] The exact dosage and duration of treatment depend on the disease to be treated and can be determined experimentally using known test protocols or by testing the composition in a model system known in the art and extrapolating from there.Controlled clinical trials can also be conducted.Dosage can also vary depending on the severity of the condition to be alleviated.Pharmaceutical compositions are generally formulated and administered to exert a therapeutically useful effect while minimizing undesirable side effects.The composition can be administered once or divided into several smaller doses that are administered at time intervals.For any particular subject, the specific dosage regimen can be adjusted over time according to individual needs.

[0189]

[0199] As used herein, the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise. For example, the term "an immunoglobulin" or "at least one immunoglobulin" can include a plurality of immunoglobulins, including mixtures thereof.

[0190]

[0200] Throughout this application, various embodiments may be presented in a range format. It should be understood that the description in range format is for convenience and brevity only and should not be construed as an inflexible limitation on the scope of the invention. Accordingly, the description of a range should be considered to include all the possible subranges specifically disclosed as well as individual numerical values ​​within that range. For example, a description of a range such as 1 to 6 should be considered to include specifically disclosed subranges such as 1 to 3, 1 to 4, 1 to 5, 2 to 4, 2 to 6, 3 to 6, etc., as well as individual numerical values ​​within that range, e.g., 1, 2, 3, 4, 5, and 6. This applies regardless of the breadth of the range.

[0191]

[0201] Embodiments of IL-8 antibodies and their uses include the following.

[0192]

[0202] An isolated anti-IL-8 antibody comprising an antibody antigen-binding domain comprising a heavy chain variable region (VH) and a light chain variable region (VL), wherein the amino acid sequences of the VH-VL pairs are selected from the pair sequences set forth in SEQ ID NOs: 2 and 3, SEQ ID NOs: 4 and 5, SEQ ID NOs: 6 and 7, SEQ ID NOs: 8 and 9, SEQ ID NOs: 10 and 11, SEQ ID NOs: 12 and 13, SEQ ID NOs: 14 and 15, SEQ ID NOs: 16 and 17, SEQ ID NOs: 18 and 19, SEQ ID NOs: 20 and 21, SEQ ID NOs: 22 and 23, SEQ ID NOs: 24 and 25, SEQ ID NOs: 26 and 27, SEQ ID NOs: 28 and 29, SEQ ID NOs: 30 and 31, SEQ ID NOs: 32 and 33, and SEQ ID NOs: 34 and 35.

[0193]

[0203] A composition comprising: an isolated anti-IL-8 antibody comprising an antibody antigen-binding domain including a heavy chain variable region (VH) and a light chain variable region (VL), wherein the amino acid sequence of the VH-VL pair is selected from the pair sequences set forth in SEQ ID NOs: 2 and 3, SEQ ID NOs: 4 and 5, SEQ ID NOs: 6 and 7, SEQ ID NOs: 8 and 9, SEQ ID NOs: 10 and 11, SEQ ID NOs: 12 and 13, SEQ ID NOs: 14 and 15, SEQ ID NOs: 16 and 17, SEQ ID NOs: 18 and 19, SEQ ID NOs: 20 and 21, SEQ ID NOs: 22 and 23, SEQ ID NOs: 24 and 25, SEQ ID NOs: 26 and 27, SEQ ID NOs: 28 and 29, SEQ ID NOs: 30 and 31, SEQ ID NOs: 32 and 33, and SEQ ID NOs: 34 and 35; and a pharmaceutically acceptable carrier.

[0194]

[0204] Isolated anti-IL-8 antibodies having the complementarity determining region (CDR) sequences set forth in Table 1F, wherein each antibody comprises a heavy chain variable region having heavy chain complementarity determining region (HCDR) 1, HCDR2, and HCDR3, and a light chain variable region having light chain complementarity determining region (LCDR) 1, LCDR2, and LCDR3, and wherein the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 for each of the antibodies comprise the amino acid sequences set forth in Table 1F.

[0195]

[0205] An isolated anti-IL-8 antibody, including an IgG, Fv, scFv, Fab, F(ab')2, minibody, diabody, triabody, nanobody, single domain antibody, multispecific antibody, bispecific antibody, trispecific antibody, single chain antibody, heavy chain antibody, chimeric antibody, or humanized antibody.

[0196]

[0206] An isolated anti-IL-8 antibody comprising IgG1, IgG2, IgG3, or IgG4.

[0197]

[0207] A composition comprising isolated anti-IL-8 antibodies having complementarity determining region (CDR) sequences set forth in Table 1F, wherein each antibody comprises a heavy chain variable region having heavy chain complementarity determining region (HCDR) 1, HCDR2, and HCDR3, and a light chain variable region having light chain complementarity determining region (LCDR) 1, LCDR2, and LCDR3, and the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 for each of the antibodies comprise the amino acid sequences set forth in Table 1F; and a pharmaceutically acceptable carrier.

[0198]

[0208] 1. An isolated polynucleotide sequence encoding a heavy chain variable region (VH) of an anti-IL-8 antibody and a light chain variable region (VL) of an anti-IL-8 antibody, wherein the amino acid sequences of the VH-VL pairs are selected from the paired sequences set forth in SEQ ID NOs: 2 and 3, SEQ ID NOs: 4 and 5, SEQ ID NOs: 6 and 7, SEQ ID NOs: 8 and 9, SEQ ID NOs: 10 and 11, SEQ ID NOs: 12 and 13, SEQ ID NOs: 14 and 15, SEQ ID NOs: 16 and 17, SEQ ID NOs: 18 and 19, SEQ ID NOs: 20 and 21, SEQ ID NOs: 22 and 23, SEQ ID NOs: 24 and 25, SEQ ID NOs: 26 and 27, SEQ ID NOs: 28 and 29, SEQ ID NOs: 30 and 31, SEQ ID NOs: 32 and 33, and SEQ ID NOs: 34 and 35.

[0199]

[0209] 1. An isolated polynucleotide sequence encoding a heavy chain variable region (VH) of an anti-IL-8 antibody comprising a VH complementarity determining region (HCDR) as set forth in Table 1F and a light chain variable region (VL) of an anti-IL-8 antibody comprising a VL complementarity determining region (LCDR) as set forth in Table 1F, wherein the heavy chain variable region comprises heavy chain complementarity determining region (HCDR) 1, HCDR2, and HCDR3, and the light chain variable region comprises light chain complementarity determining region (LCDR) 1, LCDR2, and LCDR3, and the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 for the VH and CL, respectively, are the amino acid sequences set forth in Table 1F: (a) SEQ ID NO:41, SEQ ID NO:44, SEQ ID NO:53, SEQ ID NO:60, SEQ ID NO:67, and SEQ ID NO:76; (b) SEQ ID NO:81, SEQ ID NO:84, SEQ ID NO:53, SEQ ID NO:93, SEQ ID NO:100, and SEQ ID NO:109; (c) SEQ ID NO:40, SEQ ID NO:43, SEQ ID NO:50, SEQ ID NO:57, SEQ ID NO:64, and SEQ ID NO:73; (d) SEQ ID NO:40, SEQ ID NO:43, SEQ ID NO:51, SEQ ID NO:58, SEQ ID NO:65, and SEQ ID NO:74; (e) SEQ ID NO:41, SEQ ID NO:44, SEQ ID NO:52, SEQ ID NO:59, SEQ ID NO:66, and SEQ ID NO:75; (f) SEQ ID NO:40, SEQ ID NO:43, SEQ ID NO:50, SEQ ID NO:57, SEQ ID NO:68, and SEQ ID NO:73; (g) SEQ ID NO:40, SEQ ID NO:43, SEQ ID NO:50, SEQ ID NO:57, SEQ ID NO:69, and SEQ ID NO:73; (h) SEQ ID NO:41, SEQ ID NO:45, SEQ ID NO:53, SEQ ID NO:61, SEQ ID NO:70, and SEQ ID NO:77; (i) SEQ ID NO:40, SEQ ID NO:46, SEQ ID NO:54, SEQ ID NO:62, SEQ ID NO:71, and SEQ ID NO:78; (j) SEQ ID NO:42, SEQ ID NO:47, SEQ ID NO:53, SEQ ID NO:63, SEQ ID NO:72, and SEQ ID NO:79; (k) SEQ ID NO:41, SEQ ID NO:48, SEQ ID NO:53, SEQ ID NO:60, SEQ ID NO:67, and SEQ ID NO:76; (l) SEQ ID NO:41, SEQ ID NO:49, SEQ ID NO:53, SEQ ID NO:60, SEQ ID NO:67, and SEQ ID NO:76; (m) SEQ ID NO:41, SEQ ID NO:44, SEQ ID NO:55, SEQ ID NO:60, SEQ ID NO:67, and SEQ ID NO:76; (n) SEQ ID NO:41, SEQ ID NO:44, SEQ ID NO:56, SEQ ID NO:60, SEQ ID NO:67, and SEQ ID NO:76; (o) SEQ ID NO:41, SEQ ID NO:48, SEQ ID NO:55, SEQ ID NO:60, SEQ ID NO:67, and SEQ ID NO:76; (p) SEQ ID NO:41, SEQ ID NO:48, SEQ ID NO:56, SEQ ID NO:60, SEQ ID NO:67, and SEQ ID NO:76; (q) SEQ ID NO:41, SEQ ID NO:49, SEQ ID NO:55, SEQ ID NO:60, SEQ ID NO:67, and SEQ ID NO:76; (r) SEQ ID NO:41, SEQ ID NO:49, SEQ ID NO:56, SEQ ID NO:60, SEQ ID NO:67, and SEQ ID NO:76; (s) SEQ ID NO: 80, SEQ ID NO: 83, SEQ ID NO: 50, SEQ ID NO: 90, SEQ ID NO: 97, and SEQ ID NO: 106; (t) SEQ ID NO:80, SEQ ID NO:83, SEQ ID NO:51, SEQ ID NO:91, SEQ ID NO:98, and SEQ ID NO:107; (u) SEQ ID NO:81, SEQ ID NO:84, SEQ ID NO:52, SEQ ID NO:92, SEQ ID NO:99, and SEQ ID NO:108; (v) SEQ ID NO:80, SEQ ID NO:83, SEQ ID NO:50, SEQ ID NO:90, SEQ ID NO:101, and SEQ ID NO:106; (w) SEQ ID NO:80, SEQ ID NO:83, SEQ ID NO:50, SEQ ID NO:90, SEQ ID NO:102, and SEQ ID NO:106; (x) SEQ ID NO:81, SEQ ID NO:85, SEQ ID NO:53, SEQ ID NO:94, SEQ ID NO:103, and SEQ ID NO:110; (y) SEQ ID NO:80, SEQ ID NO:86, SEQ ID NO:54, SEQ ID NO:95, SEQ ID NO:104, and SEQ ID NO:111; (z) SEQ ID NO:82, SEQ ID NO:87, SEQ ID NO:53, SEQ ID NO:93, SEQ ID NO:105, and SEQ ID NO:112; (aa) SEQ ID NO: 81, SEQ ID NO: 88, SEQ ID NO: 53, SEQ ID NO: 93, SEQ ID NO: 100, and SEQ ID NO: 109; (bb) SEQ ID NO:81, SEQ ID NO:89, SEQ ID NO:53, SEQ ID NO:93, SEQ ID NO:100, and SEQ ID NO:109; (cc) SEQ ID NO:81, SEQ ID NO:84, SEQ ID NO:55, SEQ ID NO:93, SEQ ID NO:100, and SEQ ID NO:109; (dd) SEQ ID NO:81, SEQ ID NO:84, SEQ ID NO:56, SEQ ID NO:93, SEQ ID NO:100, and SEQ ID NO:109; (ee) SEQ ID NO:81, SEQ ID NO:88, SEQ ID NO:55, SEQ ID NO:93, SEQ ID NO:100, and SEQ ID NO:109; (ff) SEQ ID NO:81, SEQ ID NO:88, SEQ ID NO:56, SEQ ID NO:93, SEQ ID NO:100, and SEQ ID NO:109; (gg) SEQ ID NO:81, SEQ ID NO:89, SEQ ID NO:55, SEQ ID NO:93, SEQ ID NO:100, and SEQ ID NO:109; or (hh) SEQ ID NO: 81, SEQ ID NO: 89, SEQ ID NO: 56, SEQ ID NO: 93, SEQ ID NO: 100, and SEQ ID NO: 109 An isolated polynucleotide sequence comprising:

[0200]

[0210] An isolated polynucleotide sequence comprising two polynucleotide sequences, wherein a first polynucleotide sequence encodes a VH of an anti-IL-8 antibody and a second polynucleotide sequence encodes a VL of the anti-IL-8 antibody.

[0201]

[0211] A vector comprising a polynucleotide sequence encoding the heavy chain variable region (VH) of an anti-IL-8 antibody and the light chain variable region (VL) of an anti-IL-8 antibody.

[0202]

[0212] A host cell comprising the vector according to claim 1 , which comprises a polynucleotide sequence encoding the heavy chain variable region (VH) of an anti-IL-8 antibody and the light chain variable region (VL) of an anti-IL-8 antibody.

[0203]

[0213] A method for producing an anti-IL-8 antibody comprising a heavy chain variable region (VH) and a light chain variable region (VL), the method comprising the steps of culturing a host cell containing a vector comprising a polynucleotide sequence encoding the heavy chain variable region (VH) of an anti-IL-8 antibody and the light chain variable region (VL) of an anti-IL-8 antibody under conditions promoting expression of the vector in the host cell, and expressing the polynucleotide sequence contained in the vector, thereby producing an anti-IL-8 antibody comprising the VH and VL.

[0204]

[0214] 1. A method for inhibiting tumor or cancer formation or growth, or a combination thereof, in a subject in need thereof, the method comprising the step of administering to the subject an anti-IL-8 antibody comprising an antibody antigen-binding domain comprising a heavy chain variable region (VH) and a light chain variable region (VL), thereby inhibiting tumor formation or growth, or a combination thereof, in the subject, wherein the amino acid sequences of the VH-VL pairs are selected from the paired sequences set forth in SEQ ID NOs: 2 and 3, SEQ ID NOs: 4 and 5, SEQ ID NOs: 6 and 7, SEQ ID NOs: 8 and 9, SEQ ID NOs: 10 and 11, SEQ ID NOs: 12 and 13, SEQ ID NOs: 14 and 15, SEQ ID NOs: 16 and 17, SEQ ID NOs: 18 and 19, SEQ ID NOs: 20 and 21, SEQ ID NOs: 22 and 23, SEQ ID NOs: 24 and 25, SEQ ID NOs: 26 and 27, SEQ ID NOs: 28 and 29, SEQ ID NOs: 30 and 31, SEQ ID NOs: 32 and 33, and SEQ ID NOs: 34 and 35.

[0205]

[0215] 1. A method of inhibiting tumor or cancer formation or growth, or a combination thereof, in a subject in need thereof, the method comprising the step of administering to the subject anti-IL-8 antibodies having complementarity determining region (CDR) sequences as set forth in Table 1F, thereby inhibiting tumor formation or growth, or a combination thereof, in the subject, wherein each antibody comprises a heavy chain variable region having heavy chain complementarity determining region (HCDR) 1, HCDR2, and HCDR3, and a light chain variable region having light chain complementarity determining region (LCDR) 1, LCDR2, and LCDR3, wherein the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 for each of the antibodies have the amino acid sequences set forth in Table 1F: (a) SEQ ID NO:41, SEQ ID NO:44, SEQ ID NO:53, SEQ ID NO:60, SEQ ID NO:67, and SEQ ID NO:76; (b) SEQ ID NO:81, SEQ ID NO:84, SEQ ID NO:53, SEQ ID NO:93, SEQ ID NO:100, and SEQ ID NO:109; (c) SEQ ID NO:40, SEQ ID NO:43, SEQ ID NO:50, SEQ ID NO:57, SEQ ID NO:64, and SEQ ID NO:73; (d) SEQ ID NO:40, SEQ ID NO:43, SEQ ID NO:51, SEQ ID NO:58, SEQ ID NO:65, and SEQ ID NO:74; (e) SEQ ID NO:41, SEQ ID NO:44, SEQ ID NO:52, SEQ ID NO:59, SEQ ID NO:66, and SEQ ID NO:75; (f) SEQ ID NO:40, SEQ ID NO:43, SEQ ID NO:50, SEQ ID NO:57, SEQ ID NO:68, and SEQ ID NO:73; (g) SEQ ID NO:40, SEQ ID NO:43, SEQ ID NO:50, SEQ ID NO:57, SEQ ID NO:69, and SEQ ID NO:73; (h) SEQ ID NO:41, SEQ ID NO:45, SEQ ID NO:53, SEQ ID NO:61, SEQ ID NO:70, and SEQ ID NO:77; (i) SEQ ID NO:40, SEQ ID NO:46, SEQ ID NO:54, SEQ ID NO:62, SEQ ID NO:71, and SEQ ID NO:78; (j) SEQ ID NO:42, SEQ ID NO:47, SEQ ID NO:53, SEQ ID NO:63, SEQ ID NO:72, and SEQ ID NO:79; (k) SEQ ID NO:41, SEQ ID NO:48, SEQ ID NO:53, SEQ ID NO:60, SEQ ID NO:67, and SEQ ID NO:76; (l) SEQ ID NO:41, SEQ ID NO:49, SEQ ID NO:53, SEQ ID NO:60, SEQ ID NO:67, and SEQ ID NO:76; (m) SEQ ID NO:41, SEQ ID NO:44, SEQ ID NO:55, SEQ ID NO:60, SEQ ID NO:67, and SEQ ID NO:76; (n) SEQ ID NO:41, SEQ ID NO:44, SEQ ID NO:56, SEQ ID NO:60, SEQ ID NO:67, and SEQ ID NO:76; (o) SEQ ID NO:41, SEQ ID NO:48, SEQ ID NO:55, SEQ ID NO:60, SEQ ID NO:67, and SEQ ID NO:76; (p) SEQ ID NO:41, SEQ ID NO:48, SEQ ID NO:56, SEQ ID NO:60, SEQ ID NO:67, and SEQ ID NO:76; (q) SEQ ID NO:41, SEQ ID NO:49, SEQ ID NO:55, SEQ ID NO:60, SEQ ID NO:67, and SEQ ID NO:76; (r) SEQ ID NO:41, SEQ ID NO:49, SEQ ID NO:56, SEQ ID NO:60, SEQ ID NO:67, and SEQ ID NO:76; (s) SEQ ID NO: 80, SEQ ID NO: 83, SEQ ID NO: 50, SEQ ID NO: 90, SEQ ID NO: 97, and SEQ ID NO: 106; (t) SEQ ID NO:80, SEQ ID NO:83, SEQ ID NO:51, SEQ ID NO:91, SEQ ID NO:98, and SEQ ID NO:107; (u) SEQ ID NO:81, SEQ ID NO:84, SEQ ID NO:52, SEQ ID NO:92, SEQ ID NO:99, and SEQ ID NO:108; (v) SEQ ID NO:80, SEQ ID NO:83, SEQ ID NO:50, SEQ ID NO:90, SEQ ID NO:101, and SEQ ID NO:106; (w) SEQ ID NO:80, SEQ ID NO:83, SEQ ID NO:50, SEQ ID NO:90, SEQ ID NO:102, and SEQ ID NO:106; (x) SEQ ID NO:81, SEQ ID NO:85, SEQ ID NO:53, SEQ ID NO:94, SEQ ID NO:103, and SEQ ID NO:110; (y) SEQ ID NO:80, SEQ ID NO:86, SEQ ID NO:54, SEQ ID NO:95, SEQ ID NO:104, and SEQ ID NO:111; (z) SEQ ID NO:82, SEQ ID NO:87, SEQ ID NO:53, SEQ ID NO:93, SEQ ID NO:105, and SEQ ID NO:112; (aa) SEQ ID NO: 81, SEQ ID NO: 88, SEQ ID NO: 53, SEQ ID NO: 93, SEQ ID NO: 100, and SEQ ID NO: 109; (bb) SEQ ID NO:81, SEQ ID NO:89, SEQ ID NO:53, SEQ ID NO:93, SEQ ID NO:100, and SEQ ID NO:109; (cc) SEQ ID NO:81, SEQ ID NO:84, SEQ ID NO:55, SEQ ID NO:93, SEQ ID NO:100, and SEQ ID NO:109; (dd) SEQ ID NO:81, SEQ ID NO:84, SEQ ID NO:56, SEQ ID NO:93, SEQ ID NO:100, and SEQ ID NO:109; (ee) SEQ ID NO:81, SEQ ID NO:88, SEQ ID NO:55, SEQ ID NO:93, SEQ ID NO:100, and SEQ ID NO:109; (ff) SEQ ID NO:81, SEQ ID NO:88, SEQ ID NO:56, SEQ ID NO:93, SEQ ID NO:100, and SEQ ID NO:109; (gg) SEQ ID NO:81, SEQ ID NO:89, SEQ ID NO:55, SEQ ID NO:93, SEQ ID NO:100, and SEQ ID NO:109; or (hh) SEQ ID NO: 81, SEQ ID NO: 89, SEQ ID NO: 56, SEQ ID NO: 93, SEQ ID NO: 100, and SEQ ID NO: 109 A method comprising:

[0206]

[0216] 1. A method of inhibiting tumor or cancer formation or growth, or a combination thereof, in a subject in need thereof, comprising administering to the subject an anti-IL-8 antibody, wherein said inhibition inhibits neutrophil or monocyte activation, or a combination thereof, within the tumor microenvironment.

[0207]

[0217] 1. A method of inhibiting tumor or cancer formation or growth, or a combination thereof, in a subject in need thereof, comprising administering to the subject an anti-IL-8 antibody, wherein said inhibition reduces activation of neutrophils or monocytes, or a combination thereof, within the tumor microenvironment.

[0208]

[0218] 1. A method of inhibiting tumor or cancer formation or growth, or a combination thereof, in a subject in need thereof, comprising administering to the subject an anti-IL-8 antibody, wherein said inhibition reduces the viability of pre-cancerous stem cells or tumor cells. In one embodiment, pre-cancerous stem cells comprise pre-leukemic stem cells.

[0209]

[0219] 1. A method of inhibiting tumor or cancer formation or growth, or a combination thereof, in a subject in need thereof, comprising administering to said subject an anti-IL-8 antibody, wherein the cancer or tumor comprises a hematological cancer.

[0210]

[0220] 1. A method of inhibiting tumor or cancer formation or growth, or a combination thereof, in a subject in need thereof, comprising the step of administering to the subject an anti-IL-8 antibody, wherein the cancer comprises a hematological cancer, including leukemia, lymphoma, myeloma, acute myeloid leukemia (AML), acute promyelocytic leukemia, erythroleukemia, mixed B-myelomonocytic leukemia, or myelodysplastic syndrome (MDS).

[0211]

[0221] 1. A method of inhibiting tumor or cancer formation or growth, or a combination thereof, in a subject in need thereof, comprising administering to said subject an anti-IL-8 antibody, wherein said cancer or tumor comprises a solid cancer or tumor.

[0212]

[0222] 1. A method of inhibiting tumor or cancer formation or growth, or a combination thereof, in a subject in need thereof, comprising the step of administering to the subject an anti-IL-8 antibody, wherein the cancer or tumor comprises a solid cancer or tumor, wherein the solid cancer or tumor comprises a sarcoma, osteosarcoma, squamous cell carcinoma of the head and neck, non-small cell lung cancer, bladder cancer, pancreatic cancer, or pancreatic ductal adenocarcinoma.

[0213]

[0223] A method of inhibiting tumor or cancer formation or growth, or a combination thereof, in a subject in need thereof, comprising administering to said subject an anti-IL-8 antibody, wherein said subject is a human.

[0214]

[0224] 1. A method of inhibiting tumor or cancer formation or growth, or a combination thereof, in a subject in need thereof, comprising administering to the subject an anti-IL-8 antibody, wherein said inhibition inhibits neutrophil or monocyte activation, or a combination thereof, within the tumor microenvironment.

[0215]

[0225] 1. A method of inhibiting tumor or cancer formation or growth, or a combination thereof, in a subject in need thereof, comprising administering to the subject an anti-IL-8 antibody, wherein said inhibition inhibits activation of neutrophils or monocytes within the tumor microenvironment, or a combination thereof.

[0216]

[0226] 1. A method of inhibiting tumor or cancer formation or growth, or a combination thereof, in a subject in need thereof, comprising administering to the subject an anti-IL-8 antibody, wherein said inhibition reduces the viability of pre-cancerous stem cells or tumor cells. In one embodiment, pre-cancerous stem cells comprise pre-leukemic stem cells.

[0217]

[0227] 1. A method of inhibiting tumor or cancer formation or growth, or a combination thereof, in a subject in need thereof, comprising administering to said subject an anti-IL-8 antibody, said subject being a human, wherein: (a) the inhibition inhibits neutrophil or monocyte activation, or a combination thereof, within the tumor microenvironment; or (b) the inhibition inhibits activation of neutrophils or monocytes, or a combination thereof, within the tumor microenvironment; or (c) the inhibition reduces the viability of pre-cancerous stem cells or tumor cells; or (d) any combination thereof.

[0218]

[0228] 1. A method for treating a subject suffering from a disease, the method comprising the step of administering to the subject an anti-IL-8 antibody comprising an antibody antigen-binding domain comprising a heavy chain variable region (VH) and a light chain variable region (VL), thereby treating the disease in the subject, wherein the disease comprises cancer or a tumor or a disease associated with viral infection or inflammation, or a combination thereof, and the amino acid sequences of the VH-VL pairs are selected from the paired sequences set forth in SEQ ID NOs: 2 and 3, SEQ ID NOs: 4 and 5, SEQ ID NOs: 6 and 7, SEQ ID NOs: 8 and 9, SEQ ID NOs: 10 and 11, SEQ ID NOs: 12 and 13, SEQ ID NOs: 14 and 15, SEQ ID NOs: 16 and 17, SEQ ID NOs: 18 and 19, SEQ ID NOs: 20 and 21, SEQ ID NOs: 22 and 23, SEQ ID NOs: 24 and 25, SEQ ID NOs: 26 and 27, SEQ ID NOs: 28 and 29, SEQ ID NOs: 30 and 31, SEQ ID NOs: 32 and 33, and SEQ ID NOs: 34 and 35.

[0219]

[0229] 1. A method of treating a subject suffering from a disease, the method comprising the step of administering to the subject anti-IL-8 antibodies having complementarity determining region (CDR) sequences set forth in Table 1F, thereby treating the disease in the subject, wherein the disease comprises cancer or a tumor or a disease associated with a viral infection or inflammation, or a combination thereof, and wherein each antibody comprises a heavy chain variable region having heavy chain complementarity determining region (HCDR) 1, HCDR2, and HCDR3, and a light chain variable region having light chain complementarity determining region (LCDR) 1, LCDR2, and LCDR3, wherein the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 for each of the antibodies have the amino acid sequences set forth in Table 1F: (a) SEQ ID NO:41, SEQ ID NO:44, SEQ ID NO:53, SEQ ID NO:60, SEQ ID NO:67, and SEQ ID NO:76; (b) SEQ ID NO:81, SEQ ID NO:84, SEQ ID NO:53, SEQ ID NO:93, SEQ ID NO:100, and SEQ ID NO:109; (c) SEQ ID NO:40, SEQ ID NO:43, SEQ ID NO:50, SEQ ID NO:57, SEQ ID NO:64, and SEQ ID NO:73; (d) SEQ ID NO:40, SEQ ID NO:43, SEQ ID NO:51, SEQ ID NO:58, SEQ ID NO:65, and SEQ ID NO:74; (e) SEQ ID NO:41, SEQ ID NO:44, SEQ ID NO:52, SEQ ID NO:59, SEQ ID NO:66, and SEQ ID NO:75; (f) SEQ ID NO:40, SEQ ID NO:43, SEQ ID NO:50, SEQ ID NO:57, SEQ ID NO:68, and SEQ ID NO:73; (g) SEQ ID NO:40, SEQ ID NO:43, SEQ ID NO:50, SEQ ID NO:57, SEQ ID NO:69, and SEQ ID NO:73; (h) SEQ ID NO:41, SEQ ID NO:45, SEQ ID NO:53, SEQ ID NO:61, SEQ ID NO:70, and SEQ ID NO:77; (i) SEQ ID NO:40, SEQ ID NO:46, SEQ ID NO:54, SEQ ID NO:62, SEQ ID NO:71, and SEQ ID NO:78; (j) SEQ ID NO:42, SEQ ID NO:47, SEQ ID NO:53, SEQ ID NO:63, SEQ ID NO:72, and SEQ ID NO:79; (k) SEQ ID NO:41, SEQ ID NO:48, SEQ ID NO:53, SEQ ID NO:60, SEQ ID NO:67, and SEQ ID NO:76; (l) SEQ ID NO:41, SEQ ID NO:49, SEQ ID NO:53, SEQ ID NO:60, SEQ ID NO:67, and SEQ ID NO:76; (m) SEQ ID NO:41, SEQ ID NO:44, SEQ ID NO:55, SEQ ID NO:60, SEQ ID NO:67, and SEQ ID NO:76; (n) SEQ ID NO:41, SEQ ID NO:44, SEQ ID NO:56, SEQ ID NO:60, SEQ ID NO:67, and SEQ ID NO:76; (o) SEQ ID NO:41, SEQ ID NO:48, SEQ ID NO:55, SEQ ID NO:60, SEQ ID NO:67, and SEQ ID NO:76; (p) SEQ ID NO:41, SEQ ID NO:48, SEQ ID NO:56, SEQ ID NO:60, SEQ ID NO:67, and SEQ ID NO:76; (q) SEQ ID NO:41, SEQ ID NO:49, SEQ ID NO:55, SEQ ID NO:60, SEQ ID NO:67, and SEQ ID NO:76; (r) SEQ ID NO:41, SEQ ID NO:49, SEQ ID NO:56, SEQ ID NO:60, SEQ ID NO:67, and SEQ ID NO:76; (s) SEQ ID NO: 80, SEQ ID NO: 83, SEQ ID NO: 50, SEQ ID NO: 90, SEQ ID NO: 97, and SEQ ID NO: 106; (t) SEQ ID NO:80, SEQ ID NO:83, SEQ ID NO:51, SEQ ID NO:91, SEQ ID NO:98, and SEQ ID NO:107; (u) SEQ ID NO:81, SEQ ID NO:84, SEQ ID NO:52, SEQ ID NO:92, SEQ ID NO:99, and SEQ ID NO:108; (v) SEQ ID NO:80, SEQ ID NO:83, SEQ ID NO:50, SEQ ID NO:90, SEQ ID NO:101, and SEQ ID NO:106; (w) SEQ ID NO:80, SEQ ID NO:83, SEQ ID NO:50, SEQ ID NO:90, SEQ ID NO:102, and SEQ ID NO:106; (x) SEQ ID NO:81, SEQ ID NO:85, SEQ ID NO:53, SEQ ID NO:94, SEQ ID NO:103, and SEQ ID NO:110; (y) SEQ ID NO:80, SEQ ID NO:86, SEQ ID NO:54, SEQ ID NO:95, SEQ ID NO:104, and SEQ ID NO:111; (z) SEQ ID NO:82, SEQ ID NO:87, SEQ ID NO:53, SEQ ID NO:93, SEQ ID NO:105, and SEQ ID NO:112; (aa) SEQ ID NO: 81, SEQ ID NO: 88, SEQ ID NO: 53, SEQ ID NO: 93, SEQ ID NO: 100, and SEQ ID NO: 109; (bb) SEQ ID NO:81, SEQ ID NO:89, SEQ ID NO:53, SEQ ID NO:93, SEQ ID NO:100, and SEQ ID NO:109; (cc) SEQ ID NO:81, SEQ ID NO:84, SEQ ID NO:55, SEQ ID NO:93, SEQ ID NO:100, and SEQ ID NO:109; (dd) SEQ ID NO:81, SEQ ID NO:84, SEQ ID NO:56, SEQ ID NO:93, SEQ ID NO:100, and SEQ ID NO:109; (ee) SEQ ID NO:81, SEQ ID NO:88, SEQ ID NO:55, SEQ ID NO:93, SEQ ID NO:100, and SEQ ID NO:109; (ff) SEQ ID NO:81, SEQ ID NO:88, SEQ ID NO:56, SEQ ID NO:93, SEQ ID NO:100, and SEQ ID NO:109; (gg) SEQ ID NO:81, SEQ ID NO:89, SEQ ID NO:55, SEQ ID NO:93, SEQ ID NO:100, and SEQ ID NO:109; or (hh) SEQ ID NO: 81, SEQ ID NO: 89, SEQ ID NO: 56, SEQ ID NO: 93, SEQ ID NO: 100, and SEQ ID NO: 109 A method comprising:

[0220]

[0230] A method for treating a subject suffering from a disease, the method comprising the step of administering to the subject an anti-IL-8 antibody comprising an antibody antigen-binding domain comprising a heavy chain variable region (VH) and a light chain variable region (VL), wherein the disease comprises cancer, tumor, or a disease associated with viral infection or inflammation, or a combination thereof, and the cancer or tumor comprises a blood cancer.

[0221]

[0231] 1. A method for treating a subject suffering from a disease, the method comprising the step of administering to the subject an anti-IL-8 antibody comprising an antibody antigen-binding domain comprising a heavy chain variable region (VH) and a light chain variable region (VL), wherein the disease comprises cancer, or tumor, or a disease associated with viral infection or inflammation, or a combination thereof, and wherein the blood cancer comprises leukemia, lymphoma, myeloma, acute myeloid leukemia (AML), acute promyelocytic leukemia, erythroleukemia, mixed B-myelomonocytic leukemia, or myelodysplastic syndrome (MDS).

[0222]

[0232] A method for treating a subject suffering from a disease, the method comprising administering to the subject an anti-IL-8 antibody comprising an antibody antigen-binding domain comprising a heavy chain variable region (VH) and a light chain variable region (VL), wherein the disease comprises cancer or tumor, a disease associated with viral infection or inflammation, or a combination thereof, and the cancer or tumor comprises a solid cancer or tumor. In one embodiment, the method treats solid cancers or tumors, including sarcoma, osteosarcoma, squamous cell carcinoma of the head and neck, non-small cell lung cancer, bladder cancer, pancreatic cancer, or pancreatic ductal adenocarcinoma.

[0223]

[0233] A method for treating a subject suffering from a disease, the method comprising the step of administering to the subject an anti-IL-8 antibody comprising an antibody antigen-binding domain comprising a heavy chain variable region (VH) and a light chain variable region (VL), the disease comprising cancer, or a tumor, or a disease associated with viral infection or inflammation, or a combination thereof, and the subject is human.

[0224]

[0234] 1. An isolated polynucleotide sequence encoding a heavy chain variable region (VH) of an anti-IL-8 antibody and a light chain variable region (VL) of an anti-IL-8 antibody, wherein the polynucleotide sequence encoding the VH-VL pair is selected from the paired sequences set forth in SEQ ID NOs: 113 and 114, SEQ ID NOs: 115 and 116, SEQ ID NOs: 117 and 118, SEQ ID NOs: 119 and 120, SEQ ID NOs: 121 and 122, SEQ ID NOs: 123 and 124, SEQ ID NOs: 125 and 126, SEQ ID NOs: 127 and 128, SEQ ID NOs: 129 and 130, SEQ ID NOs: 131 and 132, SEQ ID NOs: 133 and 134, SEQ ID NOs: 135 and 136, SEQ ID NOs: 137 and 138, SEQ ID NOs: 139 and 140, SEQ ID NOs: 141 and 142, SEQ ID NOs: 36 and 37, and SEQ ID NOs: 38 and 39.

[0225]

[0235] 1. An isolated polynucleotide sequence encoding a heavy chain variable region (VH) of an anti-IL-8 antibody and a light chain variable region (VL) of an anti-IL-8 antibody, the polynucleotide sequence comprising two polynucleotide sequences, a first polynucleotide sequence encoding the VH of the anti-IL-8 antibody and a second polynucleotide sequence encoding the VL of the anti-IL-8 antibody.

[0226]

[0236] An isolated polynucleotide sequence encoding a heavy chain variable region (VH) and a light chain variable region (VL) of an anti-IL-8 antibody, wherein the anti-IL-8 antibody comprises an IgG, Fv, scFv, Fab, F(ab')2, minibody, diabody, triabody, nanobody, single domain antibody, multispecific antibody, bispecific antibody, triabody, single chain antibody, heavy chain antibody, chimeric antibody, or humanized antibody. In one embodiment, the IgG comprises IgG1, IgG2, IgG3, or IgG4.

[0227]

[0237] A vector comprising a polynucleotide sequence encoding a heavy chain variable region (VH) of an anti-IL-8 antibody and a light chain variable region (VL) of an anti-IL-8 antibody, wherein the polynucleotide sequence encoding the VH-VL pair is selected from the pair sequences set forth in SEQ ID NOs: 113 and 114, SEQ ID NOs: 115 and 116, SEQ ID NOs: 117 and 118, SEQ ID NOs: 119 and 120, SEQ ID NOs: 121 and 122, SEQ ID NOs: 123 and 124, SEQ ID NOs: 125 and 126, SEQ ID NOs: 127 and 128, SEQ ID NOs: 129 and 130, SEQ ID NOs: 131 and 132, SEQ ID NOs: 133 and 134, SEQ ID NOs: 135 and 136, SEQ ID NOs: 137 and 138, SEQ ID NOs: 139 and 140, SEQ ID NOs: 141 and 142, SEQ ID NOs: 36 and 37, and SEQ ID NOs: 38 and 39.

[0228]

[0238] A host cell comprising a vector comprising a polynucleotide sequence encoding a heavy chain variable region (VH) of an anti-IL-8 antibody and a light chain variable region (VL) of an anti-IL-8 antibody, wherein the polynucleotide sequence encoding the VH-VL pair is selected from the pair sequences set forth in SEQ ID NOs: 113 and 114, SEQ ID NOs: 115 and 116, SEQ ID NOs: 117 and 118, SEQ ID NOs: 119 and 120, SEQ ID NOs: 121 and 122, SEQ ID NOs: 123 and 124, SEQ ID NOs: 125 and 126, SEQ ID NOs: 127 and 128, SEQ ID NOs: 129 and 130, SEQ ID NOs: 131 and 132, SEQ ID NOs: 133 and 134, SEQ ID NOs: 135 and 136, SEQ ID NOs: 137 and 138, SEQ ID NOs: 139 and 140, SEQ ID NOs: 141 and 142, SEQ ID NOs: 36 and 37, and SEQ ID NOs: 38 and 39.

[0229]

[0239] 1. A method for producing an anti-IL-8 antibody comprising a heavy chain variable region (VH) and a light chain variable region (VL), the method comprising culturing a host cell comprising a vector comprising polynucleotide sequences encoding the heavy chain variable region (VH) of an anti-IL-8 antibody and the light chain variable region (VL) of an anti-IL-8 antibody under conditions conducive to expression of the vector in the host cell, wherein the polynucleotide sequences encoding the VH-VL pair are selected from the group consisting of SEQ ID NOs: 113 and 114, SEQ ID NOs: 115 and 116, SEQ ID NOs: 117 and 118, SEQ ID NOs: 119 and 120, SEQ ID NOs: 121 and 122, and expressing the polynucleotide sequence contained in the vector, thereby producing an anti-IL-8 antibody comprising a VH and a VL.

[0230]

[0240] 1. A method of inhibiting tumor or cancer formation or growth, or a combination thereof, in a subject in need thereof, the method comprising the step of administering to the subject a polynucleotide encoding an anti-IL-8 antibody comprising an antibody antigen-binding domain comprising a heavy chain variable region (VH) and a light chain variable region (VL), thereby inhibiting tumor formation or growth, or a combination thereof, in the subject, wherein the polynucleotide sequence encoding the VH-VL pair is selected from the pair sequences set forth in SEQ ID NOs: 113 and 114, SEQ ID NOs: 115 and 116, SEQ ID NOs: 117 and 118, SEQ ID NOs: 119 and 120, SEQ ID NOs: 121 and 122, SEQ ID NOs: 123 and 124, SEQ ID NOs: 125 and 126, SEQ ID NOs: 127 and 128, SEQ ID NOs: 129 and 130, SEQ ID NOs: 131 and 132, SEQ ID NOs: 133 and 134, SEQ ID NOs: 135 and 136, SEQ ID NOs: 137 and 138, SEQ ID NOs: 139 and 140, SEQ ID NOs: 141 and 142, SEQ ID NOs: 36 and 37, and SEQ ID NOs: 38 and 39.

[0231]

[0241] 1. A method of inhibiting tumor or cancer formation or growth, or a combination thereof, in a subject in need thereof, said method comprising administering to said subject a polynucleotide encoding an anti-IL-8 antibody comprising an antibody antigen-binding domain comprising a heavy chain variable region (VH) and a light chain variable region (VL), wherein said inhibition inhibits neutrophil or monocyte activation within the tumor microenvironment, or a combination thereof.

[0232]

[0242] 1. A method of inhibiting tumor or cancer formation or growth, or a combination thereof, in a subject in need thereof, said method comprising administering to said subject a polynucleotide encoding an anti-IL-8 antibody comprising an antibody antigen-binding domain comprising a heavy chain variable region (VH) and a light chain variable region (VL), wherein said inhibition reduces activation of neutrophils or monocytes within the tumor microenvironment, or a combination thereof.

[0233]

[0243] 1. A method of inhibiting tumor or cancer formation or growth, or a combination thereof, in a subject in need thereof, said method comprising administering to said subject a polynucleotide encoding an anti-IL-8 antibody comprising an antibody antigen-binding domain comprising a heavy chain variable region (VH) and a light chain variable region (VL), wherein said inhibition reduces the viability of pre-cancerous stem cells or tumor cells.

[0234]

[0244] 1. A method for inhibiting tumor or cancer formation or growth, or a combination thereof, in a subject in need thereof, the method comprising administering to the subject a polynucleotide encoding an anti-IL-8 antibody comprising an antibody antigen-binding domain comprising a heavy chain variable region (VH) and a light chain variable region (VL), and the pre-cancerous stem cells comprise pre-leukemic stem cells.

[0235]

[0245] A method for inhibiting tumor or cancer formation or growth, or a combination thereof, in a subject in need thereof, the method comprising administering to the subject a polynucleotide encoding an anti-IL-8 antibody comprising an antibody antigen-binding domain comprising a heavy chain variable region (VH) and a light chain variable region (VL), wherein the cancer or tumor comprises a hematological cancer. In one embodiment, the hematological cancer comprises leukemia, lymphoma, myeloma, acute myeloid leukemia (AML), acute promyelocytic leukemia, erythroleukemia, mixed B-myelomonocytic leukemia, or myelodysplastic syndrome (MDS).

[0236]

[0246] 1. A method for inhibiting tumor or cancer formation or growth, or a combination thereof, in a subject in need thereof, the method comprising administering to the subject a polynucleotide encoding an anti-IL-8 antibody comprising an antibody antigen-binding domain comprising a heavy chain variable region (VH) and a light chain variable region (VL), wherein the cancer or tumor comprises a hematological cancer. In one embodiment, the solid cancer or tumor comprises sarcoma, osteosarcoma, squamous cell carcinoma of the head and neck, non-small cell lung cancer, bladder cancer, pancreatic cancer, or pancreatic ductal adenocarcinoma.

[0237]

[0247] 1. A method for inhibiting tumor or cancer formation or growth, or a combination thereof, in a subject in need thereof, the method comprising administering to the subject a polynucleotide encoding an anti-IL-8 antibody comprising an antibody antigen-binding domain comprising a heavy chain variable region (VH) and a light chain variable region (VL), wherein the subject is a human.

[0238]

[0248] A method of treating a subject suffering from a disease, the method comprising the step of administering to the subject an anti-IL-8 antibody comprising an antibody antigen-binding domain comprising a heavy chain variable region (VH) and a light chain variable region (VL), thereby treating the disease in the subject, wherein the disease comprises cancer or tumor or a disease associated with viral infection or inflammation, or a combination thereof, and wherein the polynucleotide sequences encoding the VH-VL pair are selected from the group consisting of SEQ ID NOs: 113 and 114, SEQ ID NOs: 115 and 116, SEQ ID NOs: 117 and 118, SEQ ID NOs: 119 and 120, SEQ ID NOs: 121 and 122, SEQ ID NOs: 123 and 124, SEQ ID NOs: 125 and 126, SEQ ID NOs: 127 and 128, SEQ ID NOs: 129 and 130, SEQ ID NOs: 131 and 132, SEQ ID NOs: 133 and 134, SEQ ID NOs: 135 and 136, SEQ ID NOs: 137 and 138, SEQ ID NOs: 139 and 140, SEQ ID NOs: 141 and 142, SEQ ID NOs: 143 and 144, SEQ ID NOs: 145 and 146, SEQ ID NOs: 147 and 148, SEQ ID NOs: 149 and 150, SEQ ID NOs: 151 and 152, SEQ ID NOs: 153 and 154, SEQ ID NOs: 155 and 156, SEQ ID NOs: 157 and 158, SEQ ID NOs: 159 and 160, SEQ ID NOs: 161 and 162, SEQ ID NO The method of claim 1, wherein the pair of sequences is selected from the pair of sequences set forth in SEQ ID NOs: 117 and 118, SEQ ID NOs: 119 and 120, SEQ ID NOs: 121 and 122, SEQ ID NOs: 123 and 124, SEQ ID NOs: 125 and 126, SEQ ID NOs: 127 and 128, SEQ ID NOs: 129 and 130, SEQ ID NOs: 131 and 132, SEQ ID NOs: 133 and 134, SEQ ID NOs: 135 and 136, SEQ ID NOs: 137 and 138, SEQ ID NOs: 139 and 140, SEQ ID NOs: 141 and 142, SEQ ID NOs: 36 and 37, and SEQ ID NOs: 38 and 39.

[0239]

[0249] A method for treating a subject suffering from a disease, the method comprising administering to the subject an anti-IL-8 antibody comprising an antibody antigen-binding domain comprising a heavy chain variable region (VH) and a light chain variable region (VL), wherein the disease comprises cancer, tumor, viral infection- or inflammation-related disease, or a combination thereof, and the cancer or tumor comprises a blood cancer. In one embodiment, the blood cancer comprises leukemia, lymphoma, myeloma, acute myeloid leukemia (AML), acute promyelocytic leukemia, erythroleukemia, mixed B-myelomonocytic leukemia, or myelodysplastic syndrome (MDS).

[0240]

[0250] A method for treating a subject suffering from a disease, the method comprising administering to the subject an anti-IL-8 antibody comprising an antibody antigen-binding domain comprising a heavy chain variable region (VH) and a light chain variable region (VL), wherein the disease comprises a cancer or tumor, a disease associated with a viral infection or inflammation, or a combination thereof, and the cancer or tumor comprises a blood cancer. In one embodiment, the solid cancer or tumor comprises sarcoma, osteosarcoma, squamous cell carcinoma of the head and neck, non-small cell lung cancer, bladder cancer, pancreatic cancer, or pancreatic ductal adenocarcinoma.

[0241]

[0251] A method for treating a subject suffering from a disease, the method comprising the step of administering to the subject an anti-IL-8 antibody comprising an antibody antigen-binding domain comprising a heavy chain variable region (VH) and a light chain variable region (VL), the disease comprising cancer, or a tumor, or a disease associated with viral infection or inflammation, or a combination thereof, and the subject is human.

[0242]

[0252] Whenever a numerical range is given herein, it is intended to include any recited numerical value (fractional or integral) within the given range. The phrases "ranging between / ranging between" a first indicated number and a second indicated number and "ranging from / ranging from" a first indicated number "to" a second indicated number given herein are used interchangeably herein and are intended to include the first and second indicated numbers and all fractional and integral numbers therebetween. [Example]

[0243] Example Example 1: Materials and Methods

[0253] Immunization, hybridoma generation and antibody recovery:

[0254] Recombinant human IL-8 (aa 28-99; SEQ ID NO: 1) (Sino Biological, Cat# 10098-HNCH2) was used for immunization after confirmed binding to anti-IL-8 antibody (R&D, Cat# mAb208) by ELISA. A group of 24 Alivamab mice (AMM-KL) (Ablexis transgenic mice—Ablexis, LLC, San Francisco, Calif.) were immunized with human IL-8 according to the AMMPD-4 immunization protocol (ADS), and titers were assessed twice. Additionally, 14 AMM-KL and 3 AMM-LO Alivamab mice were immunized according to the AMMPD-2 immunization protocol (ADS), and titers were assessed once at the end of the process. High titers were observed on day 10, and samples (lymph nodes and spleens) from a total of 41 mice were collected and pooled into groups based on immunization procedure, mouse strain, and titer results. Samples were processed for B cell enrichment using magnetic negative selection, and hybridomas were generated using electrofusion. Hybridomas were plated into 30 x 384-well plates at concentrations ranging from one to three hybridomas per well, and the remaining material was cryopreserved. Growing hybridoma colonies were counted on day 7, and saturated supernatants were collected for screening on day 8. Positive hybridomas were expanded to 96-well plates on day 11.

[0244]

[0255] The anti-IL-8 mAb clone used throughout the following examples was a recombinant IgG1 antibody containing the heavy and light chain CDR regions present in the positive hybridoma.

[0245]

[0256] IL-8 binding antibodies by ELISA:

[0257] Recombinant human or cynomolgus IL-8 (Sino Biological, Cat# 10098-HNCH2 and Kingfisher, Cat# RP1321Y-100, respectively) was coated at 1 μg / mL in PBS pH 7.4 in a Nunc Maxisorp high-binding black 384-well plate (Thermo Fisher, Cat# 460518) overnight at 4°C. After washing three times with wash buffer (1x PBS, pH 7.5, 0.05% Tween-20), the plate was blocked (1% BSA in PBS) for 1 hour at room temperature. The plate was then washed four times and incubated with a dilution series of monoclonal antibody (mAb) (in PBS, 0.02% Tween-20) for 1 hour at room temperature. mAbs were tested at a highest concentration of 100 nM, followed by seven half-log dilutions. After four washes, plates were incubated with anti-human-Fc-HRP detection antibody (Chimerigen, Cat# HF-2208, 1:10,000, in 0.02% Tween-20 in PBS). Plates were then washed five times, Supersignal ELISA Pico substrate (Thermofisher, Cat# 37069) was added, and the chemiluminescent signal was read in a Sprectramax L for 200 ms / well.

[0246]

[0258] Octet IL-8 binding antibodies:

[0259] The mAbs were loaded in a three-point dilution series (25 nM, 12.5 nM, 6.25 nM) against either human or cynomolgus IL-8 (Sino Biological, Cat#10098-H01H1 and Kingfisher, Cat#RP1321Y, respectively) onto a 16-channel antibody-human-Fc capture sensor (Fortebio, Cat#1810172). Human or cynomolgus IL-8 was titrated at 1:1 dilutions starting at the highest concentration of 50 nM for four or three points, respectively. The experimental parameters followed to determine the rate constants were baseline for 60 seconds, loading (antibody to sensor) for 180 seconds, binding (analyte to antibody) for 120 seconds, dissociation for 1200 seconds, and regeneration for 4 × 30 seconds.

[0247]

[0260] Interaction of IL-8 with its receptor by flow cytometry:

[0261] The mAb was mixed with IL-8-Fc (aa28-99 SEQ ID NO: 1, Sino Biological, Cat# 10098-H01H) in serial dilutions. The highest concentration of mAb tested was 1 x 10 -7 The mixture was diluted 1:3.3 with IL-8 at a concentration of 10 nM. The mixture was coated onto a 96-well plate and incubated at room temperature for 15 minutes. Separately, HEK293-CXCR2 cells were dissociated using a cell stripper, filtered through a 40 μm filter, and washed once with FACS buffer (1% BSA / 2 mM EDTA / 0.01% NAN3 in PBS pH 7.4). 4 × 10 cells were then added to the plate. 6 Cell suspensions (25 μl) at a concentration of 1 / ml were mixed with each mAb dilution and incubated first at room temperature for 15 minutes, followed by 30 minutes at 4° C. Cells were then washed with FACS buffer and stained with anti-huFc647 (Jackson, Code#109607008, 1:2000) and HelixBlue (1:1000) for 30 minutes at 4° C. After two washes, cells were resuspended in FACS buffer (25 μl) and samples were analyzed by flow cytometry using FlowJo software.

[0248]

[0262] IL-8-inducible NFkB reporter assay:

[0263] The cDNA sequences of CXCR1 and CXCR2 (R&D systems, Cat# RDC0025 and Cat# RDC0027, respectively) were cloned into an expression vector (ADS), and the constructs were transfected into HEK293 cells. Cells were placed under antibiotic selection (hygromycin, 50 μg / ml), and CXCR1 and CXCR2 expression was confirmed by flow cytometry (anti-CXCR1 antibody, Biolegend, Cat# 320605 and anti-CXCR2 antibody, Biolegend, Cat# 149305). Cells recombinantly expressing CXCR1 and CXCR2 were cloned to establish stable cell lines, and IL-8 binding was confirmed by flow cytometry. CXCR1-NFκB reporter cells were generated after transduction of a lentiviral NFκB-dependent luciferase construct (G&P Biosciences, Cat# LTR004) into stable CXCR1 clones. Cells were placed under antibiotic selection (hygromycin, 50 μg / ml and puromycin, 2.5 μg / ml), and the NFκB reporter system was validated by monitoring IL-8-dependent luciferase activity with Bright-Glo (Promega, Cat# E2620).

[0249]

[0264] HEK293-CXCR1-NFκB reporter cells were cultured at 1 × 10 in 1% FBS / OptiMEM (50 μl) in 96-well plates. 4 The cells were plated at a density of 1 x 10 cells / well and incubated overnight in a humidified incubator at 37°C / 5% CO2. Separately, mAbs were mixed with IL-8 (aa28-99-SEQ ID NO: 1, Sino Biological, Cat#10098-HNCH2) in serial dilutions for 15 minutes at room temperature. The highest concentration of each mAb tested was 1 x 10 -7The final concentration of IL-8 in the mixture was 10 nM, followed by 11 serial dilutions (1:2). The mixture was added to the cells, and the plate was incubated for 5.5 hours in a humidified incubator at 37°C / 5% CO2. The supernatant from each well (50 μl) was mixed with an equal volume of Bright-Glo, and the luminescence signal was measured using a SpectraMax L.

[0250]

[0265] Detection of CXCR1 and CXCR2 on the cell surface by flow cytometry:

[0266] Count the cells and measure 1 x 10 6 Cells were collected into round-bottom polystyrene tubes. Cells were washed with 0.5% BSA in PBS and blocked with FcR blocking reagent (Miltenyi Biotec, Inc., Cat. #130-059-901) according to the manufacturer's recommendations. Cells were then stained with anti-CXCR1 or anti-CXCR2 antibodies (Biolegend, Cat. #320605 and Bioolegend, Cat. #320714, 5:100, respectively) for 30 minutes at 4°C in the dark. Expression of CXCR1 and CXCR2 receptors on the cell surface was assessed by flow cytometry on a BD Facs Canto II or BD Facs Aria, and data were analyzed using FlowJo software.

[0251]

[0267] Detection of CXCR1 and CXCR2 on the cell surface by immunohistochemistry:

[0268] Cells were collected at 0.03 x 10 6Cells were seeded at a concentration of 500 μl per well onto Nunc™ lab-Tek™ II 8-well chamber slides™ (ThermoFisher, Cat# 154534PK) and incubated overnight in a humidified incubator at 37°C / 5% CO2. Cells were then pre-fixed with 250 μl of culture medium and an equal volume of pre-chilled 4% paraformaldehyde (Fisher Scientific, Cat# AAJ19943K2) for 10 minutes on ice. Next, cells were washed twice with pre-chilled PBS, fixed with 250 μl of pre-chilled 4% paraformaldehyde for 10 minutes on ice, and washed three times again with pre-chilled PBS. All samples were analyzed in duplicate using a mouse and rabbit-specific HRP / DAB (ABC) detection IHC kit (Abcam, Cat# ab64264). Cells were covered with a drop of hydrogen peroxide and incubated for 10 minutes at room temperature. After two washes (in PBS), a protein block drop was applied for 10 minutes at room temperature. Next, cells were washed four times and incubated with anti-CXCR1 or anti-CXCR2 antibodies (Boster Cat# PA2080, 1:500 for CXCR1; Abcam Cat# 89251, 1:200 for CXCR1; and ProteinTech Cat# 20634-1-AP, 1:200 for CXCR2) in 1% BSA in TBS overnight at 4°C. Cells were then washed four times and incubated with biotinylated goat anti-polyvalent antibody for 10 minutes at room temperature. Next, cells were washed four times and incubated with streptavidin peroxidase for 10 minutes at room temperature. Slides were rinsed four times with PBS and then incubated with DAB chromogen for 1-10 minutes. Cells were then rinsed four times with PBS and incubated with hematoxylin (Abcam Cat#220365, 1:2 in ddH2O) for 1-5 min. Finally, cells were rinsed with tap water, mounting media was applied to slides, coverslips were added, and images were viewed using a Cytation™ Cell Imaging Multi-Mode Reader (BioTek).

[0252]

[0269] IL-8-induced CXCR2 internalization:

[0270] Count THP-1 cells and count 1 x 10 for each condition. 6 Cells were collected into round-bottom polystyrene tubes. Cells were treated with IL-8 alone (12.5 nM) or together with three different concentrations of each mAb (1, 10, or 100 nM) for 30 minutes in a humidified incubator at 37°C / 5% CO2. After incubation, cells were collected, washed with 0.5% BSA in PBS, and blocked with FcR reagent (Miltenyi Biotec, Inc., Cat# 130-059-901) according to the manufacturer's recommendations. Cells were then stained with anti-CXCR2 antibody (Biologend, Cat# 320714, 5 μl of antibody per million cells per 100 μl) for 30 minutes on ice in the dark. CXCR2 expression was detected by flow cytometry using a BD Facs Canto, and data were analyzed using FlowJo software.

[0253]

[0271] Antibody evaluation by SDS-PAGE:

[0272] mAb samples (2 μg) were mixed with either 4x loading buffer, N-ethylmaleimide (1 μl) for non-reducing conditions, or DTT (1 μl of 1 M) for reducing conditions. Samples prepared under reducing conditions were boiled at 95°C for 5 min and cooled to 4°C before loading onto RunBlue SDS Gel 4-20% (Expedeon, Cat# NXG42012). All samples were electrophoresed using diluted RunBlue 20x SDS running buffer (Expedeon, Cat# NXB50500) at a constant 200 V for 50 min. The gel was then washed with water for 1 min and stained with InstantBlue (Expedeon, Cat# ISB1L) for 6 h. The gel was then placed in water, and images were captured using an Azure Biosystems c200 in visible light equipment.

[0254]

[0273] Antibody size exclusion analysis:

[0274] Size exclusion analysis was performed on an HPLC (Agilent 1260) equilibrated in isocratic running buffer (0.1 M Na2PO4 and 0.1 M Na2O4, pH 6.7). mAb samples (10 μg) were injected onto a TSKgel SuperSW3000 (TOSOH, Cat# 18675) at a flow rate of 0.35 ml / min using a refrigerated autosampler and a 40 μl injection loop. Absorbance was measured at 280 nm, 254 nm, and 215 nm using a diode array detector, and the % main peak was calculated by AUC in Chemstation software.

[0255]

[0275] Detection of IL-8 secretion levels in cancer cell lines by ELISA:

[0276] 1 x 10 cells 6Cells were seeded at a density of 1000 cells / well in 12-well plates (USA Scientific, Cat# CC7682-7512) and cultured in 1 ml of medium for 72 hours in a humidified incubator at 37°C / 5% CO2. The culture medium was removed from the wells, transferred to a 1.5 ml microcentrifuge tube, and centrifuged at 10,000 rpm for 5 minutes at room temperature. The supernatant was collected in a new 1.5 ml microcentrifuge tube and either analyzed immediately by ELISA or stored in a -80°C freezer. A high-boiling-point 96-well plate was coated with capture antibody (1:250 dilution) in PBS overnight at 4°C. The plate was then washed once (0.02% Tween-80 in PBS) and blocked with assay diluent / blocking buffer (BD, Cat# 555213) for 30 minutes at room temperature. The plate was washed three times again and incubated with cell supernatant (undiluted or diluted 1:10) for 2 hours at room temperature. IL-8 standards were used as references, with the highest concentration of 200 pg / ml followed by a two-fold dilution series. Next, the plate was washed four times and incubated with primary and detection antibodies (both at 1:250 dilution) for 1 hour at room temperature. The capture antibody, primary / secondary mixture, and IL-8 standard were provided in the IL-8 detection kit (BD OptEIA, Cat#555244). Finally, the plate was washed five times and incubated with TMB substrate (Sigma, Cat#T0440, 100 μl / well). After color development, sulfuric acid (0.16 M, 50 μl) was added to stop the reaction, and the plate was read at 450 nm using a Cytation5 (Biotek) plate reader.

[0256]

[0277] Cell viability assay for patient-derived AML samples:

[0278] Primary patient-derived acute myeloid leukemia (AML) cells were seeded at a density of 20,000 cells / well in 96-well plates on study day 0. The enriched medium (75 μl) contained StemSpan™ SFEM medium (STEMCELL Technologies, Cat# 09650), 2% FBS (heat-inactivated), StemSpan™ CC110 (STEMCELL Technologies, Cat# 02697), and recombinant human IL3 (R&D Systems, Cat# 203 / IL-010 / CF). Cells were treated with mAb at a concentration of 40 μg / ml on day 0, followed by 5-fold serial dilutions. Plates were incubated at 37°C / 5% CO in a humidified incubator. 2 The medium was kept at RT and not changed for the duration of the assay. On day 6, the plates were removed from the incubator and allowed to equilibrate to room temperature for a maximum of 30 minutes. CellTiter-Glo was then added to the wells (100 μl), and the plates were mixed on a plate rocker for 2 minutes, followed by incubation at room temperature for 10 minutes. Luminescence signals were recorded using a Tecan plate reader.

[0257]

[0279] Cell line viability assay:

[0280] Omniscreen cell lines were cultured at 4 × 10 in five 96-well plates (Corning, Cat# 3340) (Plates A, B, C, D, and E). 3Cells were seeded at a density of 1000 cells / well (90 μl) and incubated overnight at 37°C / 5% CO2 in a humidified incubator. At time 0, culture medium (10 μl) was added to each well of plate A and equilibrated at room temperature for 30 minutes. CellTiter-Glo reagent (Promega, Cat# G7572, 50 μl) was then added to each well, mixed on an orbital shaker for 5 minutes, and incubated at room temperature for 20 minutes. Luminescence was measured using an Envision Multi Label Reader (Perkin Elmer, Equip ID: TAREA0020). Cells in plates B, C, and D were treated in triplicate with antibody (10 μl) at a concentration of 200 μg / ml, followed by 5-fold serial dilutions to obtain nine test concentrations. Cells in plate E were treated with the reference control. After 72 hours of incubation at 37°C / 5% CO2 in a humidified incubator, CellTiter-Glo Reagent was added and plates were read as described for the first time point.

[0258] Example 2: Characterization of anti-IL-8 mAb clones

[0281] Objective: To investigate the physical properties of the generated recombinant anti-IL-8 mAb and characterize the mAb clones.

[0259]

[0282] Methods: mAbs were generated as described in Example 1. Briefly, antibodies were generated after immunization of AlivaMab mice and selected for high-affinity functional antibodies by hybridoma fusion and screening. Recombinant human IgG1 antibodies with either human kappa or lambda light chains were generated. Methods for analyzing the physical properties of recombinant anti-IL-8 antibodies were well known in the art, including gel migration under non-reducing and reducing conditions and HPLC size exclusion analysis. The recombinant antibody clones used throughout these examples are fully human IgG1 antibodies, in which the CDR domains of the monoclonal antibodies are generated in Alivamab mice.

[0260]

[0283] The variable heavy (VH) and variable light (VL) regions were sequenced using techniques well known in the art to generate nucleotide sequences encoding the VH and VL amino acid sequences of the anti-IL-8 clones.

[0261]

[0284] Results: Exemplary amino acid sequences of variable heavy domain (VH) and variable light domain (VL) pairs are shown below in Table 1A.

[0262] [Table 1]

[0263] [Table 2]

[0264] [Table 3]

[0265]

[0286] The complementarity-determining regions (CDRs) for each of the recombinant mAb clones were calculated using two different methods. Tables 1B and 1C show the modeled CDR determinations according to Kabat with slight modifications based on internal modeling (Kabat, EA et al., In: Sequences of Proteins of Immunological Interest, NIH Publication, 91-3242 (1991)). (Table 1B (light chain CDRs) and Table 1C (heavy chain CDRs)). Tables 1D and 1E show the modeled CDR determinations according to information found on the Immuno Gene Tics website (http: / / www.imgt.org / IMGTScientificChart / Numbering / IMGTnumbering.html) (Table 1D (light chain CDRs) and Table 1E (heavy chain CDRs)).

[0266] [Table 4]

[0267] [Table 5]

[0268] [Table 6]

[0269] [Table 7]

[0270]

[0291] The light chains of antibodies can be classified as either kappa (κ) or lambda (λ) types. Tables 1B and 1D identify for each clone whether the light chain is of the kappa or lambda type. The set of six CDR regions present in each recombinant mAb can be determined based on the CDR sequences shown in Tables 1B and 1C or Tables 1D and 1E, and is shown in Table 1F herein below.

[0271] [Table 8]

[0272] [Table 9]

[0273] [Table 10]

[0274] [Table 11]

[0275] [Table 12]

[0276] [Table 13]

[0277] [Table 14]

[0278]

[0294] Table 1G shows the nucleotide sequences encoding the VH and VL regions of the anti-IL-8 clones, where the CDR regions are indicated as follows: (a) Kabat-modeled CDRs are encoded by the bolded sequences, and (b) IMGT-modeled CDRs are encoded by the italicized sequences (5'-3' CDR1, CDR2, CDR3 for each of the VH and VL listed above).

[0279]

[0295] Expression of antibody molecules was examined under non-reducing and reducing conditions. Intact antibody migrating at approximately 150 kDa, typical of IgG, was detected in all antibody samples electrophoresed under non-reducing conditions (Figure 1 - left lane). A band migrating at 50 kDa, typical of the IgG heavy chain, and a lower band at 25 kDa, typical of the IgG light chain, were detected in all antibody samples electrophoresed under reducing conditions (Figure 1 - right lane).

[0280]

[0296] Size exclusion analysis was performed by HPLC. The antibody profile showed a symmetric main peak with a retention time of 7.6-7.9 minutes, typical of intact IgG monomers (Figure 2). All samples showed >95% of the main peak as determined by AUC, indicating minimal soluble aggregates or clipping degradation by size exclusion chromatography.

[0281]

[0297] The generated recombinant Ab clones have different light chains, where clones STLX3, STLX5 and STLX35 have κ (kappa) light chains, and STLX14, STLX18, STLX31 and STLX38 have λ (lambda) light chains.

[0282] Example 3: Binding of mAbs to human and cynomolgus IL-8

[0298] Objective: To analyze the binding kinetics of the generated mAbs for binding to human and cynomolgus monkey IL-8. Analysis of cross-reactivity with monkey IL-8 is important because it allows toxicity studies to be performed using non-human primates directly with clinical candidates rather than in chimpanzees or using surrogate molecules. Such toxicity studies can be used to provide relevant safety assessments.

[0283]

[0299] Methods: Binding assays were performed using ELISA or Octect as described above.

[0284]

[0300] result:

[0301] The results presented here are for mAb clones STLX3, STLX5, STLX14, STLX18, STLX31, and STLX35. Table 2 (below) and Figure 3 demonstrate the binding of the mAbs to human and cynomolgus IL-8 based on ELISA results. Table 2 shows the binding kinetics of the mAbs to human and cynomolgus IL-8 as determined by cell-free ELISA. EC 50 Values ​​are calculated and given as pM ranges.

[0285] [Table 15]

[0286]

[0303] The data shown in Figure 3 and Table 3 demonstrate binding of the mAbs to human and cynomolgus IL-8 in a dose-dependent manner.

[0287]

[0304] Table 3 (below) shows the binding rates of mAbs to human and cynomolgus IL-8 as determined by Octect, where the affinity of each antibody is represented by its Kd value. The data shown in Table 3 include binding on and off rates.

[0288] [Table 16]

[0289]

[0306] Summary: The analyzed mAb clones exhibit a range of IL-8 binding affinities from 1 to 600E-12M as demonstrated by the Octet method. On-rates for the tested mAb clones were greater than 1.0E+6 l / s and off-rates were up to 8.6E-4 l / s. Comparison of binding to human and cynomolgus IL-8 showed EC50 values ​​that differed by less than two-fold.

[0290] Example 4: Inhibition of IL-8 binding to cell surface receptors and IL-8-induced NFκB activity by mAb clones

[0307] Objective: To analyze the functional activity of the mAb clones.

[0291]

[0308] Methods: IL-8 interaction with the CXCR2 receptor in the presence of mAb clones was measured as described in Example 1. An IL-8-inducible NKκB reporter assay was used to determine IL-8-induced NFκB activity in the presence of different mAb clones as described in Example 1. IL-8-induced CXCR2 internalization was measured by flow cytometry as described in Example 1.

[0292]

[0309] result:

[0310] Table 4 and Figure 4 present data demonstrating the ability of the mAb clones to block the binding of IL-8 to its cell surface receptor, CXCR2.

[0293] [Table 17]

[0294]

[0312] The effect of the mAbs on blocking IL-8-Fc binding to HEK293 cells stably expressing CXCR2 was measured in a cell-based assay by flow cytometry. IC50 values ​​were calculated and are shown in Table 4 in the nM range. Figure 4 shows that the mAb clones promoted blocking of IL-8 binding to CXCR2 in a dose-dependent manner.

[0295]

[0313] Not only did the mAb clones block IL-8 binding to the CXCR2 receptor, but Table 5 and Figure 5 present data showing that these antibodies also had the ability to inhibit IL-8-induced NFκB activity in tissue culture cells.

[0296]

[0314] The effect of mAb clones on blocking IL-8-induced NFκB activity was monitored in a HEK293-CXCR1 cell-based reporter assay. The IC50 value of each antibody was calculated and the data are shown in Table 5, where the IC50 values ​​were in the nM range.

[0297] [Table 18]

[0298]

[0316] Figure 5 shows data from an analysis of mAb blocking of IL-8-induced NFκB activity as measured in a HEK293-CXCR1 cell-based reporter assay. The data show that the mAb promoted blocking of NFκB activity in a dose-dependent manner.

[0299]

[0317] Summary: The mAb clones were able to affect IL-8 binding and activity at both the CXCR1 and CXCR2 receptors, where blocking potency for the CXCR2 receptor as measured by a flow-based assay yielded IC50 values ​​ranging from 3 to 26 nM, while for CXCR1, the NFκB reporter assay yielded IC50 values ​​ranging from 5 to 66 nM.

[0300] Example 5: Expression of CXCR1 and CXCR2 receptors on cancer cells and inhibition of IL-8-induced CXCR2 internalization and cancer cell proliferation by mAb clones

[0318] Objective: To analyze the effects of mAb clones on cancer cells (primary acute myeloid leukemia (AML) cancer cells and hematological and solid tumor cell lines).

[0301]

[0319] Methods: Detection of CXCR1 and CXCR2 on the cell surface of cancer cells was performed using flow cytometry and immunohistochemistry as described in Example 1. Detection of IL-8 secretion by cancer cell lines was performed using ELISA as described in Example 1. Additional viability assays used are those described in Example 1.

[0302]

[0320] result:

[0321] Cell surface expression analysis showed detection of the CXCR2 receptor but not the CXCR1 receptor in the majority of AML cell lines analyzed (THP-1, OCI-AML5, MOLM-13, MOLM-14, HL-60, HEL, OCI-AML2, OCI-AML3, MV411, and KG-1a) (Table 6), whereas osteosarcoma cell lines (143B, SaOS-2, U-2 OS, MG-63, OS17) and pancreatic cancer cell line (PANC-1) expressed both CXCR2 and CXCR1 receptors (Table 7; Figure 6).

[0303] [Table 19]

[0304]

[0323] Table 6 shows that expression of the CXCR1 receptor was not detected in the cell lines analyzed by flow cytometry, as indicated by the (-) symbol. Expression of the CXCR2 receptor was detected by flow cytometry in most cell lines at various levels, as indicated by the (+) symbol. The highest expression level is indicated by (+++) and the lowest expression level is indicated by (+).

[0305] [Table 20]

[0306]

[0325] Staining of solid tumor cell lines (osteosarcoma and pancreatic tumor) showed that expression of CXCR1 and CXCR2 receptors was detected at various levels by IHC in osteosarcoma and pancreatic tumor cell lines (Figure 6). No primary antibody conditions were used as negative controls. Table 7 lists the expression of CXCR1 and CXCR2 receptors detected on the cell surface by IHC in all osteosarcoma and pancreatic cell lines tested, at various levels as indicated by the (+) symbol. The highest expression level is indicated by (+++), and the lowest expression level is indicated by (+).

[0307]

[0326] Recognizing that IL-8 can function in both paracrine and autocrine modes, follow-up analyses examined IL-8 expression in different solid tumor and hematological tumor cell lines. Table 8 and Figure 7 provide data demonstrating IL-8 secretion in both osteosarcoma and AML cell lines. Figure 7 shows the results of monitoring IL-8 secretion by ELISA in the supernatants of MG-63, U2-OS, SAOS-2, and 143-B osteosarcoma cells. IL-8 secretion was monitored in MG-63, U2-OS, and 143-B cells in a time-dependent manner. The lowest level of IL-8 secretion was detected in SAOS-2 cells and did not increase over the 96-hour time course. The presence of a higher percentage of FBS in the growth medium correlated with higher IL-8 secretion levels. Table 8 shows the IL-8 data for both osteosarcoma and AML cell lines.

[0308] [Table 21]

[0309]

[0328] Table 8 shows that IL-8 secretion was detected by ELISA in all cell lines tested at various levels as indicated by the (+) symbol, with the highest secretion level indicated by (+++) and the lowest secretion level indicated by (+).

[0310]

[0329] Based on the data that CXCR2 receptors were present in acute monocytic leukemia (AML) cells, the inhibition of IL-8-induced CXCR2 internalization by mAb clones was examined in AML cells (Figure 8). Figure 8 shows that in THP-1 cells, CXCR2 receptor internalization was induced by IL-8 treatment, while CXCR2 internalization was blocked in a dose-dependent manner by co-treatment with mAb clones STLX3, STLX5, STLX14, STLX18, STLX31, STLX35, and STLX38. The percentage of control is [1-(X-min) / (max-min)]. * The values ​​were calculated by 100 and shown in the graph, where IL-8 and antibody are represented by light grey, no stimulation is represented as max, and IL-8 stimulation alone is represented as min.

[0311]

[0330] The ability of the mAbs to inhibit cancer cell growth was then measured. Table 9 presents data demonstrating the inhibition of patient-derived AML cells by the mAbs. As used throughout, in some embodiments, mAb clones are identified by the letters STX followed by a number, and in other embodiments, by STLX followed by a number. Those skilled in the art will understand that for each mAb clone, STX3 and STLX3 identify the same clone. This pattern applies to all of the clones listed herein.

[0312] [Table 22]

[0313]

[0332] The effect of mAbs on inhibiting the proliferation of patient-derived AML cells was monitored by cell proliferation assay and measured by Cell-Titer Glo. Patient-derived AML cells were treated with each antibody at a concentration of 20 μg / ml for 6 days, and the percentage of growth inhibition was calculated and represented by different colors. Inhibition greater than 30% is indicated by black circles, inhibition between 3% and 30% is indicated by gray, and inhibition less than 3% is indicated by white.

[0314]

[0333] The ability of mAbs to inhibit cancer cell growth in hematological and solid tumor cell lines will also be measured. The effect of mAbs on inhibiting the growth of hematological and solid tumor cell lines will be monitored by cell proliferation assays and measured by CellTiter-Glo. Cell lines of bladder, leukemia, lymphoma, myeloma, bone, HN / pharynx, or lung origin (squamous cell carcinoma of the head and neck (SCCHN) and non-small cell lung cancer (NSCLC)) will be treated with each antibody at a concentration of 200 μg / ml for 3 days, and the percentage of growth inhibition will be calculated and presented in the table.

[0315]

[0334] Summary: The results demonstrate the presence of CXCR1 and CXCR2 receptors on cancer cells and the ability of the generated mAb clones to inhibit IL-8-induced CXCR2 internalization in AML cancer cell lines. Furthermore, the mAbs inhibited proliferation in patient-derived AML cancer cells.

[0316]

[0335] The results presented in Examples 2-5 demonstrate that fully human monoclonal antibodies against IL-8 have been generated and provide functional activity. These mAbs block the ability of IL-8 to bind to its cell surface receptor, thus interfering with IL-8's ability to transduce cell signals. Use of these mAbs also inhibits the growth of AML primary cancer cells, demonstrating their relevance for cancer therapy.

[0317] Example 6: STLX18 inhibits IL-8-induced AKT and ERK phosphorylation in neutrophils

[0336] Objective: To investigate the effect of IL-8 antibody on IL-8-induced signaling in human neutrophils.

[0318]

[0337] Methods: Human neutrophils isolated from whole blood were activated with IL-8 in the presence of an IL-8 neutralizing antibody (STLX18) or a control IgG1 antibody. Cell lysates were examined by Western blot, and IL-8-induced downstream signaling was monitored using antibodies against phosphorylated specific ERK and AKT. Signals were detected by chemiluminescence following exposure to autoradiography film. An antibody against actin was used as a loading control.

[0319]

[0338] result:

[0339] Figure 9 shows that the STLX18 monoclonal antibody inhibited IL-8-induced downstream signaling in human neutrophils, as monitored by Western blot using phospho-specific antibodies against ERK and AKT. There was no background signaling in the absence of IL-8. The IgG1 control has the same backbone as the STLX18 antibody but does not target IL-8.

[0320]

[0340] Summary: The results demonstrate that the IL-8 neutralizing antibody STLX18 inhibited IL-8-induced phosphorylation signaling in primary human neutrophils, suggesting that STLX18 inhibited IL-8 signaling via its cell surface receptor.

[0321] Example 7: STLX18 inhibits human neutrophil migration in response to IL-8 in vitro

[0341] Objective: To investigate the in vitro activity of IL-8 antibody on neutrophil chemotaxis.

[0322]

[0342] Methods: Human neutrophils isolated from whole blood were placed in the upper chamber of a multiwell insert, and IL-8 was placed in the lower chamber. The lower chamber also contained either an IL-8 neutralizing antibody (STLX18) or a control IgG1 antibody. Neutrophil migration was assessed either by live-cell imaging software or by cell counting.

[0323]

[0343] result:

[0344] Human neutrophil migration across a porous membrane toward an IL-8 source was inhibited by the STLX18 antibody. Addition of IL-8 to the lower chamber of a transwell plate increased human neutrophil migration across the membrane, which was inhibited by co-incubation of the STLX18 antibody and IL-8 in the lower chamber (FIG. 10). Neutrophil migration was assessed using the Incucyte® Live-Cell Analysis System.

[0324]

[0345] Summary: These results demonstrate that STLX18 antibody inhibited neutrophil migration toward IL-8 in an in vitro chemotaxis assay, suggesting that STLX18 may offer therapeutic benefits in neutrophil-driven inflammatory diseases, including cancer.

[0325] Example 8: STLX18 inhibits knee swelling in a rabbit model of gouty knee inflammation

[0346] Objective: To analyze the in vivo activity of IL-8 antibody in a gouty knee inflammation model.

[0326]

[0347] Methods: Monosodium urate (MSU) crystals injected into the knee joint caused swelling and inflammation. Rabbit knees were pretreated with either saline or an IL-8 neutralizing antibody (STLX18). The right knee of each rabbit was injected with MSU crystals, and the left knee was injected with saline. Knee diameters were measured at various time points using calipers.

[0327]

[0348] result:

[0349] Binding analysis of the STLX18 antibody to rabbit IL-8 showed that the STLX18 antibody recognized rabbit IL-8 (data not shown).

[0328]

[0350] Rabbit knee joints injected with MSU crystals increased in diameter immediately after injection. Treatment with STLX18 antibody reduced swelling in MSU-injected joints (FIG. 11B), while control joints injected with saline did not change in size (FIG. 11A). Knee diameters were determined by caliper measurement at the indicated time points after injection.

[0329]

[0351] summary:

[0352] These results indicate that IL-8 antibodies can reduce knee swelling induced by MSU crystals and may provide a therapeutic treatment for diseases and conditions associated with inflammation.

[0330]

[0353] While particular features of IL-8 antibodies and their uses have been exemplified and described herein, many modifications, substitutions, variations, and equivalents will now occur to those skilled in the art. It is therefore to be understood that the appended claims are intended to cover all such modifications and variations that fall within the true spirit of the invention.

Claims

1. An isolated anti-IL-8 antibody comprising a heavy chain variable region having a heavy chain complementarity determining region (HCDR) 1, HCDR2, and HCDR3, and a light chain variable region having a light chain complementarity determining region (LCDR) 1, LCDR2, and LCDR3, the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 are the amino acid sequences set forth in SEQ ID NO: 60, SEQ ID NO: 67, SEQ ID NO: 76, SEQ ID NO: 41, SEQ ID NO: 44, and SEQ ID NO: 53, respectively; or Amino acid sequences set forth in SEQ ID NO:93, SEQ ID NO:100, SEQ ID NO:109, SEQ ID NO:81, SEQ ID NO:84, and SEQ ID NO:53, respectively An isolated anti-IL-8 antibody comprising:

2. 2. The isolated anti-IL-8 antibody of claim 1, wherein the antibody comprises a heavy chain variable region (VH)-light chain variable region (VL) pair, and the amino acid sequences of the VH-VL pair are the sequences set forth in SEQ ID NOs: 8 and 9.

3. A composition comprising the isolated anti-IL-8 antibody of claim 1 or claim 2 and a pharmaceutically acceptable carrier.

4. An isolated polynucleotide sequence encoding a heavy chain variable region (VH) of an anti-IL-8 antibody and a light chain variable region (VL) of an anti-IL-8 antibody, wherein the VH comprises heavy chain complementarity determining region (HCDR) 1, HCDR2, and HCDR3, and the VL comprises light chain complementarity determining region (LCDR) 1, LCDR2, and LCDR3, and the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 have the amino acid sequences set forth in SEQ ID NO:60, SEQ ID NO:67, SEQ ID NO:76, SEQ ID NO:41, SEQ ID NO:44, and SEQ ID NO:53, respectively; Amino acid sequences set forth in SEQ ID NO:93, SEQ ID NO:100, SEQ ID NO:109, SEQ ID NO:81, SEQ ID NO:84, and SEQ ID NO:53, respectively An isolated polynucleotide comprising:

5. The isolated polynucleotide of claim 4, wherein the polynucleotide encodes a heavy chain variable region (VH)-light chain variable region (VL) pair, and the amino acid sequence of the VH-VL pair is the sequence set forth in SEQ ID NOs: 8 and 9.

6. 6. The isolated polynucleotide of claim 5, wherein the isolated polynucleotide encoding the VH-VL pair comprises the nucleotide sequence set forth in SEQ ID NOs: 119 and 120.

7. 7. The isolated polynucleotide of any one of claims 4 to 6, wherein the VH is encoded by one polynucleotide and the VL is encoded by another polynucleotide.

8. A vector comprising the polynucleotide according to any one of claims 4 to 7.

9. A host cell comprising the vector of claim 8.

10. 10. A method of producing the anti-IL-8 antibody of claim 1, comprising culturing the host cell of claim 9 under conditions conducive to expression of said anti-IL-8 antibody, whereby the antibody has a complementarity determining region (CDR) sequence as set forth below: SEQ ID NO:60, SEQ ID NO:67, SEQ ID NO:76, SEQ ID NO:41, SEQ ID NO:44, and SEQ ID NO:53 (HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3, respectively); or SEQ ID NO:93, SEQ ID NO:100, SEQ ID NO:109, SEQ ID NO:81, SEQ ID NO:84, and SEQ ID NO:53 (HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3, respectively) The amino acid sequence of The method comprises producing the anti-IL-8 antibody having the formula:

11. The method of claim 10, wherein the antibody comprises a heavy chain variable region (VH)-light chain variable region (VL) pair, and the VH-VL pair comprises the amino acid sequences set forth in SEQ ID NOs: 8 and 9, or the polynucleotide sequence encoding the heavy chain variable region (VH)-light chain variable region (VL) pair comprises the sequences set forth in SEQ ID NOs: 119 and 120.

12. 3. The isolated anti-IL-8 antibody of claim 1 or 2, wherein the antibody comprises an IgG, Fv, scFv, Fab, F(ab')2, minibody, diabody, triabody, multispecific antibody, bispecific antibody, triabody, single-chain antibody, chimeric antibody, or humanized antibody.

13. 12. Use of the isolated anti-IL-8 antibody of any one of claims 1-2 and 12, or the composition of claim 3, or the isolated polynucleotide of any one of claims 4-7, or the anti-IL-8 antibody produced by the method of any one of claims 10-11, in the preparation of a medicament for inhibiting the formation or growth of a tumor or cancer, or a combination thereof, in a human subject in need thereof.

14. wherein the inhibition (a) inhibiting neutrophil or monocyte activation, or neutrophil and monocyte activation, within the tumor microenvironment; (b) reducing neutrophil or monocyte activation, or neutrophil and monocyte activation, within the tumor microenvironment; or (c) reducing the viability of pre-cancerous stem cells or tumor cells; or (d) any combination thereof.

15. Use of the isolated anti-IL-8 antibody of any one of claims 1 to 2 and 12, or the composition of claim 3, or the isolated polynucleotide of any one of claims 4 to 7, or the anti-IL-8 antibody produced by the method of any one of claims 10 to 11, in the preparation of a medicament for treating a human subject suffering from a disease comprising cancer or a tumor or a disease associated with viral infection or inflammation, or a combination thereof.

Images