Allendoerfer R, Maresca B, Deepe G S
Division of Infectious Diseases, Department of Medicine, University of Cincinnati College of Medicine, Ohio 45267-0560, USA.
Infect Immun. 1996 Oct;64(10):4123-8. doi: 10.1128/iai.64.10.4123-4128.1996.
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.
几种微生物的热休克蛋白(hsp)70在哺乳动物中具有抗原性。在本研究中,我们对荚膜组织胞浆菌编码该蛋白的基因进行了测序和表达,以研究其免疫活性。该基因推导的氨基酸序列与人类和酿酒酵母的hsp70分别有71%和76%的同一性。通过逆转录PCR合成了cDNA,并在大肠杆菌中进行表达。重组蛋白与针对人类hsp70产生的小鼠单克隆抗体发生反应。用佐剂乳化的重组hsp70免疫的C57BL/6小鼠的脾细胞,而非酵母细胞,在体外对抗原产生反应。重组hsp70在用蛋白或活酵母细胞免疫的小鼠中引发了皮肤迟发型超敏反应。给小鼠注射重组hsp70,然后经鼻用亚致死剂量的酵母细胞接种物进行攻击。在该模型中,疫苗接种未提供保护作用。因此,重组hsp70可诱导细胞介导的免疫反应,但不诱导保护性反应。