Antibodies, uses and methods

Abstract

To provide anti-human OX40L (hOX40L) antibodies, new medical uses thereof, and methods of treating or preventing hOX40L mediated diseases or conditions.SOLUTION: Provided are antibodies or fragments thereof that specifically bind to hOX40L and compete with antibodies hOX40L for binding to said 02D10, wherein said antibodies or fragments thereof comprise a VH domain comprising a HCDR3 comprising the motif VRGXYYY, wherein X is any amino acid sequence.SELECTED DRAWING: Figure 1

Description

[Technical Field]

[0001] The present invention relates to anti-human OX40L antibodies, new medical uses and methods. [Background technology]

[0002] OX40 ligand (OX40L) is a 34 kDa type II transmembrane protein, a member of the TNF family. The crystallized complex of human OX40 and OX40L consists of one OX40L (trimer) and three OX40 monomers. The human extracellular domain is 42% identical to mouse OX40L.

[0003] OX40L is not constitutively expressed but can be induced on professional APCs such as B cells, dendritic cells (DCs), and macrophages. Other cell types, such as Langerhans cells, endothelial cells, smooth muscle cells, mast cells, and natural killer (NK) cells, can be induced to express OX40L. T cells can also express OX40L. The OX40L receptor, OX40, is expressed on activated T cells (CD4 and CD8 T cells, Th2, Th1, and Th17 cells) and CD4 T cells, even in the absence of activation. + Foxp3 + It is expressed on cells.

[0004] The interaction between OX40 and OX40L occurs during T cell-DC interactions, 2 or 3 days after antigen recognition. After leaving DCs, OX40-expressing T cells can interact with non-DC cells expressing OX40L and receive OX40 signals from these cells, which may provide essential signals for generating memory T cells, enhancing Th2 responses, and prolonging inflammatory responses. OX40 signals to recipient T cells render them resistant to Treg-mediated suppression.

[0005] Graft-versus-host disease (GVHD) is the leading cause of mortality after allogeneic bone marrow transplantation. In the acute form of the disease, mature T cells present in the bone marrow graft recognize donor tissue as foreign in the environment of the injured tissue, which, via host APCs, triggers the activation and proliferation of donor T cells and their subsequent migration to the liver, spleen, gastrointestinal tract, skin, and lungs, resulting in CTL effector responses and the release of proinflammatory cytokines / chemokines. The onset of acute disease usually occurs within the first 100 days after transplantation (Hill-Ferrara, Blood May 1, 2000 vol. 95 no. 9 2754-275; Reddy-Ferrara Blood, Volume 17, Issue 4, December 2003).

[0006] Chronic GvHD usually appears 100 days after transplantation and is thought to involve several factors, including thymic damage caused by preceding acute GvHD, which leads to reduced clearance of pathogenic T cells (Zhang et al., September 1, 2007 vol.179 no. 5 3305-3314), and upregulation of TGF-β, which leads to fibrosis (McCormick et al., J Immuno, November 15, 1999 vol.163 no. 10 5693-5699), as well as elevated B cell components driven by B cell activating factor (BAFF) (Sarantopoulos et al, Clin Cancer Res October 15, 2007 13; 6107) and autoantibodies against platelet-derived growth factor receptor (PDR) (Svegliati et al, Blood July 1, 2007 Vol. 110 No. 1 237-241).

[0007] Clinical studies have shown that OX40 is effective in treating acute (Morante et al, Clinical and Experimental Immunology, 145:36-43) and chronic (Kotani et al, Blood November 15, 2001) vol.98 no. 10 3162-3164) have shown that OX40L is upregulated in both GvHD and OX40L-dependent leukemia. Administration of antagonistic anti-OX40L enhanced survival in a lethal acute mouse model of GvHD, with a 70% survival rate in the treated group compared with the untreated group, all of which died by day 43 (Tsukada et al., Blood, 1 April 2000, Volume 95, Number 7), whereas treatment with antagonistic anti-OX40 Ab promoted disease and mortality (Blazar et al. Blood May 1, 2003 vol. 101 no. 9 3741-3748). Disruption of OX40-OX40L interaction has been shown to be effective in several other inflammatory diseases, using anti-OX40L Abs used to treat mouse models of colitis (Totsuka et al., AJP - GI April 1, 2003 vol. 284 no. 4 G595-G603), and anti-OX40L Abs can prevent the development of diabetes in NOD mice (Pakala et al., European Journal of Immunology Volume 34, Issue 11, pages 3039-3046, November 2004).

[0008] References Lamb, LS, Abhyankar, SA, Hazlett, L., O'Neal, W., Folk, RS, Vogt, S., Parrish, RS, Bridges, K., Henslee-Downey, P.J. and Gee, A. P. (1999), Expression of CD134 (0X-40) on T-cells during the first 100 days following allogeneic bone marrow transplantation as a marker for lymphocyte activation and therapy-resistant graft-versus-host disease. Cytometry, 38: 238-243。

[0009] Xupeng Ge, Julia Brown, Megan Sykes, Vassiliki A. Boussiotis, CD134-Allodepletion Allows Selective Elimination of Alloreactive Human T-cells without Loss of Virus-Specific and Leukemia-Specific Effectors, Biology of Blood and Marrow Transplantation, Volume 14, Issue 5, May 2008, Pages 518 - 530。

[0010] Naoto Ishii, Takeshi Takahashi, Pejman Soroosh, Kazuo Sugamura, Chapter 3 - OX40-OX40 Ligand Interaction in T-Cell-Mediated Immunity and Immunopathology, In: Frederick W. Alt, Editor(s), Advances in Immunology, Academic Press, 2010, Volume 105, Pages 63 - 98。

[0011] Croft, M., So, T., Duan, W. and Soroosh, P. (2009), The significance of OX40 and OX40L to T-cell biology and immune disease. Immunological Reviews, 229: 173-191. Summary of the Invention

[0012] The present invention provides anti-human OX40L (hOX40L) antibodies and fragments and novel medical applications for the treatment or prevention of hOX40L-mediated diseases or conditions in humans.

[0013] First configuration The antibody or fragment is administered to a human, and the antibody or fragment a. secretion of a cytokine selected from TNF-alpha, IL-2, IL-3, IL-4, IL-5, IL-6, IL-8, IL-9, IL-10, IL-13, IL-17, RANTES, and interferon gamma in humans; b. proliferation of human white blood cells, and c. An antibody or fragment thereof that specifically binds to hOX40L for treating or preventing an hOX40L-mediated disease or condition in a human, wherein the method is for treating or preventing an hOX40L-mediated disease or condition in a human by reducing one, more than one, or all of the binding of hOX40L expressed by endothelial cells to an hOX40 receptor expressed by human T cells.

[0014] Second Configuration An antibody or fragment thereof that specifically binds to hOX40L and competes for binding to said hOX40L with an antibody selected from the group consisting of 02D10, 10A07, 09H04, and 19H01.

[0015] Third Configuration Use of an antibody or fragment thereof, a. secretion of a cytokine selected from TNF-alpha, IL-2, IL-3, IL-4, IL-5, IL-6, IL-8, IL-9, IL-10, IL-13, IL-17, RANTES, and interferon gamma in humans; b. proliferation of human white blood cells, and c. Use of an antibody or fragment thereof that specifically binds to hOX40L in the manufacture of a medicament for administration to a human to treat or prevent a hOX40L-mediated disease or condition in the human by reducing one, more than one, or all of the binding of hOX40L expressed by endothelial cells to a hOX40 receptor expressed by human T cells.

[0016] Fourth Configuration a. secretion of a cytokine selected from TNF-alpha, IL-2, IL-3, IL-4, IL-5, IL-6, IL-8, IL-9, IL-10, IL-13, IL-17, RANTES, and interferon gamma in humans; b. proliferation of human white blood cells, and c. A method of treating or preventing a hOX40L-mediated disease or condition in a human by reducing one, more than one, or all of the following: binding of hOX40L expressed by endothelial cells to a hOX40 receptor expressed by human T cells; The method comprises administering to the human a therapeutically effective amount of an antibody or fragment that specifically binds to hOX40L.

[0017] Fifth Configuration 1. An antibody or fragment thereof that specifically binds to hOX40L and competes with antibody 02D10 for binding to said hOX40L, wherein the antibody or fragment thereof comprises a VH domain comprising an HCDR3 that comprises the motif VRGXYYY, where X is any amino acid.

[0018] Sixth Configuration An antibody or fragment thereof that specifically binds to hOX40L and competes with antibody 02D10 for binding to said hOX40L, wherein the antibody or fragment comprises a VH domain comprising the HCDR3 sequence of SEQ ID NO: 40 or 46, or the HCDR3 sequence of SEQ ID NO: 40 or 46 containing fewer than five amino acid substitutions.

[0019] Seventh Configuration A human antibody or fragment thereof comprising an HCDR3 of 16 to 27 amino acids and derived from the recombination of a human VH gene segment, a human D gene segment, and a human JH gene segment, wherein the human JH gene segment is IGHJ6 that specifically binds to hOX40L for treating or preventing an autoimmune disease selected from an autoimmune disease or condition, a systemic inflammatory disease or condition, or a transplant rejection.

[0020] Eighth Configuration Use of a human antibody or fragment thereof comprising an HCDR3 of 16 to 27 amino acids and derived from the recombination of a human VH gene segment, a human D gene segment, and a human JH gene segment, wherein the human JH gene segment is IGHJ6 that specifically binds to hOX40L, in the manufacture of a pharmaceutical for administration to a human to treat or prevent an hOX40L-mediated disease or condition in the human, selected from an autoimmune disease or condition, a systemic inflammatory disease or condition, or a transplant rejection.

[0021] 9th Configuration A method for treating or preventing an hOX40L-mediated disease or condition selected from an autoimmune disease or condition, a systemic inflammatory disease or condition, or a transplant rejection, comprising administering to said human a therapeutically effective amount of a human antibody or fragment thereof, said antibody comprising an HCDR3 of 16 to 27 amino acids and derived from the recombination of a human VH gene segment, a human D gene segment, and a human JH gene segment, wherein the human JH gene segment is IGHJ6 that specifically binds to hOX40L, thereby treating or preventing the hOX40L-mediated disease or condition.

[0022] The present invention also provides pharmaceutical compositions, kits, nucleic acids, vectors, and hosts. [Brief explanation of the drawings]

[0023] [Figure 1] Profiling of fully human recombinant anti-OX40L antibodies in an HTRF ligand / receptor neutralization assay. Data shown are representative of three replicate experiments. [Figure 2] 10. Determination of the effect of anti-OX40L antibodies in allogeneic PBMC / T-mixed lymphocyte reactions. Data shown are from three independent donor pairs, each donor being a different individual. DETAILED DESCRIPTION OF THE INVENTION

[0024] The present invention provides the following aspects 1 to 113.

[0025] The present invention is useful, for example, for the treatment or prevention of transplant rejection, such as graft-versus-host disease (GvHD) or allograft rejection. The present invention is also useful, for example, for the treatment or prevention of inflammatory bowel disease, such as UC or CD, or for the treatment or prevention of airway inflammatory diseases or conditions. In one example, this embodiment is useful for the treatment or prevention of asthma. The present invention is also useful, for example, for the treatment or prevention of fibrosis. The present invention is also useful, for example, for the treatment or prevention of diabetes. The present invention is also useful, for example, for the treatment or prevention of uveitis. The present invention is also useful, for example, for the treatment or prevention of pyoderma gangrenosum. The present invention is also useful, for example, for the treatment or prevention of giant cell arteritis. The present invention is also useful, for example, for the treatment or prevention of Schnitzler syndrome. The present invention is also useful, for example, for the treatment or prevention of non-infectious scleritis.

[0026] 1. The antibody or fragment is administered to a human, and the antibody or fragment: a. secretion of a cytokine selected from TNF-alpha, IL-2, IL-3, IL-4, IL-5, IL-6, IL-8, IL-9, IL-10, IL-13, IL-17, RANTES, and interferon gamma in humans; b. proliferation of human white blood cells, and c. An antibody or fragment thereof that specifically binds to hOX40L for treating or preventing an hOX40L-mediated disease or condition in a human, wherein the method is for treating or preventing an hOX40L-mediated disease or condition in a human by reducing one, more than one, or all of the binding of hOX40L expressed by endothelial cells to an hOX40 receptor expressed by human T cells.

[0027] The inventors have therefore, for the first time, identified the reduction of (a), (b), and (c) as a method for treating and / or preventing OX40L-mediated diseases and conditions in humans, and provide antibodies and antibody fragments for this purpose.

[0028] In one example, the secretion is leukocyte secretion. In one example, (a) is indicated by significantly elevated levels of cytokine(s) in human blood, plasma, or serum.

[0029] In one example, the cytokine is selected from (i) TNF-alpha, (ii) IL-2, and (iii) interferon gamma. In an example, the cytokine is TNF-alpha. In an example, the cytokine is IL-2. In one example, the cytokine is interferon gamma. In one example, the cytokines are (i) and (ii), or (i) and (iii), or (ii) and (iii), or (i) through (iii).

[0030] In one example, the reduction in (a), (b), or (c), or any other reduction disclosed herein, is at least a 10 or 20% reduction compared to the level in a human at risk for or suffering from an hOX40L-mediated disease or condition. In one example, the latter is a human as recited in embodiment 1 prior to administration of the antibody or fragment; in another example, the latter human is a different human. In one example, the reduction is at least 10, 20, 30, 40, 50, or 60%.

[0031] (i) In one example, the antibody or fragment can affect a reduction in the secretion of a relevant cytokine from leukocytes (e.g., human T cells) in an in vitro assay (described further below), such that administration of such an antibody or fragment to a human leads to a reduction in (a).

[0032] (ii) In one example, the antibody or fragment can affect a reduction in proliferation of leukocytes (e.g., human PBMCs and / or human T cells) in an in vitro assay (described further below), such that administration of such antibody or fragment to a human leads to a reduction in (b).

[0033] (iii) In one example, the antibody or fragment can affect a reduction in binding of hOX40 receptors expressed by human T cells to hOX40L expressed by endothelial cells in an in vitro assay (described further below), such that administration of such an antibody or fragment to a human leads to a reduction in (c).

[0034] In one example, (i) and (ii), or (i) and (iii), or (ii) and (iii), or (i) through (iii) apply.

[0035] Additionally or alternatively, the reduction can be assessed using a sample from the treated human (see, for example, J. Clin. Immunol., 2004 Jan., 24(1):74-85; "Increased expression of CCL20 in human inflammatory bowel disease"); See Kaser A et al. This publication provides an example of a generally applicable technique using tissue biopsies to read decreased cytokine levels, indicating decreased cytokine secretion after in vivo antibody treatment. Similar methods can be used to determine decreased secretion of one or more cytokines in humans who have received an antibody of the invention. Those skilled in the art will be familiar with techniques for assessing cytokine levels in patients and patient samples, for example, by using one or more of tissue biopsies, immunohistochemistry, immunofluorescence, tissue staining, cytokine mRNA quantification (e.g., using PCR such as Taqman™ PCR), and cytokine protein detection and quantification (e.g., using cytokine-specific tool antibodies and quantification, such as ELISA or another standard protein quantification technique). For example, if the disease or condition is of the digestive tract (e.g., IBD), a biopsy of relevant gastrointestinal tissue from a patient who has received an antibody of the invention can be performed, followed by quantification of cytokine mRNA and / or cytokine protein (e.g., using quantitative PCR). The results can be compared to cytokine quantification in biopsied relevant tissues from the same patient before antibody administration, or to another human patient suffering from the same disease or condition but not receiving anti-OX40L treatment or treatment for the disease or condition. In this way, one skilled in the art can determine that the antibodies of the present invention reduce cytokine secretion in a human recipient. Instead of assessing gastrointestinal tissue levels, different tissues or samples from a human patient can be used instead, depending on the nature and location of the disease or condition. For example, if the disease or condition is in one of the respiratory tracts (e.g., the lungs), lung or other respiratory tract tissue samples can be collected for cytokine assessment. Alternatively, bronchoalveolar lavage (BAL) samples can be used, as will be apparent to one skilled in the art. In another example, for some diseases or conditions, cytokine reduction can be assessed in blood, serum, or plasma samples collected from a human receiving an antibody of the present invention, and then compared to levels before receiving the antibody or to levels in an untreated human, as discussed above.

[0036] As known in the art, the term "leukocyte" includes, for example, one or more of lymphocytes, polymorphonuclear leukocytes, and monocytes. As will be readily apparent to those skilled in the art, the term "monocyte" also includes, for example, peripheral blood mononuclear cells (PBMCs) or monocyte-derived cells, such as dendritic cells (DCs). See, e.g., Immunobiology, 2013 Nov, 218(11):1392-401. doi: 10.1016 / j.imbio.2013.07.005. Epub 2013 Jul 25; "Leukoreduction system chambers are an efficient, effective, and economic source of functional monocyte-derived dendritic cells and lymphocytes," Pfeiffer IA et al.

[0037] The proliferation of leukocytes, e.g., lamina propria lymphocytes (LPL), can be assessed using tissue biopsy, staining, and histology, as will be apparent to those skilled in the art. Hematoxylin and eosin staining (H&E staining or HE staining) is commonly used in histology to look for, for example, infiltrating lymphocytes in all types of human tissues and is one of the primary stains in histology. This is the most widely used stain in medical diagnostics and is often the gold standard, and therefore can be used to assess leukocyte proliferation as in the present invention. For example, gastrointestinal tissue (e.g., gastrointestinal tissue) from a human suffering from or at risk for a hOX40L-mediated disease or condition can be harvested, stained, and assessed for the degree of LPL infiltration. Comparisons can be made between such tissue from a human receiving an antibody of the present invention and the degree of infiltration in tissue harvested from the same human before administration of the antibody, or from another human not receiving treatment and at risk for or suffering from a disease or condition. For example, comparisons can be made between human gastrointestinal tissues taken from the same (or different) humans suffering from IBD.

[0038] For example, standard binding assays known to those skilled in the art, such as ELISA or SPR, can be used to determine whether an antibody or fragment can reduce the binding of hOX40 receptors expressed by human T cells to endothelial cells expressed by hOX40L.

[0039] Inflammatory bowel disease (IBD) is a chronic inflammatory disorder affecting the gastrointestinal tract with a seemingly ever-increasing incidence and tendency toward more severe clinical phenotypes. The disease is characterized by an exaggerated immune response to the luminal microbiota, suggesting that a deficiency in the barrier function of the gut microbiota may be involved, and studies support this view (Cucchiara et al., 2012; Jostins et al., 2012; Manichanh et al., 2012; Salzman et al., 2007, all from Deuring et al., "The cell biology of the intestinal epithelium and its relation to inflammatory bowel disease", The International Journal of Inflammatory Bowel Disease, Vol. 1, No. 1, 2012). (cited in Journal of Biochemistry & Cell Biology 45 (2013) 798- 806). IBD comprises two main groups: Crohn's disease (CD) and ulcerative colitis (UC). CD patients can have inflammatory lesions throughout their digestive tract, while lesions in UC patients are limited to the colon. See also Hisamatsu et al. ("Immune aspects of the pathogenesis of inflammatory bowel disease", Pharmacology & Therapeutics 137 (2013) 283-297) and documents cited therein.

[0040] Granuloma formation is one of the most important pathological features of human Crohn's disease. Mizoguchi et al. +It has been shown that dendritic cells (DCs) produce IL-23 and contribute to granuloma formation in a mouse colitis model (Mizoguchi et al., 2007). Th1 immune responses are dominant in Crohn's disease. Indeed, CD4 + T cells expressed T-bet and produced large amounts of interferon (IFN)-γ (Matsuoka et al., 2004). Sakuraba et al. showed that DCs in the mesenteric lymph nodes of patients with Crohn's disease strongly promoted Th1 and Th17 immune responses (Sakuraba et al., 2009). Mesenteric lymph node DCs contribute to the pathogenesis of IBD, especially Crohn's disease.

[0041] The role of cytokines in diseases and conditions Muzes et al, World J Gastroenterol 2012 November 7; 18(41): 5848-5861 ISSN 1007-9327 (print) ISSN 2219-2840 (online), see "Changes of the cytokine profile in inflammatory bowel diseases".

[0042] Cytokines are essential signals for the mucosal immune system, essential for maintaining normal gastrointestinal homeostasis. An imbalance in these cytokine profiles favoring the initiation of inflammation can lead to disease states such as those observed in inflammatory bowel disease (IBD), e.g., Crohn's disease (CD) and ulcerative colitis (UC). The role of pro-inflammatory cytokines, such as IL-1α, IL-1β, IL-2, -6, -8, -12, -17, -23, IFN-gamma, or TNF-alpha, in IBD is related to the initiation and progression of UC and CD. CD is often described as a prototype T helper (Th)1-mediated disease, because the primary inflammatory mediators are Th1 cytokines, such as interleukin (IL)-12, interferon (IFN)-γ, and tumor necrosis factor (TNF)-α.

[0043] Binding of TNF-like ligands to their receptors triggers intracellular pathways directly involved in cell proliferation, differentiation, and survival. Most members of the TNF / TNF receptor protein superfamily are expressed on immune cells and play important roles in multiple components of the immune response. TNF-α is a master cytokine in the pathogenesis of IBD. It exerts its pleiotropic effects through the expression of adhesion molecules, fibroblast proliferation, and procoagulant factors, as well as through the initiation of cytotoxicity, apoptosis, and acute-phase responses. The source of TNF-α in IBD is in part immune cells such as macrophages or monocytes, as well as differentiated Th1 cells. Serum levels of TNF-α correlate with the clinical activity of UC and CD

[31] . It plays a regulatory role in colonic inflammation in IBD. The role of TNF-α in CD has been widely investigated. Binding of TNF-α to serum soluble TNF receptors 1 and 2 (sTNFR1 and 2) initiates pro-inflammatory signaling. Levels of sTNFR1 and 2 are elevated in CD.

[0044] Tumor necrosis factor-like factor 1A (TL1A), another member of the TNF family, stimulates IFN-γ secretion by binding to death receptor 3 (DR3). DR3 is expressed by a high proportion of cells from mucosal biopsies of UC and CD, and elevated IFN-γ levels have been observed in IBD patients with disease activity. The TL1A / DR3 axis is involved in the pathogenesis of CD. Lamina propria macrophages are the primary producers of TL1A, and its expression is significantly enhanced in CD. TL1A and IL-23 have been found to synergistically promote IFN-γ production by mucosal T cells. IFN-γ is produced by TH1 T cells. Upon inflammation initiation, IFN-γ is produced and subsequently acts through various molecules and pathways in the immune system to intensify the inflammatory process. A large body of literature has extensively documented the pro-inflammatory properties of IFN-γ, leading to the mainstream opinion that IFN-γ is the major pro-inflammatory cytokine in inflammatory and autoimmune diseases. Interferon gamma is causally involved in experimental inflammatory bowel disease in mice (Ito et al, Clinical and Experimental Immunology (2006), 146:330-338). - / - The results clearly showed that the mice exhibited attenuated colitis after DSS challenge in terms of the degree of weight loss, DAI, histological score, and MPO activity. IFN-γ production was enhanced in the colon of DSS-treated WT mice, which exhibited severe IBD-like symptoms.

[0045] Interleukin-2 (IL-2) is produced by T cells and is essential for their differentiation into effector T cells. IL-2 is also important for T cell proliferation. This is important in IBD because effector T cells are thought to be the primary cell type responsible for causing damage in IBD.

[0046] IL-8 (interleukin 8, also known as CXCL8) primarily mediates neutrophil activation and migration from peripheral blood to tissues and to sites of inflammation. IL-8 tissue levels have been found to be higher in active UC compared to normal colonic tissue, and its serum concentration is associated with the endoscopic and histological severity of UC. IL-8 is important in the inflammatory environment and cancer (see, e.g., "The Chemokine CXCL8 in Carcinogenesis and Drug Response", ISRN Oncol. 2013 Oct 9;2013:859154; Gales D et al., and Future Oncol., 2010 Jan;6(1):111-6. doi: 10.2217 / fon.09.128; "CXCL8 and its cognate receptors in melanoma progression and metastasis", Singh S et al.). In cancer in particular, IL-8 is also thought to contribute by supporting angiogenesis.

[0047] In any of the configurations, embodiments, or examples herein, the antibody or fragment antagonizes the binding of hOX40L to the OX40 receptor.

[0048] In any configuration, embodiment, or example herein, the antibody or fragment antagonizes the binding of hOX40L to OX40.

[0049] In any configuration, aspect, or example herein, the OX40L receptor can be human OX40.

[0050] In any of the configurations, aspects, or examples herein, the human has or is at risk for asthma, and the antibody or fragment reduces IgE in the human.

[0051] In any of the configurations, aspects, or examples herein, the human has or is at risk of having asthma, and the antibody or fragment is for reducing IgE in the human.

[0052] 2. The antibody or fragment of embodiment 1, wherein the antibody or fragment reduces binding of hOX40 receptors expressed by human T cells to hOX40L expressed by endothelial cells and reduces proliferation of human T cells, and wherein the antibody or fragment is for treating or preventing an hOX40L-mediated disease or condition by reducing secretion of a cytokine selected from TNF-alpha, IL-2, IL-3, IL-4, IL-5, IL-6, IL-8, IL-9, IL-10, IL-13, IL-17, RANTES, and interferon gamma.

[0053] In one example, the cytokine is selected from (i) TNF-alpha, (ii) IL-2, and (iii) interferon gamma. In one example, the cytokine is TNF-alpha. In one example, the cytokine is IL-2. In one example, the cytokine is interferon gamma. In one example, the cytokines are (i) and (ii), or (i) and (iii), or (ii) and (iii), or (i) through (iii).

[0054] 3. The antibody or fragment of embodiment 1, wherein the leukocyte is selected from the group consisting of polymorphonuclear leukocytes, monocytes, peripheral blood mononuclear cells (PBMCs), lymphocytes, T cells, antigen-presenting cells (APCs), dendritic cells (DC cells), and natural killer cells (NK cells).

[0055] In one embodiment, the leukocytes are peripheral blood mononuclear cells (PBMCs) and T cells (eg, PBMCs).

[0056] 4. The antibody or fragment of embodiment 3, wherein the leukocytes comprise lamina propria lymphocytes (LPLs) and the disease or condition is a disease or condition of the gastrointestinal tract (GI tract).

[0057] 5. The antibody or fragment of any preceding aspect, wherein the epithelial cell comprises a cell selected from the group consisting of a gastrointestinal cell, a colon cell, an intestinal cell, and an airway (eg, lung) epithelial cell.

[0058] In another embodiment, the epithelial cells comprise cells selected from the group consisting of gastrointestinal cells, colonic cells, intestinal cells, ocular cells, and respiratory tract (e.g., lung) epithelial cells. In another embodiment, the epithelial cells comprise cells selected from the group consisting of gastrointestinal cells, colonic cells, intestinal cells, and ocular cells. In a further embodiment, the epithelial cells comprise ocular cells.

[0059] 6. The antibody or fragment of any preceding aspect for treating or preventing said hOX40L-mediated disease or condition in said human by reducing the proliferation of T cells in said human.

[0060] In one example, the antibody or fragment is capable of affecting a decrease in T cell proliferation in an in vitro assay (e.g., a human DC cell / T cell in vitro assay, such as that described further below), and therefore, administration of such an antibody or fragment to a human leads to a decrease in T cell proliferation in the human.

[0061] 7. The antibody or fragment of any preceding aspect for treating or preventing an hOX40L-mediated disease or condition in a human by antagonizing the interaction between hOX40L and human leukocytes, thereby reducing leukocyte proliferation.

[0062] In one example, the antibody or fragment is capable of affecting a decrease in the proliferation of white blood cells (e.g., mononuclear cells) in an in vitro assay (e.g., in an MLR in vitro assay, such as that described further below), and therefore, administration of such an antibody or fragment to a human leads to a decrease in the proliferation of white blood cells in the human.

[0063] 8. The antibody or fragment of any preceding aspect for treating or preventing said hOX40L-mediated disease or condition in said human by antagonizing OX40L / OX40L receptor interaction mediated by T cells in said human, thereby reducing proliferation of white blood cells in said human.

[0064] In one example, the antibody or fragment is capable of affecting a decrease in proliferation of leukocytes (e.g., monocytes) in an in vitro assay in which the antibody or fragment antagonizes T cell-mediated OX40L / OX40L receptor interaction in the assay, such that administration of such an antibody or fragment to a human leads to a decrease in proliferation of leukocytes in the human.

[0065] 9. The antibody or fragment of any preceding aspect for treating or preventing said hOX40L-mediated disease or condition in a human by reducing secretion in said human of a cytokine selected from TNF-alpha, IL-2, and interferon-gamma.

[0066] In one example, the antibody or fragment is for treating or preventing the hOX40L-mediated disease, condition, or epithelial cell damage in a human by reducing the secretion of (i) IL-2 and interferon gamma, (ii) IL-2 and TNF alpha, or (iii) interferon gamma and TNF alpha in the human.

[0067] In one example, the antibody or fragment is capable of affecting a decrease in the secretion of a cytokine selected from IL-2, TNF-alpha, and interferon-gamma in an in vitro assay (e.g., an MLR in vitro assay, such as that described further below), and thus administration of such an antibody or fragment to a human leads to a decrease in the secretion of the selected cytokine(s) in the human.

[0068] In one example, the antibody or fragment is capable of affecting a decrease in the secretion of IL-8 in an in vitro assay (e.g., in an MLR in vitro assay, such as that described further below), and therefore, administration of such an antibody or fragment to a human leads to a decrease in the secretion of IL-8 in the human.

[0069] 10. The antibody or fragment of embodiment 9 for treating or preventing said disease or condition in a human by reducing secretion of said cytokine mediated by dendritic cell (DC cell) interaction with T cells.

[0070] In one example, the antibody or fragment is capable of affecting a decrease in the secretion of said cytokine(s) in a DC cell / T cell in vitro assay (e.g., as further described below), and therefore, administration of such an antibody or fragment to a human leads to a decrease in the secretion of said cytokine(s) in said human.

[0071] 11. The antibody or fragment of any preceding aspect, wherein gastrointestinal cell, colon cell, enterocyte, or airway (e.g., lung) cell damage is a symptom or cause of the disease or condition in humans.

[0072] In another embodiment, the epithelial cells comprise cells selected from the group consisting of gastrointestinal cells, colonic cells, intestinal cells, ocular cells, and respiratory tract (e.g., lung) epithelial cells. In another embodiment, the epithelial cells comprise cells selected from the group consisting of gastrointestinal cells, colonic cells, intestinal cells, and ocular cells. In a further embodiment, the epithelial cells comprise ocular cells.

[0073] 12. The antibody or fragment of any preceding aspect, wherein the human is suffering from or at risk of inflammatory bowel disease (IBD), allograft rejection, graft-versus-host disease (GvHD), diabetes, or airway inflammation, and the method treats or prevents IBD, allograft rejection, GvHD, diabetes, or airway inflammation in the human.

[0074] 12a. The antibody or fragment of any of the preceding aspects, wherein the human is suffering from or at risk of inflammatory bowel disease (IBD), allograft rejection, graft-versus-host disease (GvHD), uveitis, pyoderma gangrenosum, giant cell arteritis, Schnitzler syndrome, non-infectious scleritis, diabetes, or airway inflammation, and the method treats or prevents IBD, allograft rejection, GvHD, uveitis, pyoderma gangrenosum, giant cell arteritis, Schnitzler syndrome, non-infectious scleritis, diabetes, or airway inflammation in a human.

[0075] In an example according to any of the preceding aspects, the human suffers from, is at risk for, or has been diagnosed with an inflammatory or autoimmune disease or condition.

[0076] In one example, the autoimmune disease or condition is selected from the following: Acute disseminated encephalomyelitis (ADEM) Addison's disease Allergic granulomatosis and vasculitis or Churg-Strauss syndrome (CSS) Alopecia or alopecia areata (AA) ankylosing spondylitis Autoimmune chronic active hepatitis (CAH) autoimmune hemolytic anemia Autoimmune pancreatitis (AIP) Autoimmune retinopathy (AR) (see Retinitis) Autoimmune thrombocytopenic purpura Autoimmune neutropenia Autoimmune inner ear disease (AIED) Antiphospholipid syndrome (APS) Autoimmune lymphoproliferative syndrome (ALPS) Behçet syndrome Bullous pemphigoid Celiac disease Churg-Strauss syndrome (CSS) or allergic granulomatous vasculitis Chronic bullous disease of childhood Chronic inflammatory demyelinating polyneuropathy (CIDP) Cicatricial pemphigoid (CP) Central nervous system vasculitis Crohn's disease Cryoglobulinemia Dermatitis herpetiformis (DH) Discoid lupus erythematosus (DLE) Encephalomyelitis Epidermolysis bullosa acquired (EBA) Giant cell arteritis (see Temporal Arteritis) Graft-versus-host disease Graves' disease Guillain-Barré syndrome Annaud's syndrome (see Primary Biliary Cirrhosis) Hashimoto's thyroiditis (also called autoimmune thyroiditis and chronic lymphocytic thyroiditis) Hypersensitivity vasculitis (HV) or small-vessel vasculitis Immune-Mediated Infertility inflammatory bowel disease Insulin-dependent diabetes Isolated central nervous system or CNS vasculitis Isaacs syndrome: neuromyotonia Kawasaki disease (KD) Lambert-Eaton Myasthenic Syndrome (LEMS) Linear IgA disease Lupus (see Systemic Lupus Erythematosus) Meniere's disease Microscopic polyangiitis (MPA) Mixed connective tissue disease or MCTD Monoclonal gammopathy myasthenia gravis Multiple sclerosis Multifocal motor neuropathy Neuromyotonia or Isaacs syndrome Neutropenia (see Autoimmune Neutropenia) oophoritis Opsoclonus-myoclonus syndrome Orchitis Neurological paraneoplastic disorders Pemphigus vulgaris Pemphigus foliaceus (PF) Pemphigoid gestationis (PG) Pernicious anemia Paraneoplastic pemphigus (PNP) Polyangiitis (see Microscopic Polyangiitis) Polyarteritis nodosa (PAN) Polymyositis / dermatomyositis Polymyalgia rheumatica Primary biliary cirrhosis (PBC) (also known as Annaud's syndrome) Primary sclerosing cholangitis (PSC) Raynaud's phenomenon Recoverin-associated retinitis (RAR) (see Retinitis) Reactive arthritis (formerly known as Reiter's syndrome) retinitis Rheumatoid arthritis (RA) sarcoidosis Sclerosing cholangitis (see Primary Sclerosing Cholangitis) Sjögren's syndrome Systemic necrotizing vasculitis Stiff-man syndrome or Mersch-Voltmann syndrome Systemic lupus erythematosus Systemic sclerosis (scleroderma) Temporal arteritis or giant cell arteritis (GCV) Takayasu arteritis Thromboangiitis obliterans or Buerger's disease Thyroiditis with hypothyroidism Thyroiditis with hyperthyroidism Polyglandular autoimmune syndrome type 1 (PAS) Polyglandular autoimmune syndrome type 2 vasculitis Wegener's granulomatosis.

[0077] In one example of any aspect, configuration, or embodiment, the human suffers from uveitis. For example, the uveitis may be non-infectious and / or autoimmune in nature, i.e., non-infectious uveitis, or autoimmune uveitis. For example, the non-infectious / autoimmune uveitis may be caused by and / or associated with Behçet's disease, Fuchs iridocyclitis, polyangiogranulomatosis, HLA-B27-associated uveitis, juvenile idiopathic arthritis, sarcoidosis, spondyloarthritis, sympathetic ophthalmia, tubulointerstitial nephritis, or uveitis syndrome. In one example, the uveitis is systemic in nature, i.e., systemic uveitis. For example, systemic uveitis can be caused by and / or associated with ankylosing spondylitis, Behçet's disease, chronic granulomatous disease, enthesitis, inflammatory bowel disease, juvenile rheumatoid arthritis, Kawasaki disease, multiple sclerosis, polyarteritis nodosa, psoriatic arthritis, reactive arthritis, sarcoidosis, systemic lupus erythematosus, Vogt-Koyanagi-Harada syndrome, or Whipple's disease.

[0078] In one example of any aspect, configuration, or embodiment, the human is suffering from pyoderma gangrenosum, giant cell arteritis, Schnitzler's syndrome, or non-infectious scleritis. In one example, the human is suffering from pyoderma gangrenosum. In one example, the human is suffering from giant cell arteritis. In one example, the human is suffering from Schnitzler's syndrome. In one example, the human is suffering from non-infectious scleritis.

[0079] In one example of any aspect, configuration, or embodiment, the human suffers from an hOX40L-mediated disease or condition selected from an autoimmune disease or condition, a systemic inflammatory disease or condition, or transplant rejection, e.g., inflammatory bowel disease (IBD), Crohn's disease, rheumatoid arthritis, transplant rejection, allograft rejection, graft-versus-host disease (GvHD), ulcerative colitis, systemic lupus erythematosus (SLE), diabetes, uveitis, ankylosing spondylitis, contact hypersensitivity, multiple sclerosis, and atherosclerosis, particularly GvHD. In another embodiment, the human suffers from or is at risk for multiple organ transplant rejection.

[0080] 13. An antibody or fragment thereof that specifically binds to hOX40L and competes for binding to said hOX40L with an antibody selected from the group consisting of 02D10, 10A07, 09H04, and 19H01.

[0081] In one example of any aspect, configuration, or embodiment, competition is determined by surface plasmon resonance (SPR), a technique that will be readily apparent to one of skill in the art. SPR can be performed using Biacore™, Proteon™, or another standard SPR technique. Such competition can be due, for example, to antibodies / fragments that bind to the same or overlapping epitopes of hOX40L. In one example of any aspect, configuration, or embodiment, competition is determined by ELISA, a technique that will be readily apparent to one of skill in the art. In one example of any aspect, configuration, or embodiment, competition is determined by homogeneous time-resolved fluorescence (HTRF), a technique that will be readily apparent to one of skill in the art. In one example of any aspect, configuration, or embodiment, competition is determined by fluorescence-activated cell sorting (FACS), a technique that will be readily apparent to one of skill in the art. In one aspect, HTRF, ELISA, and / or FACS methods are performed as described in the Examples below.

[0082] 14. The antibody or fragment of embodiment 13, wherein the antibody or fragment is according to any one of embodiments 1 to 12.

[0083] 15. The antibody or fragment of any preceding aspect, comprising a lambda light chain variable domain (optionally human).

[0084] In one example of any aspect, configuration, or embodiment of the invention, the variable domain of the antibody or fragment is human or humanized. Additionally, optionally, the antibody or fragment further comprises a human or humanized constant region (e.g., human Fc and / or human CL). In one example of any aspect of the invention, the variable domain of the antibody or fragment is produced by a transgenic animal (e.g., a rodent, mouse, rat, rabbit, chicken, sheep, camel, or shark). In one example of any aspect of the invention, the variable domain of the antibody or fragment is produced or identified by phage display, ribosome display, or yeast display.

[0085] In one example of any aspect, configuration, or embodiment of the invention, the antibody or fragment is recombinant.

[0086] In one example of any aspect, configuration, or embodiment of the invention, the antibody or fragment is produced by a recombinant mammalian, bacterial, insect, plant, or yeast cell. In one example, the mammalian cell is a CHO or HEK293 cell, and the antibody or fragment comprises CHO or HEK293 cell glycosylation.

[0087] In one example of any aspect, configuration or embodiment of the invention, the antibody or fragment is isolated.

[0088] 16. The antibody or fragment of any preceding embodiment, a.02D10 (the antibody or fragment competes with 02D10 for binding to the hOX40L), b. 10A07 (antibody or fragment competes with 10A07 for binding to the hOX40L); c. 09H04 (the antibody or fragment competes with 09H04 for binding to the hOX40L), and d. An antibody or fragment comprising a VH domain comprising an HCDR1 sequence selected from the group consisting of the HCDR1 of 19H01 (wherein the antibody or fragment competes with 19H01 for binding to said hOX40L).

[0089] 17. The antibody or fragment of any preceding embodiment, a.02D10 (the antibody or fragment competes with 02D10 for binding to the hOX40L), b. 10A07 (antibody or fragment competes with 10A07 for binding to the hOX40L); c. 09H04 (the antibody or fragment competes with 09H04 for binding to the hOX40L), and d. An antibody or fragment comprising a VH domain comprising an HCDR2 sequence selected from the group consisting of the HCDR2 of 19H01, wherein the antibody or fragment competes with 19H01 for binding to said hOX40L.

[0090] 18. The antibody or fragment of any preceding embodiment, a.02D10 (the antibody or fragment competes with 02D10 for binding to the hOX40L), b. 10A07 (antibody or fragment competes with 10A07 for binding to the hOX40L); c. 09H04 (the antibody or fragment competes with 09H04 for binding to the hOX40L), and d. An antibody or fragment comprising a VH domain comprising an HCDR3 sequence selected from the group consisting of the HCDR3 of 19H01, wherein the antibody or fragment competes with 19H01 for binding to said hOX40L.

[0091] 19. The antibody or fragment of any preceding aspect, comprising the sequences of (i) CDR1 and 2, (ii) CDR1 and 3, (iii) CDR2 and 3, or (iv) CDR1, 2, and 3: a. As recited in (a) of embodiments 16 to 18, wherein the antibody or fragment competes with 02D10 for binding to said hOX40L; b. As recited in (b) of embodiments 16-18, wherein the antibody or fragment competes with 10A07 for binding to said hOX40L; c. As recited in (c) of embodiments 16-18, wherein the antibody or fragment competes with 09H04 for binding to said hOX40L; or d. An antibody or fragment comprising a VH domain comprising one listed in (d) of any of embodiments 16 to 18, wherein the antibody or fragment competes with 19H01 for binding to said hOX40L.

[0092] 20. The antibody or fragment of any preceding aspect, comprising a VH domain comprising an amino acid sequence selected from the group consisting of the VH amino acid sequences in the Sequence Listing.

[0093] In one aspect, the invention provides an anti-hOX40L antibody or fragment (optionally according to other aspects recited herein) comprising a VH domain comprising an amino acid sequence selected from the group consisting of the VH amino acid sequences in the Sequence Listing. In one aspect, the VH domain comprises an amino acid sequence selected from SEQ ID NO:2, SEQ ID NO:34, SEQ ID NO:66, SEQ ID NO:94, SEQ ID NO:122, SEQ ID NO:124, SEQ ID NO:126, SEQ ID NO:128, SEQ ID NO:132, or SEQ ID NO:134.

[0094] In another example of the invention, the antibody or fragment comprises a VH domain amino acid sequence set out in the sequence listing below. Additionally or alternatively, the antibody or fragment comprises an HCDR1 domain amino acid sequence set out in the sequence listing below (i.e., SEQ ID NO:4, SEQ ID NO:10, SEQ ID NO:36, SEQ ID NO:42, SEQ ID NO:68, SEQ ID NO:74, SEQ ID NO:96, or SEQ ID NO:102, in particular SEQ ID NO:36 or SEQ ID NO:42). Additionally or alternatively, the antibody or fragment comprises an HCDR2 domain amino acid sequence set out in the sequence listing below (i.e., SEQ ID NO:6, SEQ ID NO:12, SEQ ID NO:38, SEQ ID NO:44, SEQ ID NO:70, SEQ ID NO:76, SEQ ID NO:98, or SEQ ID NO:104, in particular SEQ ID NO:38 or SEQ ID NO:44). Additionally or alternatively, the antibody or fragment comprises an HCDR3 domain amino acid sequence set out in the sequence listing below (i.e., SEQ ID NO:8, SEQ ID NO:14, SEQ ID NO:40, SEQ ID NO:46, SEQ ID NO:72, SEQ ID NO:78, SEQ ID NO:100, or SEQ ID NO:106, in particular SEQ ID NO:40 or SEQ ID NO:46).

[0095] In one example of the invention, the antibody or fragment comprises a VL domain amino acid sequence set forth in the sequence listing below. Additionally or alternatively, the antibody or fragment comprises an LCDR1 domain amino acid sequence set forth in the sequence listing below (i.e., SEQ ID NO:18, SEQ ID NO:24, SEQ ID NO:50, SEQ ID NO:56, SEQ ID NO:82, SEQ ID NO:88, SEQ ID NO:110, or SEQ ID NO:116, in particular SEQ ID NO:50 or SEQ ID NO:56). Additionally or alternatively, the antibody or fragment comprises an LCDR2 domain amino acid sequence set forth in the sequence listing below (i.e., SEQ ID NO:20, SEQ ID NO:26, SEQ ID NO:52, SEQ ID NO:58, SEQ ID NO:84, SEQ ID NO:90, SEQ ID NO:112, or SEQ ID NO:118, in particular SEQ ID NO:52 or SEQ ID NO:58). Additionally or alternatively, the antibody or fragment comprises an LCDR3 domain amino acid sequence set forth in the sequence listing below (i.e., SEQ ID NO:22, SEQ ID NO:28, SEQ ID NO:54, SEQ ID NO:60, SEQ ID NO:86, SEQ ID NO:92, SEQ ID NO:114, or SEQ ID NO:120, in particular SEQ ID NO:54 or SEQ ID NO:60).

[0096] In one example of any of the embodiments herein, the antibody or fragment comprises a heavy chain comprising a constant region selected from the group consisting of a heavy chain constant region SEQ ID NO: 126, 128, 132, or 134 in the sequence listing (i.e., any of SEQ ID NO: 126, 128, 132, or 134, particularly the constant region of SEQ ID NO: 128), and optionally a VH domain listed in embodiment 19 or 20. In one example, the antibody or fragment comprises two copies of such a heavy chain. In another example, the heavy chain comprises a rodent, rat, mouse, human, rabbit, chicken, camel, sheep, cow, non-human primate, or shark constant region (e.g., Fc), particularly a mouse constant region.

[0097] In one example of any aspect herein, the antibody or fragment comprises a heavy chain comprising a gamma (e.g., human gamma) constant region, e.g., a human gamma 1 constant region. In another example of any aspect herein, the antibody fragment comprises a human gamma 4 constant region. In another embodiment, the heavy chain constant region does not bind to an Fc-γ receptor and comprises, for example, a Leu235Glu mutation (i.e., a mutation of the wild-type leucine residue to a glutamic acid residue). In another embodiment, the heavy chain constant region comprises a Ser228Pro mutation to increase stability. In another embodiment, the heavy chain constant region is an IgG4 comprising both a Leu235Glu mutation and a Ser228Pro mutation. This heavy chain constant region is referred to herein as "IgG4-PE."

[0098] In one example of any of the embodiments herein, the antibody or fragment is chimeric, eg, it comprises a human variable domain and a non-human (e.g., rodent, such as murine, mouse, or rat) constant region.

[0099] 21. The antibody or fragment of any one of aspects 16-20, comprising a first and a second copy of said VH domain.

[0100] 22. The antibody or fragment of any preceding embodiment, a.02D10 (the antibody or fragment competes with 02D10 for binding to the hOX40L), b. 10A07 (antibody or fragment competes with 10A07 for binding to the hOX40L); c. 09H04 (the antibody or fragment competes with 09H04 for binding to the hOX40L), and d. An antibody or fragment comprising a VL domain comprising a LCDR1 sequence selected from the group consisting of the LCDR1 of 19H01 (wherein the antibody or fragment competes with 19H01 for binding to said hOX40L).

[0101] 23. The antibody or fragment of any preceding embodiment, a.02D10 (the antibody or fragment competes with 02D10 for binding to the hOX40L), b. 10A07 (antibody or fragment competes with 10A07 for binding to the hOX40L); c. 09H04 (the antibody or fragment competes with 09H04 for binding to the hOX40L), and d. An antibody or fragment comprising a VL domain comprising a LCDR2 sequence selected from the group consisting of the LCDR2 of 19H01 (which antibody or fragment competes with 19H01 for binding to said hOX40L).

[0102] 24. The antibody or fragment of any preceding embodiment, a.02D10 (the antibody or fragment competes with 02D10 for binding to the hOX40L), b. 10A07 (antibody or fragment competes with 10A07 for binding to the hOX40L); c. 09H04 (the antibody or fragment competes with 09H04 for binding to the hOX40L), and d. An antibody or fragment comprising a VL domain comprising a LCDR3 sequence selected from the group consisting of the LCDR3 of 19H01 (which antibody or fragment competes with 19H01 for binding to said hOX40L).

[0103] 25. The antibody or fragment of any preceding aspect, comprising the sequences of (i) CDR1 and 2, (ii) CDR1 and 3, (iii) CDR2 and 3, or (iv) CDR1, 2, and 3: a. As recited in (a) of embodiments 22 to 24, wherein the antibody or fragment competes with 02D10 for binding to said hOX40L; b. As recited in (b) of embodiments 22-24, wherein the antibody or fragment competes with 10A07 for binding to said hOX40L; c. As recited in (c) of embodiments 22-24, wherein the antibody or fragment competes with 09H04 for binding to said hOX40L; or d. An antibody or fragment comprising a VL domain comprising one listed in (d) of any of embodiments 22 to 24, wherein the antibody or fragment competes with 19H01 for binding to said hOX40L. 26. The antibody or fragment of any preceding aspect, comprising a VL domain comprising an amino acid sequence selected from the group consisting of the VL amino acid sequences in the Sequence Listing.

[0104] In one aspect of the present invention, an anti-hOX40L antibody or fragment (optionally in accordance with any other aspect herein) is provided, comprising a VL domain comprising an amino acid sequence selected from the group consisting of the VL amino acid sequences in the sequence listing (i.e., SEQ ID NO: 16, SEQ ID NO: 48, SEQ ID NO: 80, or SEQ ID NO: 108, particularly SEQ ID NO: 48).

[0105] In one example of any of the embodiments herein, the antibody or fragment comprises a light chain (e.g., a lambda light chain) comprising a constant region selected from the group consisting of the light chain constant region sequences in the Sequence Listing (i.e., SEQ ID NO:136, SEQ ID NO:138, SEQ ID NO:140, SEQ ID NO:142, SEQ ID NO:144, SEQ ID NO:146, SEQ ID NO:148, SEQ ID NO:152, SEQ ID NO:154, SEQ ID NO:156, SEQ ID NO:158, SEQ ID NO:160, SEQ ID NO:162, SEQ ID NO:164, or SEQ ID NO:166), and optionally a VL domain (e.g., a lambda VL) listed in embodiment 25 or 26. In one example, the antibody or fragment comprises two copies of such a light chain (and optionally two copies of the heavy chain described above). In another example, the light chain comprises a rodent, rat, mouse, human, rabbit, chicken, camel, sheep, bovine, non-human primate, or shark constant region.

[0106] In one example of any of the embodiments herein, the antibody or fragment comprises a light chain (e.g., a kappa light chain) comprising a constant region selected from the group consisting of the light chain constant region sequences in the Sequence Listing (i.e., SEQ ID NO:136, SEQ ID NO:138, SEQ ID NO:140, SEQ ID NO:142, SEQ ID NO:144, SEQ ID NO:146, SEQ ID NO:148, SEQ ID NO:152, SEQ ID NO:154, SEQ ID NO:156, SEQ ID NO:158, SEQ ID NO:160, SEQ ID NO:162, SEQ ID NO:164, or SEQ ID NO:166), and optionally a VL domain (e.g., a kappa VL) listed in embodiment 25 or 26. In one example, the antibody or fragment comprises two copies of such a light chain (and optionally two copies of the heavy chain described above). In another example, the light chain comprises a rodent, rat, mouse, human, rabbit, chicken, camel, sheep, bovine, non-human primate, or shark constant region.

[0107] In one example, the antibody or fragment comprises a lambda light chain comprising a constant region selected from the group consisting of a light chain constant region sequence in the sequence listing (i.e., SEQ ID NO:146, SEQ ID NO:148, SEQ ID NO:152, SEQ ID NO:154, SEQ ID NO:156, SEQ ID NO:158, SEQ ID NO:160, SEQ ID NO:162, SEQ ID NO:164, or SEQ ID NO:166), and optionally a lambda VL domain.

[0108] In one example, the antibody or fragment comprises a kappa light chain comprising a constant region selected from the group consisting of a light chain constant region sequence in the sequence listing (i.e., SEQ ID NO:136, SEQ ID NO:138, SEQ ID NO:140, SEQ ID NO:142, or SEQ ID NO:144), and optionally a kappa VL domain.

[0109] In one example, the VL domain of the antibody or fragment is a lambda light chain variable domain. In one example, the VL domain of the antibody or fragment is a kappa light chain variable domain.

[0110] 27. The antibody or fragment of any one of aspects 22-26, comprising a first and a second copy of said VL domain.

[0111] 28. The antibody or fragment of any preceding aspect, wherein the hOX40L is hOX40L surface-expressed on a human cell, such as on an endothelial cell (eg, an airway or gastrointestinal endothelial cell).

[0112] In another embodiment, the epithelial cells comprise cells selected from the group consisting of gastrointestinal cells, colonic cells, intestinal cells, ocular cells, and respiratory tract (e.g., lung) epithelial cells. In another embodiment, the epithelial cells comprise cells selected from the group consisting of gastrointestinal cells, colonic cells, intestinal cells, and ocular cells. In a further embodiment, the epithelial cells comprise ocular cells.

[0113] 29. The antibody or fragment of any preceding embodiment, wherein the antibody or fragment reduces the proliferation of human PBMCs or T cells in the presence of hOX40L in an in vitro mixed lymphocyte reaction (MLR) assay by at least 20, 30, 40, 50, or 60%, compared to the proliferation of human PBMCs or T cells in the presence of hOX40L in a control in vitro MLR assay in the absence of an antibody specific for hOX40L. A description of a suitable assay is provided in the Examples below.

[0114] 30. The antibody or fragment of embodiment 29, wherein the hOX40L in the assay is surface-expressed on human dendritic cells (DC cells).

[0115] A description of a suitable assay is provided in the Examples below.

[0116] 31. The antibody or fragment of any preceding embodiment, wherein the antibody or fragment reduces NF-κB activity in human HT-1080 cells expressing the hOX40 receptor in vitro in the presence of hOX40L.

[0117] In one example, antibody or fragment, reduction of NF-κB activity is determined by detecting a reduction in IL-8 secretion by HT-1080 cells (ATCC® CCL-121) in vitro (optionally transfected with the hOX40 receptor in the presence of hOX40).

[0118] 32. The antibody or fragment of any preceding aspect, which reduces IL-8 secretion from human HT-1080 cells expressing the hOX40 receptor in vitro in the presence of hOX40L.

[0119] 33. The antibody or fragment of embodiment 32, wherein the antibody or fragment reduces IL-8 secretion by at least 20, 30, 40, 50, or 60% compared to IL-8 production by HT-1080 cells expressing the hOX40 receptor in vitro in the presence of hOX40L in the absence of an antibody specific for hOX40L.

[0120] 34. The antibody or fragment of any preceding embodiment, wherein the antibody or fragment reduces hOX40L-stimulated human T cell proliferation in vitro.

[0121] 35. The antibody or fragment of any preceding embodiment, wherein the antibody or fragment reduces hOX40L-stimulated IL-2 secretion from human T cells in vitro.

[0122] 36. The antibody or fragment of any preceding embodiment, wherein the antibody or fragment reduces cytokine secretion mediated by human dendritic cell (DC cell) interaction with human T cells, and the cytokines are selected from one, two, more, or all of TNF-alpha, IL-2, IL-3, IL-4, IL-5, IL-6, IL-8, IL-9, IL-10, IL-13, IL-17, RANTES, and interferon gamma.

[0123] This can be assessed, for example, using an MLR in vitro assay (e.g., a DC / T cell MLR in vitro assay). A description of a suitable assay is provided in the Examples below.

[0124] In one example, the DC cells are mismatched, e.g., MHC-mismatched, with the T cells, as is possible when the DC cells are from a human source that is different from the human source of the T cells. In one example, the DC cells are produced by in vitro induction of human monocytes with human GMCSF and IL-4.

[0125] 37. The antibody or fragment of any preceding embodiment, wherein the antibody or fragment reduces interferon gamma secretion by at least 20, 30, 40, 50, or 60% compared to interferon gamma production mediated by human dendritic cell (DC cell) interaction with human T cells in the absence of an antibody specific for hOX40L.

[0126] 38. The antibody or fragment of any preceding embodiment, wherein the antibody or fragment reduces TNF-alpha secretion by at least 20, 30, 40, 50, or 60% compared to TNF-alpha production mediated by human dendritic cell (DC cell) interaction with human T cells in the absence of an antibody specific for hOX40L.

[0127] 39. The antibody or fragment of any preceding embodiment, wherein the antibody or fragment reduces IL-2 secretion by at least 10, 20, 30, 40, 50, or 60% compared to IL-2 production mediated by human dendritic cell (DC cell) interaction with human T cells in the absence of an antibody specific for hOX40L.

[0128] 40. The antibody or fragment of any preceding aspect, wherein the antibody or fragment reduces cytokine secretion (e.g., leukocyte cytokine secretion) in a human peripheral blood mononuclear cell (PBMC) mixed lymphocyte (MLR) assay, and the cytokine is selected from one, two, more, or all of TNF-alpha, IL-2, IL-4, IL-3, IL-6, IL-8, IL-10, IL-17, RANTES, and interferon gamma.

[0129] 41. The antibody or fragment of any preceding embodiment, wherein the antibody or fragment reduces interferon gamma secretion by at least 20, 30, 40, 50, or 60%, compared to interferon gamma production in a human PBMC MLR assay in the absence of an antibody that is specific for hOX40L.

[0130] In one embodiment, the comparison is made to interferon gamma production in a human PBMC MLR assay in the absence of antibody.

[0131] 42. The antibody or fragment of any preceding embodiment, wherein the antibody or fragment reduces TNF-alpha secretion by at least 20, 30, 40, 50, or 60% compared to TNF-alpha production in a human PBMC MLR assay in the absence of an antibody specific for hOX40L.

[0132] 43. The antibody or fragment of any preceding embodiment, wherein the antibody or fragment reduces IL-2 secretion by at least 10, 20, 30, 40, 50, or 60% compared to IL-2 production in a human PBMC MLR assay in the absence of an antibody specific for hOX40L.

[0133] 44. The antibody or fragment of any one of aspects 36-43, wherein the cell is a primary cell.

[0134] "Primary cells" refers to cells in a human or cells taken from a patient for in vitro binding to an antibody or fragment of the invention (e.g., which may be useful in methods of diagnosing OX40L status or disease / condition status in humans). Primary cells, as used herein, are typically not cells of a human cell line that have undergone extensive in vitro culturing. In this embodiment, the ability of an antibody or fragment of the invention to specifically inhibit hOX40L binding to its receptor is advantageous because it provides a direct indication of the availability of treating cells in a human patient suffering from or at risk for an hOX40L-mediated disease or condition.

[0135] 45. The antibody or fragment has a 1 x 10 -8 The following IC 50 The antibody or fragment of any preceding embodiment, wherein the antibody or fragment inhibits binding of hOX40L to an hOX40L receptor (eg, hOX40).

[0136] In one example, IC50 is 1 x 10 -8 ~1×10 -11 range, or 1×10 -9 ~1×10 -10 The range is.

[0137] 46. ​​A pharmaceutical composition for treating and / or preventing an OX40L-mediated condition or disease, comprising the antibody or fragment of any preceding aspect and a diluent, excipient, or carrier, and optionally further comprising an anti-inflammatory agent.

[0138] In one example, the anti-inflammatory agents are independently selected from the group consisting of corticosteroids (e.g., methylprednisolone), anti-IL12 / IL-23 antibodies (e.g., ustekinumab), anti-VLA4 antibodies (e.g., natalizumab), anti-LFA1 antibodies, anti-complement C5 antibodies (e.g., eculizumab), anti-a4b7 integrin antibodies (e.g., vedolizumab), anti-IL6 antibodies (e.g., tocilizumab), anti-IL2R antibodies (e.g., basilixumab), or anti-TNFα antibodies / TNFα-Fc molecules (e.g., etanercept, adalimumab, infliximab, golimumab, cetolizumab pegol). In one example, the anti-inflammatory agents are independently selected from the group consisting of corticosteroids (e.g., methylprednisolone) and anti-LFA1 antibodies.

[0139] 47. A pharmaceutical composition or kit for treating and / or preventing an OX40L-mediated condition or disease, comprising an antibody or fragment of the invention (and optionally an anti-inflammatory agent), optionally in combination with a label or instructions for use in treating and / or preventing the disease or condition in a human, optionally wherein the label or instructions includes a marketing authorization number (e.g., an FDA or EMA approval number), and optionally wherein the kit includes an IV or injection device containing the antibody or fragment.

[0140] 48. A nucleic acid encoding the HCDR3 of an antibody recited in any one of embodiments 1 to 45.

[0141] In one embodiment, the HCDRs herein follow the Kabat nomenclature, hi another embodiment, the HCDRs herein follow the IMGT nomenclature.

[0142] 49. The nucleic acid of embodiment 48, comprising a nucleotide sequence that is at least 80, 85, 90, 95, 96, 97, 98, or 99% identical, or 100% identical, to an HCDR3 sequence in the sequence listing.

[0143] In one aspect, the invention provides a nucleic acid comprising a nucleotide sequence encoding the VH domain of an anti-hOX40L antibody, wherein the nucleotide sequence comprises an HCDR3 sequence that is at least 80, 85, 90, 95, 96, 97, 98, or 99% identical, or 100% identical, to an HCDR3 sequence in the sequence listing. Optionally, the antibody is according to any other aspect herein.

[0144] In another embodiment, there is provided a nucleic acid of aspect 48, comprising a nucleotide sequence that is 100% identical to an HCDR3 sequence in the sequence listing, except for one, two, or three nucleotide substitutions, each of which produces no amino acid change or produces a conservative amino acid change in the corresponding protein sequence (i.e., the nucleotide substitutions are synonymous substitutions). Those skilled in the art will be familiar with conservative amino acid changes.

[0145] Amino acid substitutions include modifications in which an amino acid is replaced with a different naturally occurring amino acid residue. Such substitutions may be classified as "conservative," in which an amino acid residue contained in a polypeptide is replaced with another naturally occurring amino acid having similar characteristics in terms of either polarity, side chain functional group, or size. Such conservative substitutions are known in the art. Substitutions encompassed by the present invention may also be "non-conservative," in which an amino acid residue present in a peptide is replaced with an amino acid having different properties, such as a naturally occurring amino acid from a different group (e.g., replacing a charged or hydrophobic amino acid with alanine), or a naturally occurring amino acid is replaced with a non-conventional amino acid.

[0146] Additionally or alternatively, there is provided a nucleic acid of embodiment 49 comprising a nucleotide sequence that is 100% identical to an HCDR3 sequence in the sequence listing except for 1, 2, 3, 4, 5, 6, or 7 synonymous nucleotide substitutions and 0, 1, 2, or 3 nucleotide substitutions that produce conservative amino acid changes in the corresponding protein sequence.

[0147] 50. A nucleic acid encoding the HCDR2 of an antibody as recited in any one of embodiments 1 to 45, optionally wherein the nucleic acid is according to embodiment 48 or 49.

[0148] 51. The nucleic acid of embodiment 50, comprising a nucleotide sequence that is at least 80, 85, 90, 95, 96, 97, 98, or 99% identical, or 100% identical, to an HCDR2 sequence in the sequence listing.

[0149] In one aspect, the invention provides a nucleic acid comprising a nucleotide sequence encoding the VH domain of an anti-hOX40L antibody, wherein the nucleotide sequence comprises an HCDR2 sequence that is at least 80, 85, 90, 95, 96, 97, 98, or 99% identical, or 100% identical, to an HCDR2 sequence in the sequence listing. Optionally, the antibody is according to any other aspect herein.

[0150] In another embodiment, there is provided a nucleic acid of aspect 51, comprising a nucleotide sequence that is 100% identical to an HCDR2 sequence in the sequence listing, except for one, two, or three nucleotide substitutions, each of which produces no amino acid change or produces a conservative amino acid change in the corresponding protein sequence (i.e., the nucleotide substitutions are synonymous substitutions). Those skilled in the art will be familiar with conservative amino acid changes.

[0151] Additionally or alternatively, there is provided a nucleic acid of embodiment 50 comprising a nucleotide sequence that is 100% identical to an HCDR2 sequence in the sequence listing, except for 1, 2, 3, 4, 5, 6, or 7 synonymous nucleotide substitutions, and 0, 1, 2, or 3 nucleotide substitutions that produce conservative amino acid changes in the corresponding protein sequence.

[0152] 52. A nucleic acid encoding the HCDR1 of an antibody as recited in any one of embodiments 1 to 45, optionally wherein the nucleic acid is according to any one of embodiments 48 to 51.

[0153] 53. The nucleic acid of embodiment 52, comprising a nucleotide sequence that is at least 80, 85, 90, 95, 96, 97, 98, or 99% identical, or 100% identical, to an HCDR1 in the sequence listing.

[0154] In one aspect, the present invention provides a nucleic acid comprising a nucleotide sequence encoding the VH domain of an anti-hOX40L antibody, wherein the nucleotide sequence comprises an HCDR1 sequence that is at least 80, 85, 90, 95, 96, 97, 98, or 99% identical, or 100% identical, to an HCDR1 sequence in the sequence listing. Optionally, the antibody is according to any other aspect herein.

[0155] In another embodiment, there is provided a nucleic acid of aspect 52, comprising a nucleotide sequence that is 100% identical to the HCDR1 sequence in the sequence listing, except for one, two, or three nucleotide substitutions, each of which produces no amino acid change or produces a conservative amino acid change in the corresponding protein sequence (i.e., the nucleotide substitutions are synonymous substitutions). Those skilled in the art will be familiar with conservative amino acid changes.

[0156] Additionally or alternatively, there is provided a nucleic acid of embodiment 52, comprising a nucleotide sequence that is 100% identical to an HCDR1 sequence in the sequence listing, except for 1, 2, 3, 4, 5, 6, or 7 synonymous nucleotide substitutions, and 0, 1, 2, or 3 nucleotide substitutions that produce conservative amino acid changes in the corresponding protein sequence.

[0157] 54. A nucleic acid encoding the VH domain and / or the VL domain of an antibody recited in any one of aspects 1 to 45.

[0158] 55. The nucleic acid of embodiment 54, comprising a nucleotide sequence that is at least 80, 85, 90, 95, 96, 97, 98, or 99% identical, or 100% identical, to a VH domain nucleotide sequence in the sequence listing.

[0159] In another embodiment, a nucleic acid of aspect 54 is provided, comprising a nucleotide sequence that is 100% identical to a VH domain nucleotide sequence in the sequence listing, except for one, two, or three nucleotide substitutions, each of which produces no amino acid change or produces a conservative amino acid change in the corresponding protein sequence (i.e., the nucleotide substitutions are synonymous substitutions). Those of skill in the art will be familiar with conservative amino acid changes.

[0160] Additionally or alternatively, there is provided a nucleic acid of embodiment 54 comprising a nucleotide sequence that is 100% identical to a VH domain nucleotide sequence in the sequence listing, except for 1, 2, 3, 4, 5, 6, or 7 synonymous nucleotide substitutions and 0, 1, 2, or 3 nucleotide substitutions that produce conservative amino acid changes in the corresponding protein sequence.

[0161] 56. The nucleic acid of embodiments 54 to 55, comprising a nucleotide sequence that is at least 80, 85, 90, 95, 96, 97, 98, or 99% identical, or 100% identical, to a VL domain nucleotide sequence in the sequence listing.

[0162] In another embodiment, there is provided a nucleic acid of aspect 54 or 55, comprising a nucleotide sequence that is 100% identical to a VL domain nucleotide sequence in the sequence listing, except for one, two, or three nucleotide substitutions, each of which produces no amino acid change or produces a conservative amino acid change in the corresponding protein sequence (i.e., the nucleotide substitutions are synonymous substitutions). Those skilled in the art will be familiar with conservative amino acid changes.

[0163] Additionally or alternatively, there is provided a nucleic acid of embodiment 54 or 55, comprising a nucleotide sequence that is 100% identical to a VL domain nucleotide sequence in the sequence listing, except for 1, 2, 3, 4, 5, 6, or 7 synonymous nucleotide substitutions, and 0, 1, 2, or 3 nucleotide substitutions that produce conservative amino acid changes in the corresponding protein sequence.

[0164] 57. A nucleic acid encoding a heavy or light chain of an antibody according to any one of embodiments 1 to 45.

[0165] 58. The nucleic acid of embodiment 57, comprising a nucleotide sequence as recited in any one of embodiments 48 to 56.

[0166] 59. A vector (e.g., a mammalian expression vector) comprising the nucleic acid of any one of embodiments 48-58, optionally wherein the vector is a CHO or HEK293 vector. In one example, the vector is a yeast vector, e.g., a Saccharomyces or Pichia vector.

[0167] 60. A host comprising a nucleic acid of any one of aspects 48 to 58, or a vector of aspect 59. In one example, the host is a mammalian (e.g., human, e.g., CHO or HEK293) cell line, or a yeast or bacterial cell line.

[0168] 61. a. secretion of a cytokine selected from TNF-alpha, IL-2, IL-3, IL-4, IL-5, IL-6, IL-8, IL-9, IL-10, IL-13, IL-17, RANTES, and interferon gamma in humans; b. proliferation of human white blood cells, and c. Use of an antibody or fragment thereof that specifically binds to hOX40L in the manufacture of a medicament for administration to a human to treat or prevent an hOX40L-mediated disease or condition in the human by reducing one, more than one, or all of the binding of hOX40L expressed by endothelial cells to a hOX40 receptor expressed by human T cells.

[0169] Optionally, features of any of the preceding aspects, configurations, examples, or embodiments apply mutatis mutandis to this use.

[0170] In one example, the human has or is at risk of having asthma, and the antibody or fragment is for treating, preventing, or reducing asthma in the human by reducing IgE in the human.

[0171] 62. a. secretion of a cytokine selected from TNF-alpha, IL-2, IL-3, IL-4, IL-5, IL-6, IL-8, IL-9, IL-10, IL-13, IL-17, RANTES, and interferon gamma in humans; b. proliferation of human white blood cells, and c. A method of treating or preventing a hOX40L-mediated disease or condition in a human by reducing one, more than one, or all of the following: binding of hOX40L expressed by endothelial cells to a hOX40 receptor expressed by human T cells; The method comprises administering to the human a therapeutically effective amount of an antibody or fragment that specifically binds to hOX40L.

[0172] The features of any of the preceding aspects, examples or embodiments apply mutatis mutandis to this method.

[0173] The methods of the invention treat or prevent the disease or condition in a human. A "therapeutically effective amount" of an antibody or fragment is that amount (administered once or in several doses, which may be spaced apart over time, e.g., substantially monthly) effective to effect the treatment or prevention. This will be readily apparent to one of skill in the art and may vary according to the particular human patient and the disease or condition being addressed.

[0174] In one example, the human has or is at risk for asthma, and the antibody or fragment treats, prevents, or reduces asthma in the human by reducing IgE in the human.

[0175] 63. The method or use of embodiments 61-62 for treating or preventing said hOX40L-mediated disease, condition, or epithelial cell damage in a human by reducing proliferation of T cells in said human.

[0176] 64. The method or use of any one of aspects 61 to 63 for treating or preventing an hOX40L-mediated disease, condition, or epithelial cell injury in a human by antagonizing the interaction between hOX40L and human leukocytes, thereby reducing leukocyte proliferation.

[0177] 65. The method or use of any one of aspects 61-64 for treating or preventing said hOX40L-mediated disease, condition, or epithelial cell damage in a human by antagonizing OX40L / OX40L receptor interaction mediated by T cells in said human, thereby reducing the proliferation of human leukocytes.

[0178] 66. The method or use of any one of aspects 61-65 for treating or preventing said hOX40L-mediated disease, condition, or epithelial cell damage in a human by reducing the secretion of IL-8 cytokine in said human.

[0179] 67. The method of embodiment 66 for treating or preventing the disease, condition, or epithelial cell damage in a human by reducing the IL-8 mediated by dendritic cell (DC) interaction with T cells.

[0180] 68. The method or use of any one of aspects 61-67, wherein gastrointestinal cell, colon cell, enterocyte, or airway (e.g., lung) cell damage is a symptom or cause of the disease or condition in a human.

[0181] In another embodiment, the epithelial cells comprise cells selected from the group consisting of gastrointestinal cells, colonic cells, intestinal cells, ocular cells, and respiratory tract (e.g., lung) epithelial cells. In another embodiment, the epithelial cells comprise cells selected from the group consisting of gastrointestinal cells, colonic cells, intestinal cells, and ocular cells. In a further embodiment, the epithelial cells comprise ocular cells.

[0182] 69. The method of any one of embodiments 61-68, wherein the human is suffering from or at risk of inflammatory bowel disease (IBD), allograft rejection, graft-versus-host disease (GvHD), diabetes, or airway inflammation, and the method treats or prevents IBD, allograft rejection, GvHD, diabetes, or airway inflammation in the human.

[0183] 69a. The method or use of any one of aspects 61-68, wherein the human is suffering from or at risk of inflammatory bowel disease (IBD), allograft rejection, graft-versus-host disease (GvHD), uveitis, pyoderma gangrenosum, giant cell arteritis, Schnitzler syndrome, non-infectious scleritis, diabetes, or airway inflammation, and the method treats or prevents IBD, allograft rejection, GvHD, uveitis, pyoderma gangrenosum, giant cell arteritis, Schnitzler syndrome, non-infectious scleritis, diabetes, or airway inflammation in the human.

[0184] In any aspect, configuration, or embodiment, the human has or is at risk for an hOX40L-mediated disease or condition selected from an autoimmune disease or condition, a systemic inflammatory disease or condition, or transplant rejection, e.g., inflammatory bowel disease (IBD), Crohn's disease, rheumatoid arthritis, transplant rejection, allograft rejection, graft-versus-host disease (GvHD), ulcerative colitis, systemic lupus erythematosus (SLE), diabetes, uveitis, ankylosing spondylitis, contact hypersensitivity, multiple sclerosis, and atherosclerosis, particularly GvHD.

[0185] 70. The method or use of any one of aspects 61 to 69a, wherein the antibody or fragment is according to any one of aspects 1 to 45, or any example, configuration, aspect, or embodiment described herein.

[0186] 71. The antibody, fragment, composition, kit, method, or use of any preceding aspect for treating or preventing an inflammatory or autoimmune disease or condition in a human, or for reducing or preventing angiogenesis in a human.

[0187] 72. The antibody, fragment, composition, kit, method, or use of any preceding aspect, wherein the disease or condition is selected from the group consisting of inflammatory bowel disease (IBD), Crohn's disease, rheumatoid arthritis, psoriasis, bronchiolitis, gingivitis, graft rejection, allograft rejection, graft versus host disease (GvHD), asthma, adult respiratory distress syndrome (ARDS), septic shock, ulcerative colitis, Sjogren's syndrome, airway inflammation, systemic lupus erythematosus (SLE), diabetes, contact hypersensitivity, multiple sclerosis, and atherosclerosis.

[0188] 72a. The antibody, fragment, composition, kit, method, or use of any preceding aspect, wherein the disease or condition is selected from the group consisting of inflammatory bowel disease (IBD), Crohn's disease, rheumatoid arthritis, psoriasis, bronchiolitis, gingivitis, graft rejection, allograft rejection, graft versus host disease (GvHD), asthma, adult respiratory distress syndrome (ARDS), septic shock, ulcerative colitis, Sjogren's syndrome, airway inflammation, systemic lupus erythematosus (SLE), uveitis, pyoderma gangrenosum, giant cell arteritis, Schnitzler syndrome, non-infectious scleritis, diabetes, contact hypersensitivity, multiple sclerosis, and atherosclerosis.

[0189] In any aspect, configuration, or embodiment, the human has or is at risk for an hOX40L-mediated disease or condition selected from an autoimmune disease or condition, a systemic inflammatory disease or condition, or transplant rejection, e.g., inflammatory bowel disease (IBD), Crohn's disease, rheumatoid arthritis, transplant rejection, allograft rejection, graft-versus-host disease (GvHD), ulcerative colitis, systemic lupus erythematosus (SLE), diabetes, uveitis, ankylosing spondylitis, contact hypersensitivity, multiple sclerosis, and atherosclerosis, particularly GvHD.

[0190] In one example, the disease or condition is an OX40L-mediated disease or condition disclosed in US Pat. No. 7,812,133 or EP Pat. No. 1,791,869.

[0191] In one example, the disease or condition is an inflammatory or autoimmune disease or condition, hi one example, the disease or condition is transplant rejection.

[0192] As used herein, inflammatory disease or condition refers to a pathological condition that causes inflammation, for example, caused by neutrophil chemotaxis.Examples of such disorders include inflammatory skin diseases, including psoriasis; inflammatory bowel disease-related responses (such as Crohn's disease and ulcerative colitis); ischemia-reperfusion; adult respiratory distress syndrome; dermatitis; meningitis; encephalitis; uveitis; autoimmune diseases, such as rheumatoid arthritis, Sjögren's syndrome, and vasculitis; diseases associated with leukocyte leakage; central nervous system (CNS) inflammatory disorders, multiple organ injury syndrome secondary to sepsis or trauma; alcoholic hepatitis, bacterial pneumonia, antigen-antibody complex-mediated disease; pulmonary inflammation, including pleuritis, alveolitis, vasculitis, pneumonia, chronic bronchitis, bronchiectasis, and cystic fibrosis.Preferred indications include bacterial pneumonia and inflammatory bowel diseases, such as ulcerative colitis. The present invention is therefore in its various forms provided for treating or preventing any one or more of such conditions.

[0193] In one example, the disease or condition is cancer.

[0194] In one example, the disease is uveitis, such as systemic uveitis or autoimmune / non-infectious uveitis.

[0195] 73. An antibody or fragment thereof that specifically binds to hOX40L and competes with antibody 02D10 for binding to said hOX40L, wherein the antibody or fragment thereof comprises a VH domain comprising an HCDR3 that comprises the motif VRGXYYY, where X is any amino acid.

[0196] The antibody characteristics of any of the aspects, configurations, examples, or embodiments described herein optionally apply mutatis mutandis to these antibodies, e.g., the antibodies may be human or chimeric antibodies with functional characteristics described herein. Competition may be determined, e.g., by SPR, ELISA, HTRF, or FACS, as described in any of the aspects, embodiments, examples, or configurations described herein.

[0197] In one embodiment, the antibody or fragment competes with the variable region of 02D10 (e.g., competes with an antibody comprising the heavy chain variable region of SEQ ID NO: 34 and the light chain variable region of SEQ ID NO: 48). In another embodiment, the antibody or fragment competes with 02D10 IgG4-PE having a heavy chain amino acid sequence of SEQ ID NO: 62 and a light chain amino acid sequence of SEQ ID NO: 64. Thus, for example, the ability of an antibody or fragment to compete with antibody 02D10 for binding to hOX40L can be determined by SPR (as described herein) using an IgG4-PE antibody having the heavy chain amino acid sequence of SEQ ID NO: 62 and the light chain amino acid sequence of SEQ ID NO: 64 as the reference 02D10 antibody.

[0198] In another embodiment, the antibody or fragment additionally or alternatively competes with 10A7. In one embodiment, the antibody or fragment competes with the variable region of 10A7 (e.g., competes with an antibody comprising the heavy chain variable region of SEQ ID NO: 2 and the light chain variable region of SEQ ID NO: 16). In another embodiment, the antibody or fragment competes with 02D10 IgG4-PE, which has a heavy chain amino acid sequence of SEQ ID NO: 30 and a light chain amino acid sequence of SEQ ID NO: 32.

[0199] In one embodiment, the amino acid is any naturally occurring amino acid.

[0200] 74. The antibody or fragment according to embodiment 73, wherein X is a neutral amino acid, optionally P or G.

[0201] In one embodiment, X is P or G. In one embodiment, X is selected from P, N, A, or G. In another embodiment, X is selected from P, G, or N. In another embodiment, X is selected from P, G, or A.

[0202] 75. An antibody or fragment thereof optionally according to claim 1 or 2 that specifically binds to hOX40L and competes with antibody 02D10 for binding to said hOX40L, wherein the antibody or fragment comprises a VH domain comprising the HCDR3 sequence of SEQ ID NO: 40 or 46, or the HCDR3 sequence of SEQ ID NO: 40 or 46 with fewer than 5 amino acid substitutions.

[0203] The antibody characteristics of any of the aspects, configurations, examples, or embodiments described herein optionally apply mutatis mutandis to these antibodies, e.g., the antibodies may be human or chimeric antibodies with functional characteristics described herein. Competition may be determined, e.g., by SPR, ELISA, HTRF, or FACS, as described in any of the aspects, embodiments, examples, or configurations described herein.

[0204] In one embodiment, the HCDR3 sequence of SEQ ID NO: 40 or 46 comprises fewer than four amino acid substitutions (i.e., three or fewer). In one embodiment, the HCDR3 sequence of SEQ ID NO: 40 or 46 comprises fewer than three amino acid substitutions (i.e., two or one substitution). In one embodiment, the HCDR3 sequence of SEQ ID NO: 40 or 46 comprises fewer than two amino acid substitutions (i.e., one substitution).

[0205] In one embodiment, the antibody or fragment competes with the variable region of 02D10 (e.g., competes with an antibody comprising the heavy chain variable region of SEQ ID NO: 34 and the light chain variable region of SEQ ID NO: 48). In another embodiment, the antibody or fragment competes with 02D10 IgG4-PE, which has a heavy chain amino acid sequence of SEQ ID NO: 62 and a light chain amino acid sequence of SEQ ID NO: 64.

[0206] In another embodiment, the antibody or fragment additionally or alternatively competes with 10A7. In one embodiment, the antibody or fragment competes with the variable region of 10A7 (e.g., competes with an antibody comprising the heavy chain variable region of SEQ ID NO: 2 and the light chain variable region of SEQ ID NO: 16). In another embodiment, the antibody or fragment competes with 02D10 IgG4-PE, which has a heavy chain amino acid sequence of SEQ ID NO: 30 and a light chain amino acid sequence of SEQ ID NO: 32.

[0207] 76. The antibody or fragment of any one of aspects 73 to 75, wherein the VH domain comprises an HCDR3 of 16 to 27 amino acids and is derived from a recombination of a human VH gene segment, a human D gene segment, and a human JH gene segment, wherein the human JH gene segment is IGHJ6 (e.g., IGHJ6*02).

[0208] In one embodiment, the human JH gene segment is selected from IGHJ6*01, IGHJ6*02, IGHJ6*03, and IGHJ6*04. In another embodiment, the human JH gene segment is selected from IGHJ6*01, IGHJ6*02, and IGHJ6*04. In another embodiment, the JH gene segment is IGHJ6*02.

[0209] In a further embodiment, the human VH gene segment is IGHV3-23, e.g., selected from IGHV3-23*01, IGHV3-23*02, IGHV3-23*03, IGHV3-23*04, or IGHV3-23*05. In another embodiment, the human VH gene segment is IGHV3-23*01 or IGHV3-23*04, particularly IGHV3-23*04.

[0210] In a further embodiment, the human DH gene segment is IGHD3-10, e.g., selected from IGHD3-10*01 or IGHD3-10*02. In one embodiment, the human DH gene segment is IGHD3-10*01. In one embodiment, the human DH gene segment is IGHD3-10*02.

[0211] 77. The antibody or fragment according to any one of aspects 73 to 76, wherein the VH domain comprises the HCDR1 sequence of SEQ ID NO: 36 or 42, or the HCDR1 sequence of SEQ ID NO: 36 or 42 comprising fewer than 4 amino acid substitutions.

[0212] In one embodiment, the HCDR1 sequence of SEQ ID NO: 36 or 42 comprises fewer than three amino acid substitutions (i.e., two or one substitution). In one embodiment, the HCDR1 sequence of SEQ ID NO: 36 or 42 comprises fewer than two amino acid substitutions (i.e., one substitution).

[0213] 78. The antibody or fragment according to any one of aspects 73 to 77, wherein the VH domain comprises the HCDR2 sequence of SEQ ID NO: 38 or 44, or the HCDR2 sequence of SEQ ID NO: 38 or 44 comprising fewer than 5 amino acid substitutions.

[0214] In one embodiment, the HCDR2 sequence of SEQ ID NO: 38 or 44 comprises fewer than four amino acid substitutions (i.e., three or fewer). In one embodiment, the HCDR2 sequence of SEQ ID NO: 38 or 44 comprises fewer than three amino acid substitutions (i.e., two or one substitution). In one embodiment, the HCDR2 sequence of SEQ ID NO: 38 or 44 comprises fewer than two amino acid substitutions (i.e., one substitution).

[0215] 79. The antibody or fragment according to any one of aspects 73 to 78, wherein the VH domain comprises the amino acid sequence of SEQ ID NO: 34, or a heavy chain variable domain amino acid sequence which is at least 80% (e.g. at least 85%) identical to SEQ ID NO: 34.

[0216] In one embodiment, the heavy chain variable domain amino acid sequence is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO:34.

[0217] 80. The antibody or fragment of any one of aspects 73-79, comprising a first and a second copy of said VH domain.

[0218] 81. The antibody or fragment of any one of aspects 73-80, comprising a VL domain comprising the LCDR1 sequence of SEQ ID NO: 54 or 60, or the LCRD3 sequence of SEQ ID NO: 54 or 60 comprising fewer than 5 amino acid substitutions.

[0219] In one embodiment, the LCRD3 sequence of SEQ ID NO: 54 or 60 contains fewer than four amino acid substitutions (i.e., three or fewer). In one embodiment, the LCRD3 sequence of SEQ ID NO: 54 or 60 contains fewer than three amino acid substitutions (i.e., two or one substitution). In one embodiment, the LCRD3 sequence of SEQ ID NO: 54 or 60 contains fewer than two amino acid substitutions (i.e., one substitution).

[0220] 82. The antibody or fragment according to any one of aspects 73 to 81, wherein the antibody or fragment comprises a VL domain or said VL domain, wherein the VL domain comprises the LCDR2 sequence of SEQ ID NO: 52 or 58, or the LCRD2 sequence of SEQ ID NO: 52 or 58 comprising fewer than two amino acid substitutions.

[0221] 83. The antibody or fragment according to any one of aspects 73 to 82, wherein the antibody or fragment comprises a VL domain or said VL domain, wherein the VL domain comprises the LCDR1 sequence of SEQ ID NO: 54 or 60, or the LCRD1 sequence of SEQ ID NO: 54 or 60 comprising fewer than 4 amino acid substitutions.

[0222] In one embodiment, the LCRD1 sequence of SEQ ID NO: 54 or 60 contains fewer than three amino acid substitutions (i.e., two or one substitution). In one embodiment, the LCDR1 sequence of SEQ ID NO: 54 or 60 contains fewer than two amino acid substitutions (i.e., one substitution).

[0223] 84. The antibody or fragment according to any one of aspects 73 to 83, wherein the antibody or fragment comprises a VL domain or a VL domain, wherein the VL domain comprises the amino acid sequence of SEQ ID NO: 48, or a light chain variable domain amino acid sequence that is at least 80% (e.g., at least 85%) identical to SEQ ID NO: 48.

[0224] In one embodiment, the light chain variable domain amino acid sequence is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO:48.

[0225] 85. The antibody or fragment according to any one of aspects 81 to 84, comprising a first and a second copy of said VL domain.

[0226] 86. The antibody or fragment according to any one of aspects 81-85, wherein the antibody or fragment comprises a kappa light chain.

[0227] In another embodiment, the VL domain is a kappa VL domain. In one embodiment, the kappa VL domain is derived from the recombination of a human VL gene segment and a human JL gene segment, and the human VL gene segment is IGKV1D-39. In another embodiment, the VL gene segment is IGKV1D-39*01.

[0228] In a further embodiment, the human JL gene segment is IGKJ1 or IGKJ3. In another embodiment, the JL gene segment is IGKJ1*01. In another embodiment, the JL gene segment is IGKJ3*01.

[0229] 87. The amino acid substitution is a conservative amino acid substitution, and optionally the conservative substitution is: 1) Alanine (A), serine (S), threonine (T), 2) Aspartic acid (D), glutamic acid (E), 3) Asparagine (N), Glutamine (Q), 4) Arginine (R), Lysine (K), 5) Isoleucine (I), Leucine (L), Methionine (M), Valine (V), and 6) The antibody or fragment of any one of aspects 75 to 86, wherein the amino acid is from one of six groups selected from phenylalanine (F), tyrosine (Y), and tryptophan (W), each group containing amino acids that are conservative substitutions for one another.

[0230] In one embodiment, conservative amino acid substitutions are as described herein. For example, substitutions may be: Y for F, T for S or K, P for A, E for D or Q, N for D or G, R for K, G for N or A, T for S or K, D for N or E, I for L or V, F for Y, S for T or A, R for K, G for N or A, K for R, A for S, K, or P. In another embodiment, conservative amino acid substitutions may be Y replaced by F, T replaced by A or S, I replaced by L or V, W replaced by Y, M replaced by L, N replaced by D, G replaced by A, T replaced by A or S, D replaced by N, I replaced by L or V, F replaced by Y or L, S replaced by A or T, and A replaced by S, G, T, or V.

[0231] 88. The antibody or fragment according to any one of aspects 73-87, wherein the antibody or fragment comprises a constant region, such as an IgG4 constant region, and optionally the constant region is IgG4-PE (SEQ ID NO: 128).

[0232] In another example of any aspect herein, the antibody fragment comprises a human gamma 4 constant region. In another embodiment, the heavy chain constant region does not bind to an Fc-γ receptor and comprises, for example, a Leu235Glu mutation (i.e., a mutation of the wild-type leucine residue to a glutamic acid residue). In another embodiment, the heavy chain constant region comprises a Ser228Pro mutation to increase stability.

[0233] 89. The antibody of any one of aspects 73 to 78, wherein the antibody comprises a heavy chain and a light chain, wherein the heavy chain amino acid sequence consists of the sequence of SEQ ID NO: 62 and the light chain amino acid sequence consists of the sequence of SEQ ID NO: 64.

[0234] 90. The antibody or fragment as defined in any one of aspects 73 to 89, 98, 99, 101, or 102 for use in the treatment or prevention of an hOX40L-mediated disease or condition selected from an autoimmune disease or condition, a systemic inflammatory disease or condition, or transplant rejection, such as inflammatory bowel disease (IBD), Crohn's disease, rheumatoid arthritis, transplant rejection, allograft rejection, graft-versus-host disease (GvHD), ulcerative colitis, systemic lupus erythematosus (SLE), diabetes, uveitis, ankylosing spondylitis, contact hypersensitivity, multiple sclerosis, or atherosclerosis, in particular GvHD.

[0235] The antibody features and hOX40L-mediated diseases of any of the aspects, configurations, examples, or embodiments described herein optionally apply mutatis mutandis to this use. Any compositions, dosing schedules, or modes of administration described in any of the aspects, configurations, examples, or embodiments herein apply mutatis mutandis to this use.

[0236] 91. Use of the antibody or fragment as defined in any one of aspects 73 to 89, 98, 99, 101, or 102 in the manufacture of a medicament for administration to a human to treat or prevent a hOX40L-mediated disease or condition in a human selected from an autoimmune disease or condition, a systemic inflammatory disease or condition, or graft / host rejection, such as inflammatory bowel disease (IBD), Crohn's disease, rheumatoid arthritis, graft rejection, allograft rejection, graft-versus-host disease (GvHD), ulcerative colitis, systemic lupus erythematosus (SLE), diabetes, uveitis, ankylosing spondylitis, contact hypersensitivity, multiple sclerosis, or atherosclerosis, in particular GvHD.

[0237] The antibody features and hOX40L-mediated diseases of any of the aspects, configurations, examples, or embodiments described herein optionally apply mutatis mutandis to this use. Any compositions, dosing schedules, or modes of administration described in any of the aspects, configurations, examples, or embodiments herein apply mutatis mutandis to this use.

[0238] 92. A method for treating or preventing a hOX40L-mediated disease or condition selected from an autoimmune disease or condition, a systemic inflammatory disease or condition, or transplant rejection, e.g., inflammatory bowel disease (IBD), Crohn's disease, rheumatoid arthritis, transplant rejection, allograft rejection, graft-versus-host disease (GvHD), ulcerative colitis, systemic lupus erythematosus (SLE), diabetes, uveitis, ankylosing spondylitis, contact hypersensitivity, multiple sclerosis, or atherosclerosis, particularly GvHD in a human, comprising administering to said human a therapeutically effective amount of the antibody or fragment defined in any one of aspects 73 to 89, 98, 99, 101, or 102, whereby the hOX40L-mediated disease or condition is treated or prevented.

[0239] The antibody features and hOX40L-mediated diseases of any of the aspects, configurations, examples, or embodiments described herein optionally apply mutatis mutandis to this method. Any of the compositions, dosing schedules, or modes of administration described in any of the aspects, configurations, examples, or embodiments herein apply mutatis mutandis to this method.

[0240] 93. The antibody or fragment according to aspect 90, the use according to aspect 91, or the method according to claim 92, wherein the hOX40L-mediated disease or condition is GvHD.

[0241] In another embodiment, the antibody or fragment is capable of treating or preventing GvHD.

[0242] 94. The antibody or fragment, use, or method of any one of aspects 90-93, wherein the antibody is administered prophylactically.

[0243] In one embodiment, prevention prevents the onset of a disease or condition, or symptoms of a disease or condition. In one embodiment, prophylactic treatment prevents the worsening or onset of a disease or condition. In one embodiment, prophylactic treatment prevents the worsening of a disease or condition.

[0244] In another embodiment, the antibody is administered intravenously. In another embodiment, the antibody is administered at a dose of about 5-10 mg / kg (e.g., about 8 mg / kg). In another embodiment, the antibody is administered at a dose selected from about 0.1 mg / kg, about 0.5 mg / kg, about 1 mg / kg, 3 mg / kg, 5 mg / kg, about 10 mg / kg, about 15 mg / kg, about 20 mg / kg, about 25 mg / kg, about 30 mg / kg, about 40 mg / kg, about 50 mg / kg, about 60 mg / kg, about 70 mg / kg, about 80 mg / kg, about 90 mg / kg, or about 100 mg / kg, particularly about 1 mg / kg or about 3 mg / kg.

[0245] In another embodiment, the antibody is administered 1-4 days prior to transplant, e.g., 1-3 days prior to transplant or 1-2 days prior to transplant. In another embodiment, the antibody is administered weekly, biweekly, or monthly, e.g., every other week, after transplant. In a further embodiment, the antibody is administered prophylactically intravenously at a dose of about 5-10 mg / kg (e.g., about 8 mg / kg) 1-3 days prior to transplant, and then intravenously at a dose of about 5-10 mg / kg (e.g., about 8 mg / kg) every other week thereafter.

[0246] In another embodiment, patients are monitored periodically after transplantation for the presence of biomarkers predictive of the development of GvHD (e.g., acute GvHD), and when the biomarker levels reach a level that determines the patient is at risk for developing GvHD (e.g., acute GvHD), an anti-OX40L antibody of the invention is administered. This strategy may avoid unnecessary administration of drugs and unnecessary suppression of the immune system. Examples of biomarkers that may be useful as predictive biomarkers for acute GvHD may be those identified in Levine et al., "A prognostic score for acute graft-versus-host disease based on biomarkers: a multicentre study," Lancet Haematol 2015;2:e21-29. These biomarkers include, but are not limited to, TNFR1, ST-2, elafin, and IL2Rα and Reg3α. 95. A human antibody or fragment thereof comprising an HCDR3 of 16 to 27 amino acids and derived by recombination of a human VH gene segment, a human D gene segment, and a human JH gene segment, wherein the human JH gene segment is IGHJ6 (e.g., IGHJ6*02), which specifically binds to hOX40L for treating or preventing an hOX40L-mediated disease or condition selected from an autoimmune disease or condition, a systemic inflammatory disease or condition, or a transplant rejection, e.g., inflammatory bowel disease (IBD), Crohn's disease, rheumatoid arthritis, transplant rejection, allograft rejection, graft-versus-host disease (GvHD), ulcerative colitis, systemic lupus erythematosus (SLE), diabetes, uveitis, ankylosing spondylitis, contact hypersensitivity, multiple sclerosis, or atherosclerosis, particularly GvHD (e.g., the antibody is for preventing GvHD).

[0247] The antibody features and hOX40L-mediated diseases of any of the aspects, configurations, examples, or embodiments optionally apply mutatis mutandis to this use. Any compositions, dosing schedules, or modes of administration described in any of the aspects, configurations, examples, or embodiments herein apply mutatis mutandis to this use.

[0248] 96. Use of a human antibody or fragment thereof comprising an HCDR3 of 16 to 27 amino acids and derived by recombination of a human VH gene segment, a human D gene segment, and a human JH gene segment, wherein the human JH gene segment is an IGHJ6 (e.g., IGHJ6*02) that specifically binds hOX40L, in the manufacture of a medicament for administration to a human to treat or prevent an hOX40L-mediated disease or condition in a human selected from an autoimmune disease or condition, a systemic inflammatory disease or condition, or a transplant rejection, e.g., inflammatory bowel disease (IBD), Crohn's disease, rheumatoid arthritis, transplant rejection, allograft rejection, graft-versus-host disease (GvHD), ulcerative colitis, systemic lupus erythematosus (SLE), diabetes, uveitis, ankylosing spondylitis, contact hypersensitivity, multiple sclerosis, or atherosclerosis, particularly GvHD.

[0249] The antibody features and hOX40L-mediated diseases of any of the aspects, configurations, examples, or embodiments optionally apply mutatis mutandis to this use. Any compositions, dosing schedules, or modes of administration described in any of the aspects, configurations, examples, or embodiments herein apply mutatis mutandis to this use.

[0250] 97. A hOX40L-mediated disease or condition selected from an autoimmune disease or condition, a systemic inflammatory disease or condition, or transplant rejection, e.g., inflammatory bowel disease (IBD), Crohn's disease, rheumatoid arthritis, transplant rejection, allograft rejection, graft-versus-host disease (GvHD), ulcerative colitis, systemic lupus erythematosus (SLE), diabetes, uveitis, ankylosing spondylitis, contact hypersensitivity, multiple sclerosis, or atherosclerosis, particularly GvHD in humans. A method for treating or preventing a hOX40L-mediated disease or condition, comprising administering to a human a therapeutically effective amount of a human antibody or fragment thereof, the antibody comprising an HCDR3 of 16 to 27 amino acids and derived from the recombination of a human VH gene segment, a human D gene segment, and a human JH gene segment, wherein the human JH gene segment is IGHJ6 (e.g., IGHJ6*02) that specifically binds to hOX40L, thereby treating or preventing a hOX40L-mediated disease or condition.

[0251] The antibody features and hOX40L-mediated diseases of any of the aspects, configurations, examples, or embodiments optionally apply mutatis mutandis to this method. Any compositions, dosing schedules, or modes of administration described in any of the aspects, configurations, examples, or embodiments herein apply mutatis mutandis to this method.

[0252] In an embodiment of any one of aspects 95 to 97, the human JH gene segment is selected from IGHJ6*01, IGHJ6*02, IGHJ6*03, and IGHJ6*04. In a further aspect of any one of aspects 95 to 97, the human JH gene segment is selected from IGHJ6*01, IGHJ6*02, and IGHJ6*04. In a further aspect of any one of aspects 95 to 97, the JH gene segment is IGHJ6*02.

[0253] In a further embodiment of any one of aspects 95 to 97, the human VH gene segment is IGHV3-23, for example selected from IGHV3-23*01, IGHV3-23*02, IGHV3-23*03, IGHV3-23*04, or IGHV3-23*05. In another embodiment of any one of aspects 95 to 97, the human VH gene segment is IGHV3-23*01 or IGHV3-23*04, in particular IGHV3-23*04.

[0254] In a further embodiment of any one of aspects 95 to 97, the human DH gene segment is IGHD3-10, for example selected from IGHD3-10*01 or IGHD3-10*02. In one embodiment of any one of aspects 95 to 97, the human DH gene segment is IGHD3-10*01. In one embodiment of any one of aspects 95 to 97, the human DH gene segment is IGHD3-10*02.

[0255] In an embodiment of any one of aspects 90 to 97, the antibody is capable of treating or preventing GvHD. In another embodiment of any one of aspects 90 to 97, the antibody or fragment is used to treat or prevent a disease other than GvD, but the antibody or fragment is capable of treating or preventing GvHD.

[0256] 98. The antibody or fragment according to aspect 86, the antibody or fragment according to aspect 95, the use according to aspect 96, or the method according to aspect 97, wherein the antibody or fragment comprises a kappa light chain, e.g. wherein the VL domain of the light chain is derived from a recombination of a human VL gene segment and a human JL gene segment, wherein the human VL gene segment is IGKV1D-39 (e.g. IGKV1D-39*01), and optionally the human JL gene segment is IGKJ1 (e.g. IGKJ1*01) or IGKJ3 (e.g. IGKJ3*01).

[0257] In another embodiment, the VL domain is a kappa VL domain. In one embodiment, the kappa VL domain is derived from the recombination of a human VL gene segment and a human JL gene segment, and the human VL gene segment is IGKV1D-39. In another embodiment, the VL gene segment is IGKV1D-39*01.

[0258] In a further embodiment, the human JL gene segment is IGKJ1. In another embodiment, the JL gene segment is IGKJ1*01. In a further embodiment, the human JL gene segment is IGKJ3. In another embodiment, the JL gene segment is IGKJ3*01.

[0259] 99. The antibody or fragment according to any one of aspects 73 to 89, 98, 101, or 102, or the antibody or fragment, use, or method according to any one of aspects 90 to 98, wherein the antibody or fragment enables greater than 80% stem cell donor chimerism by day 12 in a rhesus monkey model of haploidentical hematopoietic stem cell transplantation, and optionally the antibody is for the prevention of GvHD.

[0260] In another aspect, there is provided the antibody or fragment, use, or method according to any one of aspects 95 to 98, wherein the antibody or fragment is for treating or preventing graft rejection (e.g., GvHD) in a human after donor human hematopoietic stem cell transplantation by enabling greater than 80% stem cell donor chimerism in the human by day 12.

[0261] In another embodiment there is provided the antibody or fragment of any one of aspects 73 to 89, 98, 101, or 102, wherein the antibody or fragment enables greater than 80% stem cell donor chimerism by day 12 in a rhesus monkey model of haploidentical hematopoietic stem cell transplantation.

[0262] In one embodiment, chimerism is achieved by T cells (CD3 + / CD20 -) chimerism. In another embodiment, the chimerism is peripheral blood chimerism. In another embodiment, the chimerism is peripheral blood or T cell (CD3 + / CD20 - ) It is a chimera phenomenon.

[0263] In one embodiment, stem cell donor chimerism (e.g., peripheral blood or T cell (CD3 + / CD20 - ) Chimerism) is determined using divergent donor- and recipient-specific MHC-binding microsatellite markers by comparing the peak heights of donor- and recipient-specific amplicons. In another embodiment, stem cell donor chimerism is determined using the method described in Kean, LS, et al., "Induction of chimerism in rhesus macaques through stem In another embodiment, stem cell donor chimerism is determined as described in Example 7.

[0264] In one embodiment, the haploidentical hematopoietic stem cell rhesus monkey model is performed with transplant (HSCT) recipient animals that undergo a conditioning procedure with anti-OX40L antibody administration, followed by an infusion of peripheral blood products isolated from a half-sibling donor animal, followed by weekly dosing with the anti-OX40L antibodies of the present invention, and blood samples are taken and analyzed for chimerism.

[0265] In another embodiment, in an HSCT model, recipient animals receive conditioning radiation of 1020 cGy divided into four doses over two days (experimental days -2 and -1) to eliminate the host's hematopoietic system (intravenous administration on day -2, followed by intravenous administration on days 5, 12, 19, 26, 33, 40, and 47) prior to intravenous administration of an anti-OX40L antibody of the invention, and a transplant of leukocyte- and stem cell-enriched peripheral blood from an MHC-semimatched (half-sibling) donor animal to reconstitute the recipient's immune system, along with ongoing supportive care, blood sampling, and monitoring for signs of GVHD.

[0266] In one embodiment the antibody or fragment, use or method is for the prevention of GvHD.

[0267] In one embodiment, the anti-hOX40L antibodies of the invention are administered prophylactically. In one embodiment, prophylactic treatment prevents the worsening or onset of a disease or condition.

[0268] In another embodiment, the antibody is administered intravenously. In another embodiment, the antibody is administered at a dose of about 5-10 mg / kg (e.g., at about 8 mg / kg). In another embodiment, the antibody is administered intravenously. In another embodiment, the antibody is administered at a dose of about 5-10 mg / kg (e.g., at about 8 mg / kg). In another embodiment, the antibody is administered at a dose selected from about 0.1 mg / kg, about 0.5 mg / kg, about 1 mg / kg, 3 mg / kg, 5 mg / kg, about 10 mg / kg, about 15 mg / kg, about 20 mg / kg, about 25 mg / kg, about 30 mg / kg, about 40 mg / kg, about 50 mg / kg, about 60 mg / kg, about 70 mg / kg, about 80 mg / kg, about 90 mg / kg, or about 100 mg / kg, particularly about 1 mg / kg or about 3 mg / kg.

[0269] In another embodiment, the antibody is administered 1-4 days prior to transplant, e.g., 1-3 days prior to transplant or 1-2 days prior to transplant. In another embodiment, the antibody is administered weekly, biweekly, or monthly, e.g., every other week, after transplant. In a further embodiment, the antibody is administered prophylactically intravenously at a dose of about 5-10 mg / kg (e.g., about 8 mg / kg) 1-3 days prior to transplant, and then intravenously at a dose of about 5-10 mg / kg (e.g., about 8 mg / kg) every other week thereafter.

[0270] In another embodiment, patients are monitored periodically after transplantation for the presence of biomarkers predictive of the development of GvHD (e.g., acute GvHD), and when the biomarker levels reach a level that determines the patient is at risk for developing GvHD (e.g., acute GvHD), an anti-OX40L antibody of the invention is administered. This strategy may avoid unnecessary administration of drugs and unnecessary suppression of the immune system. Examples of biomarkers that may be useful as predictive biomarkers for acute GvHD may be those identified in Levine et al., "A prognostic score for acute graft-versus-host disease based on biomarkers: a multicentre study," Lancet Haematol 2015;2:e21-29. These biomarkers include, but are not limited to, TNFR1, ST-2, elafin, and IL2Rα and Reg3α.

[0271] In a further embodiment, the HSCT model is described in Miller, Weston P., et al. "GVHD after haploidentical transplantation: a novel, MHC-defined rhesus macaque model identifies CD28 - CD8 +"T cells as a reservoir of breakthrough T-cell proliferation during costimulation blockade and sirolimus-based immunosuppression." Blood, 116, 24(2010):5403-5418. In a further embodiment, the HSCT model is performed as described in Example 7.

[0272] 100. The antibody or fragment, use, or method according to any one of embodiments 95 to 99, wherein the antibody is as defined in any one of embodiments 73 to 89, 98, 99, 101, or 102.

[0273] 101. The antibody or fragment according to any one of aspects 73 to 89, 98, 99, or 102, or the antibody or fragment, use, or method according to any one of aspects 90 to 100, wherein the antibody or fragment is expressed as a stably transfected pool in Lonza GS-Xceed™ at a level of greater than 1.5 g / L in an overgrowth fed-batch culture using the Lonza version 8 feed system with an overgrowth period of 14 days.

[0274] In one embodiment, the expression level is greater than 1.0 g / L, greater than 1.1 g / L, greater than 1.2 g / L, greater than 1.3 g / L, or greater than 1.4 g / L.

[0275] 102. The antibody or fragment inhibited total CD4+ T cells at day 12 in a rhesus macaque model of haploidentical hematopoietic stem cell transplantation. + CD4 >20% of the T cell population + The antibody or fragment according to any one of embodiments 73 to 89, 98, 99 or 101, or the antibody or fragment, use or method according to any one of embodiments 90 to 101, which maintains a naive population of T cells.

[0276] In another aspect there is provided an antibody or fragment according to any one of aspects 73 to 89, 98, 99 or 101, or the antibody or fragment, use or method according to any one of aspects 90 to 101, wherein the antibody or fragment is + Donor CD4 >20% of the T cell population + For treating or preventing transplant rejection in said human by maintaining a naive population of T cells.

[0277] In one embodiment, the HSCT model is as described in any of the embodiments contemplated above, eg, as described in relation to aspect 99.

[0278] In another embodiment, the naive population is measured by assessing the relative proportions of specific T cell phenotypes using flow cytometry, where cell subsets are labeled with fluorescent antibody probes to identify naive CD4 or CD8 T cells as CD4 T cells, respectively. + / CD28 + / CD95 - or CD8 + / CD28 + / CD95 - and central memory CD4 or CD8 T cells are labeled CD4 + / CD28 + / CD95 + or CD8 + / CD28 + / CD95 + and effector memory CD4 or CD8 T cells are labeled CD4 + / CD28 - / CD95 + or CD8 + / CD28 - / CD95 + It is identified by being labeled as such.

[0279] 103. The method further comprising administering to the human an additional therapeutic agent, optionally wherein the additional therapeutic agent is independently selected from the group consisting of rapamycin (sirolimus), lacrolimus, cyclosporine, corticosteroids (e.g., methylprednisolone), methotrexate, mycophenolate mofetil, anti-CD28 antibodies, anti-IL12 / IL-23 antibodies (e.g., ustekinumab), anti-CD20 antibodies (e.g., rituximab), anti-CD3 0 antibodies (e.g., brentuximab), CTLA4-Fc molecules (e.g., abatacept), CCR5 receptor antagonists (e.g., maraviroc), anti-CD40L antibodies, anti-VLA4 antibodies (e.g., natalizumab), anti-LFA1 antibodies, fludarabine, anti-CD52 antibodies (e.g., alemtuzumab), anti-CD45 antibodies, cyclophosphamide, anti-thymocyte globulin, anti-complement C5 antibodies (e.g., eculizumab), anti-a4b7 integrin antibodies (e.g., vedolizumab), anti-IL6 antibodies (e.g., tocilizumab), anti-IL2R antibodies (e.g., basilixumab), anti-CD25 antibodies (e.g., daclizumab), anti-TNFα / TNFα-Fc molecules (e.g., etanercept, adalimumab, infliximab, golimumab, or cetolizumab pegol), and vorinostat, in particular rapamycin (sirolimus), lacrolimus, cyclosporin, 103. The antibody or fragment, use, or method of any one of aspects 90 to 102, wherein the therapeutic agent is selected from the group consisting of: sporin, corticosteroids (e.g., methylprednisolone), methotrexate, mycophenolate mofetil, anti-CD28 antibodies, CTLA4-Fc molecules (e.g., abatacept), anti-CD40L antibodies, anti-LFA1 antibodies, anti-CD52 antibodies (e.g., alemtuzumab), cyclophosphamide, and anti-thymocyte globulin.

[0280] In one embodiment, the additional therapeutic agent is an anti-inflammatory agent. In another embodiment, the anti-inflammatory agent is independently selected from the group consisting of a corticosteroid (e.g., methylprednisolone), an anti-IL12 / IL-23 antibody (e.g., ustekinumab), an anti-VLA4 antibody (e.g., natalizumab), an anti-LFA1 antibody, an anti-complement C5 antibody (e.g., eculizumab), an anti-a4b7 integrin antibody (e.g., vedolizumab), an anti-IL6 antibody (e.g., tocilizumab), an anti-IL2R antibody (e.g., basilixumab), or an anti-TNFα antibody / TNFα-Fc molecule (e.g., etanercept, adalimumab, infliximab, golimumab, cetolizumab pegol). In one example, the anti-inflammatory agent is independently selected from the group consisting of a corticosteroid (e.g., methylprednisolone) and an anti-LFA1 antibody.

[0281] 104. The antibody or fragment, use, or method of embodiment 103, wherein the further therapeutic agent is administered sequentially or simultaneously with the anti-hOX40L antibody or fragment.

[0282] 105. A method for treating a rheumatoid arthritis comprising administering to a subject a therapeutically effective amount of an antibody fragment as defined in any one of aspects 73 to 89, 98, 99, 101, or 102, and a pharmaceutically acceptable excipient, diluent, or carrier, optionally, independently, rapamycin (sirolimus), lacrolimus, cyclosporine, a corticosteroid (e.g., methylprednisolone), methotrexate, mycophenolate mofetil, an anti-CD28 antibody, an anti-IL12 / IL-23 antibody (e.g., uracil), or a combination thereof. tekinumab), anti-CD20 antibodies (e.g., rituximab), anti-CD30 antibodies (e.g., brentuximab), CTLA4-Fc molecules (e.g., abatacept), CCR5 receptor antagonists (e.g., maraviroc), anti-CD40L antibodies, anti-VLA4 antibodies (e.g., natalizumab), anti-LFA1 antibodies, fludarabine, anti-CD52 antibodies (e.g., alemtuzumab), anti-CD45 antibodies, cyclophosphamide, anti-thymocyte globulin, anti-complement C5 antibodies (e.g., eculizumab), anti-a4b7 integrin antibodies (e.g., vedolizumab), anti-IL6 antibodies (e.g., tocilizumab), anti-IL2R antibodies (e.g., basilixumab), anti-CD25 antibodies (e.g., daclizumab), anti-TNFα / TNFα-Fc molecules (e.g., etanercept, adalimumab, infliximab, golimumab, or cetolizumab pegol), and vorinostat, especially rapamycin (cilostazol). abatacept), anti-CD40L antibody, anti-LFA1 antibody, anti-CD52 antibody (e.g., alemtuzumab), cyclophosphamide, and anti-thymocyte globulin.

[0283] The pharmaceutically acceptable excipients, diluents, or carriers described herein apply mutatis mutandis to these compositions.

[0284] In one embodiment, the additional therapeutic agent is an anti-inflammatory agent. In another embodiment, the anti-inflammatory agent is independently selected from the group consisting of a corticosteroid (e.g., methylprednisolone), an anti-IL12 / IL-23 antibody (e.g., ustekinumab), an anti-VLA4 antibody (e.g., natalizumab), an anti-LFA1 antibody, an anti-complement C5 antibody (e.g., eculizumab), an anti-a4b7 integrin antibody (e.g., vedolizumab), an anti-IL6 antibody (e.g., tocilizumab), an anti-IL2R antibody (e.g., basilixumab), or an anti-TNFα antibody / TNFα-Fc molecule (e.g., etanercept, adalimumab, infliximab, golimumab, cetolizumab pegol). In one example, the anti-inflammatory agent is independently selected from the group consisting of a corticosteroid (e.g., methylprednisolone) and an anti-LFA1 antibody.

[0285] 106. A pharmaceutical composition according to aspect 105, or a kit comprising a pharmaceutical composition as defined in aspect 105, wherein the composition is for the treatment and / or prevention of a hOX40L-mediated condition or disease selected from an autoimmune disease or condition, a systemic inflammatory disease or condition, or graft rejection, such as inflammatory bowel disease (IBD), Crohn's disease, rheumatoid arthritis, graft rejection, allograft rejection, graft versus host disease (GvHD), ulcerative colitis, systemic lupus erythematosus (SLE), diabetes, uveitis, ankylosing spondylitis, contact hypersensitivity, multiple sclerosis, and atherosclerosis, in particular GvHD.

[0286] Any one of the hOX40L-mediated diseases aspects, configurations, examples or embodiments described herein optionally apply mutatis mutandis to this combination.

[0287] 107. The pharmaceutical composition of embodiment 105 or embodiment 106, or the kit of embodiment 106 comprising a label or instructions for use in the treatment and / or prevention of said disease or condition in a human, wherein optionally the label or instructions comprises a marketing authorization number (e.g., an FDA or EMA approval number), and optionally the kit comprises an IV or injection device comprising the antibody or fragment.

[0288] The labels, instructions, hOX40L-mediated diseases, and conditions of any one of the aspects, compositions, examples, or embodiments described herein optionally apply mutatis mutandis to this combination.

[0289] 108. A nucleic acid encoding the HCDR3 of the antibody or fragment defined in any one of embodiments 73 to 89, 98, 99, 101, or 102.

[0290] 109. A nucleic acid encoding the VH domain and / or the VL domain of an antibody or fragment as defined in any one of aspects 73 to 89, 98, 99, 101, or 102.

[0291] 110. The nucleic acid according to embodiment 109, comprising a nucleotide sequence which is at least 80% identical to the sequence of SEQ ID NO: 33 and / or SEQ ID NO: 47.

[0292] In one example, the nucleotide sequence is at least 85% identical, at least 90% identical, at least 95% identical, at least 96% identical, at least 97% identical, at least 98% identical, or at least 99% identical to the sequence of SEQ ID NO:33 and / or SEQ ID NO:47.

[0293] 111. A nucleic acid encoding a heavy or light chain of an antibody of any one of embodiments 73-89, 98, 99, 101, or 102.

[0294] 112. A vector comprising a nucleic acid according to any one of aspects 108-111, optionally wherein the vector is a CHO or HEK293 vector.

[0295] 113. A host comprising the nucleic acid of any one of embodiments 108-111, or the vector of claim 112.

[0296] As described in the Examples, the inventors have devised a set of criteria that are particularly useful for identifying antibodies and fragments of the invention. These criteria are: (a) the ability of the antibody or fragment to bind to cell surface hOX40L on CHO-S cells (optionally transfected with full-length human OX40L) and / or to bind to recombinant hOX40L in an HTRF assay; (b) the ability of the antibody or fragment to neutralize human OX40 (e.g., neutralize human OX40L binding to the human OX40 receptor) in a receptor neutralization HTRF assay and / or a flow cytometry receptor neutralization assay; (c) The ability of the antibody or fragment to specifically bind to both human OX40L and rhesus OX40L (useful so that PK, PD, efficacy, and other parameters of the antibody or fragment can be assessed in a rhesus model as a surrogate for humans).

[0297] Thus, in one example of the invention, the antibody or fragment satisfies criteria (a), (b), and (c).

[0298] In one example, criterion (a) is set such that the antibody or fragment exhibits less than 70% receptor binding to hOX40L expressed by CHO-S cells by FACS.

[0299] In one example, criterion (a) is set so that the antibody or fragment exhibits less than 90% receptor binding to OX40L in an HTRF assay.

[0300] In one example, criterion (a) is set so that the antibody or fragment exhibits at least 20% efficacy in an HTRF assay.

[0301] In one example, OX40 is used in criterion (b).

[0302] In one embodiment, assays or tests of antibodies or fragments of the invention are carried out at pH 7 or substantially at pH 7 (eg, for in vitro tests and assays) and at or substantially at rtp.

[0303] Optionally, the antibody or fragment specifically binds to hOX40L with an affinity (apparent affinity, Kd) of less than 1 microM, 1000 nM to 100 nM, 100 nM to 10 nM, 10 nM to 1 nM, 1000 pM to 500 pM, 500 pM to 200 pM, less than 200 pM, 200 pM to 150 pM, 200 pM to 100 pM, 100 pM to 10 pM, 10 pM to 1 pM, e.g., in the range of 1 mM to 1 pM (e.g., 1 mM to 100 pM, 10 nM to 100 pM, 1 nM to 10 pM, or 100 pM to 1 pM), as determined by SPR, e.g., under the SPR conditions disclosed herein. Additionally or alternatively, the antibody or fragment specifically binds to rhesus OX40L with an affinity (apparent affinity, Kd) of less than 1 microM, 1000 nM to 100 nM, 100 nM to 10 nM, 10 nM to 1 nM, 1000 pM to 500 pM, 500 pM to 200 pM, less than 200 pM, 200 pM to 150 pM, 200 pM to 100 pM, 100 pM to 10 pM, 10 pM to 1 pM, e.g., in the range of 1 mM to 1 pM (e.g., 1 mM to 100 pM, 10 nM to 100 pM, 1 nM to 10 pM, or 100 pM to 1 pM), as determined by SPR, e.g., under the SPR conditions disclosed herein. Such binding measurements can be performed using a variety of binding assays known in the art, for example, using surface plasmon resonance (SPR) such as Biacore™, or using a ProteOn XPR36™ (Bio-Rad®), using KinExA® (Sapidyne Instruments, Inc.), or using a ForteBio Octet (Pall ForteBio Corp.).

[0304] OX40L binding capacity, specificity, and affinity (Kd, K off and / or K. on ) can be determined by any method routine in the art, for example, by surface plasmon resonance (SPR). The term "Kd," as used herein, is intended to refer to the equilibrium dissociation constant of a particular antibody-antigen interaction.

[0305] In one embodiment, surface plasmon resonance (SPR) is performed at 25° C. In another embodiment, SPR is performed at 37° C.

[0306] In one embodiment, SPR is performed at physiological pH, such as about pH 7, or at about pH 7.6 (eg, using Hepes-buffered saline (also called HBS-EP) at pH 7.6).

[0307] In one embodiment, SPR is performed at physiological salt concentrations, for example, 150 mM NaCl.

[0308] In one embodiment, SPR is carried out at a detergent concentration of 0.05% or less by volume, for example, in the presence of 0.05% P20 (Polysorbate 20, e.g., Tween-20™) and 3 mM EDTA.

[0309] In one example, SPR is performed in a pH 7.6 buffer, 150 mM NaCl, 0.05% detergent (e.g., P20), and 3 mM EDTA at 25° C. or 37° C. The buffer may contain 10 mM Hepes. In one example, SPR is performed in HBS-EP at 25° C. or 37° C. HBS-EP is available from Teknova Inc. (California, catalog number H8022).

[0310] In one example, the affinity of an antibody or fragment can be determined using SPR: 1. Attaching anti-mouse (or other relevant human, rat, or non-human vertebrate antibody constant region species-matched) IgG (e.g., Biacore™ BR-1008-38) to a biosensor chip (e.g., a GLM chip), such as by primary amine coupling; 2. Exposing anti-mouse IgG (or other matching species antibody) to the test IgG antibody to capture the test antibody on the chip; 3. Passing the test antigen over the capture surface of the chip at 0 nM (i.e., buffer only), 1024 nM, 256 nM, 64 nM, 16 nM, 4 nM; 4. The binding affinity of the test antibody to the test antigen is then determined using surface plasmon resonance, e.g., by determining under the SPR conditions discussed above (e.g., in physiological buffer at 25°C). SPR can be performed using any standard SPR instrument, such as a Biacore™, or using a ProteOn XPR36™ (Bio-Rad®).

[0311] Regeneration of the capture surface can be performed using 10 mM glycerol at pH 1.7, which removes the captured antibody and allows the surface to be used for another interaction. Binding data can be fitted to a 1:1 intrinsic model using standard techniques, for example, using the intrinsic model in ProteOn XPR36™ analysis software.

[0312] In one example, the antibody or fragment of the invention is contained in a medical container, e.g., a vial, syringe, IV container, or injection device (e.g., an intraocular or intravitreal injection device). In one example, the antibody or fragment is in vitro, e.g., in a sterile container. In one example, the invention provides a kit comprising an antibody or fragment of the invention, packaging, and instructions for use in treating or preventing or diagnosing an OX40L-mediated disease or condition in a human. In one example, the instructions indicate that before administering the antibody or fragment to the human, the human should be genotyped for an OX40L variant sequence of the invention. In one example, the instructions indicate that before administering the antibody or fragment to the human, the human should be phenotyped for an OX40L variant sequence of the invention. In one example, the human is of Chinese (e.g., Han CHS) ethnicity, and the instructions are in Chinese (e.g., Mandarin).

[0313] In one example, the binding site(s) of the antibody or fragment are selected from a plurality (e.g., a library) of binding sites, e.g., the plurality of binding sites comprises or consists of a plurality of four-chain antibodies or fragments thereof, e.g., dAbs, Fabs, or scFvs. Suitable methods for generating multiple binding sites for screening include phage display (to generate a phage display library of antibody binding sites), ribosome display (to generate a ribosome display library of antibody binding sites), yeast display (to generate a yeast display library of antibody binding sites), or immunization of a non-human vertebrate (e.g., a rodent, e.g., a mouse or rat, e.g., Velocimouse™, Kymouse™, Xenomouse™, Aliva Mouse™, HuMab Mouse™, Omnimouse™, Omnirat™, or MeMo Mouse™) with hOX40L or an hOX40L epitope and a repertoire of antibody-producing cells (e.g., a B cell, plasma cell, or plasmablast repertoire) and / or a repertoire of isolated antibodies, fragments, or binding sites.

[0314] The term "epitope" is the region of an antigen to which an antibody or fragment binds. Epitopes can be defined as structural or functional. Functional epitopes are generally a subset of structural epitopes and contain residues that directly contribute to the affinity of the interaction. Epitopes can also be conformational, meaning that they can be composed of nonlinear amino acids. In certain embodiments, epitopes can include determinants that are chemically active surface groupings of molecules, such as amino acids, sugar side chains, phosphoryl groups, or sulfonyl groups, and in certain embodiments, can have specific three-dimensional structural characteristics and / or specific charge characteristics.

[0315] The term "isolated" with respect to any embodiment of the invention, e.g., an isolated antibody or fragment, means that the subject antibody, fragment, etc. (1) is free from at least some other proteins with which it is normally found, (2) is essentially free from other proteins from the same source, e.g., the same species, (3) is expressed by cells from a different species, (4) is separated from at least about 50 percent of the polynucleotides, lipids, carbohydrates, or other materials with which it is naturally associated, (5) is operably associated with polypeptides with which it is not naturally associated, or (6) is not naturally occurring. Typically, an "isolated" antibody, fragment, etc., constitutes at least about 5%, at least about 10%, at least about 25%, or at least about 50%, 60%, 70%, 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99%, or more than 99% of a given sample. Genomic DNA, cDNA, mRNA, or other RNA, or any combination thereof, of synthetic origin, may encode such an isolated antibody, fragment, etc. Preferably, an isolated antibody, fragment, etc. is substantially free of proteins or polypeptides or other contaminants found in its natural environment that may interfere with its therapeutic, diagnostic, prophylactic, research, or other uses.

[0316] For example, an "isolated" antibody is one that has been identified, separated, and / or recovered from components of its production environment (e.g., natural or recombinant). Preferably, an isolated polypeptide is free from association with all other components from its production environment, such as, for example, an antibody that has been isolated to FDA-approvable or approved standards. Contaminating components of its production environment, such as those resulting from recombinantly transfected cells, are substances that may typically interfere with research, diagnostic, or therapeutic uses for the antibody and may include enzymes, hormones, and other proteinaceous or non-proteinaceous solutes. In preferred embodiments, the polypeptide will be purified (1) to greater than 95% by weight, and in some embodiments, greater than 99% by weight, of the antibody, as determined, for example, by the Lowry method; (2) sufficiently to obtain at least 15 residues of N-terminal or internal amino acid sequence by use of a spinning cup sequenator; or (3) to homogeneity by SDS-PAGE under non-reducing or reducing conditions using Coomassie blue, preferably silver staining. Isolated antibody includes the antibody in situ within recombinant cells since at least one component of the antibody's natural environment will not be present. Generally, an isolated polypeptide or antibody will be prepared by at least one purification step.

[0317] immunoconjugate The present invention encompasses antibodies or fragments conjugated to a therapeutic moiety ("immunoconjugates"), such as a cytotoxin, a chemotherapeutic agent, an immunosuppressant, or a radioisotope. Cytotoxic agents include any agent that harms cells. Examples of suitable cytotoxic agents and chemotherapeutic agents for forming immunoconjugates are known in the art; see, for example, WO 05 / 103081, which is incorporated herein by reference in its entirety.

[0318] dual specificity The antibodies and fragments of the present invention may be monospecific, bispecific, or multispecific. Multispecific mAbs may be specific for different epitopes of a single target polypeptide or may contain antigen-binding domains specific for more than one target polypeptide. See, e.g., Tutt et al., (1991) J. Immunol. 147:60-69. A human anti-hOX40L antibody or fragment may be conjugated to or co-expressed with another functional molecule, e.g., another peptide or protein. For example, an antibody or fragment thereof may be functionally linked (e.g., by chemical coupling, genetic fusion, noncovalent association, etc.) to one or more other molecular entities, such as another antibody or antibody fragment, to produce a bispecific or multispecific antibody with a second binding specificity.

[0319] An exemplary bispecific antibody format that can be used in the context of the present invention involves the use of a first immunoglobulin (Ig) CH3 domain and a second Ig CH3 domain, where the first and second Ig CH3 domains differ from each other by at least one amino acid, and the at least one amino acid difference reduces binding of the bispecific antibody to protein A compared to a bispecific antibody lacking the amino acid difference. In one embodiment, the first Ig CH3 domain binds to protein A and the second Ig CH3 domain comprises a mutation that reduces or eliminates protein binding, such as an H95R modification (according to IMGT exon numbering; H435R in EU numbering). The second CH3 may further comprise a Y96F modification (according to IMGT, Y436F in EU). Additional modifications that may be found within the second CH3 include D16E, L18M, N44S, K52N, V57M, and V821 for IgG1 antibodies (by IMGT; D356E, L358M, N384S, K392N, V397M, and V422I in EU), N44S, K52N, and V82I for IgG2 antibodies (by IMGT; N384S, K392N, and V422I in EU), and Q15R, N44S, K52N, V57M, R69K, E79Q, and V82I for IgG4 antibodies (by IMGT; Q355R, N3845, K392N, V397M, R409K, E419Q, and V422I in EU). Variations on the above bispecific antibody formats are contemplated as being within the scope of the present invention.

[0320] In certain embodiments, the antibody or OX40L-binding fragment thereof comprises fewer than six CDRs. In some embodiments, the antibody or antigen-binding fragment thereof comprises or consists of one, two, three, four, or five CDRs selected from the group consisting of HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3. In certain embodiments, the antibody or antigen-binding fragment thereof comprises or consists of one, two, three, four, or five CDRs selected from the group consisting of HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 sequences in the sequence listing (i.e., for HCDR1, SEQ ID NO:4, SEQ ID NO:10, SEQ ID NO:36, SEQ ID NO:42, SEQ ID NO:68, SEQ ID NO:74, SEQ ID NO:96, or SEQ ID NO:102, particularly SEQ ID NO:36 or SEQ ID NO:42; for HCDR2, SEQ ID NO:6, SEQ ID NO:12, SEQ ID NO:38, SEQ ID NO:44, SEQ ID NO:70, SEQ ID NO:76, SEQ ID NO:98, or SEQ ID NO:104, particularly SEQ ID NO:38 or SEQ ID NO:44; for HCDR3, SEQ ID NO:8, SEQ ID NO:14, SEQ ID NO:40, for LCDR1, SEQ ID NO:18, SEQ ID NO:24, SEQ ID NO:50, SEQ ID NO:56, SEQ ID NO:82, SEQ ID NO:88, SEQ ID NO:110, or SEQ ID NO:116, in particular SEQ ID NO:50 or SEQ ID NO:56; for LCDR2, SEQ ID NO:20, SEQ ID NO:26, SEQ ID NO:52, SEQ ID NO:58, SEQ ID NO:84, SEQ ID NO:90, SEQ ID NO:112, or SEQ ID NO:118, in particular SEQ ID NO:52 or SEQ ID NO:58; and for LCDR3, SEQ ID NO:22, SEQ ID NO:28, SEQ ID NO:54, SEQ ID NO:60, SEQ ID NO:86, SEQ ID NO:92, SEQ ID NO:114, or SEQ ID NO:120, in particular SEQ ID NO:54 or SEQ ID NO:60).

[0321] In certain embodiments, the antibody of the present invention is a fully human antibody, a monoclonal antibody, a recombinant antibody, an antagonistic antibody, a hOX40L-neutralizing antibody, or any combination thereof, or the present invention provides an hOX40L-binding fragment thereof. In one example, the antibody is a chimeric antibody comprising a human variable domain and a non-human (e.g., mouse, rat, or rabbit) constant domain. In certain embodiments, the antibody is a fully human antibody, such as a fully human monoclonal antibody, or an antigen-binding fragment thereof, that specifically binds to hOX40L. In a preferred embodiment, the antibody is an antagonistic antibody. In a preferred embodiment, the antibody is a neutralizing antibody.

[0322] In one example, the antibody or fragment is a lambda-type antibody or fragment (i.e., the variable domains are lambda variable domains). Optionally, the antibody or fragment also comprises a lambda constant domain.

[0323] In certain embodiments, the antibody competes (e.g., in a dose-dependent manner) with OX40 or a fusion protein thereof (e.g., Fc:OX40) for binding to hOX40L, such as cell-surface-expressed or soluble hOX40L. Exemplary competitive blocking assays are provided in the Examples herein.

[0324] In another aspect, provided herein is an isolated nucleic acid encoding a hOX40L polypeptide (e.g., cell surface-expressed hOX40L or soluble hOX40L), a hOX40L polypeptide fragment, or an antibody that specifically binds to an hOX40L epitope. In certain embodiments, the nucleic acids encode a VH chain, a VL chain, a VH domain, a VL domain, HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 as disclosed in the sequence listing (i.e., SEQ ID NO:30 or SEQ ID NO:62 for the VH chain; SEQ ID NO:32 or SEQ ID NO:64 for the VL chain; SEQ ID NO:2, SEQ ID NO:34, SEQ ID NO:66, or SEQ ID NO:94, particularly SEQ ID NO:34, for the VH domain; SEQ ID NO:16, SEQ ID NO:48, SEQ ID NO:80, or SEQ ID NO:108, particularly SEQ ID NO:48, for HCDR1; SEQ ID NO:4, SEQ ID NO:10, SEQ ID NO:36, SEQ ID NO:42, SEQ ID NO:68, SEQ ID NO:74, SEQ ID NO:96, or SEQ ID NO:102, particularly SEQ ID NO:36 or SEQ ID NO:42, for HCDR2; SEQ ID NO:6, SEQ ID NO:12, SEQ ID NO:38, SEQ ID NO:44, SEQ ID NO:70, SEQ ID NO:76, SEQ ID NO: for HCDR3, SEQ ID NO:8, SEQ ID NO:14, SEQ ID NO:40, SEQ ID NO:46, SEQ ID NO:72, SEQ ID NO:78, SEQ ID NO:100, or SEQ ID NO:106, in particular SEQ ID NO:40 or SEQ ID NO:46; for LCDR1, SEQ ID NO:18, SEQ ID NO:24, SEQ ID NO:50, SEQ ID NO:56, SEQ ID NO:82, SEQ ID NO:88, SEQ ID NO:110, or SEQ ID NO:116, in particular SEQ ID NO:50 or SEQ ID NO:56; for LCDR2, SEQ ID NO:20, SEQ ID NO:26, SEQ ID NO:52, SEQ ID NO:58, SEQ ID NO:84, SEQ ID NO:90, SEQ ID NO:112, or SEQ ID NO:118, in particular SEQ ID NO:52 or SEQ ID NO:58; and for LCDR3, SEQ ID NO:22, SEQ ID NO:28, SEQ ID NO:54, SEQ ID NO:60, SEQ ID NO:86, SEQ ID NO:92, SEQ ID NO:114, or SEQ ID NO:120, in particular SEQ ID NO:54 or SEQ ID NO:60).

[0325] In another aspect, provided herein are vectors and host cells comprising nucleic acids encoding the antibodies or fragments of the invention.

[0326] In certain embodiments, the antibody specifically binds to one or more single nucleotide polymorphism (SNP) variants of hOX40L. In one example of any aspect of the invention, hOX40L is a trimer of monomers.

[0327] In one aspect, provided herein is a method for reducing (e.g., by at least 20, 30, 40, 50, or 60%, or 70, 80, 90, 95, or more than 90%) or completely inhibiting binding of hOX40L to OX40 in a subject (e.g., a human subject), the method comprising administering to the subject an effective amount of an antibody of the present invention or a fragment thereof that specifically binds to hOX40L (e.g., cell-surface-expressed hOX40L or soluble hOX40L).

[0328] In one aspect, provided herein is a method for treating or preventing an hOX40L-mediated disease or condition in a subject (e.g., a human subject), the method comprising administering to the subject an effective amount of an antibody of the present invention or a fragment thereof that specifically binds to hOX40L (e.g., cell-surface-expressed hOX40L or soluble hOX40L), wherein the disease or condition is treated or prevented by the antibody or fragment. In one example, the method comprises reducing or inhibiting hOX40L biological activity in the subject, such as secretion of one, more than one, or all of IL-2, IL-8, TNF-alpha, and interferon-gamma. In one example, the biological activity is selected from secretion of one, more than one, or all of IL-2, TNF-alpha, and interferon-gamma. In one example, the biological activity is selected from secretion of one, more than one, or all of IL-8, CCL20, and RANTES.

[0329] In one aspect, provided herein is a method for reducing or inhibiting hOX40L biological activity, such as the secretion of one, more than one, or all of IL-2, IL-8, TNF-alpha, and interferon-gamma in a subject (e.g., a human subject), the method comprising administering to the subject an effective amount of an antibody of the invention or a fragment thereof that specifically binds to hOX40L (e.g., cell-surface-expressed hOX40L or soluble hOX40L), wherein hOX40L biological activity is reduced by the antibody or fragment. In one example, the biological activity is selected from the secretion of one, more than one, or all of IL-2, TNF-alpha, and interferon-gamma. In one example, the biological activity is selected from the secretion of one, more than one, or all of IL-8, CCL20, and RANTES.

[0330] The term "about" or "approximately" means within 20%, preferably within 10%, and more preferably within 5% (or 4%, or 3%, or 2%, or in one example, 1% or less) of a given value or range.

[0331] As used herein, "administering" or "administration" refers to the act of injecting or otherwise physically delivering an exogenous substance (e.g., an anti-hOX40L antibody provided herein) to a patient, such as by mucosal, intradermal, intravenous, intramuscular delivery, and / or any other method of physical delivery described herein or known in the art. When a disease or symptom thereof is to be treated, administration of the substance typically occurs after the onset of the disease or symptom thereof. When a disease or symptom thereof is to be prevented, administration of the substance typically occurs before the onset of the disease or symptom thereof.

[0332] To determine the percent identity of two amino acid sequences of two nucleic acid sequences, the sequences are aligned for optimal comparison (e.g., gaps can be introduced into the first amino acid sequence or nucleic acid sequence for optimal alignment with the second amino acid or nucleic acid sequence). The amino acid residues or nucleotides at corresponding amino acid positions are then compared. If a position in the first sequence is occupied by the same amino acid residue or nucleotide as the corresponding position in the second sequence, the molecules are identical at that position. The percent identity between two sequences is a function of the number of identical positions shared by the sequences (i.e., % identity = number of identical overlapping positions / total number of positions × 100%). In one embodiment, the two sequences are the same length.

[0333] The determination of percent identity between two sequences (e.g., amino acid sequences or nucleic acid sequences) can also be achieved using a mathematical algorithm. A preferred, non-limiting example of a mathematical algorithm utilized for comparing two sequences is the algorithm of Karlin and Altschul, 1990, Proc. Natl. Acad. Sci. USA 87:2264 2268 (modified in Karlin and Altschul, 1993, Proc. Natl. Acad. Sci. USA 90:5873 5877). Such an algorithm is described in Altschul et al., 1990, J. Mol. Biol. 215:403. BLAST nucleotide searches can be performed with the NBLAST nucleotide program parameters set, for example, to score=100 and word length=12 to obtain nucleotide sequences homologous to the nucleic acid molecules of the present invention. BLAST protein searches can be performed with the XBLAST program parameters set, for example, to score 50 and word length=3 to obtain amino acid sequences homologous to the protein molecules of the present invention. To obtain gapped alignments for comparison purposes, Gapped BLAST can be utilized as described in Altschul et al., 1997, Nucleic Acids Res. 25:3389-3402. Alternatively, PSI BLAST can be used to perform an iterated search that detects distant relationships (Id.) between molecules. When utilizing BLAST, Gapped BLAST, and PSI Blast programs, the default settings of the respective programs (e.g., for XBLAST and NBLAST) can be used (see, for example, the National Center for (See National Center for Biotechnology Information (NCBI)). Another preferred, non-limiting example of a mathematical algorithm utilized for the comparison of sequences is the algorithm of Myers and Miller, 1988, CABIOS 4:11 17. Such an algorithm is incorporated into the ALIGN program (version 2.0), which is part of the GCG sequence alignment software package. When utilizing the ALIGN program for comparing amino acid sequences, a PAM120 weight residue table, a gap length penalty of 12, and a gap penalty of 4 can be used.

[0334] The percent identity between two sequences can be determined using techniques similar to those described above, with or without gaps. In calculating percent identity, typically only exact matches are counted.

[0335] As used herein, an "antagonist" or "inhibitor" of hOX40L refers to a ligand (e.g., an antibody or fragment) capable of inhibiting or reducing one or more of the biological activities of hOX40L, such as in cells expressing hOX40L or cells expressing an hOX40L ligand. For example, in certain embodiments, an antibody of the invention is an antagonistic antibody that inhibits or reduces secretion of CCL20, IL-8, and / or RANTES from cells having OX40 expressed on their surface when the antibody contacts the cells. In some embodiments, an hOX40L antagonist (e.g., an antagonistic antibody of the invention) acts, for example, by inhibiting or reducing the activation and / or cell signaling pathways of cells expressing OX40L, thereby inhibiting hOX40L-mediated biological activity compared to hOX40L-mediated biological activity in the absence of the antagonist. In certain embodiments, the antibodies provided herein are fully human antagonistic anti-hOX40L antibodies, preferably fully human monoclonal antagonistic anti-hOX40L antibodies.

[0336] The terms "antibody" and "immunoglobulin" or "Ig" may be used interchangeably herein. An antibody or fragment thereof that specifically binds to the hOX40L antigen may be cross-reactive with the antigen of interest. Preferably, an antibody or fragment thereof that specifically binds to the hOX40L antigen does not cross-react with other antigens (but may optionally cross-react with OX40L from a different species, e.g., rhesus monkey or mouse). An antibody or fragment thereof that specifically binds to the hOX40L antigen may be identified, for example, by immunoassay, BIAcore™, or other techniques known to those skilled in the art. An antibody or fragment thereof specifically binds to the hOX40L antigen when it binds to the hOX40L antigen with higher affinity than it binds to any cross-reactive antigen, as determined using experimental techniques such as radioimmunoassay (RIA) and enzyme-linked immunosorbent assay (ELISA). Typically, a specific or selective response will result in at least two times the background signal or noise, more typically more than ten times the background. See, e.g., Paul, et al., for a discussion of antibody specificity. ed., 1989, Fundamental Immunology Second Edition, Raven Press, New York, at pages 332-336.

[0337] Antibodies of the present invention include, but are not limited to, synthetic antibodies, monoclonal antibodies, recombinantly produced antibodies, multispecific antibodies (including bispecific antibodies), human antibodies, humanized antibodies, chimeric antibodies, intrabodies, single-chain Fvs (scFv) (including, e.g., monospecific, bispecific, etc.), camelized antibodies, Fab fragments, F(ab') fragments, disulfide-linked Fvs (sdFv), anti-idiotypic (anti-Id) antibodies, and epitope-binding fragments of any of the above. In particular, antibodies of the present invention include immunoglobulin molecules and immunologically active portions of immunoglobulin molecules, i.e., antigen-binding domains or molecules comprising an antigen-binding site that specifically binds to the hOX40L antigen (e.g., one or more complementarity-determining regions (CDRs) of an anti-hOX40L antibody). Antibodies of the invention can be of any type (e.g., IgG, IgE, IgM, IgD, IgA, and IgY), any class (e.g., IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2, particularly IgG4), or any subclass (e.g., IgG2a and IgG2b) of immunoglobulin molecule. In preferred embodiments, hOX40L antibodies are fully human, such as fully human monoclonal hOX40L antibodies. In certain embodiments, antibodies of the invention are IgG antibodies, or classes (e.g., human IgG1 or IgG4) or subclasses thereof. In certain embodiments, antibodies of the invention comprise a human gamma 4 constant region. In another embodiment, the heavy chain constant region does not bind to Fc-gamma receptors and comprises, for example, a Leu235Glu mutation. In another embodiment, the heavy chain constant region comprises a Ser228Pro mutation to increase stability. In another embodiment, the heavy chain constant region is IgG4-PE.

[0338] The terms "antigen-binding domain," "antigen-binding region," "antigen-binding fragment," and similar terms refer to the portion of an antibody (e.g., the complementarity-determining region (CDR)) that contains the amino acid residues that interact with an antigen and confer on the binder its specificity and affinity for the antigen. The antigen-binding region can be derived from any animal species, including rodents (e.g., rabbits, rats, or hamsters) and humans. Preferably, the antigen-binding region is of human origin.

[0339] As used herein, the term "composition" is intended to encompass a product that optionally contains specified ingredients (e.g., an antibody of the invention) in specified amounts, and any product that results directly or indirectly from the combination of specified ingredients, optionally in specified amounts.

[0340] In the context of a polypeptide, the term "derivative," as used herein, refers to a polypeptide comprising the amino acid sequence of a hOX40L polypeptide, a fragment of a hOX40L polypeptide, or an antibody that specifically binds to a hOX40L polypeptide that has been modified by the introduction of amino acid residue substitutions, deletions, or additions. The term "derivative," as used herein, also refers to a hOX40L polypeptide, a fragment of a hOX40L polypeptide, or an antibody that specifically binds to a hOX40L polypeptide that has been chemically modified, for example, by the covalent attachment of any type of molecule to the polypeptide. For example, and without limitation, a hOX40L polypeptide, a fragment of a hOX40L polypeptide, or an hOX40L antibody may be chemically modified, for example, by glycosylation, acetylation, pegylation, phosphorylation, amidation, derivatization, derivatization with known protecting / blocking groups, proteolytic cleavage, linkage to a cellular ligand or other protein, etc. A derivative is modified in a manner that differs from the native or starting peptide or polypeptide, either in the type or location of the attached molecule. A derivative further includes deletion of one or more chemical groups naturally present on the peptide or polypeptide. Derivatives of hOX40L polypeptides, hOX40L polypeptide fragments, or hOX40L antibodies may be chemically modified by chemical modification using techniques known to those skilled in the art, including, but not limited to, specific chemical cleavage, acetylation, formulation, metabolic synthesis with tunicamycin, etc. Additionally, derivatives of hOX40L polypeptides, hOX40L polypeptide fragments, or hOX40L antibodies may contain one or more non-classical amino acids. Polypeptide derivatives retain similar or identical functions as the hOX40L polypeptides, hOX40L polypeptide fragments, or hOX40L antibodies described herein.

[0341] The term "effective amount," as used herein, refers to an amount of a treatment (e.g., an antibody or pharmaceutical composition provided herein) that is sufficient to reduce and / or alleviate the severity and / or duration of a given disease and / or its associated symptoms. The term also encompasses the amount necessary to reduce or alleviate the development or progression of a given disease, reduce or alleviate recurrence, prevent the onset or onset of a given disease, and / or improve or enhance the prophylactic or therapeutic effect(s) of another treatment (e.g., a treatment other than an anti-hOX40L antibody provided herein). In some embodiments, an effective amount of an antibody of the invention is from about 0.1 mg / kg (mg of antibody per kg of subject body weight) to about 100 mg / kg. In certain embodiments, an effective amount of an antibody provided herein is about 0.1 mg / kg, about 0.5 mg / kg, about 1 mg / kg, 3 mg / kg, 5 mg / kg, about 10 mg / kg, about 15 mg / kg, about 20 mg / kg, about 25 mg / kg, about 30 mg / kg, about 35 mg / kg, about 40 mg / kg, about 45 mg / kg, about 50 mg / kg, about 60 mg / kg, about 70 mg / kg, about 80 mg / kg, about 90 mg / kg, or about 100 mg / kg (or a range therein). In some embodiments, "effective amount," as used herein, also refers to the amount of an antibody of the invention to achieve a specified result (e.g., inhibition of cellular hOX40L biological activity, such as inhibition of secretion of CCL20, IL-8, or RANTES, or INF-γ, TNF-α, or IL-2, particularly INF-γ, from the cells).

[0342] The term "epitope," as used herein, refers to a region located on the surface of an antigen, such as a hOX40L polypeptide or hOX40L polypeptide fragment, that has antigenic or immunogenic activity capable of binding to one or more antigen-binding regions of an antibody and eliciting an immune response in an animal, preferably a mammal, and most preferably a human. An epitope with immunogenic activity is a portion of a polypeptide that elicits an antibody response in an animal. An epitope with antigenic activity is a portion of a polypeptide to which an antibody specifically binds, as determined by any method known in the art, such as the immunoassay methods described herein. An antigenic epitope is not necessarily immunogenic. Epitopes usually consist of chemically active surface groupings of molecules, such as amino acids or sugar side chains, and have specific three-dimensional structural characteristics and specific charge characteristics. The region of a polypeptide that contributes to an epitope may be contiguous amino acids of the polypeptide, or an epitope may arise simultaneously from two or more non-contiguous regions of the polypeptide. An epitope may or may not be a three-dimensional surface feature of an antigen. In certain embodiments, the hOX40L epitope is a three-dimensional surface feature of the hOX40L polypeptide (e.g., a trimeric form of the hOX40L polypeptide). In other embodiments, the hOX40L epitope is a linear feature of the hOX40L polypeptide (e.g., a trimeric or monomeric form of the hOX40L polypeptide). The antibodies provided herein may specifically bind to an epitope of the monomeric (denatured) form of hOX40L, an epitope of the trimeric (native) form of hOX40L, or both the monomeric (denatured) and trimeric (native) forms of hOX40L. In certain embodiments, the antibodies provided herein specifically bind to an epitope of the trimeric form of hOX40L, but do not specifically bind to the monomeric form of hOX40L.

[0343] The term "excipient," as used herein, refers to an inert substance commonly used as a diluent, vehicle, preservative, binder, or stabilizer for a drug, including, but not limited to, proteins (e.g., serum albumin, etc.), amino acids (e.g., aspartic acid, glutamic acid, lysine, arginine, glycine, histidine, etc.), fatty acids and phospholipids (e.g., alkyl sulfonates, caprylate salts, etc.), surfactants (e.g., SDS, polysorbates, nonionic surfactants, etc.), saccharides (e.g., sucrose, maltose, trehalose, etc.), and polyols (e.g., mannitol, sorbitol, etc.). See also Remington's Pharmaceutical Sciences (1990) Mack Publishing Co., Easton, Pa., incorporated herein by reference in its entirety.

[0344] The term "fragment," as used herein in the context of a peptide or polypeptide, refers to a peptide or polypeptide that comprises less than the full-length amino acid sequence. Such fragments can arise, for example, from truncation at the amino terminus, truncation at the carboxy terminus, and / or internal deletion of a residue(s) from the amino acid sequence. Fragments can arise, for example, from alternative RNA splicing or from in vivo protease activity. In certain embodiments, an hOX40L fragment comprises a polypeptide comprising an amino acid sequence of at least 5 contiguous amino acid residues, at least 10 contiguous amino acid residues, at least 15 contiguous amino acid residues, at least 20 contiguous amino acid residues, at least 25 contiguous amino acid residues, at least 40 contiguous amino acid residues, at least 50 contiguous amino acid residues, at least 60 contiguous amino acid residues, at least 70 contiguous amino acid residues, at least 80 contiguous amino acid residues, at least 90 contiguous amino acid residues, at least 100 contiguous amino acid residues, at least 125 contiguous amino acid residues, at least 150 contiguous amino acid residues, at least 175 contiguous amino acid residues, at least 200 contiguous amino acid residues, or at least 250 contiguous amino acid residues of an hOX40L polypeptide or an antibody that specifically binds to an hOX40L polypeptide. In certain embodiments, a fragment of an hOX40L polypeptide or an antibody that specifically binds to an hOX40L antigen retains at least one, at least two, or at least three functions of the polypeptide or antibody.

[0345] The terms "fully human antibody" or "human antibody" are used interchangeably herein and refer to antibodies comprising human variable regions, most preferably human constant regions. In certain embodiments, these terms refer to antibodies comprising variable and constant regions of human origin. A "fully human" anti-hOX40L antibody may, in certain embodiments, encompass an antibody that binds to an hOX40L polypeptide and is encoded by a nucleic acid sequence that is a naturally occurring somatic variant of a human germline immunoglobulin nucleic acid sequence. In certain embodiments, the anti-hOX40L antibodies provided herein are fully human antibodies. The term "fully human antibody" includes antibodies having variable and constant regions corresponding to human germline immunoglobulin sequences, as described by Kabat et al. (See Kabat et al. (1991) Sequences of Proteins of Immunological Interest, Fifth Edition, US Department of Health and Human Services, NIH Publication No. 91-3242). Exemplary methods for producing fully human antibodies are provided, for example, in the Examples herein, although any method known in the art may be used.

[0346] The phrase "recombinant human antibody" includes human antibodies prepared, expressed, created, or isolated by recombinant means, such as antibodies expressed using a recombinant expression vector transfected into a host cell, antibodies isolated from a recombinant combinatorial human antibody library, antibodies isolated from a human immunoglobulin transgenic and / or transchromosomal animal (e.g., mouse or cow) (see, e.g., Taylor, LD et al. (1992) Nucl. Acids Res. 20:6287-6295), or antibodies prepared, expressed, created, or isolated by any other means involving splicing of human immunoglobulin gene sequences into other DNA sequences. Such recombinant human antibodies can have variable and constant regions derived from human germline immunoglobulin sequences (Kabat, EA et al. (1991) Sequences of Proteins of Immunological Interest, Fifth Edition, US Department of Health and (See, Human Services, NIH Publication No. 91-3242.) However, in certain embodiments, such recombinant human antibodies have been subjected to in vitro mutagenesis (or, when human Ig sequence transgenic animals are used, in vivo somatic mutagenesis) such that the amino acid sequences of the VH and VL regions of the recombinant antibodies are derived from and related to human germline VH and VL sequences, but are sequences that may not naturally exist in the in vivo human antibody germline repertoire.

[0347] The term "fusion protein," as used herein, refers to a polypeptide comprising the amino acid sequence of an antibody and the amino acid sequence of a heterologous polypeptide or protein (i.e., a polypeptide or protein that is not normally a part of an antibody (e.g., a non-anti-hOX40L antigen antibody)). The term "fusion," when used in reference to hOX40L or an anti-hOX40L antibody, refers to the conjugation of a peptide or polypeptide, or a fragment, variant, and / or derivative thereof, with a heterologous peptide or polypeptide. Preferably, the fusion protein retains the biological activity of the hOX40L or anti-hOX40L antibody. In certain embodiments, the fusion protein comprises a hOX40L antibody VH domain, a VL domain, a VH CDR (one, two, or three VH CDRs), and / or a VL CDR (one, two, or three VL CDRs), and the fusion protein specifically binds to an hOX40L epitope.

[0348] The term "heavy chain," when used in reference to antibodies, refers to five different types, designated alpha (α), delta (δ), epsilon (ε), gamma (γ), and mu (μ), based on the amino acid sequence of the heavy chain constant domain. These different heavy chain types are well known and give rise to five classes of antibodies: IgA, IgD, IgE, IgG, and IgM, which include the four subclasses of IgG: IgG1, IgG1, IgG3, and IgG4, respectively. The heavy chain is preferably a human heavy chain. In one example, the heavy chain is a neutralized IgG isotype, e.g., neutralized IgG4. In certain embodiments, the antibody of the invention comprises a human gamma 4 constant region. In another embodiment, the heavy chain constant region does not bind to an Fc-γ receptor and comprises, for example, a Leu235Glu mutation. In another embodiment, the heavy chain constant region comprises a Ser228Pro mutation to increase stability. In another embodiment, the heavy chain constant region is IgG4-PE.

[0349] The term "host", as used herein, refers to an animal, preferably a mammal, and most preferably a human.

[0350] The term "host cell," as used herein, refers to a particular subject cell transfected with a nucleic acid molecule and to the progeny or potential progeny of such a cell. The progeny of such a cell may not be identical to the parent cell transfected with the nucleic acid molecule due to possible mutations or environmental influences that occur later in life, or due to integration of the nucleic acid molecule into the host cell genome.

[0351] "Immunomodulatory agent," and variants thereof, including but not limited to immunomodulatory agents, as used herein, refers to an agent that modulates the host's immune system. In certain embodiments, an immunomodulatory agent is an immunosuppressant. In certain other embodiments, an immunomodulatory agent is an immunostimulatory agent. In accordance with the present invention, immunomodulatory agents used in the combination therapies of the present invention do not include anti-hOX40L antibodies or antigen-binding fragments. Immunomodulatory agents include, but are not limited to, small molecules, peptides, polypeptides, proteins, fusion proteins, antibodies, inorganic molecules, mimetics, and organic molecules.

[0352] As used herein, the term "in combination" in the context of administering other therapies refers to the use of more than one therapy.The use of the term "in combination" does not limit the order in which therapies are administered to a subject with an infectious disease.A first therapy can be administered before (e.g., 1 minute, 45 minutes, 30 minutes, 45 minutes, 1 hour, 2 hours, 4 hours, 6 hours, 12 hours, 24 hours, 48 ​​hours, 72 hours, 96 hours, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 8 weeks, or 12 weeks before), simultaneously with, or after (e.g., 1 minute, 45 minutes, 30 minutes, 45 minutes, 1 hour, 2 hours, 4 hours, 6 hours, 12 hours, 24 hours, 48 ​​hours, 72 hours, 96 hours, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 8 weeks, or 12 weeks after) the administration of a second therapy to a subject with, having, or susceptible to hOX40L-mediated disease. Any additional therapy may be administered in any order with other additional therapies. In certain embodiments, an antibody of the invention may be administered in combination with one or more therapies (e.g., a therapy other than an antibody of the invention currently being administered to prevent, treat, manage, and / or alleviate a hOX40L-mediated disease). Non-limiting examples of therapies that may be administered in combination with an antibody of the invention include an analgesic, an anesthetic, an antibiotic, or an immunomodulator, or any other agent listed in the United States Pharmacopeia and / or the Physician's Desk Reference.

[0353] An "isolated" or "purified" antibody is substantially free of cellular material or other contaminating proteins, e.g., from the cell or tissue source from which the antibody is derived, or, when chemically synthesized, substantially free of chemical precursors or other chemicals. The term "substantially free of cellular material" includes preparations of antibodies in which the antibody is separated from cellular components of the cells from which it is isolated or recombinantly produced. Thus, an antibody that is substantially free of cellular material includes preparations of antibodies having less than about 30%, 20%, 10%, or 5% (by dry weight) of heterologous proteins (also referred to herein as "contaminating proteins"). If the antibody is recombinantly produced, the antibody is also preferably substantially free of culture medium, i.e., culture medium comprises about 20%, 10%, or 5% of the volume of the protein preparation. If the antibody is produced by chemical synthesis, the antibody is preferably substantially free of chemical precursors or other chemicals, i.e., the antibody is separated from chemical precursors or other chemicals involved in protein synthesis. Accordingly, such preparations of antibodies have less than about 30%, 20%, 10%, 5% (by dry weight) of chemical precursors or compounds other than the antibody of interest. In preferred embodiments, antibodies of the invention are isolated or purified.

[0354] An "isolated" nucleic acid molecule is one that is separated from other nucleic acid molecules that are present in the natural source of the nucleic acid molecule. Furthermore, an "isolated" nucleic acid molecule, such as a cDNA molecule, may be substantially free of other cellular material, or culture medium if produced by recombinant techniques, or substantially free of chemical precursors or other chemicals if chemically synthesized. In certain embodiments, nucleic acid molecule(s) encoding an antibody of the invention are isolated or purified.

[0355] "Human OX40L," "hOX40L," or "hOX40L polypeptide," and like terms, refer to polypeptides ("polypeptide," "peptide," and "protein" are used interchangeably herein) that include the amino acid sequences in the sequence listing and related polypeptides, including SNP variants thereof. Related polypeptides include allelic variants (e.g., SNP variants); splice variants; fragments; derivatives; substitution, deletion, and insertion variants; fusion polypeptides; and interspecies homologs that preferably retain hOX40L activity and / or are sufficient to generate an anti-hOX40L immune response. Also encompassed are soluble forms of hOX40L that are sufficient to generate an anti-hOX40L immune response. As will be understood by those skilled in the art, the anti-hOX40L antibodies of the present invention may bind to hOX40L polypeptides, polypeptide fragments, antigens, and / or epitopes because the epitope is part of a larger antigen, which is part of a larger polypeptide fragment, which in turn is part of a larger polypeptide in which hOX40L can exist in a trimeric (native) or monomeric (denatured) form.

[0356] The term "Kabat numbering" and similar terms are art-recognized and refer to a numbering system for amino acid residues that are more variable (i.e., hypervariable) than other amino acid residues in an antibody heavy chain variable region, or antigen-binding portion thereof (Kabat et al. (1971) Ann. NY Acad. Sci. 190:382-391, and Kabat et al. (1991) Sequences of Proteins of (Immunological Interest, Fifth Edition, US Department of Health and Human Services, NIH Publication No. 91-3242). For the heavy chain variable region, the hypervariable region typically ranges from amino acid positions 31 to 35 for CDR1, amino acid positions 50 to 65 for CDR2, and amino acid positions 95 to 102 for CDR3.

[0357] The term "monoclonal antibody" refers to an antibody obtained from a population of homogeneous or substantially homogeneous antibodies, with each monoclonal antibody typically recognizing a single epitope on an antigen. In a preferred embodiment, a "monoclonal antibody," as used herein, is an antibody produced by a single hybridoma or other cell, which specifically binds only to the hOX40L epitope, as determined, for example, by ELISA or other antigen-binding or competitive binding assays known in the art or in the Examples provided herein. The term "monoclonal" is not limited to any particular method for producing the antibody. For example, the monoclonal antibodies of the present invention may be produced by hybridoma methods as described in Kohler et al.; Nature, 256:495 (1975), or may be isolated from a phage library, for example, using the techniques described herein. Other methods for preparing clonal cell lines and the monoclonal antibodies they express are known in the art (see, e.g., Chapter 11 in Short Protocols in Molecular Biology, (2002) 5th Ed., Ausubel et al., eds., John Wiley and Sons, New York). Other exemplary methods for producing other monoclonal antibodies are provided in the Examples herein.

[0358] The terms "native" or "natural" when used in reference to biological material, e.g., nucleic acid molecules, polypeptides, host cells, etc., refer to something that is found in nature and has not been manipulated by man.

[0359] The term "pharmaceutically acceptable," as used herein, refers to approved by a federal or state regulatory agency or listed in the United States Pharmacopoeia, the European Pharmacopoeia, or other generally recognized pharmacopeia for use in animals, and more particularly in humans.

[0360] "Polyclonal antibody," as used herein, refers to an antibody population generated in an immune response to a protein with many epitopes, and thus includes a variety of different antibodies directed thereto, or a variety of different antibodies directed to different epitopes in the protein. Methods for producing polyclonal antibodies are known in the art (e.g., Short Protocols in Molecular Biology, (2002) 5th Ed., Ausubel et al., eds., John Wiley and Sons, New York, see Chapter 11).

[0361] As used herein, the terms "polynucleotide," "nucleotide," "nucleic acid," "nucleic acid molecule," and other similar terms are used interchangeably and include DNA, RNA, mRNA, and the like.

[0362] As used herein, "prevent," "preventing," and "prevention" refer to the complete or partial inhibition of the onset, recurrence, development, or metastasis of an hOX40L-mediated disease and / or symptoms thereof resulting from the administration of a therapy or combination of therapies provided herein (e.g., a combination of prophylactic or therapeutic agents, such as an antibody of the invention).

[0363] As used herein, the term "prophylactic agent" refers to any agent that can completely or partially inhibit the onset, recurrence, development, or metastasis of an hOX40L-mediated disease and / or symptoms thereof in a subject. In certain embodiments, the term "prophylactic agent" refers to an antibody of the present invention. In certain other embodiments, the term "prophylactic agent" refers to an agent other than an antibody of the present invention. Preferably, a prophylactic agent is an agent that is known to be useful for, or has been used or is currently being used to, prevent an hOX40L-mediated disease and / or symptoms thereof, or hinder the onset, development, progression, and / or severity of an hOX40L-mediated disease and / or symptoms thereof. In certain embodiments, a prophylactic agent is a fully human anti-hOX40L antibody, such as a fully human anti-hOX40L monoclonal antibody.

[0364] In one embodiment, prevention prevents the onset of a disease or condition, or symptoms of a disease or condition. In one embodiment, prophylactic treatment prevents the worsening or onset of a disease or condition. In one embodiment, prophylactic treatment prevents the worsening of a disease or condition.

[0365] In another embodiment, an anti-OX40L antibody of the invention is administered intravenously (e.g., prior to or concurrent with a transplant, e.g., blood or organ transplant). In another embodiment, the antibody is administered at a dose of about 5-10 mg / kg (e.g., about 8 mg / kg). In another embodiment, the antibody is administered at a dose selected from about 0.1 mg / kg, about 0.5 mg / kg, about 1 mg / kg, 3 mg / kg, 5 mg / kg, about 10 mg / kg, about 15 mg / kg, about 20 mg / kg, about 25 mg / kg, about 30 mg / kg, about 40 mg / kg, about 50 mg / kg, about 60 mg / kg, about 70 mg / kg, about 80 mg / kg, about 90 mg / kg, or about 100 mg / kg, particularly about 1 mg / kg or about 3 mg / kg.

[0366] In another embodiment, the antibody is administered 1-4 days prior to (e.g., blood or organ) transplant, e.g., 1-3 days prior to transplant, or 1-2 days prior to transplant. In another embodiment, the antibody is administered weekly, biweekly, or monthly, e.g., every other week, following transplant. In a further embodiment, the antibody is administered prophylactically intravenously at a dose of about 5-10 mg / kg (e.g., about 8 mg / kg) 1-3 days prior to transplant, and then intravenously at a dose of about 5-10 mg / kg (e.g., about 8 mg / kg) every other week thereafter.

[0367] In another embodiment, patients are monitored periodically after transplant for the presence of biomarkers predictive of graft rejection or the development of GvHD (e.g., acute GvHD), and when the biomarker levels reach a level at which the patient is determined to be at risk for graft rejection or the development of GvHD (e.g., acute GvHD), an anti-OX40L antibody of the invention is administered. This strategy may avoid unnecessary administration of drugs and unnecessary suppression of the immune system. Examples of biomarkers that may be useful as predictive biomarkers for acute GvHD are described in Levine et al., "A prognostic score for acute graft-versus-host disease based on biomarkers: These biomarkers may be those identified in "A multicentre study," Lancet Haematol 2015;2:e21-29. These biomarkers include, but are not limited to, TNFR1, ST-2, elafin, and IL2Rα and Reg3α.

[0368] The region of hOX40L that contributes to an epitope may be adjacent amino acids of a polypeptide, or the epitope may simultaneously arise from two or more non-adjacent regions of a polypeptide. An epitope may or may not be a three-dimensional surface feature of an antigen. A region located on the surface of a hOX40L antigen that can elicit an immune response is an hOX40L epitope. An epitope may or may not be a three-dimensional surface feature of an antigen.

[0369] "hOX40L-mediated disease" and "hOX40L-mediated condition" are used interchangeably and refer to any disease or condition that is caused, in whole or in part, by or is a result of hOX40L. In certain embodiments, hOX40L is abnormally (e.g., highly) expressed on the surface of a cell. In some embodiments, hOX40L may be abnormally upregulated in a particular cell type. In other embodiments, normal, abnormal, or excessive cell signaling is caused by binding of hOX40L to an hOX40L ligand. In certain embodiments, the hOX40L ligand is OX40 expressed on the surface of a cell, such as, for example, a colonic epithelial cell. In certain embodiments, the hOX40L-mediated disease is inflammatory bowel disease (IBD), such as Crohn's disease (CD) or ulcerative colitis (UC). In other embodiments, the hOX40L-mediated disease is graft-versus-host disease (GVHD). In other embodiments, the hOX40L-mediated disease is selected from pyoderma gangrenosum, giant cell arteritis, Schnitzler's syndrome, non-infectious scleritis, and uveitis (non-infectious / autoimmune and / or systemic). In other embodiments, the hOX40L-mediated disease or condition is selected from an autoimmune disease or condition, a systemic inflammatory disease or condition, or transplant rejection, such as inflammatory bowel disease (IBD), Crohn's disease, rheumatoid arthritis, transplant rejection, allograft rejection, graft-versus-host disease (GvHD), ulcerative colitis, systemic lupus erythematosus (SLE), diabetes, uveitis, ankylosing spondylitis, contact hypersensitivity, multiple sclerosis, and atherosclerosis, particularly GvHD.

[0370] "hOX40L receptor" or "hOX40L-binding receptor" are used interchangeably herein and refer to a receptor polypeptide that binds to hOX40L. In certain embodiments, the hOX40L receptor is Hox40. In some embodiments, the hOX40L receptor is expressed on the surface of cells, such as colonic epithelial cells, or on a graft or transplanted tissue, or on host tissue.

[0371] As used herein, the terms "subject" and "patient" are used interchangeably. As used herein, a subject is preferably a mammal, such as a non-primate (e.g., cows, pigs, horses, cats, dogs, rats, etc.) or a primate (e.g., monkeys and humans), most preferably a human. In one embodiment, the subject is a mammal, preferably a human, having an hOX40L-mediated disease. In another embodiment, the subject is a mammal, preferably a human, at risk of developing an hOX40L-mediated disease.

[0372] As used herein, "substantially all" refers to at least about 60%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 98%, at least about 99%, or about 100%.

[0373] The term "substantially free of detergent," as used herein, refers to a preparation of an antibody that specifically binds to the hOX40L antigen, which contains less than 0.0005%, less than 0.0003%, or less than 0.0001% detergent, and / or less than 0.0005%, less than 0.0003%, or less than 0.0001% detergent.

[0374] The term "substantially free of salts," as used herein, refers to a preparation of an antibody that specifically binds to the hOX40L antigen, which contains less than 0.0005%, less than 0.0003%, or less than 0.0001% inorganic salts.

[0375] The term "surfactant," as used herein, refers to organic substances with amphiphilic structures, i.e., they are composed of groups with opposite solubility tendencies, typically an oil-soluble hydrocarbon chain and a water-soluble ionic group. Depending on the charge of the surface-active moiety, surfactants can be classified as anionic, cationic, and nonionic surfactants. Surfactants are often used as wetting agents, emulsifiers, solubilizers, and dispersants in the preparation of various pharmaceutical compositions and biological materials.

[0376] As used herein, the term "tag" refers to any type of moiety attached to a polypeptide and / or polynucleotide, e.g., encoding hOX40L or an hOX40L antibody or antigen-binding fragment thereof. For example, a polynucleotide encoding hOX40L, an hOX40L antibody, or an antigen-binding fragment thereof can include one or more additional tag-encoding nucleotide sequences, e.g., encoding a detectable moiety or a moiety useful for affinity purification. Upon translation, the tag and antibody can be in the form of a fusion protein. The terms "detectable" or "detection" with respect to a tag refer to any tag that can be visualized or otherwise the presence of the tag can be determined and / or measured (e.g., by quantification). A non-limiting example of a detectable tag is a fluorescent tag.

[0377] As used herein, the term "therapeutic agent" refers to any agent that can be used in the treatment, management, or alleviation of an hOX40L-mediated disease and / or symptoms thereof. In certain embodiments, the term "therapeutic agent" refers to an antibody of the invention. In certain other embodiments, the term "therapeutic agent" refers to an agent other than an antibody of the invention. Preferably, a therapeutic agent is an agent that is useful for, has been used, or is currently being used to treat, manage, or alleviate an hOX40L-mediated disease or one or more symptoms thereof. In certain embodiments, a therapeutic agent is a fully human anti-hOX40L antibody, such as a fully human anti-hOX40L monoclonal antibody.

[0378] A combination of therapies (e.g., the use of prophylactic or therapeutic agents) that is more effective than the additive effects of any two or more monotherapies. For example, the synergistic effect of a combination of prophylactic and / or therapeutic agents allows for the use of lower doses of one or more of the agents and / or less frequent administration of the agent to a subject with an hOX40L-mediated disease. The ability to use lower doses of prophylactic or therapeutic therapies and / or administer the therapies less frequently reduces the toxicity associated with administering the therapies to a subject without reducing the efficacy of the therapies in preventing, managing, treating, or alleviating hOX40L-mediated diseases. In addition, a synergistic effect can lead to improved efficacy of therapies in preventing, or in managing, treating, or alleviating hOX40L-mediated diseases. Finally, the synergistic effect of a combination of therapies (e.g., prophylactic or therapeutic agents) can avoid or reduce adverse or unwanted side effects associated with the use of any monotherapies.

[0379] In one embodiment, the combination includes an anti-OX40L antibody of the invention and, independently, rapamycin (sirolimus), lacrolimus, cyclosporine, corticosteroids (e.g., methylprednisolone), methotrexate, mycophenolate mofetil, anti-CD28 antibodies, anti-IL12 / IL-23 antibodies (e.g., ustekinumab), anti-CD20 antibodies (e.g., rituximab), anti-CD30 antibodies (e.g., brentuximab), CTLA4-Fc molecules (e.g., abatacept), CCR5 receptor antagonists (e.g., maraviroc), anti-CD40L antibodies, anti-VLA4 antibodies (e.g., natalizumab), anti-LFA and an additional therapeutic agent selected from the group consisting of an anti-CD1 antibody, fludarabine, an anti-CD52 antibody (e.g., alemtuzumab), an anti-CD45 antibody, cyclophosphamide, anti-thymocyte globulin, an anti-complement C5 antibody (e.g., eculizumab), an anti-a4b7 integrin antibody (e.g., vedolizumab), an anti-IL6 antibody (e.g., tocilizumab), an anti-IL2R antibody (e.g., basilixumab), an anti-CD25 antibody (e.g., daclizumab), an anti-TNFα / TNFα-Fc molecule (e.g., etanercept, adalimumab, infliximab, golimumab, or cetolizumab pegol), and vorinostat. In another embodiment, the combination comprises an anti-OX40L antibody of the invention and an additional therapeutic agent independently selected from the group consisting of rapamycin (sirolimus), lacrolimus, cyclosporine, corticosteroids (e.g., methylprednisolone), methotrexate, mycophenolate mofetil, anti-CD28 antibodies, CTLA4-Fc molecules (e.g., abatacept), anti-CD40L antibodies, anti-LFA1 antibodies, anti-CD52 antibodies (e.g., alemtuzumab), cyclophosphamide, and anti-thymocyte globulin.

[0380] As used herein, the term "therapy" refers to any protocol, method, and / or agent that can be used to prevent, manage, treat, and / or alleviate an hOX40L-mediated disease (e.g., IBD or GVHD). In certain embodiments, the terms "therapy" and "therapy" refer to biological, supportive, and / or other therapies known to those of skill in the art, such as healthcare professionals, that are useful in preventing, managing, treating, and / or alleviating an hOX40L-mediated disease.

[0381] As used herein, the terms "treat," "treatment," and "treating" refer to the reduction or alleviation of the progression, severity, and / or duration of an hOX40L-mediated disease (e.g., IBD or GVHD) resulting from the administration of one or more therapies (including, but not limited to, one or more prophylactic or therapeutic agents, such as an antibody of the invention). In certain embodiments, such terms refer to the reduction or inhibition of binding of hOX40L to OX40, the reduction or inhibition of CCL20 production or secretion from cells expressing hOX40 or hOX40L, the reduction or inhibition of IL-8 production or secretion from cells expressing hOX40 or hOX40L, the reduction or inhibition of RANTES production or secretion from cells expressing hOX40 or hOX40L, and / or the inhibition or reduction of one or more symptoms associated with an hOX40L-mediated disease, such as IBD or GVHD. In certain embodiments, such terms refer to reducing or inhibiting the binding of hOX40L to OX40, reducing or inhibiting the production or secretion of INF-γ from cells expressing hOX40 or hOX40L, reducing or inhibiting the production or secretion of TNF-α from cells expressing hOX40 or hOX40L, reducing or inhibiting the production or secretion of IL-2 from cells expressing hOX40 or hOX40L, and / or inhibiting or reducing one or more symptoms associated with an hOX40L-mediated disease, such as IBD or GVHD (particularly GvHD). In one example, the cell is a human cell. In certain embodiments, the prophylactic agent is a fully human anti-hOX40L antibody, such as a fully human anti-hOX40L monoclonal antibody.

[0382] "Variable region" or "variable domain" refers to OX40L and a portion of the heavy chain, typically the amino-terminal approximately 120-130 amino acids in the heavy chain and approximately 100-110 amino acids in the light chain, which vary significantly in sequence among antibodies and are responsible for the binding and specificity of each particular antibody to its particular antigen. Sequence variation is concentrated in regions called complementarity-determining regions (CDRs), while the more highly conserved regions in the variable domain are called framework regions (FRs). The CDRs of OX40L and the heavy chain are primarily responsible for the antibody's interaction with antigen. Amino acid position numbering follows the EU Index as in Kabat et al. (1991) Sequences of proteins of immunological interest. (US Department of Health and Human Services, Washington, DC) 5th ed. ("Kabat et al."). In a preferred embodiment, the variable region is a human variable region.

[0383] antibody Antibodies of the present invention include, but are not limited to, synthetic antibodies, monoclonal antibodies, recombinantly produced antibodies, multispecific antibodies (including bispecific antibodies), human antibodies, humanized antibodies, chimeric antibodies, intrabodies, single-chain Fvs (scFv) (including, e.g., monospecific, bispecific, etc.), camelized antibodies, Fab fragments, F(ab') fragments, disulfide-linked Fvs (sdFv), anti-idiotypic (anti-Id) antibodies, and epitope-binding fragments of any of the above.

[0384] In particular, the antibodies provided herein include immunoglobulin molecules and immunologically active portions of immunoglobulin molecules, i.e., molecules comprising an antigen-binding site that specifically binds to the hOX40L antigen. The immunoglobulin molecules provided herein can be of any type (e.g., IgG, IgE, IgM, IgD, IgA, and IgY), class (e.g., IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2), or subclass of immunoglobulin molecule. In certain embodiments, the antibodies provided herein are IgG antibodies, preferably IgG1 or IgG4. In certain embodiments, the antibodies of the present invention comprise a human gamma 4 constant region. In another embodiment, the heavy chain constant region does not bind to an Fc-gamma receptor and comprises, for example, a Leu235Glu mutation. In another embodiment, the heavy chain constant region comprises a Ser228Pro mutation to increase stability. In another embodiment, the heavy chain constant region is IgG4-PE.

[0385] Antibody variants and derivatives include antibody fragments that retain the ability to specifically bind to an epitope. Preferred fragments include Fab fragments; Fab' (an antibody fragment containing a single anti-binding domain comprising an Fab and an additional portion of a heavy chain through the hinge region); F(ab')2 (two Fab' molecules joined by an interchain disulfide bond at the hinge region of the heavy chain; the Fab' molecules can be directed against the same or different epitopes); bispecific Fab (a Fab molecule with two antigen-binding domains, each of which can be directed against a different epitope); and single-chain Fabs containing the variable region, also known as sFv. Fab chains; disulfide-linked Fvs, or dsFvs; camelized VHs of a single heavy chain of an antibody (the variable antigen-binding determining regions, in which some amino acids at the VH interface are found in the heavy chain of a native camelid antibody); bispecific sFvs (sFv or dsFv molecules with two antigen-binding domains that can each be directed against a different epitope); diabodies (the VH domain of one sFv assembles with the VL domain of a second sFv, and the VL domain of the first sFv binds to the VH domain of the second sFv). Examples of antibodies include dimerized sFvs, which are formed when an sFv assembles with a diabody; the two antigen-binding regions of the diabody can be directed against the same or different epitopes), and triabodies (trimerized sFvs, which are similar to diabodies, but in which three antigen-binding domains are created in a single complex, and these three antigen-binding domains can be directed against the same or different epitopes). Derivatives of antibodies also include one or more CDR sequences of the antibody binding site. The CDR sequences can be linked together on a scaffold when more than one CDR sequence is present. In certain embodiments, antibodies used with the present invention comprise single-chain Fvs ("scFvs"). scFvs are antibody fragments comprising the VH and VL domains of an antibody, where these domains are present in a single polypeptide chain. Generally, the scFv polypeptide further comprises a polypeptide linker between the VH and VL domains, which enables the scFv to form the desired structure for antigen binding.For a review of scFvs, see Pluckthun in The Pharmacology of Monoclonal Antibodies, vol. 113, Rosenburg and Moore eds. Springer-Verlag, New York, pp. 269-315 (1994).

[0386] Antibodies of the invention may be from any animal origin, including birds and mammals (e.g., human, murine, donkey, sheep, rabbit, goat, guinea pig, camel, horse, or chicken). In certain embodiments, antibodies of the invention are human or humanized monoclonal antibodies. As used herein, "human" antibodies include antibodies having the amino acid sequence of a human immunoglobulin, including antibodies isolated from a human immunoglobulin library or from a mouse expressing antibodies from human genes.

[0387] In a preferred embodiment, the antibody of the present invention is a fully human antibody, such as an hOX40L polypeptide, an hOX40L polypeptide fragment, or a fully human antibody that specifically binds to an hOX40L epitope. Such fully human antibodies are advantageous over fully murine (or other fully or partially non-human species antibodies), humanized, or chimeric antibodies, in that they minimize the occurrence of unwanted or undesired side effects, such as immune responses directed against non-fully human antibodies (e.g., anti-hOX40L antibodies derived from other species), upon administration to a subject.

[0388] The antibodies of the present invention may be monospecific, bispecific, trispecific, or of greater multispecificity. Multispecific antibodies may be specific for different epitopes of a hOX40L polypeptide, or may be specific for both a hOX40L polypeptide and a heterologous epitope, such as a heterologous polypeptide or solid support material. In a preferred embodiment, the antibodies provided herein are monospecific for a given epitope of a hOX40L polypeptide and do not specifically bind to other epitopes.

[0389] Also provided herein are B cells (eg, immortalized B cells) or hybridomas that produce the anti-hOX40L antibodies or fragments described herein.

[0390] In certain embodiments, provided herein are isolated antibodies that specifically bind to an hOX40L epitope, wherein binding by the antibody to the hOX40L epitope is competitively blocked (e.g., in a dose-dependent manner) by an antibody or fragment of the invention. The antibody may or may not be a fully human antibody. In preferred embodiments, the antibody is a fully human monoclonal anti-hOX40L antibody, and even more preferably a fully human monoclonal antagonistic anti-hOX40L antibody. An exemplary competitive blocking assay that can be used is provided in the Examples herein.

[0391] In some embodiments, antibodies or fragments of the invention compete with the OX40 receptor (or a fusion protein thereof) for binding to cell-surface-expressed hOX40L (e.g., in a dose-dependent manner). In other embodiments, antibodies or fragments of the invention compete with the OX40 receptor (or a fusion protein thereof) for binding to soluble hOX40L (e.g., in a dose-dependent manner). Exemplary competitive binding assays that can be used are provided in the Examples herein. In one embodiment, the antibody or fragment partially or completely inhibits binding of hOX40 to cell-surface-expressed OX40L, such as hOX40L. In another embodiment, the antibody partially or completely inhibits binding of hOX40 to soluble hOX40L. In some embodiments, the antibody or fragment partially or completely inhibits secretion of CCL20, IL-8, and / or RANTES, or INF-γ, TNF-α, or IL-2, particularly INF-γ, from cells having cell-surface-expressed OX40. In certain embodiments, the OX40-expressing cells are colonic epithelial cells.

[0392] Preferably, the antibodies of the invention are fully human monoclonal antibodies, such as fully human monoclonal antagonistic antibodies that specifically bind to hOX40L.

[0393] In some embodiments, the antibodies or fragments provided herein bind to a hOX40L epitope, which is a three-dimensional surface feature of a hOX40L polypeptide (e.g., in a trimeric form of a hOX40L polypeptide). The region of a hOX40L polypeptide contributing to the epitope may be contiguous amino acids of the polypeptide, or the epitope may arise simultaneously from two or more non-contiguous regions of the polypeptide. A hOX40L epitope may be present in (a) a trimeric form of hOX40L (a "trimeric hOX40L epitope"), (b) a monomeric form of hOX40L (a "monomeric hOX40L epitope"), (c) both the trimeric and monomeric forms of hOX40L, (d) the trimeric but not the monomeric form of hOX40L, or (e) the monomeric but not the trimeric form of hOX40L.

[0394] For example, in some embodiments, the epitope is present or available for binding in the monomeric (native) form, but not present or available for binding in the monomeric (denatured) form by an anti-hOX40L antibody. In other embodiments, the hOX40L epitope is a linear feature of the hOX40L polypeptide (e.g., a trimeric or monomeric form of the hOX40L polypeptide). The antibodies provided herein may specifically bind to (a) an epitope in the monomeric form of hOX40L, (b) an epitope in the trimeric form of hOX40L, (c) an epitope in the monomeric but not trimeric form of hOX40L, (d) an epitope in the trimeric but not monomeric form of hOX40L, or (e) both the monomeric and trimeric forms of hOX40L. In preferred embodiments, the antibodies provided herein specifically bind to an epitope in the trimeric form of hOX40L, but not to an epitope in the monomeric form of hOX40L.

[0395] The present invention also provides antibodies that specifically bind to the hOX40L epitope, including antibodies comprising derivatives of the VH domain, VH CDR, VL domain, and VL CDR described herein that specifically bind to the hOX40L antigen. The present invention also provides antibodies, including derivatives of the antibodies disclosed in the Examples, that specifically bind to the hOX40L epitope. Using standard techniques known to those skilled in the art, mutations, including site-directed mutagenesis and PCR-mediated mutagenesis, are introduced into the nucleotide sequence encoding the molecule of the present invention, resulting in amino acid substitutions. Preferably, the derivative contains fewer than 25 amino acid substitutions, fewer than 20 amino acid substitutions, fewer than 15 amino acid substitutions, fewer than 10 amino acid substitutions, fewer than 5 amino acid substitutions, fewer than 4 amino acid substitutions, fewer than 3 amino acid substitutions, or fewer than 2 amino acid substitutions relative to the original molecule. In another embodiment, the derivative contains conservative amino acid substitutions. In a preferred embodiment, the derivative contains conservative amino acid substitutions at one or more predicted non-essential amino acid residues. Alternatively, mutations can be introduced randomly along all or part of the coding sequence, such as by saturation mutagenesis, and the resulting mutants can be screened for biological activity to identify mutants that retain activity. After mutagenesis, the encoded protein can be expressed and the activity of the protein can be determined.

[0396] In another embodiment, an antibody that specifically binds to the hOX40L epitope comprises a variable domain amino acid sequence that is at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identical to the variable domain amino acid sequence of the sequence listing.

[0397] In certain embodiments, the antibody is a fully human anti-human antibody, such as a fully human monoclonal antibody. Fully human antibodies can be produced by any method known in the art. An exemplary method includes immunizing a transgenic animal (e.g., a mouse) that can produce a repertoire of human antibodies in the absence of endogenous immunoglobulin production with a hOX40L antigen (any hOX40L polypeptide that can elicit an immune response and is optionally complexed with a carrier). See, for example, Jakobovits et al., (1993) Proc. Natl. Acad. Sci., 90:2551; Jakobovits et al., (1993) Nature, 362:255 258 (1993); Bruggermann et al., (1993) Year in Immunol., 7:33. Other methods for producing fully human anti-hOX40L antibodies can be found in the examples provided herein.

[0398] Alternatively, fully human antibodies can be generated through in vitro screening of phage display antibody libraries. See, for example, Hoogenboom et al., J. Mol. Biol., 227:381 (1991); Marks et al., J. Mol. Biol., 222:581 (1991), which are incorporated herein by reference. Phage display libraries containing various antibodies have been described and can be readily prepared by those skilled in the art. Libraries can contain diverse human antibody sequences, such as human Fab, Fv, and scFv fragments, which can be screened against appropriate targets.

[0399] The antibodies and fragments of the present invention include antibodies and fragments that are chemically modified, i.e., modified by the covalent attachment of any type of molecule to the antibody. For example, and without limitation, antibody derivatives include antibodies that have been chemically modified by, for example, glycosylation, acetylation, pegylation, phosphorylation, amidation, derivatization with known protecting / blocking groups, proteolytic cleavage, linkage to cellular ligands or other proteins, etc. Any of a number of chemical modifications can be carried out by known techniques, including, but not limited to, specific chemical cleavage, acetylation, formulation, metabolic synthesis of tunicamycin, etc. In addition, the antibody may contain one or more non-classical amino acids.

[0400] The present invention also provides antibodies (e.g., human or non-human fragments) that specifically bind to the hOX40L antigen, comprising framework regions known to those skilled in the art. The framework regions may be, for example, native or consensus framework regions. Most preferably, the framework regions of the antibodies of the present invention are human (e.g., for a list of human framework regions, see Chothia et al., 1998, J. Mol. Biol. 278:457-479, the entire contents of which are incorporated herein by reference). See also Kabat et al. (1991) Sequences of Proteins of Immunological Interest (US Department of Health and Human Services, Washington, DC) 5th ed.

[0401] In certain embodiments, the present invention provides antibodies that specifically bind to the hOX40L antigen, wherein the antibodies have the amino acid sequence of one or more of the CDRs in the sequence listing (i.e., SEQ ID NO:4, SEQ ID NO:10, SEQ ID NO:36, SEQ ID NO:42, SEQ ID NO:68, SEQ ID NO:74, SEQ ID NO:96, or SEQ ID NO:102, particularly for HCDR1, SEQ ID NO:36 or SEQ ID NO:42; for HCDR2, SEQ ID NO:6, SEQ ID NO:12, SEQ ID NO:38, SEQ ID NO:44, SEQ ID NO:70, SEQ ID NO:76, SEQ ID NO:98, or SEQ ID NO:104, particularly SEQ ID NO:38 or or SEQ ID NO:44; for HCDR3, SEQ ID NO:8, SEQ ID NO:14, SEQ ID NO:40, SEQ ID NO:46, SEQ ID NO:72, SEQ ID NO:78, SEQ ID NO:100, or SEQ ID NO:106, in particular SEQ ID NO:40 or SEQ ID NO:46; for LCDR1, SEQ ID NO:18, SEQ ID NO:24, SEQ ID NO:50, SEQ ID NO:56, SEQ ID NO:82, SEQ ID NO:88, SEQ ID NO:110, or SEQ ID NO:116, in particular SEQ ID NO:50 or SEQ ID NO:56; for LCDR2, SEQ ID NO:20, SEQ ID NO:26, SEQ ID NO:52, SEQ ID NO:58, SEQ ID NO:84, SEQ ID NO:90, SEQ ID NO:112, or SEQ ID NO:118, particularly SEQ ID NO:52 or SEQ ID NO:58; and for LCDR3, SEQ ID NO:22, SEQ ID NO:28, SEQ ID NO:54, SEQ ID NO:60, SEQ ID NO:86, SEQ ID NO:92, SEQ ID NO:114, or SEQ ID NO:120, particularly SEQ ID NO:54 or SEQ ID NO:60), as well as the following residues: (a) rare framework residues that differ between the mouse antibody framework (i.e., the donor antibody framework) and the human antibody framework (i.e., the acceptor antibody framework), (b) vernier zone residues when they differ between the donor antibody framework and the acceptor antibody framework; (c) interchain packing residues at the VH / VL interface that differ between the donor antibody framework and the acceptor antibody framework; (d) canonical residues that differ between the donor antibody framework sequence and the acceptor antibody framework sequence (particularly the framework regions essential for defining the canonical classes of mouse antibody CDR loops); (e) residues adjacent to the CDR; (g) residues that can interact with antigen; (h) residues that can interact with the CDR;and (i) human framework regions with one or more amino acid substitutions at one, two, three, or more of the contact residues between the VH and VL domains. In certain embodiments, an antibody that specifically binds to the hOX40L antigen and comprises human framework regions with one or more amino acid substitutions at one, two, three, or more of the above-identified residues is an antagonistic hOX40L antibody;

[0402] The present invention encompasses antibodies that specifically bind to the hOX40L antigen, which comprise the amino acid sequence of the VH domain and / or VL domain in the Sequence Listings (i.e., for the VH domain, SEQ ID NO:2, SEQ ID NO:34, SEQ ID NO:66, or SEQ ID NO:94, particularly SEQ ID NO:34; for the VL domain, SEQ ID NO:16, SEQ ID NO:48, SEQ ID NO:80, or SEQ ID NO:108, particularly SEQ ID NO:48), but with mutations (e.g., one or more amino acid substitutions) in the framework regions. In certain embodiments, antibodies that specifically bind to the hOX40L antigen comprise the amino acid sequence of the VH domain and / or VL domain of an antibody disclosed in the Examples, or an antigen-binding fragment thereof, with one or more amino acid residue substitutions in the framework regions of the VH domain and / or VL domain.

[0403] In some embodiments, the antibodies provided herein reduce or inhibit hOX40L binding to hOX40 and / or reduce or inhibit hOX40L biological activity, such as the secretion of CCL20, IL8, and / or RANTES, or INF-γ, TNF-α, or IL-2, particularly INF-γ, in a subject (e.g., a human subject). In certain embodiments, the antibodies provided herein, such as human monoclonal anti-hOX40L antibodies, reduce or inhibit the binding of soluble or cell-surface-expressed hOX40L to hOX40 and / or reduce or inhibit the secretion of CCL20 and / or RANTES, or INF-γ, TNF-α, or IL-2, particularly INF-γ, in a subject following contact with soluble or cell-surface-expressed hOX40L. The blocking activity of the antibodies provided herein for hOX40L binding to hOX40 can be detected using the assays described in the Examples. Inhibition of the biological activity of cells expressing OX40 by the hOX40L antibodies provided herein can be detected using the assays described in the Examples.

[0404] The present invention also provides fusion proteins comprising an antibody provided herein that specifically binds to the hOX40L antigen and a heterologous polypeptide. In some embodiments, the heterologous polypeptide to which the antibody is fused is useful for targeting cells that have hOX40L expressed on their surface.

[0405] Antibody Conjugates and Fusion Proteins The following discussion of conjugates and fusion proteins also applies to fragments, so that disclosures referring to antibodies may be applied mutatis mutandis to fragments of the invention.

[0406] In some embodiments, antibodies of the invention are conjugated or recombinantly fused to a diagnostic, detectable, or therapeutic agent, or any other molecule. Conjugated or recombinantly fused antibodies may be useful for monitoring or prognosing the onset, development, progression, and / or severity of hOX40L-mediated diseases, for example, as part of a clinical trial procedure, such as determining the efficacy of a particular therapy.

[0407] Such diagnosis and detection can involve, for example, coupling antibodies to various enzymes, such as, but not limited to, horseradish peroxidase, alkaline phosphatase, beta-galactosidase, or acetylcholinesterase; prosthetic groups, such as, but not limited to, streptavidin / biotin and avidin / biotin; fluorescent materials, such as, but not limited to, umbelliferone, fluorescein, fluorescein isothiocynate, rhodamine, dichlorotriazinylamine fluorescein, dansyl chloride, or phycoerythrin; luminescent materials, such as, but not limited to, luminol; bioluminescent materials, such as, but not limited to, luciferase, luciferin, and aequorin; radioactive materials, such as, but not limited to, iodine ( 131 I, 125 I, 123 I, and 121 I), carbon ( 14 C), sulfur ( 35 S), tritium ( 3 H), indium ( 115 In, 113 In, 112 In, and 111 In), technetium ( 99 Tc), thallium ( 201 Ti), Gallium ( 68 Ga, 67 Ga), palladium ( 103 Pd), molybdenum ( 99 Mo), xenon ( 133 Xe), fluorine ( 18 F), 153 Sm, 177 Lu, 159 Gd, 149 Pm, 140La, 175 Yb, 166 Ho, 90 Y, 47 Sc, 186 Re, 188 Re, 142 Pr, 105 Rh, 97 Ru, 68 Ge, 57 Co, 65 Zn, 85 Sr, 32 P, 153 Gd, 169 Yb, 51 Cr, 54 Mn, 75 Se, 113 Sn, and 117 Sn; as well as positron-emitting metals using various positron emission tomography techniques, and non-radioactive paramagnetic metal ions.

[0408] The present invention further encompasses the use of antibodies of the invention that are conjugated or recombinantly fused to a therapeutic moiety (or more than one therapeutic moiety). The antibody may be conjugated or recombinantly fused to a therapeutic moiety, such as a cytotoxin, e.g., a cytostatic or cytocidal agent, a therapeutic agent, or a radioactive metal ion, e.g., an alpha emitter. A cytotoxin or cytotoxic agent includes any agent that is detrimental to cells. Therapeutic moieties include antimetabolites (e.g., methotrexate, 6-mercaptopurine, 6-thioguanine, cytarabine, 5-fluorouracil dacarbazine); alkylating agents (e.g., mechlorethamine, thiotepa, chlorambucil); chlorambucil), melphalan, carmustine (BCNU) and lomustine (CCNU), cyclothosphamide, busulfan, dibromomannitol, streptozotocin, mitomycin C, and cis-dichlorodiamineplatinum(II) (DDP), and cisplatin; anthracyclines (e.g., daunorubicin (formerly daunomycin) and doxorubicin), antibiotics (e.g., dactinomycin (formerly actinomycin), bleomycin, mithramycin, and anthramycin (AMC)); auristatin molecules (e.g., auristatin PHE, bryostatin 1, and solastatin 10; Woyke et al., Antimicrob. Agents Chemother. 46:3802-8 (2002); Woyke et al., Antimicrob. Agents Chemother. 45:3580-4 (2001), Mohammad et al., Anticancer Drugs 12:735-40 (2001), Wall et al., Biochem. Biophys. Res. Commun 266:76-80 (1999), Mohammad et al., Int. J. Oncol. 15:367-72 (1999); hormones (e.g., glucocorticoids, progestins, androgens, and estrogens), DNA repair enzyme inhibitors (e.g., etoposide or topotecan), kinase inhibitors (e.g., compound ST1571, imatinib mesylate (Kantarjian et al., Clin Cancer Res. 8(7):2167-76 (2002)); cytotoxic agents (e.g., paclitaxel, cytochalasin B, gramicidin D, ethidium bromide, emetine, mitomycin, etoposide, tenoposide, vincristine, vinblastine, colchicine, doxorubicin, daunorubicin, dihydroxyanthracin dione, mitoxantrone, mithramycin, actinomycin D, 1-dehydrotestosterone, glucorticoids, procaine, tetracaine, lidocaine, propranolol, and puromycin, and analogs or homologs thereof, and U.S. Pat. No. 6,629,296; No. 245,759, No. 6,399,633, No. 6,383,790, No. 6,335,156, No. 6,271,242, No. 6,242, No. 196, No. 6,218,410, No. 6,218,372, No. 6,057,300, No. 6,034,053, No. 5,985,877 , 5,958,769, 5,925,376, 5,922,844, 5,911,995, 5,872,223, 5,863,904, 5,840,745, 5,728,868, 5,648,239, 5,587,459);Farnesyltransferase inhibitors (e.g., R115777, BMS-214662, and compounds described in, e.g., U.S. Patent Nos. 6,458,935, 6,451,812, 6,440,974, 6,436,960, 6,432,959, 6,420,387, 6,414,145, 6,410,541, 6,410,539, 6,403,581, 6,39 No. 9,615, No. 6,387,905, No. 6,372,747, No. 6,369,034, No. 6,362,188, No. 6,342,765, No. 6,342,487, No. 6,300,50 No. 1, No. 6,268,363, No. 6,265,422, No. 6,248,756, No. 6,239,140, ​​No. 6,232,338, No. 6,228,865, No. 6,228,856, No. 6 ,225,322, No. 6,218,406, No. 6,211,193, No. 6,187,786, No. 6,169,096, No. 6,159,984, No. 6,143,766, No. 6,133 ,303, No. 6,127,366, No. 6,124,465, No. 6,124,295, No. 6,103,723, No. 6,093,737, No. 6,090,948, No. 6,080,870 , 6,077,853, 6,071,935, 6,066,738, 6,063,930, 6,054,466, 6,051,582, 6,051,574, and 6,040,305); topoisomerase inhibitors (e.g., camptothecin; irinotecan; SN-38; topotecan; 9-aminocamptothecin; GG-211 (GI 147211); DX-8951f; IST-622; rubitecan; pyrazoloacridine; XR-5000; saintpine; UCE6; UCE1022; TAN-1518A; TAN 1518B; KT6006; KT6528; ED-110; NB-506; ED-110; NB-506; and rebeccamycin); vulgarein; DNA minor groove binders such as Hoescht dye 33342 and Hoescht dye 33258; nitidine; fagaronine; epiberberine; coraline; beta-lapachone; BC-4-1;Bisphosphonates (e.g., alendronate, cimadronte, clodronate, tiludronate, etidronate, ibandronate, neridronate, olpandronate, risedronate, piridronate, pamidronate, zolendronate); HMG-CoA reductase inhibitors (e.g., lovastatin, simvastatin, atorvastatin, pravastatin, fluvastatin, statins, cerivastatin, lescol, lupitor, rosuvastatin, and atorvastatin); antisense oligonucleotides (e.g., U.S. Pat. Nos. 6,277,832, 5,998,596, 5,885,834, 5,734,033, and 5,618,709); adenosine deaminase inhibitors (e.g., fludarabine phosphate and 2-chlorodeoxyadenosine); ibritumomab tiuxetan (Zevalin®); tositumomab (Bexxar®), and pharmaceutically acceptable salts, solvates, clathrates, and prodrugs thereof.

[0409] Furthermore, the antibodies of the present invention may be conjugated or recombinantly fused to a therapeutic moiety that modifies a given biological response. Therapeutic or drug moieties are not limited to classical chemical therapeutic agents. For example, the drug moiety may be a protein, peptide, or polypeptide possessing a desired biological activity. Such proteins include, for example, toxins such as abrin, ricin A, pseudomonas exotoxin, cholera toxin, or diphtheria toxin; tumor necrosis factor, gamma-interferon, alpha-interferon, nerve growth factor, platelet-derived growth factor, tissue plasminogen activator, apoptotic agents such as TNF-γ, AIM I (see WO 97 / 33899), AIM II (see WO 97 / 34911), Fas ligand (Takahashi et al., 1994, J. Immunol., 6:1567-1574), and VEGF (see WO 99 / 23105), anti-angiogenic agents, e.g., angiostatin, endostatin, or proteins such as components of the coagulation pathway (e.g., tissue factor); or, for example, lymphokines (e.g., interferon gamma, interleukin-1 ("IL-1"), interleukin-2 ("IL-2"), interleukin-5 ("IL-5"), interleukin-6 ("IL-6"), interleukin-7 ("IL-7"), interleukin-9 ("IL-9"), interleukin-10 ("IL-10"), interleukin-12 ("IL-12"), interleukin-1 Examples of such therapeutic agents include biological response modifiers such as interleukin-15 ("IL-15"), interleukin-23 ("IL-23"), granulocyte-macrophage colony-stimulating factor ("GM-CSF"), and granulocyte-colony-stimulating factor ("G-CSF"), or growth factors (e.g., growth hormone ("GH")), or coagulation agents (e.g., calcium, vitamin K, tissue factor, including, but not limited to, Hageman factor (factor XII), high molecular weight kininogen (HMWK), prekallikrein (PK), coagulation proteins factor II (prothrombin), factors V, XIIa, VIII, XIIIa, XI, Xia, IX, IXa, X, phospholipids, and fibrin monomers).

[0410] The present invention encompasses antibodies of the present invention that are recombinantly fused or chemically conjugated (covalently or noncovalently conjugated) to a heterologous protein or polypeptide (or fragment thereof, preferably a polypeptide of about 10, about 20, about 30, about 40, about 50, about 60, about 70, about 80, about 90, or about 100 amino acids) to produce a fusion protein. In particular, the present invention provides fusion proteins comprising an antigen-binding fragment of an antibody of the present invention (e.g., a Fab fragment, an Fd fragment, an Fv fragment, an F(ab)2 fragment, a VH domain, a VH CDR, a VL domain, or a VL CDR) and a heterologous protein, polypeptide, or peptide. In one embodiment, the heterologous protein, polypeptide, or peptide to which the antibody is fused is useful for targeting the antibody to a specific cell type, such as a cell expressing hOX40L or the hOX40L receptor. For example, an antibody that specifically binds to a cell surface receptor expressed by a specific cell type (e.g., an immune cell) can be fused or conjugated to a modified antibody of the present invention.

[0411] A conjugate or fusion protein of the invention comprises any of the antibodies of the invention described herein and a heterologous polypeptide, hi one embodiment, a conjugate or fusion protein of the invention comprises a variable domain of an antibody disclosed in the Examples and a heterologous polypeptide.

[0412] In addition, the antibodies of the invention may be coupled to a therapeutic moiety, e.g., 131 In, 131 Lu, 131 Y, 131 Ho, 131 Useful for complexing radioactive metal ions, including but not limited to Sm, with polypeptides. 213The antibody may be conjugated to a radioactive metal ion such as Bi or an alpha emitter such as a macrocyclic chelator. In certain embodiments, the macrocyclic chelator is 1,4,7,10-tetraazacyclododecane-N,N',N",N'"-tetraacetic acid (DOTA), which may be attached to the antibody via a linker molecule. Such linker molecules are generally known in the art and are described in Denardo et al., 1998, Clin Cancer Res. 4(10):2483-90; Peterson et al., 1999, Bioconjug. Chem. 10(4):553-7, and Zimmerman et al., 1999, Nucl. Med. Biol., 26(8):943-50, each of which is incorporated herein by reference in its entirety.

[0413] Furthermore, the antibodies of the present invention can be fused to a marker sequence, such as a peptide, to facilitate purification. In a preferred embodiment, the marker amino acid sequence is a hexa-histidine peptide, such as the tag provided in the pQE vector (QIAGEN, Inc.), among others, many of which are commercially available. See, e.g., Gentz ​​et al., 1989, Proc. Natl. Acad. Sci. USA 86:821-824, hexa-histidine provides for convenient purification of the fusion protein. Other peptide tags useful for purification include, but are not limited to, the hemagglutinin "HA" tag, which corresponds to an epitope derived from the influenza hemagglutinin protein (Wilson et al., 1984, Cell 37:767), and the "FLAG" tag.

[0414] Methods for fusing or conjugating therapeutic moieties (including polypeptides) to antibodies are well known, see, for example, Arnon et al., 1999, incorporated herein by reference in its entirety. “Monoclonal Antibodies For Immunotargeting Of Drugs In Cancer Therapy”, in Monoclonal Antibodies And Cancer Therapy, Reisfeld et al. (eds.), pp. 243-56 (Alan R. Liss, Inc. 1985);Hellstrom et al., “Antibodies For Drug Delivery”, in Controlled Drug Delivery (2nd Ed.), Robinson et al. (eds.), pp. 623-53 (Marcel Dekker, Inc. 1987);Thorpe, “Antibody Carriers Of Cytotoxic Agents In Cancer Therapy: A Review”, in Monoclonal Antibodies 84: Biological And Clinical Applications, Pinchera et al. (eds.), pp. 475-506 (1985);“Analysis, Results, And Future Prospective Of The Therapeutic Use Of Radiolabeled Antibody In Cancer Therapy”, in Monoclonal Antibodies For Cancer Detection And Therapy, Baldwin et al. (eds.), pp. 303-16 (Academic Press 1985), Thorpe et al., 1982, Immunol. Rev. 62:119-58; U.S. Patent Nos. 5,336,603, 5,622,929, 5,359,046, 5,349,053 , No. 5,447,851, No. 5,723,125, No. 5,783,181, No. 5,908,626, No. 5,844,095, No. 5,112,946; European Patent Nos. 307,434, 367,166, 394,827, PCT Publication No. 91 / 06570, WO 96 / 04388, WO 96 / 22024, 97 / 34631 and 99 / 04813; Ashkenazi et al., Proc. See Natl. Acad. Sci. USA, 88: 10535-10539, 1991; Traunecker et al., Nature, 331:84-86, 1988; Zheng et al., J. Immunol., 154:5590-5600, 1995; Vil et al., Proc. Natl. Acad. Sci. USA, 89:11337-11341, 1992.

[0415] Fusion proteins may be generated, for example, by techniques of gene shuffling, motif shuffling, exon shuffling, and / or codon shuffling (collectively referred to as "DNA shuffling"). DNA shuffling may be used to alter the activity of the antibodies of the invention (e.g., antibodies with higher affinity and lower dissociation rates). See generally U.S. Patent Nos. 5,605,793, 5,811,238, 5,830,721, 5,834,252, and 5,837,458; Patten et al., 1997, Curr. Opinion Biotechnol. 8:724-33; Harayama, 1998, Trends Biotechnol. 16(2):76-82; Hansson et al., 1999, J. Mol. Biol. 287:265-76; and Lorenzo and Blasco, 1998, Biotechniques 24(2):308-313 (each of these patents and publications is incorporated herein by reference in its entirety). Antibodies or the encoded antibodies may be altered by subjecting them to random mutagenesis by error-prone PCR, random nucleotide insertion, or other methods prior to recombination. Polynucleotides encoding antibodies of the invention may be recombined with one or more components, motifs, sections, portions, fragments, etc., of one or more heterologous molecules.

[0416] The antibodies of the present invention can also be conjugated to a second antibody to form an antibody heteroconjugate, as described in US Pat. No. 4,676,980, which is incorporated herein by reference in its entirety.

[0417] The therapeutic moiety or drug conjugated or recombinantly fused to an antibody of the invention that specifically binds to the hOX40L antigen should be selected to achieve the desired prophylactic or therapeutic effect(s). In certain embodiments, the antibody is a modified antibody. The clinician or other medical professional should consider the nature of the disease, the severity of the disease, and the condition of the subject when deciding which therapeutic moiety or drug to conjugate or recombinantly fused to an antibody of the invention.

[0418] The antibodies of the present invention may also be attached to solid supports, which are particularly useful in immunoassays or purification of the target antigen. Such solid supports include, but are not limited to, glass, cellulose, polyacrylamide, nylon, polystyrene, polyvinyl chloride, or polypropylene.

[0419] Pharmaceutical Compositions The following discussion of compositions also applies to fragments, so that disclosures referring to antibodies may be applied mutatis mutandis to fragments of the invention.

[0420] Therapeutic formulations containing one or more antibodies of the invention provided herein can be prepared for storage in the form of a lyophilized formulation or aqueous solution by mixing antibodies having the desired purity with optional physiologically acceptable carriers, excipients, or stabilizers (Remington's Pharmaceutical Sciences (1990) Mack Publishing Co., Easton, Pa.). Acceptable carriers, excipients, or stabilizers are non-toxic to recipients at the dosages and concentrations employed and include buffers such as phosphate, citrate, and other organic acids; antioxidants including ascorbic acid and methionine; preservatives (such as octadecyldimethylbenzylammonium chloride; hexamethonium chloride; benzalkonium chloride, benzethonium chloride; phenol, butyl, or benzyl alcohol; alkyl parabens such as methyl or propyl paraben; catechol; resorcinol; cyclohexanol; 3-pentanol; and m-cresol); low molecular weight (less than about 10 residues) polypeptides; serum albumin; hydrophilic polymers such as polyvinylpyrrolidone; amino acids such as glycine, glutamine, asparagine, histidine, arginine, or lysine; monosaccharides, disaccharides, and other carbohydrates including glucose, mannose, or dextrins; chelating agents such as EDTA; sugars such as sucrose, mannitol, trehalose, or sorbitol; salt-forming counterions such as sodium; metal complexes (e.g., Zn-protein complexes); and / or non-ionic surfactants such as TWEEN™, PLURONICS™, or polyethylene glycol (PEG).

[0421] The antibodies of the present invention provided herein can also be formulated, for example, in liposomes. Liposomes containing the molecule of interest are prepared by methods known in the art, such as those described in Epstein et al. (1985) Proc. Natl. Acad. Sci. USA 82:3688; Hwang et al. (1980) Proc. Natl. Acad. Sci. USA 77:4030; and U.S. Patent Nos. 4,485,045 and 4,544,545. Liposomes with enhanced circulation time are disclosed in U.S. Patent No. 5,013,556.

[0422] Particularly useful immunoliposomes can be generated by reverse-phase evaporation using a lipid composition containing phosphatidylcholine, cholesterol, and PEG-derivatized phosphatidylethanolamine (PEG-PE). Liposomes are extruded through filters of defined pore size to yield liposomes of the desired diameter. Fab' fragments of the antibodies provided herein can be complexed with liposomes via a disulfide exchange reaction as described by Martin et al. (1982) J. Biol. Chem. 257:286-288. A chemotherapeutic agent (such as doxorubicin) is optionally contained within the liposome. Gabizon et al., (1989) J. National See Cancer Inst. 81(19):1484.

[0423] Formulations such as those described herein may also contain two or more active compounds as necessary for the particular indication being treated. In certain embodiments, the formulation comprises an antibody of the invention and one or more active compounds with complementary activities that do not adversely affect each other. Such molecules are suitably present in combination in amounts that are effective for the intended purpose. For example, an antibody of the invention can be combined with one or more other therapeutic agents. Such combination therapy may be administered sequentially, simultaneously, or sequentially.

[0424] In one embodiment, the combination includes an anti-OX40L antibody of the invention and, independently, rapamycin (sirolimus), lacrolimus, cyclosporine, corticosteroids (e.g., methylprednisolone), methotrexate, mycophenolate mofetil, anti-CD28 antibodies, anti-IL12 / IL-23 antibodies (e.g., ustekinumab), anti-CD20 antibodies (e.g., rituximab), anti-CD30 antibodies (e.g., brentuximab), CTLA4-Fc molecules (e.g., abatacept), CCR5 receptor antagonists (e.g., maraviroc), anti-CD40L antibodies, anti-VLA4 antibodies (e.g., natalizumab), anti-LFA and an additional therapeutic agent selected from the group consisting of an anti-CD1 antibody, fludarabine, an anti-CD52 antibody (e.g., alemtuzumab), an anti-CD45 antibody, cyclophosphamide, anti-thymocyte globulin, an anti-complement C5 antibody (e.g., eculizumab), an anti-a4b7 integrin antibody (e.g., vedolizumab), an anti-IL6 antibody (e.g., tocilizumab), an anti-IL2R antibody (e.g., basilixumab), an anti-CD25 antibody (e.g., daclizumab), an anti-TNFα / TNFα-Fc molecule (e.g., etanercept, adalimumab, infliximab, golimumab, or cetolizumab pegol), and vorinostat. In another embodiment, the combination comprises an anti-OX40L antibody of the invention and an additional therapeutic agent independently selected from the group consisting of rapamycin (sirolimus), lacrolimus, cyclosporine, corticosteroids (e.g., methylprednisolone), methotrexate, mycophenolate mofetil, anti-CD28 antibodies, CTLA4-Fc molecules (e.g., abatacept), anti-CD40L antibodies, anti-LFA1 antibodies, anti-CD52 antibodies (e.g., alemtuzumab), cyclophosphamide, and anti-thymocyte globulin.

[0425] The antibodies of the invention can also be encapsulated in microcapsules prepared, for example, by coacervation techniques or by interfacial polymerization, such as hydroxymethylcellulose or gelatin microcapsules and poly-(methyl methacrylate) microcapsules, in colloidal drug delivery systems (e.g., liposomes, albumin microspheres, microemulsions, nanoparticles, and nanocapsules) or macroemulsions, respectively. Such techniques are disclosed in Remington's Pharmaceutical Sciences (1990) Mack Publishing Co., Easton, Pa.

[0426] Formulations to be used for in vivo administration can be sterile, which is readily accomplished, for example, by filtration through sterile filtration membranes.

[0427] Sustained-release preparations can also be prepared. Suitable examples of sustained-release preparations include semipermeable matrices of solid hydrophobic polymers containing the antagonist, which matrices are in the form of shaped articles, such as films or microcapsules. Examples of sustained-release matrices include polyesters, hydrogels (e.g., poly(2-hydroxyethyl-methacrylate) or poly(vinyl alcohol)), polylactides (U.S. Pat. No. 3,773,919), copolymers of L-glutamic acid and ethyl-L-glutamate, non-degradable ethylene vinyl acetate, degradable lactic acid-glycolic acid copolymers such as LUPRON DEPOT™ (injectable microspheres composed of lactic acid-glycolic acid copolymer and leuprolide acetate), and poly-D-(-)-3-hydroxybutyric acid. While polymers such as ethylene vinyl acetate and lactic acid-glycolic acid enable release of molecules over 100 days, certain hydrogels release proteins for shorter periods of time. When encapsulated antibodies remain in the body for extended periods, they may denature or aggregate as a result of exposure to moisture at 37°C, resulting in loss of biological activity and possible changes in immunogenicity. Rational strategies for stabilization can be devised depending on the mechanism involved. For example, if the aggregation mechanism is found to be intermolecular S—S bond formation through thio-disulfide exchange, stabilization can be achieved by modifying sulfhydryl residues, lyophilization from acidic solution, controlling water content, using appropriate additives, and creating specific polymer matrix compositions.

[0428] Pharmaceutical compositions provided herein contain a therapeutically effective amount of one or more of the antibodies of the invention provided herein, and optionally one or more additional prophylactic agents, in a pharmaceutically acceptable carrier. Such pharmaceutical compositions are useful for preventing, treating, managing, or alleviating hOX40L-mediated diseases, such as inflammatory bowel disease, transplant rejection, GvHD, or one or more of these symptoms.

[0429] Pharmaceutical carriers suitable for administration of the compounds provided herein include any carriers known to those skilled in the art to be suitable for the particular mode of administration.

[0430] In addition, the antibodies of the invention may be formulated as the sole pharmaceutically active ingredient in the composition or may be combined with other active ingredients, such as one or more other prophylactic or therapeutic agents.

[0431] The composition may contain one or more antibodies of the present invention. In one embodiment, the antibody is formulated into a pharmaceutical preparation such as a solution, suspension, tablet, dispersible tablet, pill, capsule, powder, sustained-release formulation, or elixir for oral administration, a pharmaceutical composition in a sterile solution or suspension for parenteral administration, and a pharmaceutical composition such as a transdermal patch preparation and a dry powder inhaler. In one embodiment, the antibody is formulated into a pharmaceutical composition using techniques and procedures well known in the art (see, e.g., Ansel (1985) Introduction to Pharmaceutical Dosage Forms, 4th Ed., p. 126).

[0432] In the compositions, an effective concentration of one or more antibodies or derivatives thereof is mixed with a suitable pharmaceutical carrier. The concentration of the compound in the composition is effective to deliver an amount that, upon administration, will treat, prevent, or alleviate a hOX40L-mediated disease or its symptoms.

[0433] In one embodiment, the composition is formulated for single-dose administration. To formulate the composition, a weight fraction of the compound that alleviates, prevents, or relieves one or more symptoms of the condition being treated is dissolved, suspended, dispersed, or mixed in a selected carrier at an effective concentration.

[0434] The antibodies of the present invention are included in a pharmaceutically acceptable carrier in an effective amount sufficient to exert a therapeutically useful effect without undesirable side effects on the treated patient. The therapeutically effective concentration can be determined empirically by testing the compound in in vitro and in vivo systems using conventional methods, and then extrapolating from there to dosages for humans.

[0435] The concentration of antibody in the pharmaceutical composition will depend, for example, on the physicochemical characteristics of the antibody, the dosing schedule and dosage, as well as other factors known to those skilled in the art.

[0436] In one embodiment, a therapeutically effective dose produces a serum concentration of antibody from about 0.1 ng / ml to about 50-100 μg / ml. In another embodiment, the pharmaceutical composition provides a daily dose of antibody from about 0.001 mg to about 2000 mg per kilogram of body weight. Pharmaceutical unit dosage forms can be prepared to provide from about 0.01 mg, 0.1 mg, or 1 mg to about 500 mg, 1000 mg, or 2000 mg of antibody and / or other optional or essential ingredients, or a combination of essential ingredients, per unit dosage form, in one embodiment.

[0437] The antibody can be administered at once or divided into a number of smaller doses and administered at intervals of time. It is understood that the exact dosage and duration of treatment are a function of the disease being treated and can be determined empirically using known testing protocols or by extrapolation from in vivo or in vitro test data. It should be noted that concentrations and dosage values ​​may vary depending on the severity of the condition being alleviated. It should be further understood that for any particular subject, specific dosage regimens can be adjusted gradually according to the individual needs and the judgment of the professional administering or supervising the administration of the composition, and that the concentration ranges set forth herein are exemplary only and are not intended to limit the scope or practice of the claimed compositions.

[0438] After mixing or adding the antibody, the resulting mixture may be a solution, suspension, emulsion, etc. The form of the resulting mixture depends on a number of factors, including the intended mode of administration and the solubility of the compound in the selected carrier or vehicle. The effective concentration is sufficient for alleviating the symptoms of the disease, disorder, or condition being treated and may be empirically determined.

[0439] Pharmaceutical compositions are provided for administration to humans and animals in unit dosage forms, such as tablets, capsules, pills, powders, granules, sterile parenteral solutions or suspensions, and oral solutions or suspensions, as well as oil-water emulsions containing an appropriate amount of the compound or a pharmaceutically acceptable derivative thereof. In one embodiment, the antibody is formulated or administered in unit dosage form or multiple dosage form. As used herein, unit dosage form refers to a physically discrete unit suitable for human and animal subjects and packaged individually, as known in the art. Each unit dose contains a predetermined amount of antibody sufficient to produce the desired therapeutic effect, together with the required pharmaceutical carrier, vehicle, or diluent. Examples of unit dosage forms include ampoules and syringes, and individually packaged tablets or capsules. Unit dosage forms can be administered in fractions or multiples thereof. Multiple dosage forms are multiple identical unit dosage forms packaged in a single container for administration in separate unit dosage forms. Examples of multiple dosage forms include vials, bottles of tablets or capsules, or bottles of pints and gallons. Thus, multiple dosage form is a multiple of unit doses that are not separately packaged.

[0440] In a preferred embodiment, one or more anti-hOX40L antibodies of the present invention are in a liquid pharmaceutical formulation. Liquid pharmaceutically administrable compositions can be prepared, for example, by dissolving, dispersing, or mixing the active compound(s) defined above and optional pharmaceutical adjuvants in a carrier, such as water, saline, aqueous dextrose, glycerol, glycol, ethanol, or the like, to form a solution or suspension. If desired, the pharmaceutical composition to be administered can also contain minor amounts of non-toxic auxiliary substances such as wetting agents, emulsifiers, solubilizing agents, pH buffering agents, and the like, for example, acetic acid, sodium citrate, cyclodextrin derivatives, sorbitan monolaurate, triethanolamine sodium acetate, triethanolamine oleate, and other such agents.

[0441] Actual methods for preparing such dosage forms are known, or will be apparent, to those skilled in the art; see, for example, Remington's Pharmaceutical Sciences (1990) Mack Publishing Co., Easton, Pa.

[0442] Dosage forms or compositions can be prepared containing antibodies in the range of 0.005% to 100%, with the remainder consisting of non-toxic carriers. Methods for preparing these compositions are known in the art.

[0443] Oral dosage forms can be solid, gel, or liquid. Solid dosage forms include tablets, capsules, granules, and bulk powders. Types of oral tablets include compressed tablets, chewable lozenges, and tablets that may be enteric-coated, sugar-coated, or film-coated. Capsules can be hard or soft gelatin capsules, while granules and powders can be provided in non-effervescent or effervescent form in combination with other ingredients known to those skilled in the art.

[0444] In certain embodiments, the formulation is a solid dosage form. In certain embodiments, the formulation is a capsule or tablet. Tablets, pills, capsules, troches, etc. may contain one or more of the following ingredients, or compounds of a similar nature: binders, lubricants, diluents, glidants, disintegrants, colorants, sweeteners, flavorings, wetting agents, emetic coatings, and film coatings. Examples of binders include microcrystalline cellulose, tragacanth gum, glucose, acacia mucilage, gelatin solution, molasses, polyinylpyrrolidine, povidone, crospovidone, sucrose, and starch paste. Lubricants include talc, starch, magnesium or calcium stearate, lycopodium, and stearic acid. Diluents include, for example, lactose, sucrose, starch, kaolin, salt, mannitol, and dicalcium phosphate. Glidants include, but are not limited to, colloidal silicon dioxide. Disintegrants include croscarmellose sodium, sodium starch glycolate, alginic acid, corn starch, potato starch, bentonite, methylcellulose, agar, and carboxymethylcellulose. Coloring agents include, for example, any of the approved certified water-soluble FD and C dyes, mixtures thereof, and water-soluble FD and C dyes suspended on alumina hydrate. Sweeteners include sucrose, lactose, mannitol, and artificial sweeteners such as saccharin, as well as any number of spray-dried flavorings. Flavoring agents include natural flavors extracted from plants such as fruits, and synthetic blends of pleasant-tasting compounds, such as, but not limited to, peppermint and methyl salicylate. Humectants include propylene glycol monostearate, sorbitan monooleate, diethylene glycol monolaurate, and polyoxyethylene laural ether. Emetic coatings include fatty acids, fats, waxes, ammoniated shellac, and cellulose acetate phthalate.Film coatings include hydroxyethylcellulose, sodium carboxymethylcellulose, polyethylene glycol 4000, and cellulose acetate phthalate.

[0445] The antibodies of the present invention can be provided in a composition that protects them from the acidic environment of the stomach. For example, the composition can be formulated in an enteric coating that maintains its integrity in the stomach and releases the active compound in the intestine. The composition can also be formulated in combination with an antacid or other such ingredient.

[0446] When the dosage unit is a capsule, it can contain, in addition to the above-mentioned materials, liquid carriers such as fatty oils.In addition, dosage unit can contain various other materials that modify the physical form of dosage unit, such as sugar and other enteric coatings.Compound can also be administered as a component of elixir, suspension, syrup, wafer, powder, chewing gum, etc.Syrup can contain, in addition to active compound, sucrose as a sweetener and certain preservatives, dyes and colorings, and flavorings.

[0447] The antibodies can also be mixed with other active substances which do not impair the desired action, or with substances which supplement the desired activity, such as antacids, H2 blockers, and diuretics. The active ingredient is an antibody described herein or a pharmaceutically acceptable derivative thereof. Higher concentrations, up to about 98% by weight of the active ingredient, can be included.

[0448] In all embodiments, tablet and capsule formulations may be coated as known to those skilled in the art to modify or sustain dissolution of the active ingredient. Thus, for example, they may be coated with conventional enterically digestible coatings such as phenylsalicylate, waxes, and cellulose acetate phthalate.

[0449] In a preferred embodiment, the formulation is a liquid dosage form.Liquid oral dosage forms include aqueous solutions, emulsions, suspensions, solutions, and / or suspensions reconstituted from non-effervescent granules and effervescent preparations reconstituted from effervescent granules.Aqueous solutions include, for example, elixirs and syrups.Emulsions are either oil-in-water or water-in-oil.

[0450] Elixirs are clear, sweetened aqueous preparations. Pharmaceutically acceptable carriers used in elixirs include solvents. Syrups are concentrated aqueous solutions of sugars, such as sucrose, and may contain preservatives. Emulsions are two-phase systems in which one liquid is dispersed in the form of small globules throughout another liquid. Pharmaceutically acceptable carriers used in emulsions are non-aqueous liquids, emulsifiers, and preservatives. Suspensions use pharmaceutically acceptable suspending agents and preservatives.

[0451] Pharmaceutically acceptable substances used in non-effervescent granules to be reconstituted into a liquid oral dosage form include diluents, sweeteners, and wetting agents. Pharmaceutically acceptable substances used in effervescent granules to be reconstituted into a liquid oral dosage form include organic acids and a carbon dioxide source. Coloring and flavoring agents are used in all of the above dosage forms.

[0452] Solvents include glycerin, sorbitol, ethyl alcohol, and syrup. Examples of preservatives include glycerin, methyl and propyl parabens, benzoic acid, sodium benzoate, and alcohol. Examples of non-aqueous liquids used in emulsions include mineral oil and cottonseed oil. Examples of emulsifiers include gelatin, acacia, tragacanth, bentonite, and surfactants such as polyoxyethylene sorbitan monooleate. Suspending agents include sodium carboxymethylcellulose, pectin, tragacanth, Veegum, and acacia. Sweeteners include sucrose, syrup, glycerin, and artificial sweeteners such as saccharin. Humectants include propylene glycol monostearate, sorbitan monooleate, diethylene glycol monolaurate, and polyoxyethylene lauryl ether. Organic acids include citric acid and tartaric acid. Carbon dioxide sources include sodium bicarbonate and sodium carbonate. Coloring agents include any of the approved certified water soluble FD and C dyes, and mixtures thereof. Flavoring agents include natural flavors extracted from fruits and other plants, and synthetic blends of compounds which produce a pleasant taste sensation.

[0453] For a solid dosage form, the solution or suspension, for example, in propylene carbonate, vegetable oil, or triglyceride, is in one embodiment encapsulated in a gelatin capsule. Such solutions, and their preparation and encapsulation, are disclosed in U.S. Patent Nos. 4,328,245, 4,409,239, and 4,410,545. For a liquid dosage form, the solution, for example, in, for example, a polyethylene glycol, can be diluted with a sufficient quantity of a pharmaceutically acceptable liquid carrier, for example, water, to be easily measured for administration.

[0454] Alternatively, liquid or semi-solid oral formulations can be prepared by dissolving or dispersing the active compound or salt in vegetable oils, glycols, triglycerides, propylene glycol esters (e.g., propylene carbonate), and other such carriers, and encapsulating these solutions or suspensions in hard or soft gelatin capsule shells. Other useful formulations include those shown in U.S. Patent Nos. RE28,819 and 4,358,603. Briefly, such formulations include, but are not limited to, those containing one or more antioxidants such as a compound provided herein, 1,2-dimethoxymethane, diglyme, triglyme, tetraglyme, dialkylated mono- or polyalkylene glycols (350, 550, and 750 refer to the approximate average molecular weight of the polyethylene glycol), including but not limited to polyethylene glycol-350-dimethyl ether, polyethylene glycol-550-dimethyl ether, polyethylene glycol-750-dimethyl ether, butylated hydroxytoluene (BHT), butylated hydroxyanisole (BHA), propyl gallate, vitamin E, hydroquinone, hydroxycoumarin, ethanolamine, lecithin, cephalin, ascorbic acid, malic acid, sorbitol, phosphoric acid, thiodipropionic acid and its esters, and dithiocarbamates.

[0455] Other formulations include, but are not limited to, aqueous alcohol solutions containing pharmaceutically acceptable acetals.The alcohols used in these formulations are any pharmaceutically acceptable water-miscible solvents having one or more hydroxyl groups, including, but not limited to, propylene glycol and ethanol.Acetals include, but are not limited to, di(lower alkyl) acetals of acetaldehyde lower alkyl aldehydes, such as acetaldehyde diethyl acetal.

[0456] Parenteral administration, in one embodiment, is characterized by either subcutaneous injection, intramuscular injection, or intravenous injection, and is also contemplated herein. Injectables can be prepared in conventional formats, either as liquid solutions or suspensions, solid forms suitable for solution or suspension in liquid prior to injection, or as emulsions. Injectables, solutions, and emulsions also contain one or more excipients. Suitable excipients are, for example, water, saline, dextrose, glycerol, or ethanol. In addition, if desired, the administered pharmaceutical composition can also contain small amounts of non-toxic auxiliary substances such as wetting or emulsifying agents, pH buffering agents, stabilizers, solubility enhancers, and other such agents, for example, sodium acetate, sorbitan monolaurate, triethanolamine oleate, and cyclodextrins.

[0457] Injection of a delayed-release or sustained-release system such that a constant level of dosage is maintained (see, e.g., U.S. Pat. No. 3,710,795) is also contemplated herein. Briefly, the compounds provided herein are administered in a solid inner matrix, e.g., a hydrophilic polymer such as polymethyl methacrylate, polybutyl methacrylate, plasticized or unplasticized polyvinyl chloride, plasticized nylon, plasticized polyethylene terephthalate, natural rubber, polyisoprene, polyisobutylene, polybutadiene, polyethylene, ethylene-vinyl acetate copolymer, silicone rubber, polydimethylsiloxane, silicone carbonate copolymer, hydrogels of esters of acrylic and methacrylic acid, collagen, cross-linked polyvinyl alcohol, and cross-linked partially hydrophilic polymers surrounded by a polymeric outer membrane, which are insoluble in body fluids. The antibody is dispersed in a hydrolyzed polyvinyl acetate, such as polyethylene, polypropylene, ethylene / propylene copolymer, ethylene / ethyl acetate copolymer, ethylene / vinyl acetate copolymer, silicone rubber, polymethylsiloxane, neoprene rubber, chlorinated polyethylene, polyvinyl chloride, copolymers of vinyl chloride with vinyl acetate, vinylidene chloride, ethylene, and propylene, ionomer polyethylene terephthalate, butyl rubber epichlorohydrin rubber, ethylene / vinyl alcohol copolymer, ethylene / vinyl acetate / vinyl alcohol terpolymer, and ethylene / vinyloxyethanol copolymer. The antibody diffuses through the outer polymer membrane in a release rate-controlling step. The amount of antibody contained in such parenteral compositions depends largely on its specific nature, as well as the activity of the compound and the needs of the subject.

[0458] Preparations for parenteral administration include sterile solutions ready for injection, subcutaneous tablets, sterile suspensions ready for injection, sterile dry soluble products such as lyophilized powders ready to be combined with a solvent immediately before use, including sterile dry insoluble products ready to be combined with a vehicle immediately before use, and sterile emulsions. Solutions can be either aqueous or non-aqueous.

[0459] For intravenous administration, suitable carriers include saline or phosphate buffered saline (PBS), as well as solutions containing thickening and solubilizing agents such as glucose, polyethylene glycol, and polyethylene glycol, and mixtures thereof.

[0460] Pharmaceutically acceptable carriers used in parenteral preparations include aqueous vehicles, non-aqueous vehicles, antibacterial agents, isotonic agents, buffers, antioxidants, local anesthetics, suspending and dispersing agents, emulsifying agents, sequestering or chelating agents, and other pharmaceutically acceptable substances.

[0461] Examples of aqueous vehicles include sodium chloride injection, Ringer's injection, isotonic dextrose injection, sterile water injection, dextrose and lactated Ringer's injection. Non-aqueous parenteral vehicles include fixed oils of vegetable origin, cottonseed oil, corn oil, sesame oil, and peanut oil. Antibacterial agents in bacteriostatic and fungistatic concentrates can be added to parenteral preparations packaged in multi-dose containers, including phenol or cresol, mercurials, benzyl alcohol, chlorobutanol, methyl and propyl p-hydroxybenzoic acid esters, thimerosal, benzalkonium chloride, and benzethonium chloride. Isotonic agents include sodium chloride and dextrose. Buffers include phosphates and citrates. Antioxidants include sodium bisulfate. Local anesthetics include procaine hydrochloride. Suspending and dispersing agents include sodium carboxymethylcellulose, hydroxypropyl methylcellulose, and polyvinylpyrrolidone. Emulsifying agents include polysorbate 80 (TWEEN® 80). Sequestering or chelating agents for metal ions include EDTA. Pharmaceutical carriers also include ethyl alcohol, polyethylene glycol, and propylene glycol for water-miscible vehicles, and sodium hydroxide, hydrochloric acid, citric acid, or lactic acid for pH adjustment.

[0462] The concentration of the pharmaceutically active compound is adjusted so that an injection provides an amount effective to produce the desired pharmacological effect. The exact dose depends on the age, weight, and condition of the patient or animal, as is known in the art.

[0463] Unit-dose parenteral preparations can be packaged in an ampoule, a vial, or a syringe with a needle. All preparations for parenteral administration can be sterile, as is known and practiced in the art.

[0464] By way of example, intravenous or intraarterial infusion of a sterile aqueous solution containing an active compound is an effective mode of administration. Another embodiment is a sterile aqueous or oily solution or suspension containing an active material injected as necessary to produce the desired pharmacological effect.

[0465] Injectables are designed for local and systemic administration. In one embodiment, a therapeutically effective dose is formulated to contain a concentration of the active compound relative to the tissue(s) to be treated of at least about 0.1% w / w, up to about 90% w / w or more, and in certain embodiments, greater than 1% w / w.

[0466] The antibody may be suspended in micronized or other suitable form. The form of the resulting mixture will depend on several factors, including the intended mode of administration and the solubility of the compound in the selected carrier or vehicle. The effective concentration is sufficient to alleviate the symptoms of the condition and can be determined empirically.

[0467] In other embodiments, the pharmaceutical formulations are lyophilized powders that can be reconstituted for administration as solutions, emulsions, and other mixtures. They may also be reconstituted and formulated as solids or gels.

[0468] The lyophilized powder is prepared by dissolving an antibody or pharmaceutically acceptable derivative thereof provided herein in a suitable solvent. In some embodiments, the lyophilized powder is sterile. The solvent may contain an excipient that improves the stability or other pharmacological components of the powder or a reconstituted solution prepared from the powder. Excipients that may be used include, but are not limited to, dextrose, sorbital, fructose, corn syrup, xylitol, glycerin, glucose, sucrose, or other suitable agents. The solvent may also contain a buffer such as citrate, sodium, or potassium phosphate, or other such buffers known to those of skill in the art, in one embodiment at approximately neutral pH. Subsequent sterile filtration of the solution under standard conditions known to those of skill in the art, followed by lyophilization, produces the desired formulation. In one embodiment, the resulting solution is apportioned into vials for lyophilization. Each vial contains a single dose or multiple doses of the compound. The lyophilized powder can be stored under appropriate conditions, such as at about 4°C to room temperature.

[0469] Reconstitution of this lyophilized powder with water for injection provides a formulation for use in parenteral administration.For reconstitution, lyophilized powder is added to sterile water or other suitable carrier.The exact amount depends on the compound selected.Such amount can be determined experimentally.

[0470] Topical mixtures are prepared as described for local and systemic administration. The resulting mixture can be a solution, suspension, emulsion, etc., and can be formulated as a cream, gel, ointment, emulsion, solution, elixir, lotion, suspension, tincture, paste, foam, aerosol, douche, spray, suppository, bandage, skin patch, or any other formulation suitable for topical administration.

[0471] The antibodies of the invention can be formulated as aerosols for topical application, such as by inhalation (see, e.g., U.S. Pat. Nos. 4,044,126, 4,414,209, and 4,364,923, which describe aerosols for the delivery of steroids useful in the treatment of inflammatory diseases, particularly asthma). These formulations for airway administration can be in the form of aerosols or solutions for nebulization, or as ultrafine powders for insufflation, alone or in combination with an inert carrier such as lactose. In such cases, the particles of the formulation, in one embodiment, have diameters of less than 50 microns, and in one embodiment, less than 10 microns.

[0472] The compounds can be formulated in the form of gels, creams, and lotions for topical or external application, such as external application to the skin and mucous membranes, including in the eye, for ophthalmic application, or for intravesical or intrathecal application. External administration is contemplated for transdermal delivery, as well as administration to the eye or mucous membranes, or for inhalation therapy. Nasal solutions of the active compounds, alone or in combination with other pharmaceutically acceptable excipients, can also be administered.

[0473] These solutions, particularly those intended for ophthalmic use, may be formulated as 0.01% to 10% isotonic solutions at a pH of about 5 to 7, containing appropriate salts.

[0474] Other routes of administration, such as transdermal patches, including iontophoretic and electrophoretic devices, and rectal administration, are also contemplated herein.

[0475] Transdermal patches, including iontophoretic and electrophoretic devices, are well known to those skilled in the art. For example, such patches are disclosed in U.S. Patent Nos. 6,267,983, 6,261,595, 6,256,533, 6,167,301, 6,024,975, 6,010715, 5,985,317, 5,983,134, 5,948,433, and 5,860,957.

[0476] For example, pharmaceutical dosage forms for rectal administration include rectal suppositories, capsules, and tablets for systemic effects. As used herein, a rectal suppository refers to a solid object for insertion into the rectum that melts or softens at body temperature to release one or more pharmacologically or pharmaceutically active ingredients. Pharmaceutically acceptable substances utilized in rectal suppositories are bases or vehicles and agents to raise the melting point. Examples of bases include cocoa butter (theobroma oil), glycerin-gelatin, carbowax (polyethylene glycol), and suitable mixtures of mono-, di-, and triglycerides of fatty acids. Combinations of various bases may be used. Agents to raise the melting point of the suppository include spermaceti and wax. Rectal suppositories can be prepared by either compression or molding. In one embodiment, a rectal suppository weighs approximately 2 to 3 gm.

[0477] Tablets and capsules for rectal administration may be manufactured using the same pharmaceutically acceptable substance and by the same methods as for formulations for oral administration.

[0478] The antibodies and other compositions provided herein may also be formulated to target specific tissues, receptors, or other areas of the body of the subject being treated. Many such targeting methods are known to those skilled in the art. All such targeting methods are contemplated herein for use in the present compositions. For non-limiting examples of targeting methods, see, e.g., U.S. Patent Nos. 6,316,652, 6,274,552, 6,271,359, 6,253,872, 6,139,865, 6,131,570, 6,120,751, 6,071,495, 6,060,082, 6,048,736, 6,039,975, 6,004,534, 5,985,307, 5,972,366, 5,900,252, 5,840,674, 5,759,542, and 5,709,874. In some embodiments, the anti-hOX40L antibodies of the invention are targeted to (or administered to) the colon, such as in patients with or at risk of having IBD. In some embodiments, the anti-hOX40L antibodies of the invention are targeted to (or administered to) the eye, such as in patients with or at risk of having uveitis.

[0479] In one embodiment, liposome suspensions, including tissue-targeted liposomes such as tumor-targeted liposomes, may also be suitable as pharmaceutically acceptable carriers. These can be prepared according to methods known in the art. For example, liposome formulations can be prepared as described in U.S. Pat. No. 4,522,811. Briefly, liposomes such as multilamellar vesicles (MLVs) can be formed by drying egg phosphatidylcholine and brain phosphatidylserine (molar ratio 7:3) inside a flask. A solution of a compound provided herein in phosphate-buffered saline (PBS) lacking divalent cations is added, and the flask is shaken until the lipid film is dispersed. The resulting vesicles are washed to remove unencapsulated compounds, pelleted by centrifugation, and then resuspended in PBS.

[0480] Administration and Dosage Methods The present invention further provides compositions comprising one or more antibodies or fragments of the present invention for use in the prevention, management, treatment, and / or amelioration of an OX40L-mediated disease (or a symptom thereof). The discussion regarding antibodies applies mutatis mutandis to fragments of the present invention. In the alternative, the present invention further provides compositions comprising one or more antibodies or fragments of the present invention for use in the prevention, management, treatment, and / or amelioration of an OX40L-mediated disease (or a symptom thereof) in a subject, wherein the OX40L is non-human (e.g., canine, feline, equine, bovine, ovine, or porcine) and the subject is a canine, feline, equine, bovine, ovine, or porcine, respectively.

[0481] In certain embodiments, provided herein are compositions comprising one or more antibodies of the present invention for use in the prevention, management, treatment, and / or alleviation of hOX40L-mediated diseases or symptoms thereof, such as IBD (e.g., ulcerative colitis or Crohn's disease). IBD symptoms range from mild to severe and generally depend on the portion of the intestinal tract involved. Exemplary symptoms of IBD include abdominal cramps and pain, bloody diarrhea, defecation urgency, fever, loss of appetite, weight loss, anemia, fatigue, and / or pain in the lower legs, ankles, calves, thighs, and arms. Exemplary intestinal complications of IBD include severe bleeding from ulcers, intestinal perforation or rupture, strictures and obstructions, fistulas (abnormal bowel movements) and perianal disease, toxic megacolon (e.g., acute non-obstructive dilation of the colon), and / or malignancies (e.g., cancer of the colon or small intestine). Exemplary extraintestinal complications of IBD include arthritis, skin disorders, eye inflammation, liver and kidney damage, and / or bone loss, any combination of which symptoms may be prevented, managed, treated, and / or alleviated using the compositions and methods provided herein.

[0482] In certain embodiments, provided herein are compositions comprising one or more antibodies of the invention for use in the prevention, management, treatment, and / or amelioration of hOX40L-mediated diseases or symptoms thereof, such as GVHD, which generally occurs after allogeneic or matched unrelated bone marrow transplantation (BMT).

[0483] In some embodiments, the GVHD is acute GVHD. Symptoms of acute GVHD can develop rapidly and can be mild or severe. In certain cases, GVHD manifests within about three months after transplantation, such as when blood counts recover after transplantation. In certain cases, acute GVHD affects the skin, gastrointestinal (GI) tract, and / or liver. For example, in some patients, acute skin GVHD begins with a rash on the patient's palms, soles, or shoulders. However, the rash can spread and become itchy, painful, and / or blister and peel. Acute liver GVHD can affect the normal function of the liver, including liver enzymes, which can cause jaundice. Acute liver GVHD can also cause abdominal distension and pain as the liver enlarges. Finally, symptoms of acute gastrointestinal GVHD (or GVHD of the digestive system) can include diarrhea, mucus or blood in the stool, cramps or abdominal pain, indigestion, nausea, and / or loss of appetite. Other common symptoms of acute GVHD can include anemia, low-grade fever, and / or a tendency to contract infections. Any combination of these symptoms of acute GVHD can be prevented, managed, treated, and / or alleviated using the compositions and methods provided herein.

[0484] In other embodiments, the GVHD is chronic GVHD. Chronic GVHD can occur from about 3 months to about 1 year or longer after transplantation. Chronic GVHD can be mild or severe and generally includes symptoms similar to those of acute GVHD. Chronic GVHD can affect the skin and digestive system, including the liver, but can also involve other organs and the immune system (e.g., making the patient more susceptible to infections) and / or connective tissue. Symptoms of chronic skin GVHD include eczema, dry skin, tight skin, itchy skin, darkened skin, thickened skin, and / or can affect hair (e.g., hair loss, graying) or nails (e.g., hard or brittle nails). Chronic gastrointestinal GVHD can affect the digestive system, mouth, esophagus, stomach lining, and / or intestinal lining, and symptoms include diarrhea, dry or sore mouth, painful swallowing, decreased nutrient absorption by the stomach, bloating, and stomach cramps. Chronic liver GVHD can cause liver damage and scarring (cirrhosis). Chronic eye GVHD can affect the glands that produce tears, causing dry eyes, burning and pain, or intolerance to bright light. Chronic lung GVHD can cause shortness of breath, wheezing, persistent coughing, and / or increased susceptibility to lung infections. Chronic GVHD can affect the tendons that connect muscles to bones (e.g., inflammation), making it difficult to bend and straighten the arms and legs. Any combination of these symptoms of chronic GVHD can be prevented, managed, treated, and / or alleviated using the compositions and methods provided herein.

[0485] In certain embodiments, provided herein are compositions comprising one or more antibodies of the invention for use in the prevention, management, treatment, and / or alleviation of hOX40L-mediated diseases or symptoms thereof, such as uveitis.

[0486] In certain embodiments, provided herein are compositions comprising one or more antibodies of the invention for use in the prevention, management, treatment, and / or amelioration of hOX40L-mediated diseases, such as pyoderma gangrenosum, giant cell arteritis, Schnitzler's syndrome, or non-infectious scleritis.

[0487] In certain embodiments, provided herein are compositions comprising one or more antibodies of the invention for use in the prevention, management, treatment, and / or alleviation of a hOX40L-mediated disease or condition selected from an autoimmune disease or condition, a systemic inflammatory disease or condition, or transplant rejection, e.g., inflammatory bowel disease (IBD), Crohn's disease, rheumatoid arthritis, transplant rejection, allograft rejection, graft-versus-host disease (GvHD), ulcerative colitis, systemic lupus erythematosus (SLE), diabetes, uveitis, ankylosing spondylitis, contact hypersensitivity, multiple sclerosis, and atherosclerosis, particularly GvHD.

[0488] In certain embodiments, compositions for use in the prevention, management, treatment, and / or amelioration of hOX40L-mediated diseases comprise an OX40L-binding portion of an antibody of the invention, such as an antibody disclosed in the Examples.

[0489] In another embodiment, a composition for use in the prevention, management, treatment, and / or amelioration of hOX40L-mediated disease comprises one or more antibodies comprising one or more VH domains having the amino acid sequence of any one of the VH domains in the Sequence Listing (i.e., SEQ ID NO:2, SEQ ID NO:34, SEQ ID NO:66, or SEQ ID NO:94, particularly SEQ ID NO:34). In another embodiment, a composition for use in the prevention, management, treatment, and / or amelioration of hOX40L-mediated disease comprises one or more antibodies comprising one or more VH CDR1s having the amino acid sequence of any one of the VH CDR1s in the Sequence Listing (i.e., SEQ ID NO:4, SEQ ID NO:10, SEQ ID NO:36, SEQ ID NO:42, SEQ ID NO:68, SEQ ID NO:74, SEQ ID NO:96, or SEQ ID NO:102, particularly SEQ ID NO:36 or SEQ ID NO:42). In another embodiment, a composition for use in the prevention, management, treatment, and / or amelioration of hOX40L-mediated disease comprises one or more antibodies comprising one or more VH CDR2s having the amino acid sequence of any one of the VH CDR2s in the Sequence Listing (i.e., SEQ ID NO:6, SEQ ID NO:12, SEQ ID NO:38, SEQ ID NO:44, SEQ ID NO:70, SEQ ID NO:76, SEQ ID NO:98, or SEQ ID NO:104, particularly SEQ ID NO:38 or SEQ ID NO:44). In a preferred embodiment, a composition for use in the prevention, management, treatment, and / or amelioration of hOX40L-mediated disease comprises one or more antibodies comprising one or more VH CDR3s having the amino acid sequence of any one of the VH CDR3s in the Sequence Listing (i.e., SEQ ID NO:8, SEQ ID NO:14, SEQ ID NO:40, SEQ ID NO:46, SEQ ID NO:72, SEQ ID NO:78, SEQ ID NO:100, or SEQ ID NO:106, particularly SEQ ID NO:40 or SEQ ID NO:46).

[0490] In another embodiment, a composition for use in the prevention, management, treatment, and / or amelioration of hOX40L-mediated disease comprises one or more antibodies comprising one or more VL domains having the amino acid sequence of any one of the VL domains in the Sequence Listing (i.e., SEQ ID NO: 16, SEQ ID NO: 48, SEQ ID NO: 80, or SEQ ID NO: 108, particularly SEQ ID NO: 48) (optionally including the cognate VH domains presented in the Sequence Listing (i.e., SEQ ID NO: 2 / 16, SEQ ID NO: 34 / 48, SEQ ID NO: 66 / 80, or SEQ ID NO: 94 / 108, particularly SEQ ID NO: 34 / 48)). In another embodiment, a composition for use in the prevention, management, treatment, and / or amelioration of hOX40L-mediated disease comprises one or more antibodies comprising one or more VL CDR1s having the amino acid sequence of any one of the VL CDR1s in the Sequence Listing (i.e., SEQ ID NO: 18, SEQ ID NO: 24, SEQ ID NO: 50, SEQ ID NO: 56, SEQ ID NO: 82, SEQ ID NO: 88, SEQ ID NO: 110, or SEQ ID NO: 116, particularly SEQ ID NO: 50 or SEQ ID NO: 56). In another embodiment, a composition for use in the prevention, management, treatment, and / or amelioration of hOX40L-mediated disease comprises one or more antibodies comprising one or more VL CDR2s having the amino acid sequence of any one of the VL CDR2s in the Sequence Listing (i.e., SEQ ID NO:20, SEQ ID NO:26, SEQ ID NO:52, SEQ ID NO:58, SEQ ID NO:84, SEQ ID NO:90, SEQ ID NO:112, or SEQ ID NO:118, particularly SEQ ID NO:52 or SEQ ID NO:58). In a preferred embodiment, a composition for use in the prevention, management, treatment, and / or amelioration of hOX40L-mediated disease comprises one or more antibodies comprising one or more VL CDR3s having the amino acid sequence of any one of the VL CDR3s in the Sequence Listing (i.e., SEQ ID NO:22, SEQ ID NO:28, SEQ ID NO:54, SEQ ID NO:60, SEQ ID NO:86, SEQ ID NO:92, SEQ ID NO:114, or SEQ ID NO:120, particularly SEQ ID NO:54 or SEQ ID NO:60).

[0491] In another embodiment, a composition for use in the prevention, management, treatment, and / or amelioration of hOX40L-mediated disease comprises one or more antibodies comprising one or more VH domains having the amino acid sequence of any one of the VH domains in the sequence listing (i.e., SEQ ID NO: 2, SEQ ID NO: 34, SEQ ID NO: 66, or SEQ ID NO: 94, particularly SEQ ID NO: 34), and one or more VL domains having the amino acid sequence of any one of the VL domains in the sequence listing (i.e., SEQ ID NO: 16, SEQ ID NO: 48, SEQ ID NO: 80, or SEQ ID NO: 108, particularly SEQ ID NO: 48).

[0492] In another embodiment, a composition for use in the prevention, management, treatment, and / or amelioration of hOX40L-mediated disease comprises one or more antibodies comprising one or more VH CDR1s having the amino acid sequence of any one of the VH CDR1s in the sequence listing (i.e., SEQ ID NO:4, SEQ ID NO:10, SEQ ID NO:36, SEQ ID NO:42, SEQ ID NO:68, SEQ ID NO:74, SEQ ID NO:96, or SEQ ID NO:102, particularly SEQ ID NO:36 or SEQ ID NO:42), and one or more VL CDR1s having the amino acid sequence of any one of the VL CDR1s in the sequence listing (i.e., SEQ ID NO:18, SEQ ID NO:24, SEQ ID NO:50, SEQ ID NO:56, SEQ ID NO:82, SEQ ID NO:88, SEQ ID NO:110, or SEQ ID NO:116, particularly SEQ ID NO:50 or SEQ ID NO:56). In another embodiment, a composition for use in the prevention, management, treatment, and / or amelioration of hOX40L-mediated disease comprises one or more antibodies comprising one or more VH CDR1s having the amino acid sequence of any one of the VH CDR1s in the sequence listing (i.e., SEQ ID NO:4, SEQ ID NO:10, SEQ ID NO:36, SEQ ID NO:42, SEQ ID NO:68, SEQ ID NO:74, SEQ ID NO:96, or SEQ ID NO:102, particularly SEQ ID NO:36 or SEQ ID NO:42), and one or more VL CDR2s having the amino acid sequence of any one of the VL CDR2s in the sequence listing (i.e., SEQ ID NO:20, SEQ ID NO:26, SEQ ID NO:52, SEQ ID NO:58, SEQ ID NO:84, SEQ ID NO:90, SEQ ID NO:112, or SEQ ID NO:118, particularly SEQ ID NO:52 or SEQ ID NO:58). In another embodiment, a composition for use in the prevention, management, treatment, and / or amelioration of hOX40L-mediated disease comprises one or more VH CDR1s having the amino acid sequence of any one of the VH CDR1s in the sequence listing (i.e., SEQ ID NO:4, SEQ ID NO:10, SEQ ID NO:36, SEQ ID NO:42, SEQ ID NO:68, SEQ ID NO:74, SEQ ID NO:96, or SEQ ID NO:102, particularly SEQ ID NO:36 or SEQ ID NO:42). The present invention includes one or more antibodies comprising one or more CDR1s and one or more VL CDR3s having the amino acid sequence of any one of the VL CDR3s in the sequence listing (i.e., SEQ ID NO: 22, SEQ ID NO: 28, SEQ ID NO: 54, SEQ ID NO: 60, SEQ ID NO: 86, SEQ ID NO: 92, SEQ ID NO: 114, or SEQ ID NO: 120, particularly SEQ ID NO: 54 or SEQ ID NO: 60).

[0493] As discussed in more detail elsewhere herein, the compositions of the present invention may be used alone or in combination with other compounds or compositions. Again, the antibodies may also be recombinantly fused at the N- or C-terminus to heterologous polypeptides or chemically conjugated (including covalent and non-covalent conjugation) to polypeptides or other compositions. For example, the antibodies of the present invention may be recombinantly fused or conjugated to molecules useful as labels in detection assays, and effector molecules such as heterologous polypeptides, drugs, radionucleotides, or toxins. See, e.g., PCT Publication Nos. WO 92 / 08495, WO 91 / 14438, WO 89 / 12624, U.S. Pat. No. 5,314,995, and EP 396,387.

[0494] In some embodiments, provided herein are methods for reducing or inhibiting binding of hOX40L to an OX40L receptor or cognate ligand (e.g., OX40) in a subject (e.g., a human subject), the methods comprising administering to the subject an effective amount of an antibody that specifically binds to an hOX40L polypeptide (e.g., cell-surface-expressed hOX40L or soluble hOX40L). In some embodiments, hOX40L biological activity, such as secretion of CCL20, IL8, and / or RANTES, or INF-γ, TNF-α, or IL-2, particularly INF-γ, or another cytokine disclosed herein, is also reduced in the subject, e.g., by at least 10, 20, 30, 40, 50, or 60%, or 70%, or 80%, or 90%, or 95%, or more than 95%.

[0495] In certain embodiments, a method is provided for reducing or inhibiting hOX40L biological activity, such as secretion of interferon gamma, IL-2, CCL20, IL8, and / or RANTES, or other cytokines, or INF-γ, TNF-α, or IL-2, particularly INF-γ, in a subject (e.g., a human subject), the method comprising administering to the subject an effective amount of an antibody that specifically binds to an hOX40L polypeptide (e.g., hOX40L expressed on the surface of a cell), wherein the antibody reduces hOX40L biological activity.

[0496] In other embodiments, provided herein are methods for reducing or inhibiting binding of hOX40L to its OX40L receptor or cognate ligand (e.g., OX40) in cells having hOX40L expressed on their surface, by contacting the cells with an effective amount of an antibody that specifically binds to an hOX40L polypeptide (e.g., cell-surface-expressed hOX40L or soluble hOX40L), e.g., an hOX40L polypeptide, an hOX40L polypeptide fragment, or an hOX40L epitope. In some embodiments, hOX40L biological activity, such as secretion of interferon gamma, IL-2, CCL20, IL8, and / or RANTES, or INF-γ, TNF-α, or IL-2, particularly INF-γ, or other cytokines disclosed herein, is also reduced in the cells.

[0497] In certain embodiments, provided herein are methods for reducing or inhibiting hOX40L biological activity, such as secretion of interferon gamma, IL-2, CCL20, IL8, and / or RANTES, or other cytokines disclosed herein, in cells having a surface-expressed hOX40L receptor (such as OX40), by contacting the cells with an effective amount of an antibody that specifically binds to an hOX40L polypeptide (e.g., cell-surface-expressed hOX40L or soluble hOX40L), wherein the antibody reduces hOX40L biological activity.

[0498] The antibodies of the present invention can be used, for example, to purify, detect, and target hOX40L antigens in both in vitro and in vivo diagnostic and therapeutic methods. For example, the modified antibodies are useful in immunoassays for qualitatively and quantitatively measuring hOX40L levels in biological samples. See, e.g., Harlow et al., Antibodies: A Laboratory Manual, (Cold Spring Harbor Laboratory Press, 2nd ed. 1988), the entire contents of which are incorporated herein by reference.

[0499] The present invention also provides methods for preventing, managing, treating, and / or ameliorating hOX40L-mediated diseases by administering to a subject an effective amount of an antibody or a pharmaceutical composition comprising an antibody of the present invention. In one aspect, the antibody is substantially purified (i.e., substantially free from substances that limit its effectiveness or produce undesirable side effects). In a preferred embodiment, the antibody is a fully human monoclonal antibody, such as a fully human monoclonal antagonist antibody. The subject to which the therapy is administered is preferably a mammal, e.g., a non-primate (e.g., bovine, porcine, equine, feline, canine, rodent, mouse, or rat) or a primate (e.g., a monkey such as a rhesus or cynomolgus monkey, or a human). In a preferred embodiment, the subject is a human. In another preferred embodiment, the subject is a human infant or a human infant born prematurely. In another embodiment, the subject is a human with an hOX40L-mediated disease.

[0500] Various delivery systems are known and can be used to administer prophylactic or therapeutic agents (e.g., antibodies of the invention). These delivery systems include, but are not limited to, encapsulation in liposomes, microparticles, recombinant cells capable of expressing the antibody, receptor-mediated endocytosis (see, e.g., Wu and Wu, J. Biol. Chem. 262:4429-4432 (1987)), construction of a nucleic acid as part of a retrovirus or other vector, and the like. Methods of administration of prophylactic or therapeutic agents (e.g., antibodies of the invention) or pharmaceutical compositions include, but are not limited to, parenteral administration (e.g., intradermal, intramuscular, intraperitoneal, intravenous, and subcutaneous), epidural, and mucosal (e.g., intranasal and oral routes). In certain embodiments, prophylactic or therapeutic agents (e.g., antibodies of the invention) or pharmaceutical compositions are administered intranasally, intramuscularly, intravenously, or subcutaneously. The prophylactic or therapeutic agents or compositions may be administered by any convenient route, such as by infusion or bolus injection, or by absorption through epithelial or mucocutaneous linings (e.g., oral, nasal, rectal, and intestinal mucosa), or may be administered in conjunction with other biologically active agents. Administration may be systemic or local. In addition, pulmonary administration may be employed, for example, by use of an inhaler or nebulizer and formulation with an aerosolizer. See, e.g., U.S. Patent Nos. 6,019,968, 5,985,320, 5,985,309, 5,934,272, 5,874,064, 5,855,913, 5,290,540, and 4,880,078, and PCT Publication Nos. WO 92 / 19244, 97 / 32572, 97 / 44013, 98 / 31346, and 99 / 66903, each of which is incorporated by reference herein in its entirety.

[0501] In certain embodiments, it may be desirable to administer prophylactic or therapeutic agents, or pharmaceutical compositions of the invention, locally to the area in need of treatment. This may be achieved, for example, but not limited to, by local infusion, topical administration (e.g., nasal spray), injection, or by means of an implant, which may be a porous, nonporous, or gelatinous material, including membranes, such as sialastic membranes, or fibers. Preferably, when administering an antibody of the invention, care should be taken to use materials to which the antibody does not absorb.

[0502] In another embodiment, the prophylactic or therapeutic agents, or compositions of the invention, can be delivered in a vesicle, particularly a liposome (Langer, 1990, Science 249:1527-1533; Treat et al., in Liposomes in The Therapy of Infectious Disease and Cancer, Lopez-Berestein and Fidler (eds.), Liss, New York, pp. 353-365 (1989), see Lopez-Berestein, ibid., pp. 317-327; see generally ibid.).

[0503] In another embodiment, a prophylactic or therapeutic agent, or composition of the invention can be delivered in a controlled-release or sustained-release system. In one embodiment, controlled or sustained-release can be achieved using a pump (Langer, supra; Sefton, 1987, (See, e.g., Medical Applications of Controlled Release, Langer and Wise (eds.), CRC Pres., Boca Raton, Fla. (1974); Controlled Drug Bioavailability, Drug Product Design and See Performance, Smolen and Ball (eds.), Wiley, New York (1984), Ranger and Peppas, 1983, J., Macromol. Sci. Rev. Macromol. Chem. 23:61, and Levy et al., 1985, Science 228:190, During et al., 1989, Ann. al., 1989, J. Neurosurg. 7 1:105); see also U.S. Pat. No. 5,679,377, U.S. Pat. No. 5,916,597, U.S. Pat. Examples of polymers used in sustained-release formulations include, but are not limited to, poly(2-hydroxyethyl methacrylate), poly(methyl methacrylate), poly(acrylic acid), poly(ethylene-co-vinyl acetate), poly(methacrylic acid), polyglycolide (PLG), polyanhydrides, poly(N-vinylpyrrolidone), poly(vinyl alcohol), polyacrylamide, poly(ethylene glycol), polylactide (PLA), poly(lactide-co-glycolide) (PLGA), and polyorthoesters. In preferred embodiments, the polymers used in sustained-release formulations are inert, free of leachable impurities, stable on storage, sterile, and biodegradable. In yet another embodiment, a controlled or sustained-release system can be placed in close proximity to the therapeutic target, i.e., the nasal cavity or lungs, thereby requiring only a fraction of the systemic dose (see, e.g., Goodson, in Medical Applications of Controlled Release, supra, vol. 2, pp. 115-138 (1984)). Controlled-release systems are discussed in the review by Langer (1990, Science 249:1527-1533). Any technique known in the art can be used to produce sustained-release formulations comprising one or more antibodies of the invention.See, for example, U.S. Pat. No. 4,526,938, PCT Publication Nos. WO 91 / 05548 and WO 96 / 20698, and Ning et al., 1996, "Intratumoral Radioimmunotherapy of a. Human Colon Cancer bFGF Antibody for Cardiovascular Application,” Pro. Int'l. Symp. Control. Rel. Bioact. Mater. 24:853-854, and Lam et al., 1997, “Microencapsulation of Recombinant Humanized Monoclonal Antibody for Local Delivery,” Proc. See Int'l. Symp. Control Rel. Bioact. Mater. 24:759-760.

[0504] In certain embodiments, when the composition of the invention is a nucleic acid encoding a prophylactic or therapeutic agent (e.g., an antibody of the invention), the nucleic acid can be constructed as part of an appropriate nucleic acid expression vector and administered in vivo to promote expression of the encoded prophylactic or therapeutic agent, for example, by using a retroviral vector and administering it intracellularly (see U.S. Pat. No. 4,980,286), or by direct injection, or by using microparticle bombardment (e.g., a gene gun; Biolistic, DuPont), or by coating with lipids or cell surface receptors or transfecting agents, or by administering it while bound to a homeobox-like peptide known to enter the nucleus (see, e.g., Joliot et al., 1991, Proc. Natl. Acad. Sci. USA 88:1864-1868). Alternatively, the nucleic acid can be introduced intracellularly for expression by homologous recombination and integrated into host cell DNA.

[0505] In certain embodiments, compositions of the invention comprise one, two, or more antibodies or fragments of the invention. In another embodiment, compositions of the invention comprise one, two, or more antibodies or fragments of the invention and a prophylactic or therapeutic agent other than an antibody of the invention. Preferably, the agent is known to be useful, or has been used, or is currently being used to prevent, manage, treat, and / or alleviate hOX40L-mediated disease. In addition to a prophylactic or therapeutic agent, compositions of the invention may also comprise a carrier.

[0506] The compositions of the present invention include drug substance compositions useful for manufacturing pharmaceutical compositions (e.g., compositions suitable for administration to a subject or patient) that can be used in the preparation of unit dosage forms. In a preferred embodiment, the compositions of the present invention are pharmaceutical compositions. Such compositions comprise a prophylactically or therapeutically effective amount of one or more prophylactic or therapeutic agents (e.g., an antibody or other prophylactic or therapeutic agent of the present invention) and a pharmaceutically acceptable carrier. Pharmaceutical compositions are preferably formulated to be suitable for the route of administration to a subject.

[0507] In certain embodiments, the term "carrier" refers to a diluent, adjuvant (e.g., Freund's adjuvant (complete and incomplete)), excipient, or vehicle with which the therapeutic is administered. Such pharmaceutical carriers can be sterile liquids, such as water and oils, including those of petroleum, animal, vegetable, or synthetic origin, such as peanut oil, soybean oil, mineral oil, sesame oil, and the like. Water is a preferred carrier when the pharmaceutical composition is administered intravenously. Saline solutions and aqueous dextrose and glycerol solutions can also be employed as liquid carriers, particularly for injectable solutions. Suitable pharmaceutical excipients include starch, glucose, lactose, sucrose, gelatin, malt, rice, flour, chalk, silica gel, sodium stearate, glycerol monostearate, talc, sodium chloride, dried skim milk, glycerol, propylene, glycol, water, ethanol, and the like. The composition, if desired, can also contain minor amounts of wetting or emulsifying agents, or pH buffering agents. These compositions can take the form of solutions, suspensions, emulsions, tablets, pills, capsules, powders, sustained-release formulations, and the like. Oral formulations can include standard carriers such as pharmaceutical grades of mannitol, lactose, starch, magnesium stearate, sodium saccharin, cellulose, magnesium carbonate, and the like. Examples of suitable pharmaceutical carriers are described in Remington's Pharmaceutical Sciences (1990) Mack Publishing Co., Easton, Pa. Such compositions will contain a prophylactically or therapeutically effective amount of the antibody, preferably in purified form, together with an amount of carrier suitable to provide the form for proper administration to the patient. The formulation should suit the mode of administration.

[0508] In a preferred embodiment, the composition is formulated according to routine procedures as a pharmaceutical composition adapted for intravenous administration to humans. Typically, compositions for intravenous administration are solutions in sterile isotonic buffer. If necessary, the composition may also contain a solubilizing agent and a local anesthetic, such as lignocamne, to ease pain at the injection site. However, such compositions may be administered by routes other than intravenous.

[0509] Generally, the components of the composition of the present invention are supplied separately or mixed together in unit dosage form, for example, as lyophilized powder or water-free concentrate in a moisture-tight container, such as an ampoule or sachet indicating the amount of active ingredient.When the composition is administered by injection, it can be dispensed in an infusion bottle containing sterile pharmaceutical grade water or saline.When the composition is administered by injection, an ampoule of sterile water for injection or saline can be provided so that the components can be mixed before administration.

[0510] The present invention also provides that the antibody is packaged in a sealed container, such as an ampoule or sachet, indicating the quantity of antibody. In one embodiment, the antibody is supplied as a dry, sterile, lyophilized powder or anhydrous concentrate in a moisture-sealed container and can be reconstituted, for example, with water or saline, to the appropriate concentration for administration to a subject. Preferably, the antibody is supplied as a dry, sterile, lyophilized powder in a moisture-sealed container in a unit dose of at least 0.1 mg, at least 0.5 mg, at least 1 mg, at least 2 mg, or at least 3 mg, more preferably at least 5 mg, at least 10 mg, at least 15 mg, at least 25 mg, at least 30 mg, at least 35 mg, at least 45 mg, at least 50 mg, at least 60 mg, at least 75 mg, at least 80 mg, at least 85 mg, at least 90 mg, at least 95 mg, or at least 100 mg. The lyophilized antibody can be stored in its original container at 2-8°C, and the antibody can be administered within 12 hours, preferably within 6 hours, 5 hours, 3 hours, or 1 hour, after reconstitution. In an alternative embodiment, the antibody is supplied in liquid form in a moisture-tight container indicating the amount and concentration of the antibody. Preferably, the liquid form of the antibody is supplied in a moisture-tight container at a concentration of at least 0.1 mg / ml, at least 0.5 mg / ml, or at least 1 mg / ml, more preferably at least 5 mg / ml, at least 10 mg / ml, at least 15 mg / ml, at least 25 mg / ml, at least 30 mg / ml, at least 40 mg / ml, at least 50 mg / ml, at least 60 mg / ml, at least 70 mg / ml, at least 80 mg / ml, at least 90 mg / ml, or at least 100 mg / ml.

[0511] The compositions of the present invention can be formulated as neutral or salt forms. Pharmaceutically acceptable salts include salts formed with anions, such as those derived from hydrochloric acid, phosphoric acid, acetic acid, oxalic acid, tartaric acid, etc., and salts formed with cations, such as those derived from sodium, potassium, ammonium, calcium, ferric hydroxide, isopropylamine, triethylamine, 2-ethylaminoethanol, histidine, procaine, etc.

[0512] The amount of a prophylactic or therapeutic agent (e.g., an antibody of the invention), or a composition of the invention, that will be effective in the prevention, management, treatment, and / or amelioration of a hOX40L-mediated disease can be determined by standard clinical techniques.

[0513] Thus, from about 0.1 μg / ml to about 450 μg / ml, in some embodiments at least 0.1 μg / ml, at least 0.2 μg / ml, at least 0.4 μg / ml, at least 0.5 μg / ml, at least 0.6 μg / ml, at least 0.8 μg / ml, at least 1 μg / ml, at least 1.5 μg / ml, preferably at least 2 μg / ml, at least 5 μg / ml, at least 10 μg / ml, at least 15 μg / ml, at least 20 μg / ml, at least 25 μg / ml, at least 30 μg / ml, or at least Antibodies or compositions that produce a serum titer of at least 35 μg / ml, at least 40 μg / ml, at least 50 μg / ml, at least 75 μg / ml, at least 100 μg / ml, at least 125 μg / ml, at least 150 μg / ml, at least 200 μg / ml, at least 250 μg / ml, at least 300 μg / ml, at least 350 μg / ml, at least 400 μg / ml, or at least 450 μg / ml can be used in humans for the prevention, management, treatment, and / or amelioration of hOX40L-mediated diseases. Additionally, in vitro assays may optionally be employed to help identify optimal dosage ranges. The precise dose to be employed in the formulation will also depend on the route of administration and the severity of the hOX40L-mediated disease, and should be decided according to the judgment of the practitioner and each patient's circumstances.

[0514] Effective doses may be extrapolated from dose-response curves derived from in vitro or animal model test systems.

[0515] For antibodies of the present invention, the dosage administered to a patient is typically 0.1 mg / kg to 100 mg / kg of the patient's body weight. In some embodiments, the dosage administered to a patient is about 1 mg / kg to about 75 mg / kg of the patient's body weight. Preferably, the dosage administered to a patient is 1 mg / kg to 20 mg / kg of the patient's body weight, more preferably 1 mg / kg to 5 mg / kg of the patient's body weight. Generally, human antibodies have a longer half-life in the human body than antibodies from other species due to the immune response to foreign polypeptides. Therefore, lower doses and less frequent administration of human antibodies are often possible. Furthermore, the dosage and frequency of administration of antibodies of the present invention can be reduced by enhancing antibody uptake and tissue penetration through modifications such as lipidation.

[0516] In one embodiment, an antibody or fragment of the invention is administered at about 100 mg / kg or less, about 75 mg / kg or less, about 50 mg / kg or less, about 25 mg / kg or less, about 10 mg / kg or less, about 5 mg / kg or less, about 1 mg / kg or less, about 0.5 mg / kg or less, or about 0.1 mg / kg or less, five, four, three, two, or preferably one dose to manage hOX40L-mediated disease. In some embodiments, an antibody of the invention is administered about 1-12 times, and this dose may be administered as needed, e.g., weekly, biweekly, monthly, bimonthly, quarterly, etc., as determined by a physician. In some embodiments, a lower dose (e.g., 1-15 mg / kg) may be administered more frequently (e.g., 3-6 times). In other embodiments, a higher dose (e.g., 25-100 mg / kg) may be administered less frequently (e.g., 1-3 times). However, other dosages and schedules may be readily determined and are within the scope of the present invention, as will be apparent to those skilled in the art.

[0517] In certain embodiments, about 100 mg / kg, about 75 mg / kg or less, about 50 mg / kg or less, about 25 mg / kg or less, about 10 mg / kg or less, about 5 mg / kg or less, about 1 mg / kg or less, about 0.5 mg / kg or less, about 0.1 mg / kg or less of an antibody or fragment of the invention is administered to a subject, preferably a human, in a sustained release formulation to prevent, manage, treat, and / or alleviate a hOX40L-mediated disease. In another specific embodiment, about 100 mg / kg, about 75 mg / kg or less, about 50 mg / kg or less, about 25 mg / kg or less, about 10 mg / kg or less, about 5 mg / kg or less, about 1 mg / kg or less, about 0.5 mg / kg or less, or about 0.1 mg / kg or less of an antibody of the invention is administered to a subject, preferably a human, in a bolus rather than sustained release formulation to prevent, manage, treat, and / or alleviate an hOX40L-mediated disease, and after a certain period of time, about 100 mg / kg, about 75 mg / kg or less, about 50 mg / kg or less, about 25 mg / kg or less, about 10 mg / kg or less, about 5 mg / kg or less, about 1 mg / kg or less, about 0.5 mg / kg or less, or about 5 mg / kg or less of an antibody of the invention is administered to the subject (e.g., intranasally or intramuscularly) two, three, or four (preferably once) times in a sustained release manner. According to this embodiment, the specific period of time may be 1 to 5 days, 1 week, 2 weeks, or 1 month.

[0518] In some embodiments, a single dose of the antibody or fragment of the invention is administered to a patient 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, or 26 times every other week (e.g., about 14 days) over a period of one year to prevent, manage, treat, and / or ameliorate a hOX40L-mediated disease, at a dose of about 0.1 mg / kg, about 0.5 mg / kg, about 1 mg / kg, about 5 mg / kg, or about 6 mg / kg. , about 10 mg / kg, about 15 mg / kg, about 20 mg / kg, about 25 mg / kg, about 30 mg / kg, about 35 mg / kg, about 40 mg / kg, about 45 mg / kg, about 50 mg / kg, about 55 mg / kg, about 60 mg / kg, about 65 mg / kg, about 70 mg / kg, about 75 mg / kg, about 80 mg / kg, about 85 mg / kg, about 90 mg / kg, about 95 mg / kg, about 100 mg / kg, or combinations thereof (i.e., each monthly dose may or may not be the same).

[0519] In another embodiment, a single dose of an antibody of the invention is administered to a patient 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 times at intervals of about every month (e.g., about 30 days) for one year to prevent, manage, treat, and / or ameliorate a hOX40L-mediated disease, and the dose may be about 0.1 mg / kg, about 0.5 mg / kg, about 1 mg / kg, about 5 mg / kg, about 10 mg / kg, about 15 mg / kg, about 20 mg / kg, about 25 mg / kg, or about 30 mg / kg. mg / kg, about 30 mg / kg, about 35 mg / kg, about 40 mg / kg, about 45 mg / kg, about 50 mg / kg, about 55 mg / kg, about 60 mg / kg, about 65 mg / kg, a...

Claims

1. An injection device comprising an antibody or an antigen-binding fragment thereof that specifically binds to hOX40L, The antibody or antigen-binding fragment a VH domain comprising the HCDR1 sequence of SEQ ID NO: 36 or 42, the HCDR2 sequence of SEQ ID NO: 38 or 44, and the HCDR3 sequence of SEQ ID NO: 40 or 46; a V L domain comprising the LCDR1 sequence of SEQ ID NO: 50 or 56, the LCDR2 sequence of SEQ ID NO: 52 or 58, and the LCDR3 sequence of SEQ ID NO: 54 or 60; injection equipment.

2. An injection device as described in claim 1, wherein the antibody or antigen-binding fragment inhibits binding of hOX40L to the hOX40L receptor.

3. The antibody or antigen-binding fragment thereof, a VH domain comprising the amino acid sequence of SEQ ID NO: 34, and / or comprising a V L domain comprising the amino acid sequence of SEQ ID NO: 48; 10. An injection device according to claim 1.

4. An injection device as described in claim 1, wherein the antibody or antigen-binding fragment comprises a human gamma 4 constant region.

5. The injection device of claim 1, wherein the antibody or antigen-binding fragment comprises a heavy chain constant region that is IgG4-PE, and optionally a heavy chain constant region of SEQ ID NO:

128.

6. An injection device as described in claim 1, wherein the antibody or antigen-binding fragment comprises a kappa light chain.

7. The injection device described in claim 6, wherein the kappa light chain comprises a constant region selected from the group consisting of the amino acid sequences of the kappa light chain constant regions of SEQ ID NOs: 136, 138, 140, 142 and 144.

8. The injection device of claim 1, wherein the antibody or antigen-binding fragment comprises a first and a second copy of the V H domain and / or a first and a second copy of the V L domain.

9. An injection device as described in claim 1, wherein the antibody or antigen-binding fragment is a fully human antibody.

10. The antibody comprising a heavy chain and a light chain, the amino acid sequence of the heavy chain consists of the sequence of SEQ ID NO: 62; and 2. The injection device of claim 1, wherein the amino acid sequence of the light chain consists of the sequence of SEQ ID NO:

64.

11. An injection device described in any one of claims 1 to 10, used for pharmaceuticals.

12. An injection device as described in any one of claims 1 to 10, for the treatment and / or prevention of an hOX40L-mediated condition or disease selected from an autoimmune disease or condition, a systemic inflammatory disease or condition, or a transplant rejection.

13. The injection device of claim 12, wherein the hOX40L-mediated condition or disease is selected from inflammatory bowel disease (IBD), Crohn's disease, rheumatoid arthritis, allograft rejection, graft-versus-host disease (GvHD), ulcerative colitis, systemic lupus erythematosus (SLE), diabetes, uveitis, ankylosing spondylitis, contact hypersensitivity, multiple sclerosis, and atherosclerosis.

14. An injection device as described in any one of claims 1 to 10 for the treatment and / or prevention of dermatitis, asthma, systemic sclerosis or celiac disease.

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