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. 1994 Oct;32(10):2474–2480. doi: 10.1128/jcm.32.10.2474-2480.1994

Identification of an immunodominant 32-kilodalton membrane protein of Leishmania donovani infantum promastigotes suitable for specific diagnosis of Mediterranean visceral leishmaniasis.

F Tebourski 1, A el Gaied 1, H Louzir 1, R Ben Ismail 1, R Kammoun 1, K Dellagi 1
PMCID: PMC264086  PMID: 7814485

Abstract

Sera from 35 patients suffering from Mediterranean visceral leishmaniasis (caused by Leishmania donovani infantum) and 59 patients with various forms of cutaneous leishmaniasis prevalent in the sub-Mediterranean countries (caused by Leishmania major, L. donovani infantum, or Leishmania tropica) were tested by immunoblotting and enzyme-linked immunosorbent assay (ELISA) with both membrane and soluble antigens prepared from L. donovani infantum parasites. Control sera were from healthy children (n = 41), adults with nonleishmanial diseases (n = 40), and patients with Chagas' disease (n = 12). A P32 antigen present in the membrane preparation from L. donovani infantum parasites was recognized by 95% of serum specimens from patients with Mediterranean visceral leishmaniasis but not by serum specimens from patients with cutaneous leishmaniasis or sera from control individuals. An ELISA with electroeluted P32 antigen was found to have a specificity and sensitivity of 94% in the serodiagnosis of Mediterranean visceral leishmaniasis. Healthy children with asymptomatic Leishmania infection were seronegative for the P32 antigen by ELISA. These results suggest that antibodies to P32 antigen develop only in patients with visceral leishmaniasis and that the P32 ELISA may be useful in areas where the disease is endemic for discriminating between patients with this disease and those with other clinical conditions.

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Selected References

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

  1. Badaró R., Jones T. C., Lorenço R., Cerf B. J., Sampaio D., Carvalho E. M., Rocha H., Teixeira R., Johnson W. D., Jr A prospective study of visceral leishmaniasis in an endemic area of Brazil. J Infect Dis. 1986 Oct;154(4):639–649. doi: 10.1093/infdis/154.4.639. [DOI] [PubMed] [Google Scholar]
  2. Badaró R., Reed S. G., Barral A., Orge G., Jones T. C. Evaluation of the micro enzyme-linked immunosorbent assay (ELISA) for antibodies in American visceral leishmaniasis: antigen selection for detection of infection-specific responses. Am J Trop Med Hyg. 1986 Jan;35(1):72–78. doi: 10.4269/ajtmh.1986.35.72. [DOI] [PubMed] [Google Scholar]
  3. Berneman A., Rolland X., Broun G. La réponse humorale chez l'homme dans les infections à Leishmania infantum. Ann Inst Pasteur Immunol. 1988 May-Jun;139(3):267–278. doi: 10.1016/0769-2625(88)90140-7. [DOI] [PubMed] [Google Scholar]
  4. Bogdan C., Stosiek N., Fuchs H., Röllinghoff M., Solbach W. Detection of potentially diagnostic leishmanial antigens by western blot analysis of sera from patients with kala-azar or multilesional cutaneous leishmaniasis. J Infect Dis. 1990 Dec;162(6):1417–1418. doi: 10.1093/infdis/162.6.1417. [DOI] [PubMed] [Google Scholar]
  5. Bouvier J., Etges R., Bordier C. Identification of the promastigote surface protease in seven species of Leishmania. Mol Biochem Parasitol. 1987 May;24(1):73–79. doi: 10.1016/0166-6851(87)90117-4. [DOI] [PubMed] [Google Scholar]
  6. 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]
  7. Chiller T. M., Samudio M. A., Zoulek G. IgG antibody reactivity with Trypanosoma cruzi and Leishmania antigens in sera of patients with Chagas' disease and leishmaniasis. Am J Trop Med Hyg. 1990 Dec;43(6):650–656. doi: 10.4269/ajtmh.1990.43.650. [DOI] [PubMed] [Google Scholar]
  8. Daniel T. M., Debanne S. M. The serodiagnosis of tuberculosis and other mycobacterial diseases by enzyme-linked immunosorbent assay. Am Rev Respir Dis. 1987 May;135(5):1137–1151. doi: 10.1164/arrd.1987.135.5.1137. [DOI] [PubMed] [Google Scholar]
  9. Dwyer D. M. Isolation and partial characterization of surface membranes from Leishmania donovani promastigotes. J Protozool. 1980 May;27(2):176–182. doi: 10.1111/j.1550-7408.1980.tb04676.x. [DOI] [PubMed] [Google Scholar]
  10. Evans T. G., Krug E. C., Wilson M. E., Vasconcelos A. W., de Alencar J. E., Pearson R. D. Evaluation of antibody responses in American visceral leishmaniasis by ELISA and immunoblot. Mem Inst Oswaldo Cruz. 1989 Apr-Jun;84(2):157–166. doi: 10.1590/s0074-02761989000200003. [DOI] [PubMed] [Google Scholar]
  11. Galen R. S. Application of the predictive value model in the analysis of test effectiveness. Clin Lab Med. 1982 Dec;2(4):685–699. [PubMed] [Google Scholar]
  12. Guimarães M. C., Celeste B. J., Franco E. L. Diagnostic performance indices for immunofluorescent tests and enzyme immunoassays of leishmaniasis sera from northern and north-eastern Brazil. Bull World Health Organ. 1990;68(1):39–43. [PMC free article] [PubMed] [Google Scholar]
  13. 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]
  14. Heinzel F. P., Sadick M. D., Mutha S. S., Locksley R. M. Production of interferon gamma, interleukin 2, interleukin 4, and interleukin 10 by CD4+ lymphocytes in vivo during healing and progressive murine leishmaniasis. Proc Natl Acad Sci U S A. 1991 Aug 15;88(16):7011–7015. doi: 10.1073/pnas.88.16.7011. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Ho M., Leeuwenburg J., Mbugua G., Wamachi A., Voller A. An enzyme-linked immunosorbent assay (ELISA) for field diagnosis of visceral leishmaniasis. Am J Trop Med Hyg. 1983 Sep;32(5):943–946. doi: 10.4269/ajtmh.1983.32.943. [DOI] [PubMed] [Google Scholar]
  16. Hoerauf A., Andrade P. P., Andrade C. R., Solbach W., Röllinghoff M. Immunoblotting as a valuable tool to differentiate human visceral leishmaniasis from lymphoproliferative disorders and other clinically similar diseases. Res Immunol. 1992 May;143(4):375–383. doi: 10.1016/s0923-2494(05)80070-7. [DOI] [PubMed] [Google Scholar]
  17. Hommel M., Peters W., Ranque J., Quilici M., Lanotte G. The micro-ELISA technique in the serodiagnosis of visceral leishmaniasis. Ann Trop Med Parasitol. 1978 Jun;72(3):213–218. doi: 10.1080/00034983.1978.11719308. [DOI] [PubMed] [Google Scholar]
  18. 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]
  19. 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]
  20. 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]
  21. Lepay D. A., Nogueira N., Cohn Z. Surface antigens of Leishmania donovani promastigotes. J Exp Med. 1983 May 1;157(5):1562–1572. doi: 10.1084/jem.157.5.1562. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Melby P. C., Neva F. A., Sacks D. L. Profile of human T cell response to leishmanial antigens. Analysis by immunoblotting. J Clin Invest. 1989 Jun;83(6):1868–1875. doi: 10.1172/JCI114093. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Pappas M. G., McGreevy P. B., Hajkowski R., Hendricks L. D., Oster C. N., Hockmeyer W. T. Evaluation of promastigote and amastigote antigens in the indirect fluorescent antibody test for American cutaneous leishmaniasis. Am J Trop Med Hyg. 1983 Nov;32(6):1260–1267. doi: 10.4269/ajtmh.1983.32.1260. [DOI] [PubMed] [Google Scholar]
  24. Reed S. G., Badaro R., Lloyd R. M. Identification of specific and cross-reactive antigens of Leishmania donovani chagasi by human infection sera. J Immunol. 1987 Mar 1;138(5):1596–1601. [PubMed] [Google Scholar]
  25. Rolland-Burger L., Rolland X., Grieve C. W., Monjour L. Immunoblot analysis of the humoral immune response to Leishmania donovani infantum polypeptides in human visceral leishmaniasis. J Clin Microbiol. 1991 Jul;29(7):1429–1435. doi: 10.1128/jcm.29.7.1429-1435.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Sher A., Gazzinelli R. T., Oswald I. P., Clerici M., Kullberg M., Pearce E. J., Berzofsky J. A., Mosmann T. R., James S. L., Morse H. C., 3rd Role of T-cell derived cytokines in the downregulation of immune responses in parasitic and retroviral infection. Immunol Rev. 1992 Jun;127:183–204. doi: 10.1111/j.1600-065x.1992.tb01414.x. [DOI] [PubMed] [Google Scholar]
  27. 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]
  28. Zacharius R. M., Zell T. E., Morrison J. H., Woodlock J. J. Glycoprotein staining following electrophoresis on acrylamide gels. Anal Biochem. 1969 Jul;30(1):148–152. doi: 10.1016/0003-2697(69)90383-2. [DOI] [PubMed] [Google Scholar]

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