Effects of the fusion design and immunization route on the immunogenicity of Ag85A-Mtb32 in adenoviral vectored tuberculosis vaccine
Effects of the fusion design and immunization route on the immunogenicity of Ag85A-Mtb32 in adenoviral vectored tuberculosis vaccine
复制标题
DOI：
10.1080/21645515.2015.1042193
复制
发表时间：
2015-06
影响因子：
4.8
通讯作者：
Yiling Zhang;Liqiang Feng;Liang Li;Dimin Wang;Chufang Li;Caijun Sun;Pingchao Li;Xuehua Zheng;Yichu Liu;Wei Yang;Xuefeng Niu;N. Zhong;Ling Chen
中科院分区：
文献类型：
--
作者：
Yiling Zhang;Liqiang Feng;Liang Li;Dimin Wang;Chufang Li;Caijun Sun;Pingchao Li;Xuehua Zheng;Yichu Liu;Wei Yang;Xuefeng Niu;N. Zhong;Ling Chen
与单一抗原相比，含有多个抗原的疫苗可以诱导更广泛的免疫反应，并对结核分枝杆菌(Mtb)感染提供更好的保护。然而，将多个抗原整合到单个载体中的策略和免疫途径可能会影响它们的免疫原性。本研究以重组腺病毒5型(RAd5)为模型疫苗载体，以Ag85A(Rv3804c)和Mtb32(Rv0125)为模型抗原，比较评价了密码子使用优化、信号序列、融合接头和免疫途径对含多种抗原的结核疫苗在C57BL/6小鼠体内免疫原性的影响。我们发现，密码子优化的Ag85A和Mtb32与GSG连接子融合可诱导最强的全身和肺细胞免疫(CMI)反应。强烈的CMI反应的特征是产生强大的干扰素-γELISPOT反应以及抗原特异性的CD4T和CD8T细胞，它们分泌单一、双重或多种细胞因子。我们还发现，该候选疫苗的皮下(SC)和鼻腔(IN)/口服免疫分别对卡介苗(BCG)诱导的全身和肺CMI反应表现出最强的促进作用。我们的结果支持密码子优化的Ag85A和Mtb32与合适的连接子融合，并通过SC和IN/OLAR途径免疫，可以在卡介苗免疫的小鼠中产生最强的全身和肺CMI反应，这可能对含有多种抗原的结核疫苗的设计特别重要。
Vaccines containing multiple antigens may induce broader immune responses and provide better protection against Mycobacterium tuberculosis (Mtb) infection as compared to a single antigen. However, strategies for incorporating multiple antigens into a single vector and the immunization routes may affect their immunogenicity. In this study, we utilized recombinant adenovirus type 5 (rAd5) as a model vaccine vector, and Ag85A (Rv3804c) and Mtb32 (Rv0125) as model antigens, to comparatively evaluate the influence of codon usage optimization, signal sequence, fusion linkers, and immunization routes on the immunogenicity of tuberculosis (TB) vaccine containing multiple antigens in C57BL/6 mice. We showed that codon-optimized Ag85A and Mtb32 fused with a GSG linker induced the strongest systemic and pulmonary cell-mediated immune (CMI) responses. Strong CMI responses were characterized by the generation of a robust IFN-γ ELISPOT response as well as antigen-specific CD4+ T and CD8+ T cells, which secreted mono-, dual-, or multiple cytokines. We also found that subcutaneous (SC) and intranasal (IN)/oral immunization with this candidate vaccine exhibited the strongest boosting effects for Mycobacterium bovis bacille Calmette-Guérin (BCG)-primed systemic and pulmonary CMI responses, respectively. Our results supported that codon optimized Ag85A and Mtb32 fused with a proper linker and immunized through SC and IN/oral routes can generate the strongest systemic and pulmonary CMI responses in BCG-primed mice, which may be particularly important for the design of TB vaccines containing multiple antigens.