Development and Evaluation of CmeC Subunit Vaccine against Campylobacter jejuni.
Development and Evaluation of CmeC Subunit Vaccine against Campylobacter jejuni.
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DOI：
10.4172/2157-7560.1000112
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发表时间：
2010
期刊：
影响因子：
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通讯作者：
X. Zeng;Fuzhou Xu;Jun Lin
中科院分区：
文献类型：
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作者：
X. Zeng;Fuzhou Xu;Jun Lin
空肠弯曲菌是许多工业化国家引起人类肠炎的主要细菌。目前还没有针对C.到目前为止，空肠可供使用。CmeC是CmeABC多药外排泵的重要外膜成分，在C.空肠。CmeC在C.空肠菌株，并在体内显著诱导和免疫原性。本研究分析了CmeC序列同源性，检测了CmeC肽抗体的体外免疫保护性，并制备了全长重组CmeC（rCmeC），用于评价CmeC亚单位疫苗的免疫原性和保护效力。利用鸡的模型系统进行空肠。从24个不同的C. jejuni菌株进行比对，结果表明CmeC在C.空肠的同源性为97.3%~ 100%。CmeC肽抗体可抑制CmeABC外排泵的功能，增强C.胆汁盐是存在于肠道中的天然抗菌剂。在E. coli中，经一步亲和纯化，获得了高纯度、高产量的N-末端His标记的rCmeC。将纯化的rCmeC用于使用鸡C.空肠感染通过口服疫苗接种刺激CmeC特异性血清IgG应答需要用更高剂量的rCmeC（200μg）连同70μg粘膜佐剂mLT（修饰的E.大肠杆菌不耐热肠毒素）。rCmeC皮下免疫鸡可显著刺激血清IgG和伊加应答。然而，无论接种方案如何，CmeC特异性肠道分泌伊加应答均未被显著刺激，并且rCmeC疫苗接种未赋予针对C的保护。空肠感染总之，这些发现提供了进一步令人信服的证据，CmeC是一个有前途的亚单位疫苗候选人对C。空肠感染然而，CmeC疫苗接种方案应优化，以增强CmeC特异性粘膜免疫应答，以保护免受C。空肠。
Campylobacter jejuni is the leading bacterial cause of human enteritis in many industrialized countries. There is no commercial vaccine against C. jejuni available to date. CmeC is an essential outer membrane component of CmeABC multidrug efflux pump that plays a critical role in antibiotic resistance and in vivo colonization of C. jejuni. CmeC is prevalent in C. jejuni strains and is dramatically induced and immunogenic in vivo. In this study, we analyzed CmeC sequence homology, examined in vitro immune protection of CmeC peptide antibodies, and produced full-length recombinant CmeC (rCmeC) for evaluating immunogenicity and protective efficacy of the CmeC subunit vaccine against C. jejuni using chicken model system. Amino acid sequences of CmeC from 24 diverse C. jejuni strains were determined and subjected to alignment, which revealed that CmeC is highly conserved in C. jejuni with a identity ranging from 97.3% to 100%. CmeC peptide antibodies inhibited the function of CmeABC efflux pump and enhanced susceptibility of C. jejuni to bile salts, the natural antimicrobial present in the intestine. Two full-length rCmeC proteins with N- or C-terminal His tag were produced in E. coli; the N-terminal His-tagged rCmeC with high purity and yield was obtained by single step affinity purification. The purified rCmeC was used in two vaccination trials using a chicken model of C. jejuni infection. Stimulation of CmeC-specific serum IgG responses via oral vaccination required immunization with higher doses of rCmeC (200μg) together with 70μg of mucosal adjuvant mLT (modified E. coli heat-labile enterotoxin). Subcutaneous vaccination of chickens with rCmeC remarkably stimulated both serum IgG and IgA responses. However, CmeC-specific intestinal secretory IgA response was not significantly stimulated regardless of vaccination regimen and the rCmeC vaccination did not confer protection against C. jejuni infection. Together, these findings provide further compelling evidence that CmeC is a promising subunit vaccine candidate against C. jejuni infection. However, the CmeC vaccination regimen should be optimized to enhance CmeC-specific mucosal immune response in for protection against C. jejuni.