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通过噬菌体展示文库筛选出的痤疮丙酸杆菌和胸膜肺炎放线杆菌的B细胞交叉表位可有效预防胸膜肺炎放线杆菌感染。

B cell cross-epitope of Propionibacterium acnes and Actinobacillus pleuropneumonia selected by phage display library can efficiently protect from Actinobacillus pleuropneumonia infection.

作者信息

Liu Jianfang, Ma Qiuyue, Yang Feng, Zhu Rining, Gu Jingmin, Sun Changjiang, Feng Xin, Du Chongtao, Langford Paul R, Han Wenyu, Yang Junling, Lei Liancheng

机构信息

College of Veterinary Medicine, Jinlin University, Changchun, Jilin, 130062, People's Republic of China.

Section of Paediatrics, Imperial College London, St. Mary's Campus, London W2 1 PG, United Kingdom.

出版信息

Vet Microbiol. 2017 Jun;205:14-21. doi: 10.1016/j.vetmic.2017.04.026. Epub 2017 Apr 29.

Abstract

Contagious porcine pleuropneumonia (CPP), caused by Actinobacillus pleuropneumoniae (APP), is a highly transmissible and fatal respiratory illness that causes tremendous economic losses for the pig breeding industry worldwide. Propionibacterium acnes (PA) has a strong cross-reaction with anti-APP1 and anti-APP5 serum and can efficiently prevent APP infection, which was fortuitously found in researching the differential gene between the different APP serotypes. There seems to be some natural cross-protection between PA and APP. To identify the common epitope, the phage display library of a PA whole genome was constructed, whose size is 10. The DNA sequence of the positive clone was determined after three rounds of biopanning, and ten common protein types were identified and the epitope was predicted by computer software. Six peptide epitopes were selected and synthesized for further analysis. Among these epitopes, Ba1, Bb5 and C1 could bind to anti-PA serum and anti-APP1 serum and vice versa. Furthermore, the IgG and IL-4 levels and CD4/CD8 T cell ratios in the Ba1, Bb5 and C1 groups were significantly higher than that in the control group, indicating that the epitopes could trigger an immune response, which was mainly humoral immunity. Moreover, Ba1 and Bb5 equally protected 80% of mice from a fatal dose of APP1 infection compared with the control group. Mice could resist APP1 and APP5 challenge after being treated with the combination of Ba1 and Bb5, with survival rates of 80% and 90%, respectively. These findings suggest that the PA epitope confers antigenicity and can heterologously resist to the APP infection. This finding provides a novel strategy for preventing APP infection.

摘要

传染性猪胸膜肺炎(CPP)由胸膜肺炎放线杆菌(APP)引起,是一种高度传染性和致命性的呼吸道疾病,给全球养猪业造成巨大经济损失。痤疮丙酸杆菌(PA)与抗APP1和抗APP5血清有强烈的交叉反应,并且能够有效预防APP感染,这是在研究不同APP血清型之间的差异基因时偶然发现的。PA和APP之间似乎存在一些天然的交叉保护作用。为了鉴定共同表位,构建了PA全基因组的噬菌体展示文库,其库容为10。经过三轮生物淘选后确定了阳性克隆的DNA序列,鉴定出十种常见的蛋白类型,并通过计算机软件预测了表位。选择并合成了六个肽表位用于进一步分析。在这些表位中,Ba1、Bb5和C1能够与抗PA血清和抗APP1血清结合,反之亦然。此外,Ba1、Bb5和C1组的IgG和IL-4水平以及CD4/CD8 T细胞比率显著高于对照组,表明这些表位能够引发免疫反应,主要是体液免疫。此外,与对照组相比,Ba1和Bb5均能使80%的小鼠免受致死剂量的APP1感染。用Ba1和Bb5联合处理后的小鼠能够抵抗APP1和APP5的攻击,存活率分别为80%和90%。这些发现表明PA表位具有抗原性,并且能够异源抵抗APP感染。这一发现为预防APP感染提供了一种新策略。

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