利用多克隆抗体鉴定蜂房哈夫尼菌溶菌素 1-3 结构域中的 B 细胞线性表位。

Identification of B-cell linear epitopes in domains 1-3 of pyolysin of Trueperella pyogenes using polyclonal antibodies.

机构信息

College of Veterinary Medicine, Northeast Agricultural University, Harbin, Heilongjiang 150030, PR China; Northeastern Science Inspection Station, China Ministry of Agriculture Key Laboratory of Animal Pathogen Biology, PR China.

College of Animal Science and Technology, Northeast Agricultural University, Harbin, Heilongjiang 150030, PR China.

出版信息

Vet Microbiol. 2017 Oct;210:24-31. doi: 10.1016/j.vetmic.2017.08.018. Epub 2017 Aug 31.

DOI:10.1016/j.vetmic.2017.08.018

PMID:29103692

Abstract

Trueperella pyogenes is an important opportunistic pathogen. Pyolysin (PLO) makes important contributions to the pathogenicity of T. pyogenes. However, the structure and function of PLO has not been well documented. In the current study, epitopes in domain 1-3 of PLO have been mapped using rabbit anti-recombinant PLO (rPLO) polyclonal antibodies, and then the results were re-checked by using mouse and chicken anti-rPLO polyclonal antibodies, respectively. The results indicated that the region of aa 281-393 in PLO could not elicit antibodies against linear epitopes. A total of six B cell linear epitopes have been found in domain 1 of PLO. Two of the six epitopes (EP1 and EP2) were used to immunize mice and chicken. Chicken anti-EP1 and anti-EP2 serum and mouse anti-EP2 serum could react with rPLO and corresponding epitope polypeptide in western blot assay; however, only mouse anti-EP2 serum shows weak anti-hemolysis effect in the rPLO and sheep red blood system. Our results provide some new information to the research field of PLO structure and function.

摘要

化脓隐秘杆菌是一种重要的机会性病原体。溶葡球菌酶（PLO）对化脓隐秘杆菌的致病性有重要贡献。然而，PLO 的结构和功能尚未得到很好的阐明。在本研究中，使用兔抗重组 PLO（rPLO）多克隆抗体对 PLO 结构域 1-3 中的表位进行了定位，然后分别使用小鼠和鸡抗 rPLO 多克隆抗体对结果进行了重新检查。结果表明，PLO 中 aa281-393 区域不能诱导针对线性表位的抗体。在 PLO 的结构域 1 中发现了总共六个 B 细胞线性表位。这六个表位中的两个（EP1 和 EP2）被用于免疫小鼠和鸡。鸡抗 EP1 和抗 EP2 血清以及鼠抗 EP2 血清可以在 Western blot 检测中与 rPLO 和相应的表位多肽发生反应；然而，只有鼠抗 EP2 血清在 rPLO 和绵羊红细胞系统中显示出微弱的抗溶血作用。我们的研究结果为 PLO 结构和功能的研究领域提供了一些新的信息。