Skip to main content
NCBI home page
Infection and Immunity logoLink to Infection and Immunity
. 1992 Dec;60(12):5132–5138. doi: 10.1128/iai.60.12.5132-5138.1992

Characterization of a > 900,000-M(r) Cryptosporidium parvum sporozoite glycoprotein recognized by protective hyperimmune bovine colostral immunoglobulin.

C Petersen 1, J Gut 1, P S Doyle 1, J H Crabb 1, R G Nelson 1, J H Leech 1
PMCID: PMC258288  PMID: 1452347

Abstract

Cryptosporidium parvum, a zoonotic Apicomplexan pathogen, causes profound diarrhea, malnutrition, and dehydration in patients with AIDS. A less severe, self-limited disease occurs in immunocompetent individuals, particularly children, animal handlers, and residents of the developing world. Very little is known about the biology of the organism, the pathophysiology of the disease process, or the mechanism of protective immunity. There is no effective therapy for cryptosporidiosis, but hyperimmune bovine colostrum raised against Cryptosporidium oocysts and sporozoites has ameliorated infection and disease in some patients with AIDS, and a variety of monoclonal antibodies, as well as hyperimmune bovine colostrum, have significantly reduced cryptosporidial infection of mice and calves. We report here the identification and initial characterization of a > 900,000-M(r) Cryptosporodium sporozoite glycoprotein (GP900) that is a prominent antigen recognized by protective hyperimmune bovine colostral immunoglobulin. Three of six murine anticryptosporidial monoclonal antibodies reacted with GP900, indicating that the molecule is highly immunogenic in mice as well as in cows. GP900 is Triton X-100 soluble and N glycosylated. Western blotting of the N-deglycosylated protein, detected with antibodies eluted from recombinant clones expressing a partial GP900 fusion protein, suggested that the polypeptide backbone of the glycoprotein has an M(r) of < 190,000. GP900 is encoded by a single-copy gene that resides on the largest Cryptosporidium chromosome.

Full text

PDF
5132

Images in this article

5134Image
on p.5134
5134Image
on p.5134
5135Image
on p.5135
5136Image
on p.5136
5136Image
on p.5136
5137Image
on p.5137

Selected References

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

  1. Arrowood M. J., Mead J. R., Mahrt J. L., Sterling C. R. Effects of immune colostrum and orally administered antisporozoite monoclonal antibodies on the outcome of Cryptosporidium parvum infections in neonatal mice. Infect Immun. 1989 Aug;57(8):2283–2288. doi: 10.1128/iai.57.8.2283-2288.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Beall J. A., Mitchell G. F. Identification of a particular antigen from a parasite cDNA library using antibodies affinity purified from selected portions of Western blots. J Immunol Methods. 1986 Feb 12;86(2):217–223. doi: 10.1016/0022-1759(86)90456-4. [DOI] [PubMed] [Google Scholar]
  3. Bjorneby J. M., Hunsaker B. D., Riggs M. W., Perryman L. E. Monoclonal antibody immunotherapy in nude mice persistently infected with Cryptosporidium parvum. Infect Immun. 1991 Mar;59(3):1172–1176. doi: 10.1128/iai.59.3.1172-1176.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bjorneby J. M., Riggs M. W., Perryman L. E. Cryptosporidium parvum merozoites share neutralization-sensitive epitopes with sporozoites. J Immunol. 1990 Jul 1;145(1):298–304. [PubMed] [Google Scholar]
  5. Current W. L., Reese N. C., Ernst J. V., Bailey W. S., Heyman M. B., Weinstein W. M. Human cryptosporidiosis in immunocompetent and immunodeficient persons. Studies of an outbreak and experimental transmission. N Engl J Med. 1983 May 26;308(21):1252–1257. doi: 10.1056/NEJM198305263082102. [DOI] [PubMed] [Google Scholar]
  6. Danforth H. D., Campbell G. H., Leef M. F., Beaudoin R. L. Production of monoclonal antibodies by hybridomas sensitized to sporozoites of Plasmodium berghei. J Parasitol. 1982 Dec;68(6):1029–1033. [PubMed] [Google Scholar]
  7. Fayer R., Andrews C., Ungar B. L., Blagburn B. Efficacy of hyperimmune bovine colostrum for prophylaxis of cryptosporidiosis in neonatal calves. J Parasitol. 1989 Jun;75(3):393–397. [PubMed] [Google Scholar]
  8. Fayer R., Guidry A., Blagburn B. L. Immunotherapeutic efficacy of bovine colostral immunoglobulins from a hyperimmunized cow against cryptosporidiosis in neonatal mice. Infect Immun. 1990 Sep;58(9):2962–2965. doi: 10.1128/iai.58.9.2962-2965.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Fayer R., Perryman L. E., Riggs M. W. Hyperimmune bovine colostrum neutralizes Cryptosporidium sporozoites and protects mice against oocyst challenge. J Parasitol. 1989 Feb;75(1):151–153. [PubMed] [Google Scholar]
  10. Howard R. J., Lyon J. A., Uni S., Saul A. J., Aley S. B., Klotz F., Panton L. J., Sherwood J. A., Marsh K., Aikawa M. Transport of an Mr approximately 300,000 Plasmodium falciparum protein (Pf EMP 2) from the intraerythrocytic asexual parasite to the cytoplasmic face of the host cell membrane. J Cell Biol. 1987 May;104(5):1269–1280. doi: 10.1083/jcb.104.5.1269. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Kearney J. F., Radbruch A., Liesegang B., Rajewsky K. A new mouse myeloma cell line that has lost immunoglobulin expression but permits the construction of antibody-secreting hybrid cell lines. J Immunol. 1979 Oct;123(4):1548–1550. [PubMed] [Google Scholar]
  12. Kim K., Goozé L., Petersen C., Gut J., Nelson R. G. Isolation, sequence and molecular karyotype analysis of the actin gene of Cryptosporidium parvum. Mol Biochem Parasitol. 1992 Jan;50(1):105–113. doi: 10.1016/0166-6851(92)90248-i. [DOI] [PubMed] [Google Scholar]
  13. Knott T. J., Pease R. J., Powell L. M., Wallis S. C., Rall S. C., Jr, Innerarity T. L., Blackhart B., Taylor W. H., Marcel Y., Milne R. Complete protein sequence and identification of structural domains of human apolipoprotein B. Nature. 1986 Oct 23;323(6090):734–738. doi: 10.1038/323734a0. [DOI] [PubMed] [Google Scholar]
  14. Kurzban G. P., Wang K. Giant polypeptides of skeletal muscle titin: sedimentation equilibrium in guanidine hydrochloride. Biochem Biophys Res Commun. 1988 Feb 15;150(3):1155–1161. doi: 10.1016/0006-291x(88)90750-4. [DOI] [PubMed] [Google Scholar]
  15. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  16. Lasser K. H., Lewin K. J., Ryning F. W. Cryptosporidial enteritis in a patient with congenital hypogammaglobulinemia. Hum Pathol. 1979 Mar;10(2):234–240. doi: 10.1016/s0046-8177(79)80012-x. [DOI] [PubMed] [Google Scholar]
  17. Nord J., Ma P., DiJohn D., Tzipori S., Tacket C. O. Treatment with bovine hyperimmune colostrum of cryptosporidial diarrhea in AIDS patients. AIDS. 1990 Jun;4(6):581–584. doi: 10.1097/00002030-199006000-00015. [DOI] [PubMed] [Google Scholar]
  18. Perryman L. E., Riggs M. W., Mason P. H., Fayer R. Kinetics of Cryptosporidium parvum sporozoite neutralization by monoclonal antibodies, immune bovine serum, and immune bovine colostrum. Infect Immun. 1990 Jan;58(1):257–259. doi: 10.1128/iai.58.1.257-259.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Petersen C., Gut J., Leech J. H., Nelson R. G. Identification and initial characterization of five Cryptosporidium parvum sporozoite antigen genes. Infect Immun. 1992 Jun;60(6):2343–2348. doi: 10.1128/iai.60.6.2343-2348.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Petersen C., Nelson R., Leech J., Jensen J., Wollish W., Scherf A. The gene product of the Plasmodium falciparum 11.1 locus is a protein larger than one megadalton. Mol Biochem Parasitol. 1990 Sep-Oct;42(2):189–195. doi: 10.1016/0166-6851(90)90161-e. [DOI] [PubMed] [Google Scholar]
  21. Petersen C., Nelson R., Magowan C., Wollish W., Jensen J., Leech J. The mature erythrocyte surface antigen of Plasmodium falciparum is not required for knobs or cytoadherence. Mol Biochem Parasitol. 1989 Aug;36(1):61–65. doi: 10.1016/0166-6851(89)90200-4. [DOI] [PubMed] [Google Scholar]
  22. Pitlik S. D., Fainstein V., Garza D., Guarda L., Bolivar R., Rios A., Hopfer R. L., Mansell P. A. Human cryptosporidiosis: spectrum of disease. Report of six cases and review of the literature. Arch Intern Med. 1983 Dec;143(12):2269–2275. doi: 10.1001/archinte.1983.00350120059015. [DOI] [PubMed] [Google Scholar]
  23. Riggs M. W., McGuire T. C., Mason P. H., Perryman L. E. Neutralization-sensitive epitopes are exposed on the surface of infectious Cryptosporidium parvum sporozoites. J Immunol. 1989 Aug 15;143(4):1340–1345. [PubMed] [Google Scholar]
  24. Riggs M. W., Perryman L. E. Infectivity and neutralization of Cryptosporidium parvum sporozoites. Infect Immun. 1987 Sep;55(9):2081–2087. doi: 10.1128/iai.55.9.2081-2087.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Soave R., Danner R. L., Honig C. L., Ma P., Hart C. C., Nash T., Roberts R. B. Cryptosporidiosis in homosexual men. Ann Intern Med. 1984 Apr;100(4):504–511. doi: 10.7326/0003-4819-100-4-504. [DOI] [PubMed] [Google Scholar]
  26. Stedman H., Browning K., Oliver N., Oronzi-Scott M., Fischbeck K., Sarkar S., Sylvester J., Schmickel R., Wang K. Nebulin cDNAs detect a 25-kilobase transcript in skeletal muscle and localize to human chromosome 2. Genomics. 1988 Jan;2(1):1–7. doi: 10.1016/0888-7543(88)90102-4. [DOI] [PubMed] [Google Scholar]
  27. Tilley M., Fayer R., Guidry A., Upton S. J., Blagburn B. L. Cryptosporidium parvum (Apicomplexa: Cryptosporidiidae) oocyst and sporozoite antigens recognized by bovine colostral antibodies. Infect Immun. 1990 Sep;58(9):2966–2971. doi: 10.1128/iai.58.9.2966-2971.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Tilley M., Upton S. J., Fayer R., Barta J. R., Chrisp C. E., Freed P. S., Blagburn B. L., Anderson B. C., Barnard S. M. Identification of a 15-kilodalton surface glycoprotein on sporozoites of Cryptosporidium parvum. Infect Immun. 1991 Mar;59(3):1002–1007. doi: 10.1128/iai.59.3.1002-1007.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. 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]
  30. Tzipori S., Roberton D., Chapman C. Remission of diarrhoea due to cryptosporidiosis in an immunodeficient child treated with hyperimmune bovine colostrum. Br Med J (Clin Res Ed) 1986 Nov 15;293(6557):1276–1277. doi: 10.1136/bmj.293.6557.1276. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Tzipori S., Roberton D., Cooper D. A., White L. Chronic cryptosporidial diarrhoea and hyperimmune cow colostrum. Lancet. 1987 Aug 8;2(8554):344–345. doi: 10.1016/s0140-6736(87)90944-5. [DOI] [PubMed] [Google Scholar]
  32. Ungar B. L., Ward D. J., Fayer R., Quinn C. A. Cessation of Cryptosporidium-associated diarrhea in an acquired immunodeficiency syndrome patient after treatment with hyperimmune bovine colostrum. Gastroenterology. 1990 Feb;98(2):486–489. doi: 10.1016/0016-5085(90)90842-o. [DOI] [PubMed] [Google Scholar]
  33. Whitmire W. M., Harp J. A. Characterization of bovine cellular and serum antibody responses during infection by Cryptosporidium parvum. Infect Immun. 1991 Mar;59(3):990–995. doi: 10.1128/iai.59.3.990-995.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Wolfson J. S., Richter J. M., Waldron M. A., Weber D. J., McCarthy D. M., Hopkins C. C. Cryptosporidiosis in immunocompetent patients. N Engl J Med. 1985 May 16;312(20):1278–1282. doi: 10.1056/NEJM198505163122002. [DOI] [PubMed] [Google Scholar]

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

RESOURCES