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毕赤酵母表达抗副猪嗜血杆菌中和性鼠猪嵌合抗体的研制。

Development of a neutralizing mouse-pig chimeric antibody with therapeutic potential against Haemophilus parasuis in Pichia pastoris.

机构信息

State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin, China.

出版信息

FEMS Microbiol Lett. 2014 May;354(2):85-91. doi: 10.1111/1574-6968.12437. Epub 2014 Apr 16.

Abstract

Haemophilus parasuis is one of the most important bacterial diseases of pigs worldwide. The lack of a vaccine against a broad spectrum of strains and the limitation of antimicrobial susceptibility hamper the control of disease. In this study, we cloned the constant regions of gamma heavy chains and kappa light chain of pig lymphocytes in frame with the variable regions of heavy and light chains of mouse monoclonal antibody 1D8, which reacts with all 15 serotypes of H. parasuis and has neutralizing activity. The constructed mouse-pig chimeric antibody was expressed in Pichia pastoris. Results demonstrated that the expressed chimeric antibody inhibited the growth of H. parasuis in vitro. Furthermore, the experiments in mice showed that chimeric antibody increased survival rate of the mice compared with that of the control group (P < 0.05). Importantly, the chimeric antibody partially protected piglets against H. parasuis infection according to the clinical lesion scores and PCR results of H. parasuis in the tissues from piglets of the chimeric antibody-inoculated group and the PBS group. In summary, our results demonstrated that the mouse-pig chimeric antibody could be a therapeutic candidate to prevent the H. parasuis infection and control the prevalence of disease.

摘要

副猪嗜血杆菌是全球范围内猪最重要的细菌性疾病之一。缺乏针对广谱菌株的疫苗和抗菌药物敏感性的限制,阻碍了疾病的控制。在本研究中,我们将猪淋巴细胞的γ重链和 κ轻链的恒定区与小鼠单克隆抗体 1D8 的重链和轻链可变区进行框内克隆,该抗体与所有 15 种副猪嗜血杆菌血清型反应,具有中和活性。构建的鼠-猪嵌合抗体在毕赤酵母中表达。结果表明,表达的嵌合抗体在体外抑制了副猪嗜血杆菌的生长。此外,小鼠实验表明,嵌合抗体与对照组相比,提高了小鼠的存活率(P<0.05)。重要的是,根据嵌合抗体接种组和 PBS 组仔猪组织中副猪嗜血杆菌的临床病变评分和 PCR 结果,嵌合抗体部分保护了仔猪免受副猪嗜血杆菌感染。综上所述,我们的结果表明,鼠-猪嵌合抗体可能是预防副猪嗜血杆菌感染和控制疾病流行的治疗候选物。

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