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. 1985 May;75(5):1570–1577. doi: 10.1172/JCI111862

Recognition and characterization of stage-specific oocyst/sporozoite antigens of Toxoplasma gondii by human antisera.

L H Kasper, P L Ware
PMCID: PMC425497  PMID: 2581998

Abstract

Human infection with Toxoplasma gondii is presumed due to the ingestion of either tissue cysts containing bradyzoites or oocyst/sporozoites that are excreted in the feces of infected cats. The incidence of human infection in the general population by either of these routes is unknown. We have previously described unique stage-specific oocyst/sporozoite antigens identified by murine hybridoma monoclonal antibodies. We obtained acute and convalescent antitoxoplasma antisera from patients in an epidemiologically well-documented outbreak of oocyst-transmitted infection associated with the ingestion of contaminated water. An enzyme-linked immunosorbent assay comparing equal numbers of tachyzoites (invasive stage) and oocyst/sporozoite (excreted stage) indicated that these antisera recognized antigens from both life forms. Absorption of pooled antisera with purified oocyst/sporozoites reduced both the antioocyst immunoglobulin G (IgG) and immunoglobulin M (IgM) titer but had only minimal effect on the antitachyzoite titer. Absorption of the antisera with tachyzoites reduced the IgG and IgM antioocyst and antitachyzoite titer. A sodium dodecyl sulfate-polyacrylamide gel analysis of radioiodinated oocyst/sporozoites shows that the principal stage-specific surface proteins of the oocyst/sporozoite have approximate Mr of 67,000 and 25,000. Periodic acid and silver stain of purified oocyst/sporozoite identified bands of similar molecular weight not present in the tachyzoite preparation. Western blot analysis of purified parasites assayed with human antioocyst antisera identified specific oocyst/sporozoite antigens not present on the tachyzoites. At least two major stage-specific oocyst/sporozoite antigens of approximate Mr of 67,000 and 190,000 were identified by the infected patients' antisera and not by the normal controls. Reaction to these oocyst/sporozoite antigens was seen primarily in the IgM fraction of the acute phase and the IgG fraction of convalescent phase antisera. Neither absorption of the antisera with tachyzoites nor periodate treatment of the oocyst/sporozoites reduced the antibody recognition of these stage-specific antigens. These data suggest that individuals infected by a presumed oocyst-transmitted route develop antibodies against unique stage-specific oocyst/sporozoite antigens.

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Selected References

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  1. Benenson M. W., Takafuji E. T., Lemon S. M., Greenup R. L., Sulzer A. J. Oocyst-transmitted toxoplasmosis associated with ingestion of contaminated water. N Engl J Med. 1982 Sep 9;307(11):666–669. doi: 10.1056/NEJM198209093071107. [DOI] [PubMed] [Google Scholar]
  2. Dubray G., Bezard G. A highly sensitive periodic acid-silver stain for 1,2-diol groups of glycoproteins and polysaccharides in polyacrylamide gels. Anal Biochem. 1982 Jan 15;119(2):325–329. doi: 10.1016/0003-2697(82)90593-0. [DOI] [PubMed] [Google Scholar]
  3. Frenkel J. K., Dubey J. P., Miller N. L. Toxoplasma gondii in cats: fecal stages identified as coccidian oocysts. Science. 1970 Feb 6;167(3919):893–896. doi: 10.1126/science.167.3919.893. [DOI] [PubMed] [Google Scholar]
  4. Frenkel J. K., Ruiz A. Endemicity of toxoplasmosis in Costa Rica. Am J Epidemiol. 1981 Mar;113(3):254–269. doi: 10.1093/oxfordjournals.aje.a113095. [DOI] [PubMed] [Google Scholar]
  5. Ganley J. P., Comstock G. W. Association of cats and toxoplasmosis. Am J Epidemiol. 1980 Feb;111(2):238–246. doi: 10.1093/oxfordjournals.aje.a112891. [DOI] [PubMed] [Google Scholar]
  6. Handman E., Goding J. W., Remington J. S. Detection and characterization of membrane antigens of Toxoplasma gondii. J Immunol. 1980 Jun;124(6):2578–2583. [PubMed] [Google Scholar]
  7. Hughes H. P., Van Knapen F., Atkinson H. J., Balfour A. H., Lee D. L. A new soluble antigen preparation of Toxoplasma gondii and its use in serological diagnosis. Clin Exp Immunol. 1982 Jul;49(1):239–246. [PMC free article] [PubMed] [Google Scholar]
  8. Hutchison W. M., Dunachie J. F., Siim J. C., Work K. Coccidian-like nature of Toxoplasma gondii. Br Med J. 1970 Jan 17;1(5689):142–144. doi: 10.1136/bmj.1.5689.142. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Kasper L. H., Bradley M. S., Pfefferkorn E. R. Identification of stage-specific sporozoite antigens of Toxoplasma gondii by monoclonal antibodies. J Immunol. 1984 Jan;132(1):443–449. [PubMed] [Google Scholar]
  10. Kasper L. H., Crabb J. H., Pfefferkorn E. R. Isolation and characterization of a monoclonal antibody-resistant antigenic mutant of Toxoplasma gondii. J Immunol. 1982 Oct;129(4):1694–1699. [PubMed] [Google Scholar]
  11. Kasper L. H., Crabb J. H., Pfefferkorn E. R. Purification of a major membrane protein of Toxoplasma gondii by immunoabsorption with a monoclonal antibody. J Immunol. 1983 May;130(5):2407–2412. [PubMed] [Google Scholar]
  12. Lunde M. N., Jacobs L. Antigenic differences between endozoites and cystozoites of Toxoplasma gondii. J Parasitol. 1983 Oct;69(5):806–808. [PubMed] [Google Scholar]
  13. Naot Y., Guptill D. R., Mullenax J., Remington J. S. Characterization of Toxoplasma gondii antigens that react with human immunoglobulin M and immunoglobulin G antibodies. Infect Immun. 1983 Jul;41(1):331–338. doi: 10.1128/iai.41.1.331-338.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Nema H. V., Bal A., Nath K., Mathur J. S., Shukla B. R. Sulphisoxazole in eye diseases. J Indian Med Assoc. 1965 Aug 1;45(3):142–144. [PubMed] [Google Scholar]
  15. Ogata K., Kasahara T., Shioiri-Nakano K., Igarashi I., Suzuki M. Immunoenzymatic detection of three kinds of 43,000-molecular-weight antigens by monoclonal antibodies in the insoluble fraction of Toxoplasma gondii. Infect Immun. 1984 Mar;43(3):1047–1053. doi: 10.1128/iai.43.3.1047-1053.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Overdulve J. P. The identity of Toxoplasma Nicolle and Manceaux, 1909 with Isospora Schneider, 1881. I. Proc K Ned Akad Wet C. 1970;73(1):129–141. [PubMed] [Google Scholar]
  17. Pfefferkorn E. R., Kasper L. H. Toxoplasma gondii: genetic crosses reveal phenotypic suppression of hydroxyurea resistance by fluorodeoxyuridine resistance. Exp Parasitol. 1983 Apr;55(2):207–218. doi: 10.1016/0014-4894(83)90015-2. [DOI] [PubMed] [Google Scholar]
  18. Pfefferkorn E. R., Pfefferkorn L. C. Toxoplasma gondii: isolation and preliminary characterization of temperature-sensitive mutants. Exp Parasitol. 1976 Jun;39(3):365–376. doi: 10.1016/0014-4894(76)90040-0. [DOI] [PubMed] [Google Scholar]
  19. Sharma S. D., Mullenax J., Araujo F. G., Erlich H. A., Remington J. S. Western Blot analysis of the antigens of Toxoplasma gondii recognized by human IgM and IgG antibodies. J Immunol. 1983 Aug;131(2):977–983. [PubMed] [Google Scholar]
  20. Sheffield H. G., Melton M. L. Toxoplasma gondii: the oocyst, sporozoite, and infection of cultured cells. Science. 1970 Feb 6;167(3919):892–893. doi: 10.1126/science.167.3919.892. [DOI] [PubMed] [Google Scholar]
  21. Teutsch S. M., Juranek D. D., Sulzer A., Dubey J. P., Sikes R. K. Epidemic toxoplasmosis associated with infected cats. N Engl J Med. 1979 Mar 29;300(13):695–699. doi: 10.1056/NEJM197903293001302. [DOI] [PubMed] [Google Scholar]
  22. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Wray W., Boulikas T., Wray V. P., Hancock R. Silver staining of proteins in polyacrylamide gels. Anal Biochem. 1981 Nov 15;118(1):197–203. doi: 10.1016/0003-2697(81)90179-2. [DOI] [PubMed] [Google Scholar]

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