Use of in vivo-induced antigen technology (IVIAT) for the identification of Streptococcus suis serotype 2 in vivo-induced bacterial protein antigens
Use of in vivo-induced antigen technology (IVIAT) for the identification of Streptococcus suis serotype 2 in vivo-induced bacterial protein antigens
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DOI：
10.1186/1471-2180-9-201
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发表时间：
2009-09-18
期刊：
影响因子：
4.2
通讯作者：
Lu, Chengping
中科院分区：
文献类型：
--
作者：
Gu, Hongwei;Zhu, Haodan;Lu, Chengping
背景：猪链球菌血清型2(SS2)是一种人畜共患病的病原体，可引起人和猪的死亡和疾病。更好地了解SS2与宿主的分子相互作用对于了解SS2的发病机制和免疫学是至关重要的。用于研究SS2毒力因子的常规遗传和生化方法无法考虑与感染过程相关的复杂和动态的环境刺激。本研究利用免疫筛选技术体内诱导抗原技术(IVIAT)对SS2感染过程中在体内诱导或上调的免疫原性细菌蛋白进行鉴定。结果：将SS2感染猪的恢复期血清混合，吸附体外抗原，并用于筛选SS2基因组表达文库。通过对鉴定出的蛋白质的分析，我们能够对48个蛋白质中的40个蛋白质进行推测功能。这些蛋白质包括与细胞被膜结构、调节、分子合成、物质和能量代谢、运输、翻译有关的蛋白质，以及那些未知功能的蛋白质。用实时逆转录聚合酶链式反应(RT-PCR)检测了这40个基因中的10个基因在体内诱导的表达变化，结果表明在体内条件下，这10个基因中有6个基因的表达上调。用聚合酶链式反应分析了这10个基因的菌株分布，发现它们存在于最强毒力的SS2菌株中。此外，新鉴定的蛋白质的蛋白质序列比对表明，其中三个可能是与毒力相关的蛋白。结论：综上所述，我们的结果表明这些在体内诱导或上调的基因可能与SS2疾病的发生有关。我们推测，识别SS2感染过程中特异性诱导或上调的因子将有助于我们理解SS2的发病机制，并可能有助于控制SS2的暴发。此外，使用IVIAT鉴定的蛋白质可能是有用的潜在候选疫苗或毒力标记。
Background: Streptococcus suis serotype 2 (SS2) is a zoonotic agent that causes death and disease in both humans and swine. A better understanding of SS2-host molecular interactions is crucial for understanding SS2 pathogenesis and immunology. Conventional genetic and biochemical approaches used to study SS2 virulence factors are unable to take into account the complex and dynamic environmental stimuli associated with the infection process. In this study, in vivo-induced antigen technology (IVIAT), an immunoscreening technique, was used to identify the immunogenic bacterial proteins that are induced or upregulated in vivo during SS2 infection.Results: Convalescent-phase sera from pigs infected with SS2 were pooled, adsorbed against in vitro antigens, and used to screen SS2 genomic expression libraries. Upon analysis of the identified proteins, we were able to assign a putative function to 40 of the 48 proteins. These included proteins implicated in cell envelope structure, regulation, molecule synthesis, substance and energy metabolism, transport, translation, and those with unknown functions. The in vivo-induced changes in the expression of 10 of these 40 genes were measured using real-time reverse transcription (RT)-PCR, revealing that the expression of 6 of the 10 genes was upregulated in the in vivo condition. The strain distribution of these 10 genes was analyzed by PCR, and they were found in the most virulent SS2 strains. In addition, protein sequence alignments of the newly identified proteins demonstrate that three are putative virulence-associated proteins.Conclusion: Collectively, our results suggest that these in vivo-induced or upregulated genes may contribute to SS2 disease development. We hypothesize that the identification of factors specifically induced or upregulated during SS2 infection will aid in our understanding of SS2 pathogenesis and may contribute to the control SS2 outbreaks. In addition, the proteins identified using IVIAT may be useful potential vaccine candidates or virulence markers.