Skip to main content
NCBI home page
Clinical and Diagnostic Laboratory Immunology logoLink to Clinical and Diagnostic Laboratory Immunology
. 1997 Sep;4(5):530–535. doi: 10.1128/cdli.4.5.530-535.1997

Antibody response against a Leishmania donovani amastigote-stage-specific protein in patients with visceral leishmaniasis.

E Ghedin 1, W W Zhang 1, H Charest 1, S Sundar 1, R T Kenney 1, G Matlashewski 1
PMCID: PMC170587  PMID: 9302200

Abstract

The antibody response against an amastigote-specific protein (A2) from Leishmania donovani was investigated. Sera from patients with trypanosomiasis and various forms of leishmaniasis were screened for anti-A2 antibodies. Sera from patients infected only with L. donovani or Leishmania mexicana specifically recognized the A2 recombinant protein. These results were consistent with karyotype analyses which revealed that the A2 gene is conserved in L. donovani and L. mexicana strains. The potential of this antigen in diagnosis was further explored by screening a series of sera obtained from patients in regions of the Sudan and India where L. donovani is endemic. The prevalence of anti-A2 antibodies was determined by Western blotting for all samples. Enzyme-linked immunosorbent assay (ELISA) and an immunoprecipitation assay were also performed on some of the samples. Anti-A2 antibodies were detected by ELISA in 82 and 60% of the samples from individuals with active visceral leishmaniasis (kala-azar) from the Sudan and India, respectively, while the immunoprecipitation assay detected the antibodies in 92% of the samples from India. These data suggest that the A2 protein may be a useful diagnostic antigen for visceral leishmaniasis.

Full Text

The Full Text of this article is available as a PDF (3.2 MB).

Selected References

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

  1. Badaró R., Benson D., Eulálio M. C., Freire M., Cunha S., Netto E. M., Pedral-Sampaio D., Madureira C., Burns J. M., Houghton R. L. rK39: a cloned antigen of Leishmania chagasi that predicts active visceral leishmaniasis. J Infect Dis. 1996 Mar;173(3):758–761. doi: 10.1093/infdis/173.3.758. [DOI] [PubMed] [Google Scholar]
  2. Brandau S., Dresel A., Clos J. High constitutive levels of heat-shock proteins in human-pathogenic parasites of the genus Leishmania. Biochem J. 1995 Aug 15;310(Pt 1):225–232. doi: 10.1042/bj3100225. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Burns J. M., Jr, Shreffler W. G., Benson D. R., Ghalib H. W., Badaro R., Reed S. G. Molecular characterization of a kinesin-related antigen of Leishmania chagasi that detects specific antibody in African and American visceral leishmaniasis. Proc Natl Acad Sci U S A. 1993 Jan 15;90(2):775–779. doi: 10.1073/pnas.90.2.775. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Charest H., Matlashewski G. Developmental gene expression in Leishmania donovani: differential cloning and analysis of an amastigote-stage-specific gene. Mol Cell Biol. 1994 May;14(5):2975–2984. doi: 10.1128/mcb.14.5.2975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Charest H., Zhang W. W., Matlashewski G. The developmental expression of Leishmania donovani A2 amastigote-specific genes is post-transcriptionally mediated and involves elements located in the 3'-untranslated region. J Biol Chem. 1996 Jul 19;271(29):17081–17090. doi: 10.1074/jbc.271.29.17081. [DOI] [PubMed] [Google Scholar]
  6. Grondin K., Papadopoulou B., Ouellette M. Homologous recombination between direct repeat sequences yields P-glycoprotein containing amplicons in arsenite resistant Leishmania. Nucleic Acids Res. 1993 Apr 25;21(8):1895–1901. doi: 10.1093/nar/21.8.1895. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Harith A. E., Kolk A. H., Kager P. A., Leeuwenburg J., Muigai R., Kiugu S., Kiugu S., Laarman J. J. A simple and economical direct agglutination test for serodiagnosis and sero-epidemiological studies of visceral leishmaniasis. Trans R Soc Trop Med Hyg. 1986;80(4):583–536. doi: 10.1016/0035-9203(86)90149-5. [DOI] [PubMed] [Google Scholar]
  8. Jaffe C. L., Bennett E., Grimaldi G., Jr, McMahon-Pratt D. Production and characterization of species-specific monoclonal antibodies against Leishmania donovani for immunodiagnosis. J Immunol. 1984 Jul;133(1):440–447. [PubMed] [Google Scholar]
  9. Jaffe C. L., McMahon-Pratt D. Serodiagnostic assay for visceral leishmaniasis employing monoclonal antibodies. Trans R Soc Trop Med Hyg. 1987;81(4):587–594. doi: 10.1016/0035-9203(87)90418-4. [DOI] [PubMed] [Google Scholar]
  10. Jaffe C. L., Zalis M. Purification of two Leishmania donovani membrane proteins recognized by sera from patients with visceral leishmaniasis. Mol Biochem Parasitol. 1988 Jan 1;27(1):53–62. doi: 10.1016/0166-6851(88)90024-2. [DOI] [PubMed] [Google Scholar]
  11. Jaffe C. L., Zalis M. Use of purified parasite proteins from Leishmania donovani for the rapid serodiagnosis of visceral leishmaniasis. J Infect Dis. 1988 Jun;157(6):1212–1220. doi: 10.1093/infdis/157.6.1212. [DOI] [PubMed] [Google Scholar]
  12. Kar K. Serodiagnosis of leishmaniasis. Crit Rev Microbiol. 1995;21(2):123–152. doi: 10.3109/10408419509113537. [DOI] [PubMed] [Google Scholar]
  13. Magill A. J., Grögl M., Gasser R. A., Jr, Sun W., Oster C. N. Visceral infection caused by Leishmania tropica in veterans of Operation Desert Storm. N Engl J Med. 1993 May 13;328(19):1383–1387. doi: 10.1056/NEJM199305133281904. [DOI] [PubMed] [Google Scholar]
  14. Okong'o-Odera E. A., Kurtzhals J. A., Hey A. S., Kharazmi A. Measurement of serum antibodies against native Leishmania gp63 distinguishes between ongoing and previous L. donovani infection. APMIS. 1993 Aug;101(8):642–646. doi: 10.1111/j.1699-0463.1993.tb00158.x. [DOI] [PubMed] [Google Scholar]
  15. Pan A. A., McMahon-Pratt D. Monoclonal antibodies specific for the amastigote stage of Leishmania pifanoi. I. Characterization of antigens associated with stage- and species-specific determinants. J Immunol. 1988 Apr 1;140(7):2406–2414. [PubMed] [Google Scholar]
  16. Sacks D. L., Kenney R. T., Kreutzer R. D., Jaffe C. L., Gupta A. K., Sharma M. C., Sinha S. P., Neva F. A., Saran R. Indian kala-azar caused by Leishmania tropica. Lancet. 1995 Apr 15;345(8955):959–961. doi: 10.1016/s0140-6736(95)90703-3. [DOI] [PubMed] [Google Scholar]
  17. Shreffler W. G., Burns J. M., Jr, Badaró R., Ghalib H. W., Button L. L., McMaster W. R., Reed S. G. Antibody responses of visceral leishmaniasis patients to gp63, a major surface glycoprotein of Leishmania species. J Infect Dis. 1993 Feb;167(2):426–430. doi: 10.1093/infdis/167.2.426. [DOI] [PubMed] [Google Scholar]
  18. Sinha R., Sehgal S. Comparative evaluation of serological tests in Indian kala-azar. J Trop Med Hyg. 1994 Dec;97(6):333–340. [PubMed] [Google Scholar]
  19. Smith D. B., Johnson K. S. Single-step purification of polypeptides expressed in Escherichia coli as fusions with glutathione S-transferase. Gene. 1988 Jul 15;67(1):31–40. doi: 10.1016/0378-1119(88)90005-4. [DOI] [PubMed] [Google Scholar]
  20. Tebourski F., el Gaied A., Louzir H., Ben Ismail R., Kammoun R., Dellagi K. Identification of an immunodominant 32-kilodalton membrane protein of Leishmania donovani infantum promastigotes suitable for specific diagnosis of Mediterranean visceral leishmaniasis. J Clin Microbiol. 1994 Oct;32(10):2474–2480. doi: 10.1128/jcm.32.10.2474-2480.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Zhang W. W., Charest H., Ghedin E., Matlashewski G. Identification and overexpression of the A2 amastigote-specific protein in Leishmania donovani. Mol Biochem Parasitol. 1996 Jun;78(1-2):79–90. doi: 10.1016/s0166-6851(96)02612-6. [DOI] [PubMed] [Google Scholar]

Articles from Clinical and Diagnostic Laboratory Immunology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES