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Cellular immune responses to recombinant heat shock protein 70 from Histoplasma capsulatum.

机译:细胞对荚膜组织胞浆菌重组热休克蛋白70的细胞免疫应答。

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摘要

Heat shock protein (hsp) 70 from several microbes is antigenic in mammals. In this study we sequenced and expressed the gene encoding this protein from Histoplasma capsulatum to study its immunological activity. The deduced amino acid sequence of the gene demonstrated 71 and 76% identity to hsp7O from humans and Saccharomyces cerevisiae, respectively. A cDNA was synthesized by reverse transcription-PCR and was expressed in Escherichia coli. Recombinant protein reacted with a mouse monoclonal antibody raised against human hsp7O. Splenocytes from C57BL/6 mice immunized with recombinant hsp7O emulsified in adjuvant, but not yeast cells, reacted in vitro to the antigen. Recombinant hsp7O elicited a cutaneous delayed-type hypersensitivity response in mice immunized with protein or with viable yeast cells. Mice were injected with recombinant hsp7O and challenged intranasally with a sublethal inoculum of yeast cells. Vaccination did not confer protection in this model. Thus, recombinant hsp7O can induce a cell-mediated immune response but does not induce a protective response.
机译:来自几种微生物的热激蛋白(hsp)70在哺乳动物中具有抗原性。在这项研究中,我们从荚膜组织胞浆菌中测序并表达了编码该蛋白的基因,以研究其免疫活性。该基因的推导氨基酸序列分别与人和酿酒酵母中的hsp70具有71%和76%的同一性。通过逆转录PCR合成cDNA,并在大肠杆菌中表达。重组蛋白与针对人hsp7O的小鼠单克隆抗体反应。用在佐剂中乳化的重组hsp7O免疫的C57BL / 6小鼠的脾细胞(而非酵母细胞)在体外与抗原反应。重组hsp7O在用蛋白质或活酵母细胞免疫的小鼠中引起皮肤迟发型超敏反应。给小鼠注射重组hsp7O,并在鼻内用酵母细胞的亚致死接种物攻击。在该模型中,疫苗接种并未提供保护。因此,重组hsp70可诱导细胞介导的免疫反应,但不诱导保护反应。

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