Differential functional patterns of memory CD4+ and CD8+ T-cells from volunteers immunized with Ty21a typhoid vaccine observed using a recombinant Escherichia coli system expressing S. Typhi proteins.
Differential functional patterns of memory CD4+ and CD8+ T-cells from volunteers immunized with Ty21a typhoid vaccine observed using a recombinant Escherichia coli system expressing S. Typhi proteins.
复制标题
使用表达伤寒沙门氏菌蛋白的重组大肠杆菌系统观察到接受 Ty21a 伤寒疫苗免疫的志愿者的记忆 CD4 和 CD8 T 细胞的差异功能模式。
DOI：
10.1016/j.vaccine.2019.10.020
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发表时间：
2020
期刊：
影响因子：
5.5
通讯作者：
Sztein,MarceloB
中科院分区：
文献类型：
--
作者：
Salerno-Gonçalves,Rosângela;Tettelin,Hervé;Luo,David;Guo,Qin;Ardito,MatthewT;Martin,WilliamD;DeGroot,AnneS;Sztein,MarceloB
人们普遍认为，CD4+和CD8+T细胞对伤寒沙门氏菌具有重要的保护作用。伤寒沙门氏菌是伤寒的病原体。然而，这些T细胞的抗原特异性在很大程度上仍不清楚。在此之前，我们论证了利用重组大肠杆菌表达系统揭示CD4+和CD8+T细胞抗原特异性的可行性。在这里，我们扩展了这些研究，包括对12个附加物的评估。伤寒蛋白：4个外膜蛋白(omph、OmpL、OmpR、OmpX)，3个Vi-多糖生物合成蛋白(TviA、TviB、TviE)，3个冷休克蛋白(CSPA、CSPB、CSPC)，以及2个保守的假设蛋白(CHP 1和Chp2)，都是基于对推测的T细胞表位含量的生物信息学分析而选择的。用流式细胞仪检测15名成年志愿者在口服Ty21a减毒活疫苗免疫前和免疫后42 天的CD_4~+和CD_8~+T细胞的功能(即对所选抗原刺激产生的细胞因子和细胞毒性表达标记)。尽管志愿者的T细胞抗原特异性不同，但我们观察到了针对ALL的T细胞免疫反应。对伤寒蛋白进行评估。这些反应包括9种蛋白：omph、OmpR、TviA、TviE、CSPA、CSPB、CSPC、CHP 1和CHP 2，这些都是以前没有报道过的T细胞反应。有趣的是，我们还观察到，与蛋白质无关，记忆T细胞的功能模式在CD4+和CD8+T细胞之间是不同的。综上所述，这些研究证明了使用生物信息学分析和治疗的可行性。在此描述的大肠杆菌表达系统，以发现新的免疫原性T细胞蛋白，可作为生产基于蛋白质的疫苗的潜在靶点。
It is widely accepted that CD4+and CD8+T-cells play a significant role in protection againstSalmonella entericaserovar Typhi (S.Typhi), the causative agent of the typhoid fever. However, the antigen specificity of these T-cells remains largely unknown. Previously, we demonstrated the feasibility of using a recombinantEscherichia coli(E. coli) expression system to uncover the antigen specificity of CD4+and CD8+T cells. Here, we expanded these studies to include the evaluation of 12 additionalS. Typhi proteins: 4 outer membrane proteins (OmpH, OmpL, OmpR, OmpX), 3 Vi-polysaccharide biosynthesis proteins (TviA, TviB, TviE), 3 cold shock proteins (CspA, CspB, CspC), and 2 conserved hypothetical proteins (Chp 1 and Chp2), all selected based on the bioinformatic analyses of the content of putative T-cell epitopes. CD4+and CD8+T cells from 15 adult volunteers, obtained before and 42 days after immunization with oral live attenuated Ty21a vaccine, were assessed for their functionality (i.e., production of cytokines and cytotoxic expression markers in response to stimulation with selected antigens) as measured by flow cytometry. Although volunteers differed on their T-cell antigen specificity, we observed T-cell immune responses against allS. Typhi proteins evaluated. These responses included 9 proteins, OmpH, OmpR, TviA, TviE, CspA, CspB, CspC, Chp 1 and Chp 2, which have not been previously reported to elicit T-cell responses. Interestingly, we also observed that, regardless of the protein, the functional patterns of the memory T-cells were different between CD4+and CD8+T cells. In sum, these studies demonstrated the feasibility of using bioinformatic analysis and theE. coliexpressing system described here to uncover novel immunogenic T-cell proteins that could serve as potential targets for the production of protein-based vaccines.