Skip to main content
NCBI home page
Infection and Immunity logoLink to Infection and Immunity
. 1983 Jun;40(3):1052–1059. doi: 10.1128/iai.40.3.1052-1059.1983

Cellular Immunity in a Cutaneous Model of Cryptococcosis

Judith E Domer 1, Frank L Lyon 1,†, Juneann W Murphy 2
PMCID: PMC348157  PMID: 6343238

Abstract

The temporal development of cellular immune responses in mice inoculated cutaneously with viable Cryptococcus neoformans 145 was determined in vivo and in vitro by comparing several antigen preparations for their efficacy in the assays selected. Three antigens derived from C. neoformans 145, viz., a culture filtrate preparation (CneF-145), a membrane extract (B-HEX), and soluble cytoplasmic substances (SCS), were compared for their ability to detect delayed hypersensitivity (DH) in vivo in a footpad assay or to stimulate lymphocytes in vitro in a thymidine incorporation assay. DH to B-HEX could be demonstrated as early as 1 week after infection, whereas significant responses to SCS and CneF-145 were not regularly detected until 3 weeks after infection. Substantial reactions were observed to all three antigens up to 12 weeks, although they peaked at 2 to 3 weeks. Reactions to B-HEX and SCS were somewhat better than those to CneF. Differences in the efficacies of the three antigens were not obvious after the sixth week of infection, however. In vitro, lymph node cells from infected animals were stimulated significantly with all three antigens beginning at week 1. As with DH, however, responses to CneF-145 were usually less than those to SCS and B-HEX. In vitro lymphocyte responses waned after approximately 6 weeks, whereas DH responses were clearly positive through 12 weeks. In addition to the studies in infected animals, animals immunized with heat-killed cells of C. neoformans 145 or 184 were tested 6 to 8 days later for DH with CneF-145, CneF-184, or B-HEX derived from C. neoformans 145. The CneF-145 and CneF-184 were equally effective for detecting DH, regardless of the cryptococcal strain used for immunization. Likewise, the B-HEX detected equivalent responses in mice sensitized with each cryptococcal strain. Since all three antigens were soluble and easily extracted and since each elicited significant cellular immune responses in infected animals, further studies involving their specificity and the nature of their reactive components seems warranted as they may help evaluate immune responses in humans infected with this fungus.

Full text

PDF
1052

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Atkinson A. J., Jr, Bennett J. E. Experience with a new skin test antigen prepared from Cryptococcus neoformans. Am Rev Respir Dis. 1968 Apr;97(4):637–643. doi: 10.1164/arrd.1968.97.4.637. [DOI] [PubMed] [Google Scholar]
  2. BENNETT J. E., HASENCLEVER H. F., BAUM G. L. EVALUATION OF A SKIN TEST FOR CRYPTOCOCCOSIS. Am Rev Respir Dis. 1965 Apr;91:616–616. doi: 10.1164/arrd.1965.91.4.616. [DOI] [PubMed] [Google Scholar]
  3. Bennett J. E. Cryptococcal skin test antigen: preparation variables and characterization. Infect Immun. 1981 Apr;32(1):373–380. doi: 10.1128/iai.32.1.373-380.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Brummer E., Vris T. W., Lawrence H. S. A microculture system for the measurement of antigen-induced murine lymphocyte proliferation: advantages of 5% horse serum and 5 X 10(-5) M mercaptoethanol. J Immunol Methods. 1977;17(3-4):319–327. doi: 10.1016/0022-1759(77)90114-4. [DOI] [PubMed] [Google Scholar]
  5. Cauley L. K., Murphy J. W. Response of congenitally athymic (nude) and phenotypically normal mice to Cryptococcus neoformans infection. Infect Immun. 1979 Mar;23(3):644–651. doi: 10.1128/iai.23.3.644-651.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Diamond R. D., Bennett J. E. Disseminated cryptococcosis in man: decreased lymphocyte transformation in response to Cryptococcus neoformans. J Infect Dis. 1973 Jun;127(6):694–697. doi: 10.1093/infdis/127.6.694. [DOI] [PubMed] [Google Scholar]
  7. Dykstra M. A., Friedman L. Pathogenesis, lethality, and immunizing effect of experimental cutaneous cryptococcosis. Infect Immun. 1978 May;20(2):446–455. doi: 10.1128/iai.20.2.446-455.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Fromtling R. A., Blackstock R., Hall N. K., Bulmer G. S. Immunization of mice with an avirulent pseudohyphal form of Cryptococcus neoformans. Mycopathologia. 1979 Sep 28;68(3):179–181. doi: 10.1007/BF00578527. [DOI] [PubMed] [Google Scholar]
  9. Fromtling R. A., Blackstock R., Hall N. K., Bulmer G. S. Kinetics of lymphocyte transformation in mice immunized with viable avirulent forms of Cryptococcus neoformans. Infect Immun. 1979 May;24(2):449–453. doi: 10.1128/iai.24.2.449-453.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Fromtling R. A., Fromtling A. M., Staib F., Müller S. Effect of uremia on lymphocyte transformation and chemiluminescence by spleen cells of normal and Cryptococcus neoformans-infected mice. Infect Immun. 1981 Jun;32(3):1073–1078. doi: 10.1128/iai.32.3.1073-1078.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Fung P. Y., Murphy J. W. In vitro interactions of immune lymphocytes and Cryptococcus neoformans. Infect Immun. 1982 Jun;36(3):1128–1138. doi: 10.1128/iai.36.3.1128-1138.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Giger D. K., Domer J. E., McQuitty J. T., Jr Experimental murine candidiasis: pathological and immune responses to cutaneous inoculation with Candida albicans. Infect Immun. 1978 Feb;19(2):499–509. doi: 10.1128/iai.19.2.499-509.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Graybill J. R., Alford R. H. Cell-mediated immunity in Cryptococcosis. Cell Immunol. 1974 Oct;14(1):12–21. doi: 10.1016/0008-8749(74)90164-6. [DOI] [PubMed] [Google Scholar]
  14. Graybill J. R., Straus D. C., Nealon T. J., Hague M., Paque R. E. Immunogenic fractions of Cryptococcus neoformans. Mycopathologia. 1982 Apr 23;78(1):31–39. doi: 10.1007/BF00436579. [DOI] [PubMed] [Google Scholar]
  15. Graybill J. R., Taylor R. L. Host defense in cryptococcosis. I. An in vivo model for evaluating immune response. Int Arch Allergy Appl Immunol. 1978;57(2):101–113. [PubMed] [Google Scholar]
  16. Hay R. J., Reiss E. Delayed-type hypersensitivity responses in infected mice elicited by cytoplasmic fractions of Cryptococcus neoformans. Infect Immun. 1978 Oct;22(1):72–79. doi: 10.1128/iai.22.1.72-79.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Jones A. E., Reiss E., Spira T. J. A microsomal fraction of Cryptococcus neoformans induces lymphocyte blastogenesis in infected guinea pigs. Mycopathologia. 1981 Sep 11;75(3):129–138. doi: 10.1007/BF00482807. [DOI] [PubMed] [Google Scholar]
  18. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  19. Lim T. S., Murphy J. W., Cauley L. K. Host-etiological agent interactions in intranasally and intraperitoneally induced Cryptococcosis in mice. Infect Immun. 1980 Aug;29(2):633–641. doi: 10.1128/iai.29.2.633-641.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Martin M. V., Schneidau J. D., Jr A simple and reliable assimilation test for the identification of candida species. Am J Clin Pathol. 1970 Jun;53(6):875–879. doi: 10.1093/ajcp/53.6.875. [DOI] [PubMed] [Google Scholar]
  21. Moser S. A., Domer J. E. Effects of cyclophosphamide on murine candidiasis. Infect Immun. 1980 Feb;27(2):376–386. doi: 10.1128/iai.27.2.376-386.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Moser S. A., Lyon F. L., Domer J. E., Williams J. E. Immunization of mice by intracutaneous inoculation with viable virulent Cryptococcus neoformans: immunological and histopathological parameters. Infect Immun. 1982 Feb;35(2):685–696. doi: 10.1128/iai.35.2.685-696.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Muchmore H. G., Felton F. G., Salvin S. B., Rhoades E. R. Delayed hypersensitivity to cryptococcin in man. Sabouraudia. 1968 Oct;6(4):285–288. doi: 10.1080/00362176885190561. [DOI] [PubMed] [Google Scholar]
  24. Murphy J. W., Gregory J. A., Larsh H. W. Skin testing of guinea pigs and footpad testing of mice with a new antigen for detecting delayed hypersensitivity to Cryptococcus neoformans. Infect Immun. 1974 Feb;9(2):404–409. doi: 10.1128/iai.9.2.404-409.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Murphy J. W., Moorhead J. W. Regulation of cell-mediated immunity in cryptococcosis. I. Induction of specific afferent T suppressor cells by cryptococcal antigen. J Immunol. 1982 Jan;128(1):276–283. [PubMed] [Google Scholar]
  26. Murphy J. W., Pahlavan N. Cryptococcal culture filtrate antigen for detection of delayed-type hypersensitivity in cryptococcosis. Infect Immun. 1979 Jul;25(1):284–292. doi: 10.1128/iai.25.1.284-292.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Restrepo-Moreno A., Schneidau J. D., Jr Nature of the skin-reactive principle in culture filtrates prepared from Paracoccidioides brasiliensis. J Bacteriol. 1967 Jun;93(6):1741–1748. doi: 10.1128/jb.93.6.1741-1748.1967. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. SALVIN S. B., SMITH R. F. An antigen for detection of hypersensitivity to Cryptococcus neoformans. Proc Soc Exp Biol Med. 1961 Nov;108:498–501. doi: 10.3181/00379727-108-26977. [DOI] [PubMed] [Google Scholar]
  29. Schimpff S. C., Bennett J. E. Abnormalities in cell-mediated immunity in patients with Cryptococcus neoformans infection. J Allergy Clin Immunol. 1975 Jun;55(6):430–441. doi: 10.1016/0091-6749(75)90082-2. [DOI] [PubMed] [Google Scholar]
  30. Shields A. B., Ajello L. Medium for selective isolation of Cryptococcus neoformans. Science. 1966 Jan 14;151(3707):208–209. doi: 10.1126/science.151.3707.208. [DOI] [PubMed] [Google Scholar]

Articles from Infection and Immunity are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES