Abstract
Cellular immune mechanisms resulting in gamma interferon production are critical for protection against visceral leishmaniasis. Antigens stimulating T-cell responses are likely present in the intracellular amastigote form of the parasite, since this is the form found in a mammalian host. To identify T-cell antigens of Leishmania chagasi, the parasite causing South American visceral leishmaniasis, we used a double antibody-T-cell technique to screen an amastigote cDNA library. One cDNA selected (Lcr1) encodes an antigen that stimulated proliferation of splenic T lymphocytes from infected mice that were either resistant (C3H.HeJ) or susceptible (BALB/c) to L. chagasi infection. The Lcr1 cDNA contains four highly divergent 201-bp repeats homologous to the 204-bp repeat of a Trypanosoma cruzi flagellar antigen gene. Results are consistent with a single copy of the Lcr1 gene producing an mRNA of > 10 kb and a protein of > 200 kDa. Recombinant Lcr1, cloned adjacent to polyhistidine and purified on a nickel affinity column, stimulated gamma interferon but not interleukin-4 (IL-4), IL-5, or IL-10 secretion by T-cell-enriched splenocytes from either susceptible or resistant mice during L. chagasi infection. Immunization with Lcr1 partially protected BALB/c mice against challenge with L. chagasi, indicating the utility of the double screening approach in selecting relevant T-cell antigens.
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- Anders R. F. Multiple cross-reactivities amongst antigens of Plasmodium falciparum impair the development of protective immunity against malaria. Parasite Immunol. 1986 Nov;8(6):529–539. doi: 10.1111/j.1365-3024.1986.tb00867.x. [DOI] [PubMed] [Google Scholar]
- Bradley D. J., Taylor B. A., Blackwell J., Evans E. P., Freeman J. Regulation of Leishmania populations within the host. III. Mapping of the locus controlling susceptibility to visceral leishmaniasis in the mouse. Clin Exp Immunol. 1979 Jul;37(1):7–14. [PMC free article] [PubMed] [Google Scholar]
- Bretscher P. A., Wei G., Menon J. N., Bielefeldt-Ohmann H. Establishment of stable, cell-mediated immunity that makes "susceptible" mice resistant to Leishmania major. Science. 1992 Jul 24;257(5069):539–542. doi: 10.1126/science.1636090. [DOI] [PubMed] [Google Scholar]
- Burns J. M., Jr, Scott J. M., Carvalho E. M., Russo D. M., March C. J., Van Ness K. P., Reed S. G. Characterization of a membrane antigen of Leishmania amazonensis that stimulates human immune responses. J Immunol. 1991 Jan 15;146(2):742–748. [PubMed] [Google Scholar]
- Carvalho E. M., Bacellar O., Barral A., Badaro R., Johnson W. D., Jr Antigen-specific immunosuppression in visceral leishmaniasis is cell mediated. J Clin Invest. 1989 Mar;83(3):860–864. doi: 10.1172/JCI113969. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Carvalho E. M., Badaró R., Reed S. G., Jones T. C., Johnson W. D., Jr Absence of gamma interferon and interleukin 2 production during active visceral leishmaniasis. J Clin Invest. 1985 Dec;76(6):2066–2069. doi: 10.1172/JCI112209. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Carvalho E. M., Barral A., Pedral-Sampaio D., Barral-Netto M., Badaró R., Rocha H., Johnson W. D., Jr Immunologic markers of clinical evolution in children recently infected with Leishmania donovani chagasi. J Infect Dis. 1992 Mar;165(3):535–540. doi: 10.1093/infdis/165.3.535. [DOI] [PubMed] [Google Scholar]
- Champsi J., McMahon-Pratt D. Membrane glycoprotein M-2 protects against Leishmania amazonensis infection. Infect Immun. 1988 Dec;56(12):3272–3279. doi: 10.1128/iai.56.12.3272-3279.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- Fiorentino D. F., Bond M. W., Mosmann T. R. Two types of mouse T helper cell. IV. Th2 clones secrete a factor that inhibits cytokine production by Th1 clones. J Exp Med. 1989 Dec 1;170(6):2081–2095. doi: 10.1084/jem.170.6.2081. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Fiorentino D. F., Zlotnik A., Mosmann T. R., Howard M., O'Garra A. IL-10 inhibits cytokine production by activated macrophages. J Immunol. 1991 Dec 1;147(11):3815–3822. [PubMed] [Google Scholar]
- Gajewski T. F., Pinnas M., Wong T., Fitch F. W. Murine Th1 and Th2 clones proliferate optimally in response to distinct antigen-presenting cell populations. J Immunol. 1991 Mar 15;146(6):1750–1758. [PubMed] [Google Scholar]
- Galvão-Castro B., Sá Ferreira J. A., Marzochi K. F., Marzochi M. C., Coutinho S. G., Lambert P. H. Polyclonal B cell activation, circulating immune complexes and autoimmunity in human american visceral leishmaniasis. Clin Exp Immunol. 1984 Apr;56(1):58–66. [PMC free article] [PubMed] [Google Scholar]
- Ghalib H. W., Piuvezam M. R., Skeiky Y. A., Siddig M., Hashim F. A., el-Hassan A. M., Russo D. M., Reed S. G. Interleukin 10 production correlates with pathology in human Leishmania donovani infections. J Clin Invest. 1993 Jul;92(1):324–329. doi: 10.1172/JCI116570. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gorczynski R. M. Nature of resistance to leishmaniasis in experimental rodents. Dev Comp Immunol. 1982 Spring;6(2):199–207. doi: 10.1016/s0145-305x(82)80003-7. [DOI] [PubMed] [Google Scholar]
- Heinzel F. P., Sadick M. D., Holaday B. J., Coffman R. L., Locksley R. M. Reciprocal expression of interferon gamma or interleukin 4 during the resolution or progression of murine leishmaniasis. Evidence for expansion of distinct helper T cell subsets. J Exp Med. 1989 Jan 1;169(1):59–72. doi: 10.1084/jem.169.1.59. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- Ho J. L., Badaró R., Schwartz A., Dinarello C. A., Gelfand J. A., Sobel J., Barral A., Netto M. B., Carvalho E. M., Reed S. G. Diminished in vitro production of interleukin-1 and tumor necrosis factor-alpha during acute visceral leishmaniasis and recovery after therapy. J Infect Dis. 1992 Jun;165(6):1094–1102. doi: 10.1093/infdis/165.6.1094. [DOI] [PubMed] [Google Scholar]
- Howard J. G. Immunological regulation and control of experimental leishmaniasis. Int Rev Exp Pathol. 1986;28:79–116. [PubMed] [Google Scholar]
- Ibañez C. F., Affranchino J. L., Macina R. A., Reyes M. B., Leguizamon S., Camargo M. E., Aslund L., Pettersson U., Frasch A. C. Multiple Trypanosoma cruzi antigens containing tandemly repeated amino acid sequence motifs. Mol Biochem Parasitol. 1988 Jul;30(1):27–33. doi: 10.1016/0166-6851(88)90129-6. [DOI] [PubMed] [Google Scholar]
- Kaye P. M., Bancroft G. J. Leishmania donovani infection in scid mice: lack of tissue response and in vivo macrophage activation correlates with failure to trigger natural killer cell-derived gamma interferon production in vitro. Infect Immun. 1992 Oct;60(10):4335–4342. doi: 10.1128/iai.60.10.4335-4342.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kaye P. M., Curry A. J., Blackwell J. M. Differential production of Th1- and Th2-derived cytokines does not determine the genetically controlled or vaccine-induced rate of cure in murine visceral leishmaniasis. J Immunol. 1991 Apr 15;146(8):2763–2770. [PubMed] [Google Scholar]
- 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]
- Lafaille J. J., Linss J., Krieger M. A., Souto-Padrón T., de Souza W., Goldenberg S. Structure and expression of two Trypanosoma cruzi genes encoding antigenic proteins bearing repetitive epitopes. Mol Biochem Parasitol. 1989 Jun 15;35(2):127–136. doi: 10.1016/0166-6851(89)90115-1. [DOI] [PubMed] [Google Scholar]
- Liew F. Y., Millott S. M., Schmidt J. A. A repetitive peptide of Leishmania can activate T helper type 2 cells and enhance disease progression. J Exp Med. 1990 Nov 1;172(5):1359–1365. doi: 10.1084/jem.172.5.1359. [DOI] [PMC free article] [PubMed] [Google Scholar]
- McElrath M. J., Murray H. W., Cohn Z. A. The dynamics of granuloma formation in experimental visceral leishmaniasis. J Exp Med. 1988 Jun 1;167(6):1927–1937. doi: 10.1084/jem.167.6.1927. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Melby P. C. Experimental leishmaniasis in humans: review. Rev Infect Dis. 1991 Sep-Oct;13(5):1009–1017. doi: 10.1093/clinids/13.5.1009. [DOI] [PubMed] [Google Scholar]
- Mendonça S. C., Russell D. G., Coutinho S. G. Analysis of the human T cell responsiveness to purified antigens of Leishmania: lipophosphoglycan (LPG) and glycoprotein 63 (gp 63). Clin Exp Immunol. 1991 Mar;83(3):472–478. doi: 10.1111/j.1365-2249.1991.tb05663.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Mosmann T. R., Schumacher J. H., Fiorentino D. F., Leverah J., Moore K. W., Bond M. W. Isolation of monoclonal antibodies specific for IL-4, IL-5, IL-6, and a new Th2-specific cytokine (IL-10), cytokine synthesis inhibitory factor, by using a solid phase radioimmunoadsorbent assay. J Immunol. 1990 Nov 1;145(9):2938–2945. [PubMed] [Google Scholar]
- Murray H. W., Rubin B. Y., Rothermel C. D. Killing of intracellular Leishmania donovani by lymphokine-stimulated human mononuclear phagocytes. Evidence that interferon-gamma is the activating lymphokine. J Clin Invest. 1983 Oct;72(4):1506–1510. doi: 10.1172/JCI111107. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Murray H. W., Stern J. J., Welte K., Rubin B. Y., Carriero S. M., Nathan C. F. Experimental visceral leishmaniasis: production of interleukin 2 and interferon-gamma, tissue immune reaction, and response to treatment with interleukin 2 and interferon-gamma. J Immunol. 1987 Apr 1;138(7):2290–2297. [PubMed] [Google Scholar]
- Mustafa A. S., Oftung F., Deggerdal A., Gill H. K., Young R. A., Godal T. Gene isolation with human T lymphocyte probes. Isolation of a gene that expresses an epitope recognized by T cells specific for Mycobacterium bovis BCG and pathogenic mycobacteria. J Immunol. 1988 Oct 15;141(8):2729–2733. [PubMed] [Google Scholar]
- Pearson R. D., Steigbigel R. T. Mechanism of lethal effect of human serum upon Leishmania donovani. J Immunol. 1980 Nov;125(5):2195–2201. [PubMed] [Google Scholar]
- Rachamim N., Jaffe C. L. Pure protein from Leishmania donovani protects mice against both cutaneous and visceral leishmaniasis. J Immunol. 1993 Mar 15;150(6):2322–2331. [PubMed] [Google Scholar]
- Reiner S. L., Wang Z. E., Hatam F., Scott P., Locksley R. M. TH1 and TH2 cell antigen receptors in experimental leishmaniasis. Science. 1993 Mar 5;259(5100):1457–1460. doi: 10.1126/science.8451641. [DOI] [PubMed] [Google Scholar]
- Russell D. G., Alexander J. Effective immunization against cutaneous leishmaniasis with defined membrane antigens reconstituted into liposomes. J Immunol. 1988 Feb 15;140(4):1274–1279. [PubMed] [Google Scholar]
- Russo D. M., Jardim A., Carvalho E. M., Sleath P. R., Armitage R. J., Olafson R. W., Reed S. G. Mapping human T cell epitopes in leishmania gp63. Identification of cross-reactive and species-specific epitopes. J Immunol. 1993 Feb 1;150(3):932–939. [PubMed] [Google Scholar]
- Russo D. M., Turco S. J., Burns J. M., Jr, Reed S. G. Stimulation of human T lymphocytes by Leishmania lipophosphoglycan-associated proteins. J Immunol. 1992 Jan 1;148(1):202–207. [PubMed] [Google Scholar]
- 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]
- Scott P., Natovitz P., Coffman R. L., Pearce E., Sher A. Immunoregulation of cutaneous leishmaniasis. T cell lines that transfer protective immunity or exacerbation belong to different T helper subsets and respond to distinct parasite antigens. J Exp Med. 1988 Nov 1;168(5):1675–1684. doi: 10.1084/jem.168.5.1675. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Spithill T. W., Samaras N. Genomic organization, chromosomal location and transcription of dispersed and repeated tubulin genes in Leishmania major. Mol Biochem Parasitol. 1987 May;24(1):23–37. doi: 10.1016/0166-6851(87)90112-5. [DOI] [PubMed] [Google Scholar]
- Squires K. E., Schreiber R. D., McElrath M. J., Rubin B. Y., Anderson S. L., Murray H. W. Experimental visceral leishmaniasis: role of endogenous IFN-gamma in host defense and tissue granulomatous response. J Immunol. 1989 Dec 15;143(12):4244–4249. [PubMed] [Google Scholar]
- Stern J. J., Oca M. J., Rubin B. Y., Anderson S. L., Murray H. W. Role of L3T4+ and LyT-2+ cells in experimental visceral leishmaniasis. J Immunol. 1988 Jun 1;140(11):3971–3977. [PubMed] [Google Scholar]
- Tam J. P., Clavijo P., Lu Y. A., Nussenzweig V., Nussenzweig R., Zavala F. Incorporation of T and B epitopes of the circumsporozoite protein in a chemically defined synthetic vaccine against malaria. J Exp Med. 1990 Jan 1;171(1):299–306. doi: 10.1084/jem.171.1.299. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ulczak O. M., Ghadirian E., Skamene E., Blackwell J. M., Kongshavn P. A. Characterization of protective T cells in the acquired response to Leishmania donovani in genetically determined cure (H-2b) and noncure (H-2d) mouse strains. Infect Immun. 1989 Sep;57(9):2892–2899. doi: 10.1128/iai.57.9.2892-2899.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
- White A. C., Jr, McMahon-Pratt D. Prophylactic immunization against experimental Leishmania donovani infection by use of a purified protein vaccine. J Infect Dis. 1990 Jun;161(6):1313–1314. doi: 10.1093/infdis/161.6.1313. [DOI] [PubMed] [Google Scholar]
- Wilson M. E., Hardin K. K., Donelson J. E. Expression of the major surface glycoprotein of Leishmania donovani chagasi in virulent and attenuated promastigotes. J Immunol. 1989 Jul 15;143(2):678–684. [PubMed] [Google Scholar]
- Wilson M. E., Hardin K. K. The major concanavalin A-binding surface glycoprotein of Leishmania donovani chagasi promastigotes is involved in attachment to human macrophages. J Immunol. 1988 Jul 1;141(1):265–272. [PubMed] [Google Scholar]
- Xu Y. H., Macedonia J., Sher A., Pearce E., Cheever A. W. Dynamic analysis of splenic Th1 and Th2 lymphocyte functions in mice infected with Schistosoma japonicum. Infect Immun. 1991 Sep;59(9):2934–2940. doi: 10.1128/iai.59.9.2934-2940.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
- dos Santos C. N., Krieger M. A., Almeida E., Lafaille J. J., Goldenberg S., Galler R. Trypanosoma cruzi flagellar repetitive antigen expression by recombinant baculovirus: towards an improved diagnostic reagent for Chagas' disease. Biotechnology (N Y) 1992 Nov;10(11):1474–1477. doi: 10.1038/nbt1192-1474. [DOI] [PubMed] [Google Scholar]