Skip to main content
PMC home page
Infection and Immunity logoLink to Infection and Immunity
. 1997 Nov;65(11):4761–4769. doi: 10.1128/iai.65.11.4761-4769.1997

Profiles of healing and nonhealing Cryptosporidium parvum infection in C57BL/6 mice with functional B and T lymphocytes: the extent of gamma interferon modulation determines the outcome of infection.

C M Theodos 1, K L Sullivan 1, J K Griffiths 1, S Tzipori 1
PMCID: PMC175683  PMID: 9353062

Abstract

This study describes healing and nonhealing models of Cryptosporidium parvum infection with adult mice that have functional T and B lymphocytes. In our nonhealing model, mice on a C57BL/6 background which have a targeted disruption in the gamma interferon (IFN-gamma) gene (GKO mice) are utilized. C. parvum-infected GKO mice shed extremely high levels of oocysts and displayed overwhelming infection of the entire small intestine. The majority of these mice succumbed within 2 to 3 weeks due to severe acute infection and profound mucosal destruction. In our healing murine model, C57BL/6J mice treated with a single injection of the neutralizing anti-IFN-gamma monoclonal antibody XMG 1.2 prior to infection were used. These mice developed two peaks of oocyst shedding but were ultimately free of parasites on day 30 of infection. Again, the small intestine was the primary site of infection. Mesenteric lymph node (MLN) cells isolated from C. parvum-infected nonhealing GKO mice proliferated and secreted interleukin 2 (IL-2) but not IFN-gamma or IL-4 in response to ex vivo restimulation with intact C. parvum sporozoites or a C. parvum sporozoite antigen preparation. In contrast, parasite-specific MLN cells isolated from healing C57BL/6J mice secreted IL-2 and IFN-gamma but not IL-4. These results suggest that IFN-gamma, either directly or indirectly, is important for resistance to and resolution of cryptosporidiosis. Moreover, these models now allow the analysis of parasite-specific cell-mediated and humoral mucosal immune responses to determine what constitutes protective immunity to C. parvum.

Full Text

The Full Text of this article is available as a PDF (1.3 MB).

Selected References

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

  1. Aguirre S. A., Mason P. H., Perryman L. E. Susceptibility of major histocompatibility complex (MHC) class I- and MHC class II-deficient mice to Cryptosporidium parvum infection. Infect Immun. 1994 Feb;62(2):697–699. doi: 10.1128/iai.62.2.697-699.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Amiri P., Haak-Frendscho M., Robbins K., McKerrow J. H., Stewart T., Jardieu P. Anti-immunoglobulin E treatment decreases worm burden and egg production in Schistosoma mansoni-infected normal and interferon gamma knockout mice. J Exp Med. 1994 Jul 1;180(1):43–51. doi: 10.1084/jem.180.1.43. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. 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]
  4. 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]
  5. Black R. E., Brown K. H., Becker S. Effects of diarrhea associated with specific enteropathogens on the growth of children in rural Bangladesh. Pediatrics. 1984 Jun;73(6):799–805. [PubMed] [Google Scholar]
  6. Bouley D. M., Kanangat S., Wire W., Rouse B. T. Characterization of herpes simplex virus type-1 infection and herpetic stromal keratitis development in IFN-gamma knockout mice. J Immunol. 1995 Oct 15;155(8):3964–3971. [PubMed] [Google Scholar]
  7. Campbell P. N., Current W. L. Demonstration of serum antibodies to Cryptosporidium sp. in normal and immunodeficient humans with confirmed infections. J Clin Microbiol. 1983 Jul;18(1):165–169. doi: 10.1128/jcm.18.1.165-169.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Chakkalath H. R., Theodos C. M., Markowitz J. S., Grusby M. J., Glimcher L. H., Titus R. G. Class II major histocompatibility complex-deficient mice initially control an infection with Leishmania major but succumb to the disease. J Infect Dis. 1995 May;171(5):1302–1308. doi: 10.1093/infdis/171.5.1302. [DOI] [PubMed] [Google Scholar]
  9. Chen W., Harp J. A., Harmsen A. G., Havell E. A. Gamma interferon functions in resistance to Cryptosporidium parvum infection in severe combined immunodeficient mice. Infect Immun. 1993 Aug;61(8):3548–3551. doi: 10.1128/iai.61.8.3548-3551.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Chen W., Harp J. A., Harmsen A. G. Requirements for CD4+ cells and gamma interferon in resolution of established Cryptosporidium parvum infection in mice. Infect Immun. 1993 Sep;61(9):3928–3932. doi: 10.1128/iai.61.9.3928-3932.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Cheng L., Rasmussen K. R., Healey M. C., Yang S. Primary and secondary infections with Cryptosporidium parvum in immunosuppressed adult mice. Am J Trop Med Hyg. 1996 Sep;55(3):324–329. doi: 10.4269/ajtmh.1996.55.324. [DOI] [PubMed] [Google Scholar]
  12. Cooper A. M., Dalton D. K., Stewart T. A., Griffin J. P., Russell D. G., Orme I. M. Disseminated tuberculosis in interferon gamma gene-disrupted mice. J Exp Med. 1993 Dec 1;178(6):2243–2247. doi: 10.1084/jem.178.6.2243. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Current W. L., Garcia L. S. Cryptosporidiosis. Clin Microbiol Rev. 1991 Jul;4(3):325–358. doi: 10.1128/cmr.4.3.325. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Dalton D. K., Pitts-Meek S., Keshav S., Figari I. S., Bradley A., Stewart T. A. Multiple defects of immune cell function in mice with disrupted interferon-gamma genes. Science. 1993 Mar 19;259(5102):1739–1742. doi: 10.1126/science.8456300. [DOI] [PubMed] [Google Scholar]
  15. Doyle P. S., Crabb J., Petersen C. Anti-Cryptosporidium parvum antibodies inhibit infectivity in vitro and in vivo. Infect Immun. 1993 Oct;61(10):4079–4084. doi: 10.1128/iai.61.10.4079-4084.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Enriquez F. J., Sterling C. R. Cryptosporidium infections in inbred strains of mice. J Protozool. 1991 Nov-Dec;38(6):100S–102S. [PubMed] [Google Scholar]
  17. Ernest J. A., Blagburn B. L., Lindsay D. S., Current W. L. Infection dynamics of Cryptosporidium parvum (Apicomplexa: Cryptosporiidae) in neonatal mice (Mus musculus). J Parasitol. 1986 Oct;72(5):796–798. [PubMed] [Google Scholar]
  18. Flynn J. L., Chan J., Triebold K. J., Dalton D. K., Stewart T. A., Bloom B. R. An essential role for interferon gamma in resistance to Mycobacterium tuberculosis infection. J Exp Med. 1993 Dec 1;178(6):2249–2254. doi: 10.1084/jem.178.6.2249. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Flynn J. L., Goldstein M. M., Triebold K. J., Sypek J., Wolf S., Bloom B. R. IL-12 increases resistance of BALB/c mice to Mycobacterium tuberculosis infection. J Immunol. 1995 Sep 1;155(5):2515–2524. [PubMed] [Google Scholar]
  20. Graham M. B., Dalton D. K., Giltinan D., Braciale V. L., Stewart T. A., Braciale T. J. Response to influenza infection in mice with a targeted disruption in the interferon gamma gene. J Exp Med. 1993 Nov 1;178(5):1725–1732. doi: 10.1084/jem.178.5.1725. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Harp J. A., Sacco R. E. Development of cellular immune functions in neonatal to weanling mice: relationship to Cryptosporidium parvum infection. J Parasitol. 1996 Apr;82(2):245–249. [PubMed] [Google Scholar]
  22. Harp J. A., Whitmire W. M., Sacco R. In vitro proliferation and production of gamma interferon by murine CD4+ cells in response to Cryptosporidium parvum antigen. J Parasitol. 1994 Feb;80(1):67–72. [PubMed] [Google Scholar]
  23. Harty J. T., Bevan M. J. Specific immunity to Listeria monocytogenes in the absence of IFN gamma. Immunity. 1995 Jul;3(1):109–117. doi: 10.1016/1074-7613(95)90163-9. [DOI] [PubMed] [Google Scholar]
  24. Heine J., Moon H. W., Woodmansee D. B. Persistent Cryptosporidium infection in congenitally athymic (nude) mice. Infect Immun. 1984 Mar;43(3):856–859. doi: 10.1128/iai.43.3.856-859.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Kassa M., Comby E., Lemeteil D., Brasseur P., Ballet J. J. Characterization of anti-Cryptosporidium IgA antibodies in sera from immunocompetent individuals and HIV-infected patients. J Protozool. 1991 Nov-Dec;38(6):179S–180S. [PubMed] [Google Scholar]
  26. Koster F. T., Curlin G. C., Aziz K. M., Haque A. Synergistic impact of measles and diarrhoea on nutrition and mortality in Bangladesh. Bull World Health Organ. 1981;59(6):901–908. [PMC free article] [PubMed] [Google Scholar]
  27. Kuhls T. L., Mosier D. A., Abrams V. L., Crawford D. L., Greenfield R. A. Inability of interferon-gamma and aminoguanidine to alter Cryptosporidium parvum infection in mice with severe combined immunodeficiency. J Parasitol. 1994 Jun;80(3):480–485. [PubMed] [Google Scholar]
  28. McDonald V., Bancroft G. J. Mechanisms of innate and acquired resistance to Cryptosporidium parvum infection in SCID mice. Parasite Immunol. 1994 Jun;16(6):315–320. doi: 10.1111/j.1365-3024.1994.tb00354.x. [DOI] [PubMed] [Google Scholar]
  29. McDonald V., Deer R., Uni S., Iseki M., Bancroft G. J. Immune responses to Cryptosporidium muris and Cryptosporidium parvum in adult immunocompetent or immunocompromised (nude and SCID) mice. Infect Immun. 1992 Aug;60(8):3325–3331. doi: 10.1128/iai.60.8.3325-3331.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Mead J. R., Arrowood M. J., Healey M. C., Sidwell R. W. Cryptosporidial infections in SCID mice reconstituted with human or murine lymphocytes. J Protozool. 1991 Nov-Dec;38(6):59S–61S. [PubMed] [Google Scholar]
  31. Mead J. R., Arrowood M. J., Sidwell R. W., Healey M. C. Chronic Cryptosporidium parvum infections in congenitally immunodeficient SCID and nude mice. J Infect Dis. 1991 Jun;163(6):1297–1304. doi: 10.1093/infdis/163.6.1297. [DOI] [PubMed] [Google Scholar]
  32. Mead J. R., Arrowood M. J., Sterling C. R. Antigens of Cryptosporidium sporozoites recognized by immune sera of infected animals and humans. J Parasitol. 1988 Feb;74(1):135–143. [PubMed] [Google Scholar]
  33. Mead J. R., Ilksoy N., You X., Belenkaya Y., Arrowood M. J., Fallon M. T., Schinazi R. F. Infection dynamics and clinical features of cryptosporidiosis in SCID mice. Infect Immun. 1994 May;62(5):1691–1695. doi: 10.1128/iai.62.5.1691-1695.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Moss D. M., Lammie P. J. Proliferative responsiveness of lymphocytes from Cryptosporidium parvum-exposed mice to two separate antigen fractions from oocysts. Am J Trop Med Hyg. 1993 Sep;49(3):393–401. doi: 10.4269/ajtmh.1993.49.393. [DOI] [PubMed] [Google Scholar]
  35. 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]
  36. Ortega-Mora L. M., Troncoso J. M., Rojo-Vazquez F. A., Gomez-Bautista M. Identification of Cryptosporidium parvum oocyst/sporozoite antigens recognized by infected and hyperimmune lambs. Vet Parasitol. 1994 May;53(1-2):159–166. doi: 10.1016/0304-4017(94)90028-0. [DOI] [PubMed] [Google Scholar]
  37. Ortega-Mora L. M., Troncoso J. M., Rojo-Vázquez F. A., Gómez-Bautista M. Serum antibody response in lambs naturally and experimentally infected with Cryptosporidium parvum. Vet Parasitol. 1993 Oct;50(1-2):45–54. doi: 10.1016/0304-4017(93)90006-9. [DOI] [PubMed] [Google Scholar]
  38. Peeters J. E., Villacorta I., Vanopdenbosch E., Vandergheynst D., Naciri M., Ares-Mazás E., Yvoré P. Cryptosporidium parvum in calves: kinetics and immunoblot analysis of specific serum and local antibody responses (immunoglobulin A [IgA], IgG, and IgM) after natural and experimental infections. Infect Immun. 1992 Jun;60(6):2309–2316. doi: 10.1128/iai.60.6.2309-2316.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Perryman L. E., Bjorneby J. M. Immunotherapy of cryptosporidiosis in immunodeficient animal models. J Protozool. 1991 Nov-Dec;38(6):98S–100S. [PubMed] [Google Scholar]
  40. Perryman L. E., Mason P. H., Chrisp C. E. Effect of spleen cell populations on resolution of Cryptosporidium parvum infection in SCID mice. Infect Immun. 1994 Apr;62(4):1474–1477. doi: 10.1128/iai.62.4.1474-1477.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Petersen C. Cryptosporidiosis in patients infected with the human immunodeficiency virus. Clin Infect Dis. 1992 Dec;15(6):903–909. doi: 10.1093/clind/15.6.903. [DOI] [PubMed] [Google Scholar]
  42. Reperant J. M., Naciri M., Chardes T., Bout D. T. Immunological characterization of a 17-kDa antigen from Cryptosporidium parvum recognized early by mucosal IgA antibodies. FEMS Microbiol Lett. 1992 Nov 15;78(1):7–14. doi: 10.1016/0378-1097(92)90280-2. [DOI] [PubMed] [Google Scholar]
  43. Riggs M. W., Cama V. A., Leary H. L., Jr, Sterling C. R. Bovine antibody against Cryptosporidium parvum elicits a circumsporozoite precipitate-like reaction and has immunotherapeutic effect against persistent cryptosporidiosis in SCID mice. Infect Immun. 1994 May;62(5):1927–1939. doi: 10.1128/iai.62.5.1927-1939.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. 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]
  45. Répérant J. M., Naciri M., Iochmann S., Tilley M., Bout D. T. Major antigens of Cryptosporidium parvum recognised by serum antibodies from different infected animal species and man. Vet Parasitol. 1994 Oct;55(1-2):1–13. doi: 10.1016/0304-4017(94)90051-5. [DOI] [PubMed] [Google Scholar]
  46. Scott P. IFN-gamma modulates the early development of Th1 and Th2 responses in a murine model of cutaneous leishmaniasis. J Immunol. 1991 Nov 1;147(9):3149–3155. [PubMed] [Google Scholar]
  47. Sherwood D., Angus K. W., Snodgrass D. R., Tzipori S. Experimental cryptosporidiosis in laboratory mice. Infect Immun. 1982 Nov;38(2):471–475. doi: 10.1128/iai.38.2.471-475.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Taghi-Kilani R., Sekla L., Hayglass K. T. The role of humoral immunity in Cryptosporidium spp. infection. Studies with B cell-depleted mice. J Immunol. 1990 Sep 1;145(5):1571–1576. [PubMed] [Google Scholar]
  49. Tatalick L. M., Perryman L. E. Attempts to protect severe combined immunodeficient (scid) mice with antibody enriched for reactivity to Cryptosporidium parvum surface antigen-1. Vet Parasitol. 1995 Jul;58(4):281–290. doi: 10.1016/0304-4017(94)00729-v. [DOI] [PubMed] [Google Scholar]
  50. Tatalick L. M., Perryman L. E. Effect of surface antigen-1 (SA-1) immune lymphocyte subsets and naive cell subsets in protecting scid mice from initial and persistent infection with Cryptosporidium parvum. Vet Immunol Immunopathol. 1995 Jul;47(1-2):43–55. doi: 10.1016/0165-2427(94)05391-5. [DOI] [PubMed] [Google Scholar]
  51. Theodos C. M., Titus R. G. Salivary gland material from the sand fly Lutzomyia longipalpis has an inhibitory effect on macrophage function in vitro. Parasite Immunol. 1993 Aug;15(8):481–487. doi: 10.1111/j.1365-3024.1993.tb00634.x. [DOI] [PubMed] [Google Scholar]
  52. Titus R. G., Lima G. C., Engers H. D., Louis J. A. Exacerbation of murine cutaneous leishmaniasis by adoptive transfer of parasite-specific helper T cell populations capable of mediating Leishmania major-specific delayed-type hypersensitivity. J Immunol. 1984 Sep;133(3):1594–1600. [PubMed] [Google Scholar]
  53. Tzipori S. Cryptosporidiosis in perspective. Adv Parasitol. 1988;27:63–129. doi: 10.1016/S0065-308X(08)60353-X. [DOI] [PMC free article] [PubMed] [Google Scholar]
  54. Tzipori S., Rand W., Griffiths J., Widmer G., Crabb J. Evaluation of an animal model system for cryptosporidiosis: therapeutic efficacy of paromomycin and hyperimmune bovine colostrum-immunoglobulin. Clin Diagn Lab Immunol. 1994 Jul;1(4):450–463. doi: 10.1128/cdli.1.4.450-463.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  55. Tzipori S., Rand W., Theodos C. Evaluation of a two-phase scid mouse model preconditioned with anti-interferon-gamma monoclonal antibody for drug testing against Cryptosporidium parvum. J Infect Dis. 1995 Oct;172(4):1160–1164. doi: 10.1093/infdis/172.4.1160. [DOI] [PubMed] [Google Scholar]
  56. 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]
  57. 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]
  58. Ungar B. L., Burris J. A., Quinn C. A., Finkelman F. D. New mouse models for chronic Cryptosporidium infection in immunodeficient hosts. Infect Immun. 1990 Apr;58(4):961–969. doi: 10.1128/iai.58.4.961-969.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  59. Ungar B. L., Kao T. C., Burris J. A., Finkelman F. D. Cryptosporidium infection in an adult mouse model. Independent roles for IFN-gamma and CD4+ T lymphocytes in protective immunity. J Immunol. 1991 Aug 1;147(3):1014–1022. [PubMed] [Google Scholar]
  60. 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]
  61. Urban J. F., Jr, Fayer R., Chen S. J., Gause W. C., Gately M. K., Finkelman F. D. IL-12 protects immunocompetent and immunodeficient neonatal mice against infection with Cryptosporidium parvum. J Immunol. 1996 Jan 1;156(1):263–268. [PubMed] [Google Scholar]
  62. Wang Z. E., Reiner S. L., Zheng S., Dalton D. K., Locksley R. M. CD4+ effector cells default to the Th2 pathway in interferon gamma-deficient mice infected with Leishmania major. J Exp Med. 1994 Apr 1;179(4):1367–1371. doi: 10.1084/jem.179.4.1367. [DOI] [PMC free article] [PubMed] [Google Scholar]
  63. Waters W. R., Harp J. A. Cryptosporidium parvum infection in T-cell receptor (TCR)-alpha- and TCR-delta-deficient mice. Infect Immun. 1996 May;64(5):1854–1857. doi: 10.1128/iai.64.5.1854-1857.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  64. Whitmire W. M., Harp J. A. In vitro murine lymphocyte blastogenic responses to Cryptosporidium parvum. J Parasitol. 1990 Jun;76(3):450–452. [PubMed] [Google Scholar]

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

RESOURCES