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首页> 外文期刊>Infection and immunity >A recombinant protein based on the Trypanosoma cruzi metacyclic trypomastigote 82-kilodalton antigen that induces and effective immune response to acute infection.
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A recombinant protein based on the Trypanosoma cruzi metacyclic trypomastigote 82-kilodalton antigen that induces and effective immune response to acute infection.

机译：一种重组蛋白，基于克鲁斯锥虫锥虫Trypomastigote 82-千洛酮抗原，可诱导对急性感染的有效免疫反应。

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摘要

To further investigate the immunological properties of the stage-specific 82-kDa glycoprotein (gp82) of Trypanosoma cruzi metacyclic trypomastigotes, previously shown to induce antigen-specific humoral and T-cell responses in mice, we performed a series of experiments with recombinant proteins containing sequences of gp82 fused to glutathione S-transferase. Of five fusion proteins tested, only J18b and J18b1, the carboxyproximal peptides containing amino acids 224 to 516 and 303 to 516, respectively, were recognized by monoclonal antibody 3F6 as well as by various anti-T. cruzi antisera and, when administered to mice, were capable of eliciting antibodies directed to the native gp82. The amino-terminal peptide and other carboxyterminal recombinant proteins lacking the central domain of gp82 (amino acids 224 to 356), which is exposed on the surface of live metacyclic forms, did not display any of these properties. Spleen cells derived from mice immunized with any of the five recombinant proteins proliferated in vitro in the presence of native gp82.J18b was the most stimulatory, whereas J18b3, the peptide containing amino acids 408 to 516, elicited the weakest response. When BALB/c mice immunized with J18b antigen plus A1(OH)3 as adjuvant were challenged 10 5 metacyclic trypomastigotes, 85% of them resisted acute infection, in comparison with control mice that received glutathione S-transferase plus adjuvant. Antibodies induced by J18b protein lacked agglutinating or complement-dependent lytic activity and failed to neutralize parasite infectivity. On the other hand, CD4+T cells from the spleens of J18b-immunized mice displayed an intense proliferative activity upon stimulation with 1.25 microgram of native gp82 per ml, which resulted in increased production of gamma interferon, a cytokine associated with resistance to T. cruzi infection.
机译：为了进一步研究锥虫锥虫亚环锥虫的阶段特异性82 kDa糖蛋白（gp82）的免疫学特性，先前已证明在小鼠中可诱导抗原特异性体液和T细胞反应，我们对含有以下成分的重组蛋白进行了一系列实验与谷胱甘肽S-转移酶融合的gp82序列。在测试的五种融合蛋白中，只有J18b和J18b1（分别包含氨基酸224至516和303至516氨基酸的羧基近端肽）被单克隆抗体3F6以及各种抗T蛋白识别。 Cruzi抗血清，当施用于小鼠时，能够引发针对天然gp82的抗体。缺少gp82中央结构域（氨基酸224至356）的氨基末端肽和其他羧基末端重组蛋白暴露在活的元环形式的表面，没有任何这些特性。在天然gp82存在下，用五种重组蛋白中的任何一种免疫的小鼠衍生的脾细胞在体外具有增生作用.J18b具有最大的刺激性，而J18b3（含408至516位氨基酸的肽）引起的应答最弱。当用J18b抗原加A1（OH）3作为佐剂免疫的BALB / c小鼠受到10 5个间环锥虫的攻击时，与接受谷胱甘肽S-转移酶加佐剂的对照小鼠相比，其中的85％抵抗了急性感染。 J18b蛋白诱导的抗体缺乏凝集或补体依赖性的裂解活性，并且无法中和寄生虫的感染性。另一方面，经J18b免疫的小鼠脾脏中的CD4 + T细胞在以每毫升1.25微克的天然gp82刺激后表现出强烈的增殖活性，这导致γ干扰素（一种与T抗性相关的细胞因子）的产生增加。克鲁兹感染。

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