Abstract
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.
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- Adams E., Garsia R. J., Hellqvist L., Holt P., Basten A. T cell reactivity to the purified mycobacterial antigens p65 and p70 in leprosy patients and their household contacts. Clin Exp Immunol. 1990 May;80(2):206–212. doi: 10.1111/j.1365-2249.1990.tb05235.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ardeshir F., Flint J. E., Richman S. J., Reese R. T. A 75 kd merozoite surface protein of Plasmodium falciparum which is related to the 70 kd heat-shock proteins. EMBO J. 1987 Feb;6(2):493–499. doi: 10.1002/j.1460-2075.1987.tb04780.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Behr C., Sarthou J. L., Rogier C., Trape J. F., Dat M. H., Michel J. C., Aribot G., Dieye A., Claverie J. M., Druihle P. Antibodies and reactive T cells against the malaria heat-shock protein Pf72/Hsp70-1 and derived peptides in individuals continuously exposed to Plasmodium falciparum. J Immunol. 1992 Nov 15;149(10):3321–3330. [PubMed] [Google Scholar]
- Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1016/0003-2697(76)90527-3. [DOI] [PubMed] [Google Scholar]
- Britton W. J., Hellqvist L., Basten A., Inglis A. S. Immunoreactivity of a 70 kD protein purified from Mycobacterium bovis Bacillus Calmette-Guerin by monoclonal antibody affinity chromatography. J Exp Med. 1986 Sep 1;164(3):695–708. doi: 10.1084/jem.164.3.695. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Buchmeier N. A., Heffron F. Induction of Salmonella stress proteins upon infection of macrophages. Science. 1990 May 11;248(4956):730–732. doi: 10.1126/science.1970672. [DOI] [PubMed] [Google Scholar]
- Caruso M., Sacco M., Medoff G., Maresca B. Heat shock 70 gene is differentially expressed in Histoplasma capsulatum strains with different levels of thermotolerance and pathogenicity. Mol Microbiol. 1987 Sep;1(2):151–158. doi: 10.1111/j.1365-2958.1987.tb00507.x. [DOI] [PubMed] [Google Scholar]
- Deepe G. S., Jr, Durose G. G. Immunobiological activity of recombinant H antigen from Histoplasma capsulatum. Infect Immun. 1995 Aug;63(8):3151–3157. doi: 10.1128/iai.63.8.3151-3157.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Deepe G. S., Jr Protective immunity in murine histoplasmosis: functional comparison of adoptively transferred T-cell clones and splenic T cells. Infect Immun. 1988 Sep;56(9):2350–2355. doi: 10.1128/iai.56.9.2350-2355.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gomez F. J., Allendoerfer R., Deepe G. S., Jr Vaccination with recombinant heat shock protein 60 from Histoplasma capsulatum protects mice against pulmonary histoplasmosis. Infect Immun. 1995 Jul;63(7):2587–2595. doi: 10.1128/iai.63.7.2587-2595.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gomez F. J., Gomez A. M., Deepe G. S., Jr An 80-kilodalton antigen from Histoplasma capsulatum that has homology to heat shock protein 70 induces cell-mediated immune responses and protection in mice. Infect Immun. 1992 Jul;60(7):2565–2571. doi: 10.1128/iai.60.7.2565-2571.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hedstrom R., Culpepper J., Harrison R. A., Agabian N., Newport G. A major immunogen in Schistosoma mansoni infections is homologous to the heat-shock protein Hsp70. J Exp Med. 1987 May 1;165(5):1430–1435. doi: 10.1084/jem.165.5.1430. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kaufmann S. H. Heat shock proteins and the immune response. Immunol Today. 1990 Apr;11(4):129–136. doi: 10.1016/0167-5699(90)90050-j. [DOI] [PubMed] [Google Scholar]
- Kumar N., Zhao Y., Graves P., Perez Folgar J., Maloy L., Zheng H. Human immune response directed against Plasmodium falciparum heat shock-related proteins. Infect Immun. 1990 May;58(5):1408–1414. doi: 10.1128/iai.58.5.1408-1414.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lindquist S., Craig E. A. The heat-shock proteins. Annu Rev Genet. 1988;22:631–677. doi: 10.1146/annurev.ge.22.120188.003215. [DOI] [PubMed] [Google Scholar]
- MacFarlane J., Blaxter M. L., Bishop R. P., Miles M. A., Kelly J. M. Identification and characterisation of a Leishmania donovani antigen belonging to the 70-kDa heat-shock protein family. Eur J Biochem. 1990 Jun 20;190(2):377–384. doi: 10.1111/j.1432-1033.1990.tb15586.x. [DOI] [PubMed] [Google Scholar]
- Munro S., Pelham H. R. An Hsp70-like protein in the ER: identity with the 78 kd glucose-regulated protein and immunoglobulin heavy chain binding protein. Cell. 1986 Jul 18;46(2):291–300. doi: 10.1016/0092-8674(86)90746-4. [DOI] [PubMed] [Google Scholar]
- Mustafa A. S., Lundin K. E., Oftung F. Human T cells recognize mycobacterial heat shock proteins in the context of multiple HLA-DR molecules: studies with healthy subjects vaccinated with Mycobacterium bovis BCG and Mycobacterium leprae. Infect Immun. 1993 Dec;61(12):5294–5301. doi: 10.1128/iai.61.12.5294-5301.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Rees A., Scoging A., Mehlert A., Young D. B., Ivanyi J. Specificity of proliferative response of human CD8 clones to mycobacterial antigens. Eur J Immunol. 1988 Dec;18(12):1881–1887. doi: 10.1002/eji.1830181203. [DOI] [PubMed] [Google Scholar]
- Shearer G., Jr, Birge C. H., Yuckenberg P. D., Kobayashi G. S., Medoff G. Heat-shock proteins induced during the mycelial-to-yeast transitions of strains of Histoplasma capsulatum. J Gen Microbiol. 1987 Dec;133(12):3375–3382. doi: 10.1099/00221287-133-12-3375. [DOI] [PubMed] [Google Scholar]
- Smith J. G., Magee D. M., Williams D. M., Graybill J. R. Tumor necrosis factor-alpha plays a role in host defense against Histoplasma capsulatum. J Infect Dis. 1990 Dec;162(6):1349–1353. doi: 10.1093/infdis/162.6.1349. [DOI] [PubMed] [Google Scholar]
- Welch W. J. Mammalian stress response: cell physiology, structure/function of stress proteins, and implications for medicine and disease. Physiol Rev. 1992 Oct;72(4):1063–1081. doi: 10.1152/physrev.1992.72.4.1063. [DOI] [PubMed] [Google Scholar]
- Young D., Lathigra R., Hendrix R., Sweetser D., Young R. A. Stress proteins are immune targets in leprosy and tuberculosis. Proc Natl Acad Sci U S A. 1988 Jun;85(12):4267–4270. doi: 10.1073/pnas.85.12.4267. [DOI] [PMC free article] [PubMed] [Google Scholar]