Structure of CD40 Ligand in Complex with the Fab Fragment of a Neutralizing Humanized Antibody

Karpusas, Michael; Lucci, Jodie; Ferrant, Janine; Benjamin, Christopher D.; Taylor, Frederick R.; Strauch, Kathy; Garber, Ellen A.; Hsu, Yen-Ming

doi:10.1016/s0969-2126(01)00590-1

Cited by 45 publications

(49 citation statements)

References 36 publications

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“…3G. Similarly sized higher-order structures were observed for Abs in complex with other TNF family proteins, including 5c8 and other human CD40L-binding mAbs, as well as anti-TNF mAbs (27,(32)(33)(34). Confocal microscopy showed that these small soluble complexes can translate into even larger ordered structures on the cell surface, presumably driven by avidity (32).…”

Section: Mechanism Of Action Of Mcd40l Dab-fcsmentioning

confidence: 76%

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Engineering of a Novel Anti-CD40L Domain Antibody for Treatment of Autoimmune Diseases

Xie

Yamniuk

Borowski

et al. 2014

The Journal of Immunology

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CD40–CD40L interactions play a critical role in regulating immune responses. Blockade of CD40L by Abs, such as the anti-CD40L Ab 5c8, demonstrated positive clinical effects in patients with autoimmune diseases; however, incidents of thromboembolism (TE) precluded further development of these molecules. In this study, we examined the role of the Fc domain interaction with FcγRs in modulating platelet activation and potential for TE. Our results show that the interaction of the 5c8 wild-type IgG1 Fc domain with FcγRs is responsible for platelet activation, as measured by induction of PAC-1 and CD62P. A version of 5c8 with a mutated IgG1 tail was identified that showed minimal FcγR binding and platelet activation while maintaining full binding to CD40L. To address whether Fc effector function is required for immunosuppression, a potent Ab fragment, termed a “domain Ab” (dAb), against murine CD40L was identified and fused to a murine IgG1 Fc domain containing a D265A mutation that lacks Fc effector function. In vitro, this dAb–Fc demonstrated comparable potency to the benchmark mAb MR-1 in inhibiting B cell and dendritic cell activation. Furthermore, the anti-CD40L dAb–Fc exhibited a notable efficacy comparable to MR-1 in various preclinical models, such as keyhole limpet hemocyanin–induced Ab responses, alloantigen-induced T cell proliferation, “heart-to-ear” transplantation, and NZB × NZW F1 spontaneous lupus. Thus, our data show that immunosuppression and TE can be uncoupled and that a CD40L dAb with an inert Fc tail is expected to be efficacious for treating autoimmune diseases, with reduced risk for TE.

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Section: Mechanism Of Action Of Mcd40l Dab-fcsmentioning

confidence: 76%

“…The 5c8 IgG 1 (27) was converted to a F(ab9) 2 by incubation with 1% pepsin (Sigma-Aldrich) in 50 mM sodium citrate (pH 3.5) for 5 h at room temperature. The reaction was quenched by adding Tris to a final pH of 8.…”

Section: Generation Of 5c8 Ab Fragmentsmentioning

confidence: 99%

Engineering of a Novel Anti-CD40L Domain Antibody for Treatment of Autoimmune Diseases

Xie

Yamniuk

Borowski

et al. 2014

The Journal of Immunology

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“…The CD154 monomer has a typical TNF family structure, containing a sandwich of two antiparallel ␤-sheets, with a Greek key fold (11,25). The extensive trimerization interface between the subunits is formed mostly by hydrophobic residues.…”

Section: Resultsmentioning

confidence: 99%

Crystallographic and Mutational Analysis of the CD40-CD154 Complex and Its Implications for Receptor Activation

An¹,

Kim²,

Song³

et al. 2011

Journal of Biological Chemistry

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CD40 is a tumor necrosis factor receptor (TNFR) family protein that plays an important role in B cell development. CD154/ CD40L is the physiological ligand of CD40. We have determined the crystal structure of the CD40-CD154 complex at 3.5 Å resolution. The binding site of CD40 is located in a crevice formed between two CD154 subunits. Charge complementarity plays a critical role in the CD40-CD154 interaction. Some of the missense mutations found in hereditary hyper-IgM syndrome can be mapped to the CD40-CD154 interface. The CD40 interaction area of one of the CD154 subunits is twice as large as that of the other subunit forming the binding crevice. This is because cysteine-rich domain 3 (CRD3) of CD40 has a disulfide bridge in an unusual position that alters the direction of the ladder-like structure of CD40. The Ser 132 loop of CD154 is not involved in CD40 binding but its substitution significantly reduces p38-and ERK-dependent signaling by CD40, whereas JNK-dependent signaling is not affected. These findings suggest that ligand-induced di-or trimerization is necessary but not sufficient for complete activation of CD40.

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“…50 HCV translational inhibitors were also obtained by selecting for RNA aptamers against the viral internal ribosome entry site, where translation starts. 51 Combinatorial RNA therapies for HIV-1 infection may have potential for stem cell-based gene therapy…”

Section: Aptamers For Intracellular Targets Are Being Developedmentioning

confidence: 99%

Gene therapy progress and prospects: RNA aptamers

2007

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Aptamers are oligonucleotides evolved in vitro or in nature to bind target ligands with high affinity and specificity. They are emerging as powerful tools in the fields of therapeutics, drug development, target validation and diagnostics. Aptamers are attractive alternatives to antibody-and small-moleculebased therapeutics owing to their stability, low toxicity, low immunogenicity and improved safety. With the recent approval of the first aptamer drug Macugen by the US FDA, there is great impetus to develop therapeutic aptamers that can target a wide array of disease states. The recent demonstration that aptamer activity can be reversed by the administration of a simple antidote greatly enhances the potential value of aptamers as therapeutic agents.

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Structure of CD40 Ligand in Complex with the Fab Fragment of a Neutralizing Humanized Antibody

Cited by 45 publications

References 36 publications

Engineering of a Novel Anti-CD40L Domain Antibody for Treatment of Autoimmune Diseases

Engineering of a Novel Anti-CD40L Domain Antibody for Treatment of Autoimmune Diseases

Crystallographic and Mutational Analysis of the CD40-CD154 Complex and Its Implications for Receptor Activation

Gene therapy progress and prospects: RNA aptamers

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