Therapeutic anti-CD40 ligand antibodies

Abstract

Provided are anti-CD40L antibodies and antigen-binding fragments thereof, compositions comprising such antibodies or antigen-binding fragments, anti-CD40L antibodies with reduced effector function, and methods of use thereof for treating CD40L-associated diseases or disorders. [Selected Figure] Figure 1A

Description

[Technical Field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This PCT application claims the benefit of U.S. Provisional Patent Application No. 62 / 510,471, filed May 24, 2017, the disclosure of which is incorporated herein by reference in its entirety.

[0002] Sequence Listing This application incorporates by reference in its entirety the Sequence Listing entitled "224823-431206_ST25.txt", created on May 23, 2018 at 2:40 PM, 100 KB, which was submitted electronically with the application.

[0003] Technical Field Anti-CD40L antibodies, compositions comprising such antibodies, and methods of using them for treating CD40L-associated diseases or disorders. [Background technology]

[0004] The interaction between CD40 and its ligand, CD40L, plays an important role in regulating immune responses. Binding of CD40L to CD40 activates the CD40 pathway, upregulating costimulatory molecules such as CD80 and CD86. Blockade of the CD40-CD40L interaction with monoclonal antibodies has been shown to protect against autoimmunity and graft rejection in various preclinical models. Recently, in a mouse model of amyotrophic lateral sclerosis, CD40L-directed antibodies were shown to delay disease onset and extend survival after disease onset (U.S. Patent No. 8,435,514, incorporated herein by reference). In early clinical trials, the humanized anti-CD40L antibody, hu5c8, demonstrated efficacy in patients with lupus and immune thrombocytopenic purpura. However, further clinical trials were discontinued due to the occurrence of thromboembolic events in patients treated with hu5c8. Furthermore, in vitro and animal preclinical studies have confirmed that the interaction of Fc with the Fc receptor FcγRIIa causes platelet activation and subsequent aggregation, leading to thromboembolic events. Furthermore, it has been reported that Fc binding to complement can inhibit or prevent the induction of immune tolerance. Various attempts have been made to inhibit or eliminate the interaction between the Fc region of immunoglobulins and FcγRIIa and / or the interaction with complement. For example, point mutation(s) have been introduced into the Fc region to generate anti-CD40L antibodies lacking Fc effector function. Other attempts have used antibody fragments lacking the Fc region or antibodies containing multiple amino acid substitutions in the Fc region. Although the anti-CD40L antibody hu5c8 has demonstrated efficacy in human patients, no anti-CD40L antibodies are commercially available. Thus, there is a need for improved anti-CD40L antibodies for administration to humans that are effective, do not cause platelet activation or aggregation, do not bind complement, yet are stable and bind CD40L. Summary of the Invention

[0005] The novel antibody polypeptides and antigen-binding fragments thereof of the present invention provide such improved anti-CD40L antibodies. The following section merely summarizes certain aspects of the present disclosure and is not intended to be limiting in any way. These and other aspects and embodiments are described in further detail below. All references cited herein are incorporated by reference in their entirety. In the event of a conflict between the express disclosure of this specification and a reference incorporated by reference, the express disclosure of this specification shall control.

[0006] The present invention provides isolated, engineered, non-naturally occurring antibodies and antigen-binding fragments thereof that bind to human CD40L and block CD40 binding to CD40L. The antibodies and antigen-binding fragments thereof are engineered to have desired activity and binding. In some embodiments, the antibodies or antigen-binding fragments thereof are engineered to have attenuated Fc effector function relative to some prior anti-CD40L antibodies. The antibodies and antigen-binding fragments thereof disclosed in the present application are useful for treating diseases in which CD40L activation is implicated, including neurodegenerative or neuromuscular diseases or disorders, inflammatory or immune diseases or disorders, and autoimmune diseases.

[0007] In one aspect, the disclosure provides an isolated antibody or antigen-binding fragment thereof that specifically binds to CD40L, comprising: (a) a heavy chain variable region (V) comprising i) a CDRH1 domain comprising the sequence set forth in SEQ ID NO: 9; ii) a CDRH2 domain comprising the sequence set forth in SEQ ID NO: 10, 11, 12, 13, or 14; and iii) a CDRH3 domain comprising the sequence set forth in SEQ ID NO: 15. H ), and (b) a light chain variable region (V) comprising i) a CDRL1 domain comprising the sequence set forth in SEQ ID NO: 16 or 17, ii) a CDRL2 domain comprising the sequence set forth in SEQ ID NO: 18 or 19, and iii) a CDRL3 domain comprising the sequence set forth in SEQ ID NO: 20. L ) is included.

[0008] In some embodiments of the above aspects of the disclosure, an isolated antibody or antigen-binding fragment thereof that specifically binds to CD40L is provided, comprising: (a) a heavy chain variable region (V) comprising i) a CDRH1 domain consisting of the sequence set forth in SEQ ID NO:9; ii) a CDRH2 domain consisting of the sequence set forth in SEQ ID NO:10, 11, 12, 13, or 14; and iii) a CDRH3 domain consisting of the sequence set forth in SEQ ID NO:15. H ), and (b) a light chain variable region (V) comprising i) a CDRL1 domain consisting of the sequence set forth in SEQ ID NO: 16 or 17, ii) a CDRL2 domain consisting of the sequence set forth in SEQ ID NO: 18 or 19, and iii) a CDRL3 domain consisting of the sequence set forth in SEQ ID NO: 20. L ) is included.

[0009] The present disclosure also provides an isolated antibody or antigen-binding fragment thereof that specifically binds to CD40L, comprising: (a) a heavy chain variable region (V) having the amino acid sequence set forth in SEQ ID NO: 1, 2, 3, or 4; H ), and (b) a light chain variable region (V) having the amino acid sequence set forth in SEQ ID NO: 5, 6, 7, or 8. L ) is included.

[0010] In some embodiments, an isolated antibody or antigen-binding fragment thereof according to the present disclosure comprises an Fc region, wherein the Fc region has been engineered to reduce or eliminate one or more Fc effector functions. In some embodiments, the isolated antibody is of the IgG1 isotype, and the heavy chain constant region comprises the amino acid sequence set forth in SEQ ID NO: 21. In still other embodiments, the antibody comprises a heavy chain constant region comprising the amino acid sequence set forth in SEQ ID NO: 22. In yet other embodiments, the antibody comprises a heavy chain constant region comprising the amino acid sequence set forth in SEQ ID NO: 23. In another embodiment, the antibody comprises a heavy chain constant region comprising the amino acid sequence set forth in SEQ ID NO: 24.

[0011] Another aspect of the present disclosure provides a method for treating a subject having a CD40L-associated disease or disorder, comprising administering to such subject a therapeutically effective amount of an antibody or antibody fragment according to the present disclosure. In another aspect, the present disclosure provides a method for inhibiting an immune response in a subject, comprising administering to such subject a therapeutically effective amount of an antibody or antibody fragment according to the present disclosure. [Brief explanation of the drawings]

[0012] [Figure 1] Figure 1A is a bar graph showing the potency ranking of 16 antibody clones against the anti-CD40L antibody 5c8. The potency ranking is calculated by dividing the IC50 of the clone by the IC50 of 5c8 and multiplying by 100. Figure 1B is a bar graph showing the potency ranking of 16 antibody clones against the anti-CD40L antibody AT-1501. The potency ranking is calculated by dividing the IC50 of the clone by the IC50 of AT-1501 and multiplying by 100. [Figure 2] Figure 2A shows the binding curves of the antibodies of each clone and 5c8. Figure 2B shows the binding curves of the antibodies of each clone and 5c8. Figure 2C shows the binding curves of the antibodies of each clone and 5c8. Figure 2D shows the binding curves of the antibodies of each clone and 5c8. Figure 2E shows the binding curves of the antibodies of each clone and 5c8. Figure 2F shows the binding curves of the antibodies of each clone and 5c8. Figure 2G shows the binding curves of the antibodies of each clone and 5c8. Figure 2I shows the binding curves of the antibodies of each clone and 5c8. Figure 2J shows the binding curves of the antibodies of each clone and 5c8. Figure 2K shows the binding curves of the antibodies of each clone and 5c8. Figure 2L shows the binding curves of the antibodies of each clone and 5c8. Figure 2M shows the binding curves of the antibodies of each clone and 5c8. Figure 2N shows the binding curves of the antibodies of each clone and 5c8. Figure 2O shows the binding curves of the antibodies of each clone and 5c8. Figure 2P shows the binding curves of the antibodies of each clone and 5c8. Figure 2Q shows the binding curves of the antibodies of each clone and 5c8. Figure 2H shows the binding curves of AT-1501 and 5c8. [Figure 3] Figure 3A shows the binding curves of the antibodies of each clone and the binding curves of AT-1501. Figure 3B shows the binding curves of the antibodies of each clone and the binding curves of AT-1501. Figure 3C shows the binding curves of the antibodies of each clone and the binding curves of AT-1501. Figure 3D shows the binding curves of the antibodies of each clone and the binding curves of AT-1501. Figure 3E shows the binding curves of the antibodies of each clone and the binding curves of AT-1501. Figure 3F shows the binding curves of the antibodies of each clone and the binding curves of AT-1501. Figure 3G shows the binding curves of the antibodies of each clone and the binding curves of AT-1501. Figure 3I shows the binding curves of the antibodies of each clone and the binding curves of AT-1501. Figure 3J shows the binding curves of the antibodies of each clone and the binding curves of AT-1501. Figure 3K shows the binding curves of the antibodies of each clone and the binding curves of AT-1501. Figure 3L shows the binding curves of the antibodies of each clone and the binding curves of AT-1501. Figure 3M shows the binding curves of the antibodies of each clone and the binding curves of AT-1501. Figure 3N shows the binding curves of the antibodies of each clone and the binding curves of AT-1501. Figure 3O shows the binding curves of the antibodies of each clone and the binding curves of AT-1501. Figure 3P shows the binding curves of the antibodies of each clone and the binding curves of AT-1501. Figure 3Q shows the binding curves of the antibodies of each clone and the binding curves of AT-1501. Figure 3H shows the binding curves of AT-1501 and 5c8. [Figure 4] Figure 4A is a graph showing the binding of eight of the antibody clones, 5c8, AT-1501, and abatacept, to FcγRI. The only antibody with significant binding is the 5c8 antibody. Figure 4B is a graph showing the binding of eight of the antibody clones, 5c8, AT-1501, and abatacept, to FcγRI. The only antibody with significant binding is the 5c8 antibody. [Figure 5]Figure 5A is a graph showing the binding of eight of the antibody clones, 5c8, AT-1501, and abatacept, to FcγRIIa. The only antibody with significant binding is the 5c8 antibody. Figure 5B is a graph showing the binding of eight of the antibody clones, 5c8, AT-1501, and abatacept, to FcγRIIa. The only antibody with significant binding is the 5c8 antibody. [Figure 6] Figure 6A is a graph showing the binding of eight of the antibody clones, 5c8, AT-1501, and abatacept, to FcγRIIIa. None of the antibodies showed significant binding to FcγRIIIa. Figure 6B is a graph showing the binding of eight of the antibody clones, 5c8, AT-1501, and abatacept, to FcγRIIIa. None of the antibodies showed significant binding to FcγRIIIa. Figure 6C is a graph showing the binding of eight of the antibody clones, 5c8, AT-1501, and abatacept, to FcγRIIIb. None of the antibodies showed significant binding to FcγRIIIb. Figure 6D is a graph showing the binding of eight of the antibody clones, 5c8, AT-1501, and abatacept, to FcγRIIIb. None of the antibodies showed significant binding to FcγRIIIb. [Figure 7] Figure 7A is a graph showing the binding of eight of the antibody clones, 5c8, AT-1501, and abatacept, to C1q. The only antibody that showed significant binding to C1q was the 5c8 antibody. Figure 7B is a graph showing the binding of eight of the antibody clones, 5c8, AT-1501, and abatacept, to C1q. The only antibody that showed significant binding to C1q was the 5c8 antibody. DETAILED DESCRIPTION OF THE INVENTION

[0013] definition

[0014] For convenience, the meaning of certain terms and phrases employed in the specification, examples, and appended claims are provided below.

[0015] Terms such as "comprises," "comprised," "comprising," "contains," "containing," and the like, have the meaning ascribed to them in U.S. patent law, and are inclusive or open-ended and do not exclude additional, unrecited elements or method steps. Terms such as "consisting essentially of" and "consists essentially of" have the meaning ascribed to them in U.S. patent law, and permit the inclusion of additional ingredients or steps that do not materially affect the basic and novel characteristics of the claimed invention. The terms "consists of" and "consisting of" have the meaning ascribed to them in U.S. patent law, and are open-ended.

[0016] TMB is an abbreviation for 3,3',5,5'-tetramethylbenzidine.

[0017] As used herein, "CDR domain" means an antibody complementarity determining region, with or without flanking sequences.

[0018] The inventions illustratively described herein may suitably be practiced in the absence of any element(s), limitation(ies), not specifically disclosed herein.

[0019] "Effector function" refers to the biological activities attributable to the Fc region of an antibody and varies depending on the antibody isotype. Examples of antibody effector functions include C1q binding and complement-dependent cytotoxicity (CDC), Fc receptor binding, antibody-dependent cell-mediated cytotoxicity (ADCC), phagocytosis, down-regulation of cell surface receptors (e.g., B cell receptors), and B cell activation. One effector function is the ability of the Fc region, or constant region, of an antibody to bind proteins, including, but not limited to, Fc receptors (FcRs) (e.g., high-affinity IgG Fc region receptor Ia (FCγRIa) (CD64) (SEQ ID NO: 34), low-affinity immunoglobulin gamma Fc region receptor IIa (FCγRIIa) (CD32) (SEQ ID NO: 35), low-affinity immunoglobulin gamma Fc region receptor IIIa (FCγRIIIA) (CD16a) (SEQ ID NO: 36), and low-affinity immunoglobulin gamma Fc region receptor IIIb (FCγRIIIb) (CD16b) (SEQ ID NO: 37)). In embodiments of the invention in which the antibodies and antigen-binding fragments thereof comprise an Fc domain, the Fc domain has been engineered to reduce or eliminate one or more Fc effector functions. In preferred embodiments, the Fc domain has been engineered to reduce or eliminate platelet activation and / or platelet aggregation and the risk of thromboembolic complications.

[0020] As used herein, the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise. The use of "or" or "and" means "and / or" unless stated otherwise. Furthermore, the use of the term "including," as well as other forms such as "include," "includes," and "included," is not limiting.

[0021] The use of the conjunction "or" is used interchangeably with "at least one." For example, if a composition includes A or B, the method must include at least one of A and B, but may include both A and B. Similarly, if a composition includes "A, B, C, or D," the method must include at least one of the group A, B, C, and D, but may include all or any combination of A, B, C, and D.

[0022] The term "about" indicates that a stated numerical value is allowed some minimal imprecision (approximating to a certain extent the exact value; roughly or fairly near the value; approximately the value). Unless the imprecision conferred by "about" is otherwise understood in the art in contrast to this ordinary meaning, "about" as used herein encompasses a variation of up to ±10% from the specified value. Unless otherwise specified, any numbers expressing quantities of ingredients, properties such as molecular weight, reaction conditions, and the like used herein should be understood as modified by the term "about."

[0023] CD40L is also known as CD154, gp39, T-BAM, 5c8 antigen, or TNF-related activation protein (TRAP).

[0024] The terms "treat," "treatment," and the like include therapeutic and prophylactic treatment. Therapeutic treatment is treatment of a subject who has signs or symptoms of the disease, condition, or disorder to be treated. Prophylactic treatment refers to treatment of a subject who is susceptible to such disease, condition, or disorder but who does not exhibit overt signs of the disease, condition, or disorder. Thus, treatment may result in the arrest, partial or total alleviation, or suppression of signs or symptoms of a disease, and specifically includes, but is not limited to, prolongation of survival and cure.

[0025] An "effective amount" of an agent, e.g., a pharmaceutical formulation, refers to that amount, at dosages and for periods of time necessary, effective to achieve a desired therapeutic or prophylactic result.

[0026] "Affinity" refers to the overall strength of the non-covalent interaction between a single binding site of a molecule (e.g., an antibody) and its binding partner (e.g., an antigen). Unless otherwise specified, as used herein, "binding affinity" refers to intrinsic binding affinity, reflecting a 1:1 interaction between members of a binding pair (e.g., an antibody and an antigen). The affinity of a molecule X for its partner Y may generally be represented by the equilibrium dissociation constant (KD), which is the ratio of koff / kon between an antibody and its antigen. KD and affinity are inversely related. The KD value is related to the antibody concentration (the amount of antibody required for a particular experiment), so the lower the KD value (lower concentration), the higher the affinity of the antibody. Affinity may be measured by common methods known in the art, including those described herein. In a specific exemplary embodiment for measuring binding affinity, the binding affinity may be measured by radioimmunoassay (RIA), surface plasmon resonance (SPR) on a BIAcore® instrument (GE Healthcare Europe GmbH, Glattbrugg, Switzerland) by capturing the antibody on a Protein A-coupled CM5 research-grade sensor chip (GE Healthcare Europe GmbH, Glattbrugg, Switzerland; BR-1000-14) with a human CD40 ligand polypeptide used as the analyte. Other methods may include radioimmunoassay and equilibrium exclusion binding assays. The equilibrium exclusion binding assay is a versatile immunoassay platform capable of measuring the equilibrium dissociation constant, as well as the association and dissociation rate constants of antigen / antibody interactions.

[0027] As used herein, reference is made to the percent identity between polypeptide or amino acid sequences. The percent identity between two sequences is a function of the number of identical positions shared by the two sequences, taking into account the number of gaps and the length of each gap, which may need to be introduced for optimal alignment of the two sequences. Identity can be measured as either "local identity" or "global identity." Local identity refers to the degree of sequence relatedness between polypeptides and is determined by the string-to-string match of such sequences. Global identity refers to the degree of sequence relatedness of a polypeptide compared to the full length of a reference polypeptide. Unless otherwise specified, as used herein, identity refers to global identity. For purposes of this disclosure and claims, percent global identity is calculated using the Needleman and Wunsch ((1970) J. Mol. Biol. 48:444-453) algorithm using the Blossum62 scoring matrix with a gap penalty of 12, a gap extension penalty of 4, and a frameshift gap penalty of 5. There are a number of publicly available software programs that incorporate the Needleman and Wunsch algorithm, for example the GAP program in the GCG software package.

[0028] Those skilled in the art will understand that starting materials, biological and chemical materials, biological and chemical reagents, synthetic methods, purification methods, analytical methods, assay methods, and biological methods, other than those specifically exemplified, can be employed in the practice of the present invention without undue experimentation. Art-known functional equivalents of any such materials and methods are intended to be included in this disclosure.

[0029] Amino acid substitutions are conventionally designated by identifying the original amino acid, the position of such amino acid within the designated sequence, and the substituted amino acid, e.g., C11S indicates that a cysteine ​​at position 11 of the polypeptide sequence is replaced with a serine.

[0030] Humanized antibodies are antibodies produced by non-human species whose protein sequences have been altered to resemble antibody variants naturally produced in humans. The "humanization" process is typically applied to monoclonal antibodies produced for administration to humans (e.g., antibodies produced as anti-cancer drugs).

[0031] Currently, it is common to humanize non-human antibodies by inserting the relevant CDRs from antibodies generated in non-human animals into a human antibody "scaffold." The "direct" creation of humanized antibodies can be achieved by inserting the appropriate CDR-encoding segments (responsible for the desired binding properties) into a human antibody "scaffold." This may be achieved by recombinant DNA techniques using an appropriate vector for expression in mammalian cells. That is, after an antibody is generated in a mouse (or other non-human) with the desired properties, the DNA encoding the antibody can be isolated, cloned into a vector, and sequenced. The DNA sequences corresponding to the antibody's CDRs can then be determined. Once the precise sequences of the desired CDRs are known, a strategy can be devised for appropriately inserting these sequences into a construct containing DNA for the human antibody variant. The CDRs may be altered prior to insertion into the scaffold, e.g., to increase specificity.

[0032] The term "human" antibody refers to an antibody having an amino acid sequence corresponding to an antibody produced in a human and / or an antibody produced using any technique that produces human antibodies. This definition of human antibody specifically excludes humanized antibodies that comprise non-human antigen-binding residues.

[0033] "Antigen-binding antibody fragment" refers to a molecule other than an intact antibody, including a portion of an intact antibody that binds to the antigen and that binds to the antigen. Examples of antibody fragments include, but are not limited to, Fv, Fab, Fab', Fd fragments, dAb fragments, Fab'-SH, F(ab')2; diabodies; triabodies; linear antibodies; single-chain antibody molecules (e.g., scFv); and multispecific antibodies formed from an antibody fragment and a minimal recognition unit consisting of amino acid residues mimicking the hypervariable region of an antibody (e.g., an isolated complementarity-determining region (CDR) such as a CDR3 peptide). The antigen-binding fragments disclosed in the present application bind to the antigen CD40L.

[0034] Antigen-binding fragments of antibodies typically contain at least one variable domain. The variable domain may be of any size or amino acid composition and generally contains at least one CDR adjacent to or in frame with one or more framework sequences. In antigen-binding fragments having a VH domain associated with a VL domain, the VH and VL domains may be positioned in an appropriate orientation relative to each other. For example, the variable region may be dimeric and contain VH-VH, VH-VL, or VL-VL dimers. Alternatively, the antigen-binding fragment of an antibody may contain a monomeric VH or VL domain.

[0035] Isolated antibodies include bispecific antibodies in which each arm or antigen-binding fragment of the antibody binds to a different target or epitope.

[0036] The terms "full length antibody," "intact antibody," and "whole antibody" are used interchangeably herein and refer to antibodies having a structure substantially similar to a native antibody structure or having heavy chains containing an Fc region.

[0037] "5c8" refers to a mouse anti-human antibody that binds CD40L and is produced by the hybridoma described in U.S. Patent No. 5,474,771, available from the ATCC under accession number HB10916. "hu5c8" refers to the humanized version of 5c8, the sequence of which is disclosed in Karpusas, et al., Structure vol. 9, pp. 321-329, (2001).

[0038] The terms "subject" and "individual" and "patient" are used interchangeably herein to refer to a human subject, individual or patient.

[0039] All compositional percentages herein are by weight of the total composition unless otherwise specified. As used herein, the term "comprises" and variations thereof are intended to be non-limiting, and the recitation of listed items does not exclude other similar items and may be useful in the materials, compositions, devices, and methods of the present technology. Similarly, the terms "may" and "may comprise" and variations thereof are intended to be non-limiting, and when a recitation states that an embodiment may or may comprise certain elements or features, it does not exclude other embodiments of the present technology that do not contain those elements or features.

[0040] The term "isolated protein" or "isolated polypeptide" (e.g., an isolated antibody or isolated antigen-binding fragment) refers to a protein or polypeptide that, by virtue of its origin or source of derivation, is not associated with naturally associated components with which it is found in its natural state; a protein or polypeptide that is substantially free of other proteins from the same species; a protein or polypeptide expressed by cells obtained from a different species; or a protein or polypeptide that does not occur in nature. Thus, a polypeptide that is chemically synthesized or synthesized in a cellular system different from the cell from which it naturally originates is "isolated" from its naturally associated components. A protein may be rendered substantially free of naturally associated components by isolation using protein purification techniques well known in the art.

[0041] The term "mutant," as used herein, is defined as a modified or altered form of a wild-type sequence, for example, where one or more amino acids may be substituted with other amino acid(s) or non-amino acid(s) that do not substantially affect function. In some embodiments, a variant may contain an altered side chain for at least one amino acid residue.

[0042] As used herein, the term "antigen" is defined as an entity that can stimulate the production of antibodies, can specifically bind to those antibodies, and / or elicit an immune system response, such as a cell surface protein or specific linear or non-linear portion thereof. The term herein may be abbreviated as "Ag."

[0043] The term "specifically binds" and the like means that an antibody or antigen-binding fragment thereof forms a complex with an antigen that is relatively stable under physiological conditions. Specific binding may be characterized by an equilibrium dissociation constant (KD) of about 3000 nM or less (i.e., a smaller KD indicates tighter binding), about 2000 nM or less, about 1000 nM or less, about 500 nM or less, about 300 nM or less, about 200 nM or less, about 100 nM or less, about 50 nM or less, about 1 nM or less, or about 0.5 nM.

[0044] Specific binding to a particular antigen or epitope can be exhibited, for example, by an antibody having a KD for the antigen or epitope of at least about 1×10 M, at least about 1×10 M, at least about 1×10 M, at least about 1×10 M, at least about 1×10 M, or at least about 1×10 M, or in other cases at least about 1×10 M, at least about 1×10 M, or at least about 1×10 M, where KD refers to the equilibrium dissociation constant of the particular antibody-antigen interaction. Typically, an antibody that specifically binds to an antigen has a KD for a control molecule that is 20-fold, 50-fold, 100-fold, 500-fold, 1000-fold, 5,000-fold, 10,000-fold, or more times greater than that for that antigen or epitope. Specific binding to a particular antigen or epitope can also be exhibited, for example, by an antibody having a K a for that antigen or epitope that is at least 20-fold, 50-fold, 100-fold, 500-fold, 1000-fold, 5,000-fold, 10,000-fold, or more times greater than for a control epitope, where K a refers to the association rate of a particular antibody-antigen interaction.

[0045] The term "neutralizing antibody" includes antibodies that can inhibit and / or neutralize the biological activity of CD40L, such as anti-CD40L antibodies or antigenic fragments thereof that inhibit, prevent, or reduce binding of CD40L to CD40, thereby inhibiting or reducing signal transduction pathways evoked by CD40L and / or inhibiting or reducing binding of CD40L to CD40.

[0046] The terms "antagonistic antibody" or "antagonist antibody" are used equivalently herein and include antibodies that can inhibit and / or neutralize the biological signaling activity of CD40L, as described above for neutralizing antibodies.

[0047] The terms and expressions used herein are used as descriptive terms rather than as limiting terms, and the use of such terms and expressions is not intended to exclude any equivalents of the features shown and described, or portions thereof, but it is recognized that various modifications are possible within the scope of the invention as claimed. Thus, while aspects of the invention have been specifically disclosed by preferred embodiments, exemplary embodiments, and various embodiments that may include optional features, it should be understood that modifications and variations of the concepts disclosed herein may occur to those skilled in the art. Such modifications and variations are deemed to be within the scope of the embodiments of the invention, which may be described and defined in the appended claims.

[0048] Pharmaceutical compositions and methods of administration

[0049] The pharmaceutical compositions used in accordance with the method of the present disclosure can be formulated in a conventional manner using one or more physiologically acceptable carriers.Pharmacologically acceptable carriers are determined in part by the specific composition to be administered and by the specific method used to administer the composition.Therefore, the formulations of compounds useful in the method of the present disclosure are diverse (see, for example, Remington: The Science and Practice of Pharmacy, 20th ed., Gennaro et al. Eds., Lippincott Williams and Wilkins, 2000).

[0050] Formulations suitable for parenteral administration include aqueous and non-aqueous isotonic sterile injection solutions, which may contain antioxidants, buffers, bacteriostats, and solutes that render the formulation isotonic with the blood of the intended recipient, as well as aqueous and non-aqueous sterile suspensions, which may include suspending agents, solubilizers, thickening agents, stabilizers, and preservatives. Exemplary injection or infusion additives may include mannitol, citric acid monohydrate, sodium hydrogen phosphate dihydrate, sodium dihydrogen phosphate dihydrate, polysorbate 80, sodium chloride, sodium citrate, and water.

[0051] In accordance with the present disclosure, compounds can be administered by any suitable means, which may vary depending on the type of disorder being treated and the properties of the compound itself. For example, for the antibodies and antigen-binding fragments thereof of the present invention, preferred routes of administration include parenteral routes, such as intramuscular, intravenous, intraarterial, intraperitoneal, intracerebrospinal, intraspinal, epidural, subcutaneous, or via sustained-release systems or implants. Parenteral administration is preferably by injection, most preferably by intravenous, intramuscular, or subcutaneous injection.

[0052] In some embodiments, the anti-CD40L antibodies or antigen-binding fragments thereof of the present disclosure are formulated and may be lyophilized for storage and reconstituted in appropriate excipients prior to use according to art-known lyophilization and reconstitution methods.

[0053] In one exemplary pharmaceutical composition containing an anti-CD40L antibody or antigen-binding fragment thereof, the composition is formulated as a preservative-free sterile solution of the anti-CD40L antibody or antigen-binding fragment thereof for intravenous or subcutaneous administration. The formulation may be supplied as a single-use prefilled pen, a single-use prefilled glass syringe containing, for example, about 1 mL, or a single-use vial for medical use. Preferably, the pharmaceutical composition containing the anti-CD40L antibody or antigen-binding fragment thereof is clear and colorless, and has a pH ranging from about 5.0 to about 6.9, preferably from about 5.0 to about 6.5, and even more preferably from about 5.0 to about 6.0. In various embodiments, the formulation containing the pharmaceutical composition, when reconstituted and administered to a subject, may contain about 500 mg to about 1 mg, or about 400 mg to about 10 mg, or about 300 mg to about 30 mg, or about 200 mg to about 50 mg of anti-CD40L antibody or antigen-binding fragment thereof per mL of solution.

[0054] The selection of a dosing regimen for a given therapeutic agent depends on several factors, including the metabolic rate of the agent in serum or tissue, the level of symptoms, the immunogenicity of the agent, and the accessibility of target cells within the biological matrix, the patient's overall health, the patient's medical history, etc. Preferably, a dosing regimen delivers the highest amount of therapeutic agent to the patient, subject to an acceptable level of side effects. Thus, the amount delivered will depend, in part, on the individual agent and the severity of the condition being treated. It should be understood that the determination of appropriate dosage forms, doses, and routes of administration is within the level of those skilled in the pharmaceutical and medical arts.

[0055] dose

[0056] The pharmaceutical formulation of the present disclosure may contain about 0.001 to about 200 mg / kg of an anti-CD40L antibody or antigen-binding fragment thereof. For example, the anti-CD40L antibody or antigen-binding fragment thereof may be administered intravenously as a bolus dose of about 0.001 mg / kg to about 100 mg / kg, about 0.001 mg / kg to about 50 mg / kg, or about 0.001 mg / kg to about 10 mg / kg, with the remaining antibody dose administered intravenously. A predetermined dose of the anti-CD40L antibody or antigen-binding fragment thereof may be administered over a period of, for example, 1 to 2 hours to 5 hours.

[0057] In further embodiments, a portion of the dose of the antibody formulation is administered as a subcutaneous injection and / or bolus injection, with the remainder administered as an infusion. In some exemplary doses, the antibody formulation may be administered subcutaneously at a dose ranging from about 0.001 to about 200 mg / kg, e.g., about 0.001 mg / kg to about 100 mg / kg, or about 0.001 mg / kg to about 50 mg / kg, or about 0.001 mg / kg to about 10 mg / kg of the anti-CD40L antibody or antigen-binding fragment thereof may be administered intravenously. In some embodiments, the dose may be given as a bolus, with the remaining antibody dose administered subcutaneously or intravenously. A given dose of the anti-CD40L antibody or antigen-binding fragment thereof may be administered over a period of, for example, 1 hour, or 2 hours, or 3 hours, or 4 hours, or 5 hours, or longer.

[0058] Combination therapy

[0059] The antibodies or antibody fragments thereof described herein can be administered alone (monotherapy) or in combination, i.e., in combination with other agents. For example, in one embodiment, the combination therapy may include one or more additional therapeutic agents. In another embodiment, the combination therapy includes standard therapeutic treatment (consisting essentially of an antibody or antibody fragment thereof), which may or may not include additional therapeutic agents.

[0060] Adjunctive or combined administration (coadministration) includes simultaneous administration of any of the antibodies or antigen-binding fragments thereof described herein with one or more other agents in the same or different dosage forms, or separate administration (e.g., sequential administration) of the polypeptide and one or more other agents. Preferably, such combined or sequential administration results in both the polypeptide and one or more other agents being present in the treated patient at the same time.

[0061] Kits and manufactured products

[0062] Further provided are kits containing the antibodies or antigen-binding fragments thereof described herein and instructions for use. The kits typically include a package containing a combination of reagents in predetermined amounts along with instructions for use, and a label indicating the intended use of the kit contents. The term label or instructions includes any written or recorded material supplied on or with the kit, or otherwise accompanies the kit at any time during its manufacture, transport, sale, or use. It may be in a form required by a government agency regulating the manufacture, use, or sale of pharmaceuticals or biological products, and the notice reflects approval by such agency for manufacture, use, or sale for human administration or veterinary use. Labels or instructions can also include advertising booklets and pamphlets, packaging materials, and audio or video instructions.

[0063] Embodiment

[0064] All embodiments of the isolated antibody or antigen-binding fragment thereof bind to CD40L and inhibit or block binding of CD40L to CD40. As used herein, "blocks binding of CD40L to CD40" and "blocks the interaction of CD40L and CD40" are used interchangeably. Inhibition or blocking of binding can be direct or indirect. Generally, the antibody or antigen-binding fragment thereof physically prevents binding of CD40L to CD40 by directly and specifically competing with CD40 for the same binding site on CD40L, or by steric hindrance caused by the antibody or antigen-binding fragment thereof binding closely to the CD40-binding site on CD40L. In other cases, the effect is indirect, e.g., the antibody or antigen-binding fragment thereof induces an allosteric change in the conformation of CD40L, thereby inhibiting or eliminating binding of CD40L to CD40.

[0065] One embodiment is an isolated antibody that binds to CD40L and comprises a light chain and a heavy chain, wherein the light chain comprises a light chain variable region comprising an amino acid sequence that has at least 90%, or at least 91%, or at least 92%, or at least 93%, or at least 94%, or at least 95%, or at least 96%, or at least 97%, or at least 98%, or at least 99% sequence identity to SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, or SEQ ID NO:8, and the heavy chain comprises a heavy chain variable region, wherein the heavy chain variable region comprises an amino acid sequence that has at least 90%, or at least 91%, or at least 92%, or at least 93%, or at least 94%, or at least 95%, or at least 96%, or at least 97%, or at least 98%, or at least 99% sequence identity to SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, or SEQ ID NO:4.

[0066] In one embodiment, the disclosure provides an isolated antibody or antigen-binding fragment thereof that specifically binds to CD40L, comprising: a) a heavy chain variable region (VH) comprising: i) a CDRH1 domain comprising the sequence set forth in SEQ ID NO: 9; ii) a CDRH2 domain comprising the sequence set forth in SEQ ID NO: 10, 11, 12, 13, or 14; and iii) a CDRH3 domain comprising the sequence set forth in SEQ ID NO: 15; and (b) a light chain variable region (VL) comprising: i) a CDRL1 domain comprising the sequence set forth in SEQ ID NO: 16 or 17; ii) a CDRL2 domain comprising the sequence set forth in SEQ ID NO: 18 or 19; and iii) a CDRL3 domain comprising the sequence set forth in SEQ ID NO: 20.

[0067] In aspect A, the disclosure provides an isolated antibody or antigen-binding fragment thereof that specifically binds to CD40L, comprising: a) a heavy chain variable region (VH) comprising i) a CDRH1 domain consisting of the sequence set forth in SEQ ID NO: 9; ii) a CDRH2 domain consisting of the sequence set forth in SEQ ID NO: 10, 11, 12, 13, or 14; and iii) a CDRH3 domain consisting of the sequence set forth in SEQ ID NO: 15; and (b) a light chain variable region (VL) comprising i) a CDRL1 domain consisting of the sequence set forth in SEQ ID NO: 16 or 17; ii) a CDRL2 domain consisting of the sequence set forth in SEQ ID NO: 18 or 19; and iii) a CDRL3 domain consisting of the sequence set forth in SEQ ID NO: 20.

[0068] One embodiment of aspect A is an isolated antibody or antigen-binding fragment thereof comprising: (a) a heavy chain variable region (VH) comprising i) a CDRH1 domain comprising the sequence set forth in SEQ ID NO:9; ii) a CDRH2 domain comprising the sequence set forth in SEQ ID NO:10; and iii) a CDRH3 domain comprising the sequence set forth in SEQ ID NO:15; and (b) a light chain variable region (VL) comprising i) a CDRL1 domain comprising the sequence set forth in SEQ ID NO:16; ii) a CDRL2 domain comprising the sequence set forth in SEQ ID NO:18; and iii) a CDRL3 domain comprising the sequence set forth in SEQ ID NO:20. Another embodiment of aspect A is an isolated antibody or antigen-binding fragment thereof comprising: (a) a heavy chain variable region (VH) comprising i) a CDRH1 domain consisting of the sequence set forth in SEQ ID NO:9, ii) a CDRH2 domain consisting of the sequence set forth in SEQ ID NO:10, and iii) a CDRH3 domain consisting of the sequence set forth in SEQ ID NO:15; and (b) a light chain variable region (VL) comprising i) a CDRL1 domain consisting of the sequence set forth in SEQ ID NO:16, ii) a CDRL2 domain consisting of the sequence set forth in SEQ ID NO:18, and iii) a CDRL3 domain consisting of the sequence set forth in SEQ ID NO:20.

[0069] Another embodiment of aspect A is an isolated antibody or antigen-binding fragment thereof comprising: (a) a heavy chain variable region (VH) comprising i) a CDRH1 domain comprising the sequence set forth in SEQ ID NO:9; ii) a CDRH2 domain comprising the sequence set forth in SEQ ID NO:10; and iii) a CDRH3 domain comprising the sequence set forth in SEQ ID NO:15; and (b) a light chain variable region (VL) comprising i) a CDRL1 domain comprising the sequence set forth in SEQ ID NO:17; ii) a CDRL2 domain comprising the sequence set forth in SEQ ID NO:18; and iii) a CDRL3 domain comprising the sequence set forth in SEQ ID NO:20. Yet another embodiment of aspect A is an isolated antibody or antigen-binding fragment thereof comprising: (a) a heavy chain variable region (VH) comprising i) a CDRH1 domain consisting of the sequence set forth in SEQ ID NO:9, ii) a CDRH2 domain consisting of the sequence set forth in SEQ ID NO:10, and iii) a CDRH3 domain consisting of the sequence set forth in SEQ ID NO:15; and (b) a light chain variable region (VL) comprising i) a CDRL1 domain consisting of the sequence set forth in SEQ ID NO:17, ii) a CDRL2 domain consisting of the sequence set forth in SEQ ID NO:18, and iii) a CDRL3 domain consisting of the sequence set forth in SEQ ID NO:20.

[0070] Yet another embodiment of aspect A is an isolated antibody or antigen-binding fragment thereof comprising: (a) a heavy chain variable region (VH) comprising i) a CDRH1 domain comprising the sequence set forth in SEQ ID NO:9; ii) a CDRH2 domain comprising the sequence set forth in SEQ ID NO:10; and iii) a CDRH3 domain comprising the sequence set forth in SEQ ID NO:15; and (b) a light chain variable region (VL) comprising i) a CDRL1 domain comprising the sequence set forth in SEQ ID NO:16; ii) a CDRL2 domain comprising the sequence set forth in SEQ ID NO:19; and iii) a CDRL3 domain comprising the sequence set forth in SEQ ID NO:20. Another embodiment of aspect A is an isolated antibody or antigen-binding fragment thereof comprising: (a) a heavy chain variable region (VH) comprising i) a CDRH1 domain consisting of the sequence set forth in SEQ ID NO:9, ii) a CDRH2 domain consisting of the sequence set forth in SEQ ID NO:10, and iii) a CDRH3 domain consisting of the sequence set forth in SEQ ID NO:15; and (b) a light chain variable region (VL) comprising i) a CDRL1 domain consisting of the sequence set forth in SEQ ID NO:16, ii) a CDRL2 domain consisting of the sequence set forth in SEQ ID NO:19, and iii) a CDRL3 domain consisting of the sequence set forth in SEQ ID NO:20.

[0071] Another embodiment of aspect A is an isolated antibody or antigen-binding fragment thereof comprising: (a) a heavy chain variable region (VH) comprising i) a CDRH1 domain comprising the sequence set forth in SEQ ID NO:9; ii) a CDRH2 domain comprising the sequence set forth in SEQ ID NO:10; and iii) a CDRH3 domain comprising the sequence set forth in SEQ ID NO:15; and (b) a light chain variable region (VL) comprising i) a CDRL1 domain comprising the sequence set forth in SEQ ID NO:17; ii) a CDRL2 domain comprising the sequence set forth in SEQ ID NO:19; and iii) a CDRL3 domain comprising the sequence set forth in SEQ ID NO:20. Yet another embodiment of aspect A is an isolated antibody or antigen-binding fragment thereof comprising: (a) a heavy chain variable region (VH) comprising i) a CDRH1 domain consisting of the sequence set forth in SEQ ID NO:9, ii) a CDRH2 domain consisting of the sequence set forth in SEQ ID NO:10, and iii) a CDRH3 domain consisting of the sequence set forth in SEQ ID NO:15; and (b) a light chain variable region (VL) comprising i) a CDRL1 domain consisting of the sequence set forth in SEQ ID NO:17, ii) a CDRL2 domain consisting of the sequence set forth in SEQ ID NO:19, and iii) a CDRL3 domain consisting of the sequence set forth in SEQ ID NO:20.

[0072] Another embodiment of aspect A is an isolated antibody or antigen-binding fragment thereof comprising: (a) a heavy chain variable region (VH) comprising i) a CDRH1 domain comprising the sequence set forth in SEQ ID NO:9; ii) a CDRH2 domain comprising the sequence set forth in SEQ ID NO:11; and iii) a CDRH3 domain comprising the sequence set forth in SEQ ID NO:15; and (b) a light chain variable region (VL) comprising i) a CDRL1 domain comprising the sequence set forth in SEQ ID NO:16; ii) a CDRL2 domain comprising the sequence set forth in SEQ ID NO:18; and iii) a CDRL3 domain comprising the sequence set forth in SEQ ID NO:20. Yet another embodiment of aspect A is an isolated antibody or antigen-binding fragment thereof comprising: (a) a heavy chain variable region (VH) comprising i) a CDRH1 domain consisting of the sequence set forth in SEQ ID NO:9, ii) a CDRH2 domain consisting of the sequence set forth in SEQ ID NO:11, and iii) a CDRH3 domain consisting of the sequence set forth in SEQ ID NO:15; and (b) a light chain variable region (VL) comprising i) a CDRL1 domain consisting of the sequence set forth in SEQ ID NO:16, ii) a CDRL2 domain consisting of the sequence set forth in SEQ ID NO:18, and iii) a CDRL3 domain consisting of the sequence set forth in SEQ ID NO:20.

[0073] Another embodiment of aspect A is an isolated antibody or antigen-binding fragment thereof comprising: (a) a heavy chain variable region (VH) comprising i) a CDRH1 domain comprising the sequence set forth in SEQ ID NO:9; ii) a CDRH2 domain comprising the sequence set forth in SEQ ID NO:11; and iii) a CDRH3 domain comprising the sequence set forth in SEQ ID NO:15; and (b) a light chain variable region (VL) comprising i) a CDRL1 domain comprising the sequence set forth in SEQ ID NO:17; ii) a CDRL2 domain comprising the sequence set forth in SEQ ID NO:18; and iii) a CDRL3 domain comprising the sequence set forth in SEQ ID NO:20. Another embodiment of aspect A is an isolated antibody or antigen-binding fragment thereof comprising: (a) a heavy chain variable region (VH) comprising i) a CDRH1 domain consisting of the sequence set forth in SEQ ID NO:9, ii) a CDRH2 domain consisting of the sequence set forth in SEQ ID NO:11, and iii) a CDRH3 domain consisting of the sequence set forth in SEQ ID NO:15; and (b) a light chain variable region (VL) comprising i) a CDRL1 domain consisting of the sequence set forth in SEQ ID NO:17, ii) a CDRL2 domain consisting of the sequence set forth in SEQ ID NO:18, and iii) a CDRL3 domain consisting of the sequence set forth in SEQ ID NO:20.

[0074] Another embodiment of aspect A is an isolated antibody or antigen-binding fragment thereof comprising: (a) a heavy chain variable region (VH) comprising i) a CDRH1 domain comprising the sequence set forth in SEQ ID NO:9; ii) a CDRH2 domain comprising the sequence set forth in SEQ ID NO:11; and iii) a CDRH3 domain comprising the sequence set forth in SEQ ID NO:15; and (b) a light chain variable region (VL) comprising i) a CDRL1 domain comprising the sequence set forth in SEQ ID NO:16; ii) a CDRL2 domain comprising the sequence set forth in SEQ ID NO:19; and iii) a CDRL3 domain comprising the sequence set forth in SEQ ID NO:20. Another embodiment of aspect A is an isolated antibody or antigen-binding fragment thereof comprising: (a) a heavy chain variable region (VH) comprising i) a CDRH1 domain consisting of the sequence set forth in SEQ ID NO:9, ii) a CDRH2 domain consisting of the sequence set forth in SEQ ID NO:11, and iii) a CDRH3 domain consisting of the sequence set forth in SEQ ID NO:15; and (b) a light chain variable region (VL) comprising i) a CDRL1 domain consisting of the sequence set forth in SEQ ID NO:16, ii) a CDRL2 domain consisting of the sequence set forth in SEQ ID NO:19, and iii) a CDRL3 domain consisting of the sequence set forth in SEQ ID NO:20.

[0075] Another embodiment of aspect A is an isolated antibody or antigen-binding fragment thereof comprising: (a) a heavy chain variable region (VH) comprising i) a CDRH1 domain comprising the sequence set forth in SEQ ID NO:9; ii) a CDRH2 domain comprising the sequence set forth in SEQ ID NO:11; and iii) a CDRH3 domain comprising the sequence set forth in SEQ ID NO:15; and (b) a light chain variable region (VL) comprising i) a CDRL1 domain comprising the sequence set forth in SEQ ID NO:17; ii) a CDRL2 domain comprising the sequence set forth in SEQ ID NO:19; and iii) a CDRL3 domain comprising the sequence set forth in SEQ ID NO:20. Another embodiment of aspect A is an isolated antibody or antigen-binding fragment thereof comprising: (a) a heavy chain variable region (VH) comprising i) a CDRH1 domain consisting of the sequence set forth in SEQ ID NO:9, ii) a CDRH2 domain consisting of the sequence set forth in SEQ ID NO:11, and iii) a CDRH3 domain consisting of the sequence set forth in SEQ ID NO:15; and (b) a light chain variable region (VL) comprising i) a CDRL1 domain consisting of the sequence set forth in SEQ ID NO:17, ii) a CDRL2 domain consisting of the sequence set forth in SEQ ID NO:19, and iii) a CDRL3 domain consisting of the sequence set forth in SEQ ID NO:20.

[0076] Another embodiment of aspect A is an isolated antibody or antigen-binding fragment thereof comprising: (a) a heavy chain variable region (VH) comprising i) a CDRH1 domain comprising the sequence set forth in SEQ ID NO:9; ii) a CDRH2 domain comprising the sequence set forth in SEQ ID NO:12; and iii) a CDRH3 domain comprising the sequence set forth in SEQ ID NO:15; and (b) a light chain variable region (VL) comprising i) a CDRL1 domain comprising the sequence set forth in SEQ ID NO:16; ii) a CDRL2 domain comprising the sequence set forth in SEQ ID NO:18; and iii) a CDRL3 domain comprising the sequence set forth in SEQ ID NO:20. Yet another embodiment of aspect A is an isolated antibody or antigen-binding fragment thereof comprising: (a) a heavy chain variable region (VH) comprising i) a CDRH1 domain consisting of the sequence set forth in SEQ ID NO:9, ii) a CDRH2 domain consisting of the sequence set forth in SEQ ID NO:12, and iii) a CDRH3 domain consisting of the sequence set forth in SEQ ID NO:15; and (b) a light chain variable region (VL) comprising i) a CDRL1 domain consisting of the sequence set forth in SEQ ID NO:16, ii) a CDRL2 domain consisting of the sequence set forth in SEQ ID NO:18, and iii) a CDRL3 domain consisting of the sequence set forth in SEQ ID NO:20.

[0077] Another embodiment of aspect A is an isolated antibody or antigen-binding fragment thereof comprising: (a) a heavy chain variable region (VH) comprising i) a CDRH1 domain comprising the sequence set forth in SEQ ID NO:9; ii) a CDRH2 domain comprising the sequence set forth in SEQ ID NO:12; and iii) a CDRH3 domain comprising the sequence set forth in SEQ ID NO:15; and (b) a light chain variable region (VL) comprising i) a CDRL1 domain comprising the sequence set forth in SEQ ID NO:17; ii) a CDRL2 domain comprising the sequence set forth in SEQ ID NO:18; and iii) a CDRL3 domain comprising the sequence set forth in SEQ ID NO:20. Another embodiment of aspect A is an isolated antibody or antigen-binding fragment thereof comprising: (a) a heavy chain variable region (VH) comprising i) a CDRH1 domain consisting of the sequence set forth in SEQ ID NO:9, ii) a CDRH2 domain consisting of the sequence set forth in SEQ ID NO:12, and iii) a CDRH3 domain consisting of the sequence set forth in SEQ ID NO:15; and (b) a light chain variable region (VL) comprising i) a CDRL1 domain consisting of the sequence set forth in SEQ ID NO:17, ii) a CDRL2 domain consisting of the sequence set forth in SEQ ID NO:18, and iii) a CDRL3 domain consisting of the sequence set forth in SEQ ID NO:20.

[0078] Another embodiment of aspect A is an isolated antibody or antigen-binding fragment thereof comprising: (a) a heavy chain variable region (VH) comprising i) a CDRH1 domain comprising the sequence set forth in SEQ ID NO:9; ii) a CDRH2 domain comprising the sequence set forth in SEQ ID NO:12; and iii) a CDRH3 domain comprising the sequence set forth in SEQ ID NO:15; and (b) a light chain variable region (VL) comprising i) a CDRL1 domain comprising the sequence set forth in SEQ ID NO:16; ii) a CDRL2 domain comprising the sequence set forth in SEQ ID NO:19; and iii) a CDRL3 domain comprising the sequence set forth in SEQ ID NO:20. Another embodiment of aspect A is an isolated antibody or antigen-binding fragment thereof comprising: (a) a heavy chain variable region (VH) comprising i) a CDRH1 domain consisting of the sequence set forth in SEQ ID NO:9, ii) a CDRH2 domain consisting of the sequence set forth in SEQ ID NO:12, and iii) a CDRH3 domain consisting of the sequence set forth in SEQ ID NO:15; and (b) a light chain variable region (VL) comprising i) a CDRL1 domain consisting of the sequence set forth in SEQ ID NO:16, ii) a CDRL2 domain consisting of the sequence set forth in SEQ ID NO:19, and iii) a CDRL3 domain consisting of the sequence set forth in SEQ ID NO:20.

[0079] Yet another embodiment of aspect A is an isolated antibody or antigen-binding fragment thereof comprising: (a) a heavy chain variable region (VH) comprising i) a CDRH1 domain comprising the sequence set forth in SEQ ID NO:9; ii) a CDRH2 domain comprising the sequence set forth in SEQ ID NO:12; and iii) a CDRH3 domain comprising the sequence set forth in SEQ ID NO:15; and (b) a light chain variable region (VL) comprising i) a CDRL1 domain comprising the sequence set forth in SEQ ID NO:17; ii) a CDRL2 domain comprising the sequence set forth in SEQ ID NO:19; and iii) a CDRL3 domain comprising the sequence set forth in SEQ ID NO:20. Another embodiment of aspect A is an isolated antibody or antigen-binding fragment thereof comprising: (a) a heavy chain variable region (VH) comprising i) a CDRH1 domain consisting of the sequence set forth in SEQ ID NO:9, ii) a CDRH2 domain consisting of the sequence set forth in SEQ ID NO:12, and iii) a CDRH3 domain consisting of the sequence set forth in SEQ ID NO:15; and (b) a light chain variable region (VL) comprising i) a CDRL1 domain consisting of the sequence set forth in SEQ ID NO:17, ii) a CDRL2 domain consisting of the sequence set forth in SEQ ID NO:19, and iii) a CDRL3 domain consisting of the sequence set forth in SEQ ID NO:20.

[0080] Another embodiment of aspect A is an isolated antibody or antigen-binding fragment thereof comprising: (a) a heavy chain variable region (VH) comprising i) a CDRH1 domain comprising the sequence set forth in SEQ ID NO:9; ii) a CDRH2 domain comprising the sequence set forth in SEQ ID NO:13; and iii) a CDRH3 domain comprising the sequence set forth in SEQ ID NO:15; and (b) a light chain variable region (VL) comprising i) a CDRL1 domain comprising the sequence set forth in SEQ ID NO:16; ii) a CDRL2 domain comprising the sequence set forth in SEQ ID NO:18; and iii) a CDRL3 domain comprising the sequence set forth in SEQ ID NO:20. Another embodiment of aspect A is an isolated antibody or antigen-binding fragment thereof comprising: (a) a heavy chain variable region (VH) comprising i) a CDRH1 domain consisting of the sequence set forth in SEQ ID NO:9, ii) a CDRH2 domain consisting of the sequence set forth in SEQ ID NO:13, and iii) a CDRH3 domain consisting of the sequence set forth in SEQ ID NO:15; and (b) a light chain variable region (VL) comprising i) a CDRL1 domain consisting of the sequence set forth in SEQ ID NO:16, ii) a CDRL2 domain consisting of the sequence set forth in SEQ ID NO:18, and iii) a CDRL3 domain consisting of the sequence set forth in SEQ ID NO:20.

[0081] Another embodiment of aspect A is an isolated antibody or antigen-binding fragment thereof comprising: (a) a heavy chain variable region (VH) comprising i) a CDRH1 domain comprising the sequence set forth in SEQ ID NO:9; ii) a CDRH2 domain comprising the sequence set forth in SEQ ID NO:13; and iii) a CDRH3 domain comprising the sequence set forth in SEQ ID NO:15; and (b) a light chain variable region (VL) comprising i) a CDRL1 domain comprising the sequence set forth in SEQ ID NO:17; ii) a CDRL2 domain comprising the sequence set forth in SEQ ID NO:18; and iii) a CDRL3 domain comprising the sequence set forth in SEQ ID NO:20. Yet another embodiment of aspect A is an isolated antibody or antigen-binding fragment thereof comprising: (a) a heavy chain variable region (VH) comprising i) a CDRH1 domain consisting of the sequence set forth in SEQ ID NO:9, ii) a CDRH2 domain consisting of the sequence set forth in SEQ ID NO:13, and iii) a CDRH3 domain consisting of the sequence set forth in SEQ ID NO:15; and (b) a light chain variable region (VL) comprising i) a CDRL1 domain consisting of the sequence set forth in SEQ ID NO:17, ii) a CDRL2 domain consisting of the sequence set forth in SEQ ID NO:18, and iii) a CDRL3 domain consisting of the sequence set forth in SEQ ID NO:20.

[0082] Yet another embodiment of aspect A is an isolated antibody or antigen-binding fragment thereof comprising: (a) a heavy chain variable region (VH) comprising i) a CDRH1 domain comprising the sequence set forth in SEQ ID NO:9; ii) a CDRH2 domain comprising the sequence set forth in SEQ ID NO:13; and iii) a CDRH3 domain comprising the sequence set forth in SEQ ID NO:15; and (b) a light chain variable region (VL) comprising i) a CDRL1 domain comprising the sequence set forth in SEQ ID NO:17; ii) a CDRL2 domain comprising the sequence set forth in SEQ ID NO:19; and iii) a CDRL3 domain comprising the sequence set forth in SEQ ID NO:20. Another embodiment of aspect A is an isolated antibody or antigen-binding fragment thereof comprising: (a) a heavy chain variable region (VH) comprising i) a CDRH1 domain consisting of the sequence set forth in SEQ ID NO:9, ii) a CDRH2 domain consisting of the sequence set forth in SEQ ID NO:13, and iii) a CDRH3 domain consisting of the sequence set forth in SEQ ID NO:15; and (b) a light chain variable region (VL) comprising i) a CDRL1 domain consisting of the sequence set forth in SEQ ID NO:17, ii) a CDRL2 domain consisting of the sequence set forth in SEQ ID NO:19, and iii) a CDRL3 domain consisting of the sequence set forth in SEQ ID NO:20.

[0083] Another embodiment of aspect A is an isolated antibody or antigen-binding fragment thereof comprising: (a) a heavy chain variable region (VH) comprising i) a CDRH1 domain comprising the sequence set forth in SEQ ID NO:9; ii) a CDRH2 domain comprising the sequence set forth in SEQ ID NO:14; and iii) a CDRH3 domain comprising the sequence set forth in SEQ ID NO:15; and (b) a light chain variable region (VL) comprising i) a CDRL1 domain comprising the sequence set forth in SEQ ID NO:16; ii) a CDRL2 domain comprising the sequence set forth in SEQ ID NO:18; and iii) a CDRL3 domain comprising the sequence set forth in SEQ ID NO:20. Another embodiment of aspect A is an isolated antibody or antigen-binding fragment thereof comprising: (a) a heavy chain variable region (VH) comprising i) a CDRH1 domain consisting of the sequence set forth in SEQ ID NO:9, ii) a CDRH2 domain consisting of the sequence set forth in SEQ ID NO:14, and iii) a CDRH3 domain consisting of the sequence set forth in SEQ ID NO:15; and (b) a light chain variable region (VL) comprising i) a CDRL1 domain consisting of the sequence set forth in SEQ ID NO:16, ii) a CDRL2 domain consisting of the sequence set forth in SEQ ID NO:18, and iii) a CDRL3 domain consisting of the sequence set forth in SEQ ID NO:20.

[0084] Another embodiment of aspect A is an isolated antibody or antigen-binding fragment thereof comprising: (a) a heavy chain variable region (VH) comprising i) a CDRH1 domain comprising the sequence set forth in SEQ ID NO:9; ii) a CDRH2 domain comprising the sequence set forth in SEQ ID NO:14; and iii) a CDRH3 domain comprising the sequence set forth in SEQ ID NO:15; and (b) a light chain variable region (VL) comprising i) a CDRL1 domain comprising the sequence set forth in SEQ ID NO:17; ii) a CDRL2 domain comprising the sequence set forth in SEQ ID NO:18; and iii) a CDRL3 domain comprising the sequence set forth in SEQ ID NO:20. Another embodiment of aspect A is an isolated antibody or antigen-binding fragment thereof comprising: (a) a heavy chain variable region (VH) comprising i) a CDRH1 domain consisting of the sequence set forth in SEQ ID NO:9, ii) a CDRH2 domain consisting of the sequence set forth in SEQ ID NO:14, and iii) a CDRH3 domain consisting of the sequence set forth in SEQ ID NO:15; and (b) a light chain variable region (VL) comprising i) a CDRL1 domain consisting of the sequence set forth in SEQ ID NO:17, ii) a CDRL2 domain consisting of the sequence set forth in SEQ ID NO:18, and iii) a CDRL3 domain consisting of the sequence set forth in SEQ ID NO:20.

[0085] Another embodiment of aspect A is an isolated antibody or antigen-binding fragment thereof comprising: (a) a heavy chain variable region (VH) comprising i) a CDRH1 domain comprising the sequence set forth in SEQ ID NO:9; ii) a CDRH2 domain comprising the sequence set forth in SEQ ID NO:14; and iii) a CDRH3 domain comprising the sequence set forth in SEQ ID NO:15; and (b) a light chain variable region (VL) comprising i) a CDRL1 domain comprising the sequence set forth in SEQ ID NO:17; ii) a CDRL2 domain comprising the sequence set forth in SEQ ID NO:19; and iii) a CDRL3 domain comprising the sequence set forth in SEQ ID NO:20. Another embodiment of aspect A is an isolated antibody or antigen-binding fragment thereof comprising: (a) a heavy chain variable region (VH) comprising i) a CDRH1 domain consisting of the sequence set forth in SEQ ID NO:9, ii) a CDRH2 domain consisting of the sequence set forth in SEQ ID NO:14, and iii) a CDRH3 domain consisting of the sequence set forth in SEQ ID NO:15; and (b) a light chain variable region (VL) comprising i) a CDRL1 domain consisting of the sequence set forth in SEQ ID NO:17, ii) a CDRL2 domain consisting of the sequence set forth in SEQ ID NO:19, and iii) a CDRL3 domain consisting of the sequence set forth in SEQ ID NO:20.

[0086] In aspect B, the disclosure provides an isolated antibody or antigen-binding fragment thereof that specifically binds to CD40L, comprising: (a) a heavy chain variable region (VH) having the amino acid sequence set forth in SEQ ID NO: 1, 2, 3, or 4; and (b) a light chain variable region (VL) having the amino acid sequence set forth in SEQ ID NO: 5, 6, 7, or 8.

[0087] An embodiment of aspect B is an isolated antibody or antigen-binding fragment thereof, comprising a heavy chain variable region (VH) having the amino acid sequence set forth in SEQ ID NO:1 and a light chain variable region (VL) having the amino acid sequence set forth in SEQ ID NO:5.

[0088] Another embodiment of aspect B is an isolated antibody or antigen-binding fragment thereof, comprising a heavy chain variable region (VH) having the amino acid sequence set forth in SEQ ID NO:1 and a light chain variable region (VL) having the amino acid sequence set forth in SEQ ID NO:6.

[0089] Another embodiment of aspect B is an isolated antibody or antigen-binding fragment thereof, comprising a heavy chain variable region (VH) having the amino acid sequence set forth in SEQ ID NO:1 and a light chain variable region (VL) having the amino acid sequence set forth in SEQ ID NO:7.

[0090] Another embodiment of aspect B is an isolated antibody or antigen-binding fragment thereof, comprising a heavy chain variable region (VH) having the amino acid sequence set forth in SEQ ID NO:1 and a light chain variable region (VL) having the amino acid sequence set forth in SEQ ID NO:8.

[0091] Another embodiment of aspect B is an isolated antibody or antigen-binding fragment thereof, comprising a heavy chain variable region (VH) having the amino acid sequence set forth in SEQ ID NO:1 and a light chain variable region (VL) having the amino acid sequence set forth in SEQ ID NO:5.

[0092] Yet another embodiment of aspect B is an isolated antibody or antigen-binding fragment thereof, comprising a heavy chain variable region (VH) having the amino acid sequence set forth in SEQ ID NO:1 and a light chain variable region (VL) having the amino acid sequence set forth in SEQ ID NO:6.

[0093] Another embodiment of aspect B is an isolated antibody or antigen-binding fragment thereof, comprising a heavy chain variable region (VH) having the amino acid sequence set forth in SEQ ID NO:1 and a light chain variable region (VL) having the amino acid sequence set forth in SEQ ID NO:7.

[0094] Another embodiment of aspect B is an isolated antibody or antigen-binding fragment thereof, comprising a heavy chain variable region (VH) having the amino acid sequence set forth in SEQ ID NO:1 and a light chain variable region (VL) having the amino acid sequence set forth in SEQ ID NO:8.

[0095] Another embodiment of aspect B is an isolated antibody or antigen-binding fragment thereof, comprising a heavy chain variable region (VH) having the amino acid sequence set forth in SEQ ID NO:3 and a light chain variable region (VL) having the amino acid sequence set forth in SEQ ID NO:5.

[0096] Another embodiment of aspect B is an isolated antibody or antigen-binding fragment thereof, comprising a heavy chain variable region (VH) having the amino acid sequence set forth in SEQ ID NO:3 and a light chain variable region (VL) having the amino acid sequence set forth in SEQ ID NO:6.

[0097] Another embodiment of aspect B is an isolated antibody or antigen-binding fragment thereof, comprising a heavy chain variable region (VH) having the amino acid sequence set forth in SEQ ID NO:3 and a light chain variable region (VL) having the amino acid sequence set forth in SEQ ID NO:7.

[0098] Yet another embodiment of aspect B is an isolated antibody or antigen-binding fragment thereof, comprising a heavy chain variable region (VH) having the amino acid sequence set forth in SEQ ID NO:3 and a light chain variable region (VL) having the amino acid sequence set forth in SEQ ID NO:8.

[0099] Another embodiment of aspect B is an isolated antibody or antigen-binding fragment thereof, comprising a heavy chain variable region (VH) having the amino acid sequence set forth in SEQ ID NO:4 and a light chain variable region (VL) having the amino acid sequence set forth in SEQ ID NO:5.

[0100] Yet another embodiment of aspect B is an isolated antibody or antigen-binding fragment thereof, comprising a heavy chain variable region (VH) having the amino acid sequence set forth in SEQ ID NO:4 and a light chain variable region (VL) having the amino acid sequence set forth in SEQ ID NO:6.

[0101] Another embodiment of aspect B is an isolated antibody or antigen-binding fragment thereof, comprising a heavy chain variable region (VH) having the amino acid sequence set forth in SEQ ID NO:4 and a light chain variable region (VL) having the amino acid sequence set forth in SEQ ID NO:7.

[0102] Another embodiment of aspect B is an isolated antibody or antigen-binding fragment thereof, comprising a heavy chain variable region (VH) having the amino acid sequence set forth in SEQ ID NO:4 and a light chain variable region (VL) having the amino acid sequence set forth in SEQ ID NO:8.

[0103] Aspect C provides an isolated antibody of any embodiment described herein, wherein such antibody comprises an Fc region, and wherein such Fc region has been engineered to reduce or eliminate one or more Fc effector functions.

[0104] In one embodiment of aspect C, the Fc region is engineered to reduce or eliminate binding to an Fc receptor (FcR), or to reduce or eliminate binding to C1q. In a specific embodiment of aspect C, the FcR is FcγRIa (CD64), FcγRIIa (CD32), FcγRIIIa (CD16a), or FcγRIIIb (CD16b). In another embodiment of aspect C, the FcR is FcγRIIa.

[0105] An embodiment of Aspect C is an isolated antibody according to any one of the embodiments of the invention, wherein such antibody is of the IgG1, IgG2, IgG3, or IgG4 isotype, or any combination or hybrid thereof. Another embodiment of Aspect C is an isolated antibody according to any one of the embodiments of the invention, wherein such antibody is of the IgG1 isotype and the heavy chain constant region comprises the amino acid sequence set forth in SEQ ID NO: 21 (P238S and N297G). As shown in Experiment 3, none of the antibody clones provided in the present disclosure, engineered with an Fc of SEQ ID NO: 21, bound to FcγRIa, FcγRIIa, FcγRIIIa, or FcγRIIIb. As shown in Experiment 4, none of the antibody clones provided in the present disclosure, engineered with an Fc of SEQ ID NO: 21, bound to C1q.

[0106] A mutation at position 297 of the IgG backbone Fc (N297A) has been shown to abrogate IgG glycosylation and Fc gamma receptor binding. A mutation at position 265 of the IgG backbone Fc (D265A) has been shown to abrogate Fc gamma receptor binding. Accordingly, embodiments of the invention have been prepared that comprise these mutations in the Fc region. One embodiment of aspect C is an isolated antibody according to any one of the embodiments of the present disclosure, comprising a heavy chain constant region comprising the amino acid sequence set forth in SEQ ID NO: 22 (D265A and N297A).

[0107] US Patent Application Publication No. 2014 / 0294834 discloses mutations in the heavy chain constant region. Accordingly, another embodiment is an isolated antibody according to any one of the embodiments disclosed herein having an Fc region comprising a heavy chain constant region comprising the amino acid sequence set forth in SEQ ID NO: 23 (T299K-MM131).

[0108] Another embodiment is an isolated antibody according to any one of the embodiments disclosed herein having an Fc region comprising a heavy chain constant region comprising the amino acid sequence set forth in SEQ ID NO: 24 (N299K).

[0109] In certain embodiments, the isolated antibody comprises a heavy chain variable region connected to a heavy chain constant region. In one embodiment, the heavy chain variable region VH1 is directly connected to a heavy chain constant region having P238S and N297G, and the isolated antibody heavy chain consists of the amino acid sequence set forth in SEQ ID NO: 42. In another embodiment, the heavy chain variable region VH2 is directly connected to a heavy chain constant region having P238S and N297G, and the isolated antibody heavy chain consists of the amino acid sequence set forth in SEQ ID NO: 43. In yet another embodiment, the heavy chain variable region VH3 is directly connected to a heavy chain constant region having P238S and N297G, and the isolated antibody heavy chain consists of the amino acid sequence set forth in SEQ ID NO: 44. In yet another embodiment, the heavy chain variable region VH4 is directly connected to a heavy chain constant region having P238S and N297G, and the isolated antibody heavy chain consists of the amino acid sequence set forth in SEQ ID NO: 45.

[0110] In one embodiment, the heavy chain variable region VH1 is directly connected to a heavy chain constant region having D265A / N297A, and the isolated antibody heavy chain consists of the amino acid sequence set forth in SEQ ID NO: 46. In another embodiment, the heavy chain variable region VH2 is directly connected to a heavy chain constant region having D265A / N297A, and the isolated antibody heavy chain consists of the amino acid sequence set forth in SEQ ID NO: 47. In yet another embodiment, the heavy chain variable region VH3 is directly connected to a heavy chain constant region having 265A / N297A, and the isolated antibody heavy chain consists of the amino acid sequence set forth in SEQ ID NO: 48. In yet another embodiment, the heavy chain variable region V4 is directly connected to a heavy chain constant region having 265A / N297A, and the isolated antibody heavy chain consists of the amino acid sequence set forth in SEQ ID NO: 49.

[0111] In yet another embodiment, the heavy chain variable region VH1 is directly connected to a heavy chain constant region comprising an IgG1 / IgG4 hybrid, and the isolated antibody heavy chain consists of the amino acid sequence set forth in SEQ ID NO: 50. In yet another embodiment, the heavy chain variable region VH2 is directly connected to a heavy chain constant region comprising an IgG1 / IgG4 hybrid, and the isolated antibody heavy chain consists of the amino acid sequence set forth in SEQ ID NO: 51. In yet another embodiment, the heavy chain variable region VH3 is directly connected to a heavy chain constant region comprising an IgG1 / IgG4 hybrid, and the isolated antibody heavy chain consists of the amino acid sequence set forth in SEQ ID NO: 51. In another embodiment, the heavy chain variable region VH4 is directly connected to a heavy chain constant region comprising an IgG1 / IgG4 hybrid, and the isolated antibody heavy chain consists of the amino acid sequence set forth in SEQ ID NO: 53.

[0112] In another embodiment, the heavy chain variable region VH1 is directly connected to a heavy chain constant region having an IgG4 domain, and the isolated antibody heavy chain consists of the amino acid sequence set forth in SEQ ID NO: 54. In yet another embodiment, the heavy chain variable region VH2 is directly connected to a heavy chain constant region having an IgG4 domain, and the isolated antibody heavy chain consists of the amino acid sequence set forth in SEQ ID NO: 55. In yet another embodiment, the heavy chain variable region VH3 is directly connected to a heavy chain constant region having an IgG4 domain, and the isolated antibody heavy chain consists of the amino acid sequence set forth in SEQ ID NO: 56. In another embodiment, the heavy chain variable region VH4 is directly connected to a heavy chain constant region having an IgG4 domain, and the isolated antibody heavy chain consists of the amino acid sequence set forth in SEQ ID NO: 57.

[0113] Another embodiment is an isolated antibody according to any one of the embodiments, having a variable light chain and a light chain constant region comprising the sequence set forth in SEQ ID NO: 25. Another embodiment is variable light chain 1 having a light chain constant region the amino acid sequence of which is set forth in SEQ ID NO: 38. Another embodiment is variable light chain 2 having a light chain constant region the amino acid sequence of which is set forth in SEQ ID NO: 39. In yet another embodiment, variable light chain 3 having a light chain constant region the amino acid sequence of which is set forth in SEQ ID NO: 40. In yet another embodiment, variable light chain 4 having a light chain constant region the amino acid sequence of which is set forth in SEQ ID NO: 41.

[0114] Another embodiment is a method for treating a subject having a CD40L-associated disease or disorder, comprising administering to such subject a therapeutically effective amount of an antibody or antigen-binding fragment thereof disclosed in the present application.

[0115] Another embodiment is an isolated antibody having a heavy chain of the amino acid sequence set forth in SEQ ID NO:42 and a light chain sequence set forth in SEQ ID NO:38. Another embodiment is an isolated antibody having a heavy chain of the amino acid sequence set forth in SEQ ID NO:43 and a light chain sequence set forth in SEQ ID NO:38. Another embodiment is an isolated antibody having a heavy chain of the amino acid sequence set forth in SEQ ID NO:44 and a light chain sequence set forth in SEQ ID NO:38. Another embodiment is an isolated antibody having a heavy chain of the amino acid sequence set forth in SEQ ID NO:45 and a light chain sequence set forth in SEQ ID NO:38. Another embodiment is an isolated antibody having a heavy chain of the amino acid sequence set forth in SEQ ID NO:42 and a light chain sequence set forth in SEQ ID NO:39. Another embodiment is an isolated antibody having a heavy chain of the amino acid sequence set forth in SEQ ID NO:43 and a light chain sequence set forth in SEQ ID NO:39. Another embodiment is an isolated antibody having a heavy chain of the amino acid sequence set forth in SEQ ID NO:44 and a light chain sequence set forth in SEQ ID NO:39. Another embodiment is an isolated antibody having a heavy chain of the amino acid sequence set forth in SEQ ID NO:45 and a light chain sequence set forth in SEQ ID NO:39. Another embodiment is an isolated antibody having a heavy chain of the amino acid sequence set forth in SEQ ID NO: 42 and a light chain sequence set forth in SEQ ID NO: 40. Another embodiment is an isolated antibody having a heavy chain of the amino acid sequence set forth in SEQ ID NO: 43 and a light chain sequence set forth in SEQ ID NO: 40. Another embodiment is an isolated antibody having a heavy chain of the amino acid sequence set forth in SEQ ID NO: 44 and a light chain sequence set forth in SEQ ID NO: 40. Another embodiment is an isolated antibody having a heavy chain of the amino acid sequence set forth in SEQ ID NO: 45 and a light chain sequence set forth in SEQ ID NO: 40. Another embodiment is an isolated antibody having a heavy chain of the amino acid sequence set forth in SEQ ID NO: 42 and a light chain sequence set forth in SEQ ID NO: 41. Another embodiment is an isolated antibody having a heavy chain of the amino acid sequence set forth in SEQ ID NO: 43 and a light chain sequence set forth in SEQ ID NO: 41. Another embodiment is an isolated antibody having a heavy chain of the amino acid sequence set forth in SEQ ID NO: 44 and a light chain sequence set forth in SEQ ID NO: 41. Another embodiment is an isolated antibody having a heavy chain of the amino acid sequence set forth in SEQ ID NO: 45 and a light chain sequence set forth in SEQ ID NO: 41.

[0116] Another embodiment is an isolated antibody having a heavy chain of the amino acid sequence set forth in SEQ ID NO: 46 and a light chain of the amino acid sequence set forth in SEQ ID NO: 38. Another embodiment is an isolated antibody having a heavy chain of the amino acid sequence set forth in SEQ ID NO: 47 and a light chain amino acid sequence set forth in SEQ ID NO: 38. Another embodiment is an isolated antibody having a heavy chain of the amino acid sequence set forth in SEQ ID NO: 48 and a light chain amino acid sequence set forth in SEQ ID NO: 38. Another embodiment is an isolated antibody having a heavy chain of the amino acid sequence set forth in SEQ ID NO: 49 and a light chain amino acid sequence set forth in SEQ ID NO: 38. Another embodiment is an isolated antibody having a heavy chain of the amino acid sequence set forth in SEQ ID NO: 46 and a light chain amino acid sequence set forth in SEQ ID NO: 39. Another embodiment is an isolated antibody having a heavy chain of the amino acid sequence set forth in SEQ ID NO: 47 and a light chain amino acid sequence set forth in SEQ ID NO: 39. Another embodiment is an isolated antibody having a heavy chain of the amino acid sequence set forth in SEQ ID NO: 48 and a light chain amino acid sequence set forth in SEQ ID NO: 39. Another embodiment is an isolated antibody having a heavy chain of the amino acid sequence set forth in SEQ ID NO: 49 and a light chain amino acid sequence set forth in SEQ ID NO: 39. Another embodiment is an isolated antibody having a heavy chain of the amino acid sequence set forth in SEQ ID NO: 46 and a light chain amino acid sequence set forth in SEQ ID NO: 40. Another embodiment is an isolated antibody having a heavy chain of the amino acid sequence set forth in SEQ ID NO: 47 and a light chain amino acid sequence set forth in SEQ ID NO: 40. Another embodiment is an isolated antibody having a heavy chain of the amino acid sequence set forth in SEQ ID NO: 48 and a light chain amino acid sequence set forth in SEQ ID NO: 40. Another embodiment is an isolated antibody having a heavy chain of the amino acid sequence set forth in SEQ ID NO: 49 and a light chain amino acid sequence set forth in SEQ ID NO: 40. Another embodiment is an isolated antibody having a heavy chain of the amino acid sequence set forth in SEQ ID NO: 46 and a light chain amino acid sequence set forth in SEQ ID NO: 41. Another embodiment is an isolated antibody having a heavy chain of the amino acid sequence set forth in SEQ ID NO: 47 and a light chain amino acid sequence set forth in SEQ ID NO: 41. Another embodiment is an isolated antibody having a heavy chain of the amino acid sequence set forth in SEQ ID NO: 48 and a light chain amino acid sequence set forth in SEQ ID NO: 41.Another embodiment is an isolated antibody having a heavy chain amino acid sequence set forth in SEQ ID NO:49 and a light chain amino acid sequence set forth in SEQ ID NO:41.

[0117] Another embodiment is an isolated antibody having a heavy chain of the amino acid sequence set forth in SEQ ID NO:50 and a light chain amino acid sequence set forth in SEQ ID NO:38. Another embodiment is an isolated antibody having a heavy chain of the amino acid sequence set forth in SEQ ID NO:51 and a light chain amino acid sequence set forth in SEQ ID NO:38. Another embodiment is an isolated antibody having a heavy chain of the amino acid sequence set forth in SEQ ID NO:52 and a light chain amino acid sequence set forth in SEQ ID NO:38. Another embodiment is an isolated antibody having a heavy chain of the amino acid sequence set forth in SEQ ID NO:53 and a light chain amino acid sequence set forth in SEQ ID NO:38. Another embodiment is an isolated antibody having a heavy chain of the amino acid sequence set forth in SEQ ID NO:50 and a light chain amino acid sequence set forth in SEQ ID NO:39. Another embodiment is an isolated antibody having a heavy chain of the amino acid sequence set forth in SEQ ID NO:51 and a light chain amino acid sequence set forth in SEQ ID NO:39. Another embodiment is an isolated antibody having a heavy chain of the amino acid sequence set forth in SEQ ID NO:52 and a light chain amino acid sequence set forth in SEQ ID NO:39. Another embodiment is an isolated antibody having a heavy chain of the amino acid sequence set forth in SEQ ID NO:53 and a light chain amino acid sequence set forth in SEQ ID NO:39. Another embodiment is an isolated antibody having a heavy chain of the amino acid sequence set forth in SEQ ID NO:50 and a light chain amino acid sequence set forth in SEQ ID NO:40. Another embodiment is an isolated antibody having a heavy chain of the amino acid sequence set forth in SEQ ID NO:51 and a light chain amino acid sequence set forth in SEQ ID NO:40. Another embodiment is an isolated antibody having a heavy chain of the amino acid sequence set forth in SEQ ID NO:52 and a light chain amino acid sequence set forth in SEQ ID NO:40. Another embodiment is an isolated antibody having a heavy chain of the amino acid sequence set forth in SEQ ID NO:53 and a light chain amino acid sequence set forth in SEQ ID NO:40. Another embodiment is an isolated antibody having a heavy chain of the amino acid sequence set forth in SEQ ID NO:50 and a light chain amino acid sequence set forth in SEQ ID NO:41. Another embodiment is an isolated antibody having a heavy chain of the amino acid sequence set forth in SEQ ID NO:51 and a light chain amino acid sequence set forth in SEQ ID NO:41. Another embodiment is an isolated antibody having a heavy chain of the amino acid sequence set forth in SEQ ID NO:52 and a light chain amino acid sequence set forth in SEQ ID NO:41.Another embodiment is an isolated antibody having a heavy chain amino acid sequence set forth in SEQ ID NO:53 and a light chain amino acid sequence set forth in SEQ ID NO:41.

[0118] Another embodiment is an isolated antibody having a heavy chain of the amino acid sequence set forth in SEQ ID NO:54 and a light chain amino acid sequence set forth in SEQ ID NO:38. Another embodiment is an isolated antibody having a heavy chain of the amino acid sequence set forth in SEQ ID NO:55 and a light chain amino acid sequence set forth in SEQ ID NO:38. Another embodiment is an isolated antibody having a heavy chain of the amino acid sequence set forth in SEQ ID NO:56 and a light chain amino acid sequence set forth in SEQ ID NO:38. Another embodiment is an isolated antibody having a heavy chain of the amino acid sequence set forth in SEQ ID NO:57 and a light chain amino acid sequence set forth in SEQ ID NO:38. Another embodiment is an isolated antibody having a heavy chain of the amino acid sequence set forth in SEQ ID NO:54 and a light chain amino acid sequence set forth in SEQ ID NO:39. Another embodiment is an isolated antibody having a heavy chain of the amino acid sequence set forth in SEQ ID NO:55 and a light chain amino acid sequence set forth in SEQ ID NO:39. Another embodiment is an isolated antibody having a heavy chain of the amino acid sequence set forth in SEQ ID NO:56 and a light chain amino acid sequence set forth in SEQ ID NO:39. Another embodiment is an isolated antibody having a heavy chain of the amino acid sequence set forth in SEQ ID NO:57 and a light chain amino acid sequence set forth in SEQ ID NO:39. Another embodiment is an isolated antibody having a heavy chain of the amino acid sequence set forth in SEQ ID NO:54 and a light chain amino acid sequence set forth in SEQ ID NO:40. Another embodiment is an isolated antibody having a heavy chain of the amino acid sequence set forth in SEQ ID NO:55 and a light chain amino acid sequence set forth in SEQ ID NO:40. Another embodiment is an isolated antibody having a heavy chain of the amino acid sequence set forth in SEQ ID NO:56 and a light chain amino acid sequence set forth in SEQ ID NO:40. Another embodiment is an isolated antibody having a heavy chain of the amino acid sequence set forth in SEQ ID NO:57 and a light chain amino acid sequence set forth in SEQ ID NO:40. Another embodiment is an isolated antibody having a heavy chain of the amino acid sequence set forth in SEQ ID NO:54 and a light chain amino acid sequence set forth in SEQ ID NO:41. Another embodiment is an isolated antibody having a heavy chain of the amino acid sequence set forth in SEQ ID NO:55 and a light chain amino acid sequence set forth in SEQ ID NO:41. Another embodiment is an isolated antibody having a heavy chain of the amino acid sequence set forth in SEQ ID NO:56 and a light chain amino acid sequence set forth in SEQ ID NO:41.Another embodiment is an isolated antibody having a heavy chain amino acid sequence set forth in SEQ ID NO:57 and a light chain amino acid sequence set forth in SEQ ID NO:41.

[0119] Yet another embodiment is a method for treating a subject having a neurodegenerative or neuromuscular disease or disorder, an inflammatory or immune disease or disorder, or an autoimmune disease, comprising administering to such subject a therapeutically effective amount of an antibody or antigen-binding fragment thereof disclosed herein.

[0120] Another embodiment is a method for treating a subject having an autoimmune disease selected from the group consisting of systemic lupus erythematosus, type 1 diabetes, myasthenia gravis, psoriasis, Addison's disease, Crohn's disease, uveitis, multiple sclerosis, hemolytic anemia, inflammatory bowel disease, immune thrombocytopenic purpura, Graves' disease, and rheumatoid arthritis, comprising administering to the subject a therapeutically effective amount of an antibody or antigen-binding fragment thereof disclosed herein.

[0121] Yet another embodiment is a method for treating a subject having a neurodegenerative disease or neuromuscular disorder selected from the group consisting of Alzheimer's disease, Parkinson's disease, amyotrophic lateral sclerosis, multifocal motor neuropathy, primary lateral sclerosis, spinal muscular atrophy, Kennedy's disease, and spinocerebellar ataxia, comprising administering to such subject a therapeutically effective amount of an antibody or antigen-binding fragment thereof disclosed herein.

[0122] Yet another embodiment is a method for treating a subject with amyotrophic lateral sclerosis, comprising administering to such subject a therapeutically effective amount of an antibody or antigen-binding fragment thereof disclosed herein.

[0123] Another embodiment is a method for inhibiting an immune response in a subject, comprising administering to such subject a therapeutically effective amount of an antibody or antigen-binding fragment thereof disclosed in the present application.

[0124] Another embodiment is a method of inhibiting an immune response in a subject, comprising administering to such subject a therapeutically effective amount of an antibody or antigen-binding fragment thereof, wherein the immune response is graft-versus-host disease or organ transplant rejection.

[0125] Another embodiment is a method according to any of the disclosed methods, wherein the antibody or antibody fragment is administered in combination with another therapeutic agent. One embodiment is a method according to any of the disclosed methods, wherein the antibody or antibody fragment is administered in combination with a compound that blocks the interaction between CD28 and CD86 or between CD28 and CD80.

[0126] Yet another embodiment is a method according to any of the disclosed methods, wherein the antibody or antibody fragment is administered in combination with a CTLA4-Ig fusion protein, a compound that blocks the interaction between CD28 and CD86 or between CD28 and CD80.

[0127] Another embodiment is a method according to any of the disclosed methods, wherein the antibody or antibody fragment is administered in combination with abatacept or belatacept or galiximab, which are compounds that block the interaction between CD28 and CD86 or between CD28 and CD80. [Table 1-1] [Table 1-2] [Table 1-3] [Table 1-4] [Table 1-5] [Table 1-6] [Table 1-7] [Table 1-8] [Table 1-9] [Example]

[0128] Example 1 CD40L Binding Assay

[0129] To compare the CD40L binding of all 16 antibody clones with that of 5c8 or AT-1501, binding assays were performed using two clones along with 5c8 and AT-1501 on the same 96-well assay plate. A three-part sandwich ELISA assay was used to determine the binding levels of the disclosed antibodies relative to the reference antibodies 5c8-19 and AT1501. 96-well polystyrene plates were coated with 2 μg / ml of recombinant human CD40L (BioLegend catalog no. 591706) in PBS, and 50 μl / well of the plate was added to a Costar high-binding half-area 96-well assay plate (Corning 3690) and incubated overnight at 4°C. To prevent background binding, plates were blocked with (1x) PBS / 1.0% BSA (140 μl / well) for 1 hour at room temperature. Binding curves of 5C8 or AT1501 (2-fold serial dilutions starting at 2 μg / ml) were added (50 μl / well) and incubated for 1 hour at room temperature. Plates were washed and incubated with 1:10,000 dilution of HRP-(Fab2) donkey anti-human IgG (Fc specific) (Jackson Immuno. 709-036-098) (50 μl / well) for 1 hour at room temperature. Plates were washed and TMB substrate (Surmodics BioFX TMBW-1000-01) was added (50 μl / well). After 5 minutes at room temperature, development was stopped with 25 μl of 2NH2SO4. Plates were read on a Molecular Devices SpectraMax M5 microplate reader using the SoftMax Pro 6.2.2 program to measure absorbance at 450 nm.

[0130] The relative CD40L binding activity was calculated as follows: [IC50 (clone) / IC50 (5c8 or AT-1501) × 100%]. The potency rankings of the 16 clones against 5c8 are shown in Figure 1A, and their potency rankings against AT-1501 are shown in Figure 1B. The resulting binding curves are shown in Figures 2A-2Q (comparison of the 16 clones with 5c8, Figure 2H shows a comparison of AT-1501 with 5c8), and Figures 3A-3Q (comparison of the 16 clones with AT-1501, Figure 3H shows a comparison of AT-1501 with 5c8). The IC50, LCL, UCL, and relative potency of each clone are shown in Table 2 (comparison with 5c8) and Table 3 (comparison with AT-1501). [Table 2-1] [Table 2-2] [Table 3-1] [Table 3-2]

[0131] Example 2 Binding activity to human FcγRI, FcγRIIa, FcγRIIIa, and FcγRIIIb

[0132] Sixteen VH / VL antibody clones were constructed using an IgG1 Fc (SEQ ID NO: 21) with two mutations, P238S and N297G, and assayed for Fc effector function by binding to human FcγRI, FcγRIIa, and FcγRIIIa.

[0133] Anti-CD40L antibodies (including abatacept as a negative control) were diluted to 2 μg / ml in 1× PBS and added at 50 μl / well to a Costar high-binding 1 / 2 area 96-well assay plate (Corning 3690) for overnight incubation at 4°C. To prevent background binding, the plate was blocked with 1× PBS / 1.0% BSA (140 μl / well) for 1 hour at room temperature. Binding curve recombinant human FcγRI, IIa, IIIa, and IIIb (2-fold serial dilutions starting from 5 μg / ml) were added at 50 μl / well and incubated for 1 hour at room temperature. Plates were washed and incubated with 2 μg / ml mouse anti-human CD16 (anti-FcRIII), CD32 (anti-FcRIIa), or CD64 (anti-FcRI) (eBioSciences / Invitrogen 14-0168-82, 16-0329-81, 14-0649-82) (50 μl / well) for 1 hour at room temperature. Plates were washed and incubated with 1:10,000 dilution of HRP-(Fab2) goat anti-mouse IgG (Fc specific) (Jackson Immuno. 116-036-071) (50 μl / well) for 1 hour at room temperature. Plates were washed and TMB substrate (Surmodics BioFX TMBW-1000-01) was added (50 μl / well). After 5 minutes at room temperature, color development was stopped with 25 μl / well of 2NH2SO4. Plates were read on a Molecular Devices SpectraMax M5 microplate reader using the SoftMax Pro 6.2.2 program to measure absorbance at 450 nm.

[0134] As can be seen from the figures, the clones disclosed herein were negative for binding to the three Fc receptors, with 5c8 binding to FcγRI and FcγRIIa but not to FcγRIIIa or FcγRIIIb. Figures 4A and 4B show the binding of eight antibody clones to FcγRIa compared with the binding curves of 5c8, AT-1501, and abatacept. Figures 5A and 5B show the binding of eight antibody clones to FcγRIIa compared with the binding curves of 5c8, AT-1501, and abatacept. Figures 6A and 6B show the binding of eight antibody clones to FcγRIIIa compared with the binding curves of 5c8, AT-1501, and abatacept. Figures 6C and 6D show the binding of eight antibody clones to FcγRIIIb compared with the binding curves of 5c8, AT-1501, and abatacept. (Figures 4A, 5A, 6A, and 6C show the results for antibody clones 4-4, 5-3, 6-6, 7-5, 8-3, 8-4, 10-1, and 10-4, and Figures 4B, 5B, 6B, and 6D show the results for antibody clones 11-5, 12-4, 13-2, 15-1, 15-4, 16-3, 17-1, and 18-2).

[0135] Example 4 Binding activity to C1q

[0136] Anti-CD40L antibodies (including abatacept as a negative control) were diluted to 2 μg / ml in 1x PBS and added at 50 μl / well to a Costar high-binding 1 / 2-area 96-well assay plate (Corning 3690) for overnight incubation at 4°C. To prevent background binding, the plate was blocked with 1x PBS / 1.0% BSA (140 μl / well) for 1 hour at room temperature. A binding curve of native human C1q protein (Abcam ab96363) was added at 50 μl / well in serial two-fold dilutions starting from 10 μg / ml for 1 hour at room temperature. After washing the plate, HRP-sheep anti-human C1q (Abcam ab46191) was added at a dilution of 1:400 (50 μl / well) and incubated at room temperature for 1 hour. After washing the plate, TMB substrate (Surmodics BioFX TMBW-1000-01) was added (50 μl / well). After 5 minutes at room temperature, color development was stopped with 25 μl / well 2NH2SO4. Plates were read on a Molecular Devices SpectraMax M5 microplate reader using the SoftMax Pro 6.2.2 program to measure absorbance at 450 nm.

[0137] As can be seen from the figures, all antibodies from the 16 antibody clones disclosed in the present application were negative for C1q binding, except for 5c8, which showed significant binding. Figures 7A and 7B show the C1q binding of eight antibody clones compared with the binding curves of 5c8, AT-1501, and abatacept, respectively. Figure 7A shows the results for antibody clones 4-4, 5-3, 6-6, 7-5, 8-3, 8-4, 10-1, and 10-4, and Figure 7B shows the results for antibody clones 11-5, 12-4, 13-2, 15-1, 15-4, 16-3, 17-1, and 18-2.

Claims

1. (a) i) a CDRH1 domain comprising the sequence set forth in SEQ ID NO: 9; ii) a CDRH2 domain comprising the sequence set forth in SEQ ID NO: 10; and iii) a CDRH3 domain comprising the sequence set forth in SEQ ID NO: 15; and A heavy chain variable region (V Ｈ ), and (b) i) a CDRL1 domain comprising the sequence set forth in SEQ ID NO: 16; and ii) a CDRL2 domain comprising the sequence set forth in SEQ ID NO: 18; and iii) a CDRL3 domain comprising the sequence set forth in SEQ ID NO: 20; and A light chain variable region (V Ｌ ) Contains, or (a) i) a CDRH1 domain comprising the sequence set forth in SEQ ID NO: 9; ii) a CDRH2 domain comprising the sequence set forth in SEQ ID NO: 10; and iii) a CDRH3 domain comprising the sequence set forth in SEQ ID NO: 15; and A heavy chain variable region (V Ｈ ), and (b) i) a CDRL1 domain comprising the sequence set forth in SEQ ID NO: 17; and ii) a CDRL2 domain comprising the sequence set forth in SEQ ID NO: 18; and iii) a CDRL3 domain comprising the sequence set forth in SEQ ID NO: 20; and A light chain variable region (V Ｌ ) Contains, or (a) i) a CDRH1 domain comprising the sequence set forth in SEQ ID NO: 9; ii) a CDRH2 domain comprising the sequence set forth in SEQ ID NO: 10; and iii) a CDRH3 domain comprising the sequence set forth in SEQ ID NO: 15; and A heavy chain variable region (V Ｈ ), and (b) i) a CDRL1 domain comprising the sequence set forth in SEQ ID NO: 16; and ii) a CDRL2 domain comprising the sequence set forth in SEQ ID NO: 19; and iii) a CDRL3 domain comprising the sequence set forth in SEQ ID NO: 20; and A light chain variable region (V Ｌ ) Contains, or (a) i) a CDRH1 domain comprising the sequence set forth in SEQ ID NO: 9; ii) a CDRH2 domain comprising the sequence set forth in SEQ ID NO: 10; and iii) a CDRH3 domain comprising the sequence set forth in SEQ ID NO: 15; and A heavy chain variable region (V Ｈ ), and (b) i) a CDRL1 domain comprising the sequence set forth in SEQ ID NO: 17; and ii) a CDRL2 domain comprising the sequence set forth in SEQ ID NO: 19; and iii) a CDRL3 domain comprising the sequence set forth in SEQ ID NO: 20; and A light chain variable region (V Ｌ ) Contains, or (a) i) a CDRH1 domain comprising the sequence set forth in SEQ ID NO: 9; ii) a CDRH2 domain comprising the sequence set forth in SEQ ID NO: 11; and iii) a CDRH3 domain comprising the sequence set forth in SEQ ID NO: 15; and A heavy chain variable region (V Ｈ ), and (b) i) a CDRL1 domain comprising the sequence set forth in SEQ ID NO: 16; and ii) a CDRL2 domain comprising the sequence set forth in SEQ ID NO: 18; and iii) a CDRL3 domain comprising the sequence set forth in SEQ ID NO: 20; and A light chain variable region (V Ｌ ) Contains, or (a) i) a CDRH1 domain comprising the sequence set forth in SEQ ID NO: 9; ii) a CDRH2 domain comprising the sequence set forth in SEQ ID NO: 11; and iii) a CDRH3 domain comprising the sequence set forth in SEQ ID NO: 15; and A heavy chain variable region (V Ｈ ), and (b) i) a CDRL1 domain comprising the sequence set forth in SEQ ID NO: 16; and ii) a CDRL2 domain comprising the sequence set forth in SEQ ID NO: 19; and iii) a CDRL3 domain comprising the sequence set forth in SEQ ID NO: 20; and A light chain variable region (V Ｌ ) Contains, or (a) i) a CDRH1 domain comprising the sequence set forth in SEQ ID NO: 9; ii) a CDRH2 domain comprising the sequence set forth in SEQ ID NO: 11; and iii) a CDRH3 domain comprising the sequence set forth in SEQ ID NO: 15; and A heavy chain variable region (V Ｈ ), and (b) i) a CDRL1 domain comprising the sequence set forth in SEQ ID NO: 17; and ii) a CDRL2 domain comprising the sequence set forth in SEQ ID NO: 19; and iii) a CDRL3 domain comprising the sequence set forth in SEQ ID NO: 20; and A light chain variable region (V Ｌ ) Contains, or (a) i) a CDRH1 domain comprising the sequence set forth in SEQ ID NO: 9; ii) a CDRH2 domain comprising the sequence set forth in SEQ ID NO: 12; and iii) a CDRH3 domain comprising the sequence set forth in SEQ ID NO: 15; and A heavy chain variable region (V Ｈ ), and (b) i) a CDRL1 domain comprising the sequence set forth in SEQ ID NO: 16; and ii) a CDRL2 domain comprising the sequence set forth in SEQ ID NO: 18; and iii) a CDRL3 domain comprising the sequence set forth in SEQ ID NO: 20; and A light chain variable region (V Ｌ ) Contains, or (a) i) a CDRH1 domain comprising the sequence set forth in SEQ ID NO: 9; ii) a CDRH2 domain comprising the sequence set forth in SEQ ID NO: 12; and iii) a CDRH3 domain comprising the sequence set forth in SEQ ID NO: 15; and A heavy chain variable region (V Ｈ ), and (b) i) a CDRL1 domain comprising the sequence set forth in SEQ ID NO: 17; and ii) a CDRL2 domain comprising the sequence set forth in SEQ ID NO: 18; and iii) a CDRL3 domain comprising the sequence set forth in SEQ ID NO: 20; and A light chain variable region (V Ｌ ) Contains, or (a) i) a CDRH1 domain comprising the sequence set forth in SEQ ID NO: 9; ii) a CDRH2 domain comprising the sequence set forth in SEQ ID NO: 12; and iii) a CDRH3 domain comprising the sequence set forth in SEQ ID NO: 15; and A heavy chain variable region (V Ｈ ), and (b) i) a CDRL1 domain comprising the sequence set forth in SEQ ID NO: 17; and ii) a CDRL2 domain comprising the sequence set forth in SEQ ID NO: 19; and iii) a CDRL3 domain comprising the sequence set forth in SEQ ID NO: 20; and A light chain variable region (V Ｌ ) Contains, or (a) i) a CDRH1 domain comprising the sequence set forth in SEQ ID NO: 9; ii) a CDRH2 domain comprising the sequence set forth in SEQ ID NO: 13; and iii) a CDRH3 domain comprising the sequence set forth in SEQ ID NO: 15; and A heavy chain variable region (V Ｈ ), and (b) i) a CDRL1 domain comprising the sequence set forth in SEQ ID NO: 16; and ii) a CDRL2 domain comprising the sequence set forth in SEQ ID NO: 18; and iii) a CDRL3 domain comprising the sequence set forth in SEQ ID NO: 20; and A light chain variable region (V Ｌ ) Contains, or (a) i) a CDRH1 domain comprising the sequence set forth in SEQ ID NO: 9; ii) a CDRH2 domain comprising the sequence set forth in SEQ ID NO: 13; and iii) a CDRH3 domain comprising the sequence set forth in SEQ ID NO: 15; and A heavy chain variable region (V Ｈ ), and (b) i) a CDRL1 domain comprising the sequence set forth in SEQ ID NO: 17; and ii) a CDRL2 domain comprising the sequence set forth in SEQ ID NO: 18; and iii) a CDRL3 domain comprising the sequence set forth in SEQ ID NO: 20; and A light chain variable region (V Ｌ ) Contains, or (a) i) a CDRH1 domain comprising the sequence set forth in SEQ ID NO: 9; ii) a CDRH2 domain comprising the sequence set forth in SEQ ID NO: 13; and iii) a CDRH3 domain comprising the sequence set forth in SEQ ID NO: 15; and A heavy chain variable region (V Ｈ ), and (b) i) a CDRL1 domain comprising the sequence set forth in SEQ ID NO: 16; and ii) a CDRL2 domain comprising the sequence set forth in SEQ ID NO: 19; and iii) a CDRL3 domain comprising the sequence set forth in SEQ ID NO: 20; and A light chain variable region (V Ｌ ) 1. An isolated antibody that specifically binds to CD40L, comprising:

2. (a) a heavy chain variable region (V) having the amino acid sequence set forth in SEQ ID NO: 1; Ｈ ) and a light chain variable region (V Ｌ ),or (b) a heavy chain variable region (V) having the amino acid sequence set forth in SEQ ID NO: 1; Ｈ ) and a light chain variable region (V Ｌ ),or (c) a heavy chain variable region (V) having the amino acid sequence set forth in SEQ ID NO: 1 Ｈ ) and a light chain variable region (V Ｌ ),or (d) a heavy chain variable region (V) having the amino acid sequence set forth in SEQ ID NO: 1 Ｈ ) and a light chain variable region (V Ｌ ),or (e) a heavy chain variable region (V) having the amino acid sequence set forth in SEQ ID NO: 2 Ｈ ) and a light chain variable region (V Ｌ ),or (f) a heavy chain variable region (V) having the amino acid sequence set forth in SEQ ID NO: 2 Ｈ ) and a light chain variable region (V Ｌ ),or (g) a heavy chain variable region (V) having the amino acid sequence set forth in SEQ ID NO: 2 Ｈ ) and a light chain variable region (V Ｌ ),or (h) a heavy chain variable region (V) having the amino acid sequence set forth in SEQ ID NO: 3 Ｈ ) and a light chain variable region (V Ｌ ),or (i) a heavy chain variable region (V) having the amino acid sequence set forth in SEQ ID NO: 3 Ｈ ) and a light chain variable region (V Ｌ ),or (j) a heavy chain variable region (V) having the amino acid sequence set forth in SEQ ID NO: 3 Ｈ ) and a light chain variable region (V Ｌ ),or (k) a heavy chain variable region (V) having the amino acid sequence set forth in SEQ ID NO: 4 Ｈ ) and a light chain variable region (V Ｌ ),or (l) a heavy chain variable region (V) having the amino acid sequence set forth in SEQ ID NO: 4 Ｈ ) and a light chain variable region (V Ｌ ),or (m) a heavy chain variable region (V) having the amino acid sequence set forth in SEQ ID NO: 4 Ｈ ) and a light chain variable region (V Ｌ ) 1. An isolated antibody that specifically binds to CD40L, comprising:

3. (a) a heavy chain having the amino acid sequence set forth in SEQ ID NO: 42 and a light chain having the amino acid sequence set forth in SEQ ID NO: 38; or (b) a heavy chain having the amino acid sequence set forth in SEQ ID NO: 42 and a light chain having the amino acid sequence set forth in SEQ ID NO: 39; or (c) a heavy chain having the amino acid sequence set forth in SEQ ID NO: 42 and a light chain having the amino acid sequence set forth in SEQ ID NO: 40; or (d) a heavy chain having the amino acid sequence set forth in SEQ ID NO: 42 and a light chain having the amino acid sequence set forth in SEQ ID NO: 41; or (e) a heavy chain having the amino acid sequence set forth in SEQ ID NO: 43 and a light chain having the amino acid sequence set forth in SEQ ID NO: 38; or (f) a heavy chain having the amino acid sequence set forth in SEQ ID NO: 43 and a light chain having the amino acid sequence set forth in SEQ ID NO: 40; or (g) a heavy chain having the amino acid sequence set forth in SEQ ID NO: 43 and a light chain having the amino acid sequence set forth in SEQ ID NO: 41; or (h) a heavy chain having the amino acid sequence set forth in SEQ ID NO: 44 and a light chain having the amino acid sequence set forth in SEQ ID NO: 38; or (i) a heavy chain having the amino acid sequence set forth in SEQ ID NO: 44 and a light chain having the amino acid sequence set forth in SEQ ID NO: 39; or (j) a heavy chain having the amino acid sequence set forth in SEQ ID NO: 44 and a light chain having the amino acid sequence set forth in SEQ ID NO: 41; or (k) a heavy chain having the amino acid sequence set forth in SEQ ID NO: 45 and a light chain having the amino acid sequence set forth in SEQ ID NO: 38; or (l) a heavy chain having the amino acid sequence set forth in SEQ ID NO: 45 and a light chain having the amino acid sequence set forth in SEQ ID NO: 39; or (m) a heavy chain having the amino acid sequence set forth in SEQ ID NO: 45 and a light chain having the amino acid sequence set forth in SEQ ID NO: 40; 3. The isolated antibody of claim 1 or 2.

4. A pharmaceutical composition for use in a method for treating a subject having a disease or disorder associated with CD40L, comprising the antibody of any one of claims 1 to 3, the method comprising administering to the subject the antibody in a therapeutically effective amount.

5. 4. A pharmaceutical composition for use in a method for treating a subject having a neurodegenerative or neuromuscular disease or disorder, an inflammatory or immune disease or disorder, or an autoimmune disease, comprising the antibody of any one of claims 1 to 3, said method comprising administering to the subject a therapeutically effective amount of the antibody.

6. 6. The pharmaceutical composition of claim 5, wherein the autoimmune disease is selected from the group consisting of systemic lupus erythematosus, type 1 diabetes, myasthenia gravis, psoriasis, Addison's disease, Crohn's disease, uveitis, multiple sclerosis, hemolytic anemia, inflammatory bowel disease, immune thrombocytopenic purpura, Graves' disease, and rheumatoid arthritis.

7. 6. The pharmaceutical composition of claim 5, wherein the neurodegenerative or neuromuscular disease or disorder is selected from the group consisting of Alzheimer's disease, Parkinson's disease, amyotrophic lateral sclerosis, multifocal motor neuropathy, primary lateral sclerosis, spinal muscular atrophy, Kennedy's disease, and spinocerebellar ataxia.

8. 6. The pharmaceutical composition of claim 5, wherein the neurodegenerative or neuromuscular disease or disorder is amyotrophic lateral sclerosis.

9. 4. A pharmaceutical composition for use in a method for inhibiting an immune response in a subject, comprising the antibody of any one of claims 1 to 3, said method comprising administering to the subject the antibody in a therapeutically effective amount.

10. 10. The pharmaceutical composition of claim 9, wherein the immune response is graft-versus-host disease or organ transplant rejection.

11. The pharmaceutical composition of any one of claims 4 to 10, wherein the method further comprises administering the antibody in combination with another therapeutic agent.

12. The pharmaceutical composition of claim 11 , wherein the method further comprises administering the antibody in combination with a compound that blocks the interaction between CD28 and CD86 or between CD28 and CD80.

13. 13. The pharmaceutical composition of claim 12, wherein the compound that blocks the interaction between CD28 and CD86 or between CD28 and CD80 is a CTLA4-Ig fusion protein, or the compound that blocks the interaction between CD28 and CD86 or between CD28 and CD80 is abatacept, belatacept, or galiximab.

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