Efficacy of a recombinant single-chain fragment variable region, VasSF, as a new drug for vasculitis

Yosuke Kameoka1 Department of Research and Development, A-CLIP Institute, Ltd., Chiba, Japan, [email protected]Correspondence[email protected]

Fukuko Kishi1 Department of Research and Development, A-CLIP Institute, Ltd., Chiba, Japan, [email protected]

Minako Koura2 Laboratory of Animal Models for Human Diseases, National Institutes of Biomedical Innovation, Health and Nutrition, Osaka, Japan

Yoshio Yamakawa1 Department of Research and Development, A-CLIP Institute, Ltd., Chiba, Japan, [email protected]

Rora Nagasawa1 Department of Research and Development, A-CLIP Institute, Ltd., Chiba, Japan, [email protected]

Fuyu Ito3 Asia International Institute of Infectious Disease Control, Teikyo University, Tokyo, Japan

Junichiro Matsuda2 Laboratory of Animal Models for Human Diseases, National Institutes of Biomedical Innovation, Health and Nutrition, Osaka, Japan

Osamu Suzuki2 Laboratory of Animal Models for Human Diseases, National Institutes of Biomedical Innovation, Health and Nutrition, Osaka, Japan

Toshinori Nakayama4 Department of Immunology, Graduate School of Medicine, Chiba University, Chiba, Japan

Kazuo Suzuki1 Department of Research and Development, A-CLIP Institute, Ltd., Chiba, Japan, [email protected];3 Asia International Institute of Infectious Disease Control, Teikyo University, Tokyo, Japan;4 Department of Immunology, Graduate School of Medicine, Chiba University, Chiba, Japan;5 Department of Immunology, National Institute of Infectious Diseases, Tokyo, Japan

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Abstract

Background

Anti-neutrophil cytoplasmic autoantibodies (ANCA) associated vasculitis is a pauci-immune disease with the inflammation of the small blood vessels. The efficacies of antibody drugs for induction therapies of vasculitis vary among cases. Here, we developed a novel clone of a single chain Fv region (ScFv) with vasculitis-specific therapeutic potential.

Materials and methods

The clone, termed VasSF, was selected from our Escherichia coli expression library of recombinant human ScFv based on the therapeutic efficacy in an SCG/Kj mouse model of MPO-ANCA-associated vasculitis (MAAV), such as improvement of the urinary score and decreased crescent formation in glomeruli, granulomatous in lung, MPO-ANCA biomarkers, the anti-moesin antibody, and some cytokine levels.

Results

We identified vasculitis-associated apolipoprotein A-II (VAP2) as a target molecule of the clone and confirmed the independently-established VAP2 antibodies were also therapeutic in SCG/Kj mice. In MAAV, MPO-ANCA and cytokines stimulate neutrophils by facilitating heterodimer formation of VAP2 with apolipoprotein A-I in HDL.

Conclusion

VasSF would constitute a novel antibody drug for vasculitis by suppressing the heterodimer formation of the apolipoproteins.

Keywords:

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Efficacy of a recombinant single-chain fragment variable region, VasSF, as a new drug for vasculitis [Corrigendum]

Supplementary materials

Figure S1 Plasmid preparation for hScFv from the 204-clone hScFv library.

Notes: Part of the phylogenetic tree of hScF clones in our library (A). Schematic display of clones selected from our hScFv library (B).
Abbreviation: hScFv, human single chain Fv region.

Figure S2 Plasmid preparation for hScFv and the purification procedures.

Notes: The original hScFv library was constructed in the pBAD expression vector (A). The selected clones were optimized for Escherichia coli expression and inserted in pET32-a (B). Gel filtration separation profile of hScFv (C). CBB stain of isolated hScFV on SDS-PAGE (D). Western blot analysis of hScFv by anti-human Fab2 antibody (E).
Abbreviation: hScFv, human single-chain Fv regions.

Figure S3 Blood cell counts.

Notes: Counts of WBC, LYM, MON, and GRA in peripheral blood are compared among healthy control C57BL/6, solvent, hIgG-treated, and aVAP2-treated mice in panels (A and B).
Abbreviations: aVAP2, anti-VAP2 polyclonal antibodies; GRA, granulocytes; LYM, lymphocytes; MON, monocytes; WBC, white blood cells.

Figure S4 Alignment of amino acid sequence of the variable region among anti-VAP2.

Notes: Alignment of amino acid sequence of the variable regions among anti-apolipoprotein monoclonal antibodies and URq01 (A). Phylogenic analysis of amino acid sequence of the registered variable region of anti-VAP2 monoclonal antibodies (B).

Acknowledgments

This study was supported, in part, by a grant from the Gamma-globulin Project from the Japan Science and Technology Agency and the Japan Agency for Medical Research and Development in Japan. We thank Drs Toshiko Ito-Ihara and Wako Yumura for medical advice, Ms Hisae Onodera and Ms Yuko Okada of A-CLIP Institute and Ms Kaoru Tosaka and Ms Haruko Haisa of Teikyo University for their documentation.

Disclosure

The authors report no conflicts of interest in this work.