Tc52 amino-terminal-domain DNA carried by attenuated Salmonella enterica serovar Typhimurium induces protection against a Trypanosoma cruzi lethal challenge.
Tc52 amino-terminal-domain DNA carried by attenuated Salmonella enterica serovar Typhimurium induces protection against a Trypanosoma cruzi lethal challenge.
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减毒肠沙门氏菌鼠伤寒血清型携带的 Tc52 氨基末端结构域 DNA 可诱导针对克氏锥虫致命攻击的保护。
DOI：
10.1128/iai.02190-14
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发表时间：
2014
影响因子：
3.1
通讯作者：
Malchiodi,EmilioL
中科院分区：
文献类型：
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作者：
Matos,MarinaN;Cazorla,SilviaI;Bivona,AugustoE;Morales,Celina;Guzmán,CarlosA;Malchiodi,EmilioL
在这项工作中，我们用减毒沙门氏菌携带的全长Tc52或其氨基或羧基末端(分别为N-和C-末端)结构域的DNA作为DNA递送系统免疫小鼠。正如预期的那样，沙门氏菌介导的DNA传递导致了低抗体效价和主要的Th1反应，如IgG2a/IgG1特异性抗体的比率所示。尽管Tc52在类鞭毛虫中有少量的表达，但免疫后产生的抗体能够在补体存在的情况下介导类鞭毛虫的裂解，并在体外抑制它们对哺乳动物细胞的侵袭。从沙门氏菌免疫的小鼠血清中观察到最强的功能活性，携带N-Term结构域(SN-Term)，其次是Tc52(STc52)和C-Term结构域(SC-Term)。所有免疫组都出现了强烈的细胞反应，主要是Th1细胞的激活。然而，SN-Term免疫的小鼠表现出更高的白介素10(IL-10)水平，抵消了炎症反应，并强烈激活了Tc52特异性干扰素阳性(干扰素-γ+)CD8+T细胞。与此一致的是，尽管所有的原型都提供了对感染的保护，但对于所有测试的参数，SN-Term免疫比SC-Term免疫的保护作用更强，比STC52的保护略好，特别是在感染的急性阶段。我们的结论是，Tc52的N-末端结构域是该蛋白对感染提供最大保护的部分，并建议将其作为疫苗开发的一个有前景的候选。
In this work we immunized mice with DNA encoding full-length Tc52 or its amino- or carboxy-terminal (N- and C-term, respectively) domain carried by attenuated Salmonella as a DNA delivery system. As expected, Salmonella-mediated DNA delivery resulted in low antibody titers and a predominantly Th1 response, as shown by the ratio of IgG2a/IgG1-specific antibodies. Despite modest expression of Tc52 in trypomastigotes, the antibodies elicited by vaccination were able to mediate lysis of the trypomastigotes in the presence of complement and inhibit their invasion of mammal cellsin vitro. The strongest functional activity was observed with sera from mice immunized with Salmonella carrying the N-term domain (SN-term), followed by Tc52 (STc52), and the C-term domain (SC-term). All immunized groups developed strong cellular responses, with predominant activation of Th1 cells. However, mice immunized with SN-term showed higher levels of interleukin-10 (IL-10), counterbalancing the inflammatory reaction, and also strong activation of Tc52-specific gamma interferon-positive (IFN-γ+) CD8+T cells. In agreement with this, although all prototypes conferred protection against infection, immunization with SN-term promoted greater protection than that with SC-term for all parameters tested and slightly better protection than that with STc52, especially in the acute stage of infection. We conclude that the N-terminal domain of Tc52 is the section of the protein that confers maximal protection against infection and propose it as a promising candidate for vaccine development.