Enhanced efficacy of the novel recombinant clone VasSF in a mouse model of antineutrophil cytoplasmic antibody-associated vasculitis.

IF 3.4 3区 医学 Q3 IMMUNOLOGY
Minako Koura, Yosuke Kameoka, Fukuko Kishi, Yoshio Yamakawa, Fuyu Ito, Ryuichi Sugamata, Yuko Doi, Kazuko Uno, Toshinori Nakayama, Takashi Miki, Hiroshi Nakajima, Kazuo Suzuki, Osamu Suzuki
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Abstract

Based on the efficacy of intravenous immunoglobulin (IVIg) for the treatment of antineutrophil cytoplasmic antibody (ANCA)-associated vasculitis (AAV), we developed a recombinant single-chain-fragment variable clone, VasSF, therapeutic against AAV in a mouse model (SCG/Kj mice). VasSF is thought to bind to vasculitis-associated apolipoprotein A-II (APOA2) as a target molecule. VasSF is a promising new drug against AAV, but difficulties in the yield and purification of VasSF remain unresolved. We produced monomers of new VasSF molecules by modifying the plasmid structure for VasSF expression and simplifying the purification method using high-performance liquid chromatography. We compared the therapeutic effects between 5-day continuous administration of the monomers, as in IVIg treatment, and single shots of 5-day-equivalent doses. We also evaluated the life-prolonging effect of the single-shot treatment. Two-dimensional western blots were used to examine the binding of VasSF to APOA2. Our improved manufacturing method resulted in a 100-fold higher yield of VasSF than in our previous study. Monomerization of VasSF stabilized its efficacy. Single shots of a small amount (1/80 000 of IVIg) produced sufficient therapeutic effects, including decreased glomerular crescent formation, a decreasing trend of serum ANCA against myeloperoxidase (MPO-ANCA), decreases in multiple proinflammatory cytokines, and a trend toward prolonged survival. Two-dimensional western blots confirmed the binding of VasSF to APOA2. The newly produced pure VasSF monomers are stable and therapeutic for AAV with a single low-dose injection, possibly by removing vasculitis-associated APOA2. Thus, the new VasSF described herein is a promising drug against AAV.

新型重组克隆 VasSF 在抗中性粒细胞胞浆抗体相关性血管炎小鼠模型中的疗效增强。
基于静脉注射免疫球蛋白(IVIg)治疗抗中性粒细胞胞浆抗体(ANCA)相关性脉管炎(AAV)的疗效,我们在小鼠模型(SCG/Kj 小鼠)中开发了一种针对 AAV 的重组单链片段可变克隆 VasSF。VasSF 被认为能与脉管炎相关载脂蛋白 A-II (APOA2) 结合,成为一种靶分子。VasSF 是一种很有前途的抗 AAV 新药,但 VasSF 的产量和纯化方面的困难仍未解决。我们通过修改表达 VasSF 的质粒结构和使用高效液相色谱法简化纯化方法,制备了新的 VasSF 分子单体。我们比较了连续 5 天给予单体(如 IVIg 治疗)和单次注射 5 天同等剂量的治疗效果。我们还评估了单次注射治疗对延长生命的作用。我们使用二维 Western 印迹检查了 VasSF 与 APOA2 的结合情况。我们改进了生产方法,使 VasSF 的产量比以前的研究高出 100 倍。VasSF 的单体化使其药效更加稳定。单次注射少量(1/80000 IVIg)就能产生足够的治疗效果,包括肾小球新月体形成减少、血清 ANCA 抗髓过氧化物酶(MPO-ANCA)呈下降趋势、多种促炎细胞因子减少以及存活时间延长的趋势。二维 Western 印迹证实了 VasSF 与 APOA2 的结合。新生产的纯 VasSF 单体是稳定的,只需一次低剂量注射就能治疗 AAV,这可能是通过去除脉管炎相关 APOA2 实现的。因此,本文所述的新型 VasSF 是一种很有前景的抗 AAV 药物。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
CiteScore
8.40
自引率
2.20%
发文量
101
审稿时长
3-8 weeks
期刊介绍: Clinical & Experimental Immunology (established in 1966) is an authoritative international journal publishing high-quality research studies in translational and clinical immunology that have the potential to transform our understanding of the immunopathology of human disease and/or change clinical practice. The journal is focused on translational and clinical immunology and is among the foremost journals in this field, attracting high-quality papers from across the world. Translation is viewed as a process of applying ideas, insights and discoveries generated through scientific studies to the treatment, prevention or diagnosis of human disease. Clinical immunology has evolved as a field to encompass the application of state-of-the-art technologies such as next-generation sequencing, metagenomics and high-dimensional phenotyping to understand mechanisms that govern the outcomes of clinical trials.
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