Antigen characterization of vector-borne and cultured metacyclic trypomastigotes of Trypanosoma cruzi

Yoshida, N.; Teixeira, M. M. G.; Sbravate, C. A.

doi:10.1590/S0036-46651986000200003

Abstracts

Metacyclic trypomastigotes ol the CL strain of Trypanosoma cruzi obtained from triatomid vectors and from axenic cultures were comparatively analysed as to their antigen make-up and immunogenic characteristics. They were found to be similar by the various parameters examined. Thus, sera of mice immunized with either one of the two metacyclic types precipitated a 82Kd surface protein from 131I-labeled culture metacyclics. Sera of mice protected against acute T. cruzi infection by immunization with killed culture metacyclics of a different strain (G) recognized, by immunoblotting, a 77Kd protein in both types of CL strain metacyclics. A monoclonal antibody raised against G strain metacyclics, and specific for metacyclic stages of this strain, reacted with both CL strain metacyclic types. Both metacyclic forms were similarly Iysed by various anti-T. cruzi sera, in a complement-mediated reaction.

Chagas'disease; vector borne; Trypanosoma cruzi, cultured metacyclic tripomastigotes

Tripomastigotas metacíclicos da cepa CL de Trypanosoma cruzi, originários de triatomineos e de cultura axênica, foram comparativamente analisados quanto a sua composição antigênica e características ímunogênicas. Os vários parâmetros examinados mostraram que as duas formas metacíclicas sáo semelhantes. Assim, soros de camundongos imunizados com qualquer uma das formas metacíclicas precipitaram uma proteína de superfície de 82Kd de tripomastigotas metacíclicos de cultura marcados com 131I. Soros de camundongos protegidos con trn infecção aguda por T. cruzi, por imunização com formas metacíclicas de cultura inativadas, de uma cepa diferente (G), detectaram por imunoblotting uma proteína de 77Kd em ambas as formas metacíclicas da cepa CL. Um anticorpo monoclonal, produzido contra as for mas metaciclicas da cepa G, e específico para tripomastigotas metacíclicos, reagiu com formas metacíclicas da cepa CL tanto do vetor quanto de cultura. As duas formas metacíclicas mostraram suscetibilidade semelhante à lise por vários soros anti-T. cruzi, em uma reação mediada por complemento.

ORIGINAL ARTICLES

Antigen characterization of vector-borne and cultured metacyclic trypomastigotes of Trypanosoma cruzi

Caracterização antigênica de tripomastigotas metacíclicos de Trypanosoma cruzi provenientes do vetor e de cultura

N. Yoshida; M. M. G. Teixeira; C. A. Sbravate

Departamento de Microbiologia, Imunologia e Parasitologia. Escola Paulista de Medicina. São Paulo, SP, Brasil

SUMMARY

Metacyclic trypomastigotes ol the CL strain of Trypanosoma cruzi obtained from triatomid vectors and from axenic cultures were comparatively analysed as to their antigen make-up and immunogenic characteristics. They were found to be similar by the various parameters examined. Thus, sera of mice immunized with either one of the two metacyclic types precipitated a 82Kd surface protein from ¹³¹I-labeled culture metacyclics. Sera of mice protected against acute T. cruzi infection by immunization with killed culture metacyclics of a different strain (G) recognized, by immunoblotting, a 77Kd protein in both types of CL strain metacyclics. A monoclonal antibody raised against G strain metacyclics, and specific for metacyclic stages of this strain, reacted with both CL strain metacyclic types. Both metacyclic forms were similarly Iysed by various anti-T. cruzi sera, in a complement-mediated reaction.

Key-words: Chagas'disease vector borne Trypanosoma cruzi, cultured metacyclic tripomastigotes.

RESUMO

Tripomastigotas metacíclicos da cepa CL de Trypanosoma cruzi, originários de triatomineos e de cultura axênica, foram comparativamente analisados quanto a sua composição antigênica e características ímunogênicas. Os vários parâmetros examinados mostraram que as duas formas metacíclicas sáo semelhantes. Assim, soros de camundongos imunizados com qualquer uma das formas metacíclicas precipitaram uma proteína de superfície de 82Kd de tripomastigotas metacíclicos de cultura marcados com ¹³¹I. Soros de camundongos protegidos con trn infecção aguda por T. cruzi, por imunização com formas metacíclicas de cultura inativadas, de uma cepa diferente (G), detectaram por imunoblotting uma proteína de 77Kd em ambas as formas metacíclicas da cepa CL. Um anticorpo monoclonal, produzido contra as for mas metaciclicas da cepa G, e específico para tripomastigotas metacíclicos, reagiu com formas metacíclicas da cepa CL tanto do vetor quanto de cultura. As duas formas metacíclicas mostraram suscetibilidade semelhante à lise por vários soros anti-T. cruzi, em uma reação mediada por complemento.

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ACKNOWLEDGEMENT

This work was supported by Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP), Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) and FINEP. Brasil.

We thank Dr. Erney P. Camargo for reviewing the manuscript, Dr. Fidel Zavala for critical reading of the manuscript and Ms. M. Russo for excelent technical assistance.

Recebido para publicação em 8/4/1985.

1. BRENER, Z. & CHIARI, E. Variações morfológicas observadas em diferentes amostras de Trypanosoma cruzi Rev. Inst. Med. trop. S. Paulo, 5: 220-224, 1963.
2. CAMARGO, E. P. Growth and differentiation in Trypanosoma cruzi I. Origin of metacyclic trypanosomes in liquid media. Rev. Inst. Med. trop. S. Paulo, 6: 93-100, 1964.
3. CAMARGO, E. P.; MATTEI, D. M.; BARBIERI, C. L. & MOREL, C. M. Electrophoretic analysis of endonuclease-generated fragments of k-DNA, of esterase isoenzymes and surface proteins as aids for species identification of insect trypanosomatids. J. Protozool., 29: 251-258, 1982.
4. HANSON, W. L. Immune response and mechanisms of resistance in Trypanosoma cruzi Pan Amer. Hlth. Org. Scient. Publ., 347: 22-34, 1977.
5. KESSLER, S. W. Rapid isolation of antigens from cells with staphylococcal protein A antibody absorbent: parameters of the interaction of antibody antigen complexes with protein A. J. Immunol., 115: 1617-1624, 1975.
6. KIERSZENBAUM, F. & LIMA, M. F. Susceptibility of insectborne, metacyclic forms of Trypanosoma cruzi to antibody-mediated mechanisms of destruction. Amer. J. trop. Med. Hyg., 32: 1236-1241, 1983.
7. KOHLER, G. & MILSTEIN, C. Continuous cultures of fused celts secreting antibody of predefined specificity. Nature (London), 856: 495-497, 1975.
8 KRETTLI, A. U. & BRENER, Z. Protective effects of specific antibodies in Trypanosoma cruzi infection. J. Immunol., 116: 755-760, 1976.
9. KRETTLI, A. U. & BRENER, Z. Resistance against Trypanosoma cruzi associated to anti-living trypomastigote antibodies. J. Immunol., 128: 2009-2012, 1982.
10. LAEMMLI, U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature (London), 227: 680-685, 1970.
11. MARKWELL, M. A. K. & FOX, C. F. Surface-specific iodination of membrane proteins of viruses and eukaryotic cells using 1,3,4,6-tetraehloro-3a,6a-diphenylglycoluril. Biochemistry. 17: 4807-4817, 1978.
12. NOGUEIRA, N.; UNKELESS, J. & COHN, Z. Specific glycoprotein antigens on the surface of insect and mammalian stages of Trypanosoma cruzi Proc. nat. Acad. Sci. (Wash.), 79: 1259-1263, 1982.
13. POTOCNJAK, P.; YOSHIDA, N.; NUSSENZWEIG, R. S. & NUSSENZWEIG, V. - Monovalent fragments (Fab) of monoclonal antibodies to a sporozoite surface antigen (Pb44) protect mice against malarial infection. J. exp. Med., 151: 1504-1513, 1980.
14. TOWBIN, H.; STAEHELIN, T. & GORDON, J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: Procedures and some applications. Proc. natl. Acad. Scl. (Wash.), 76: 4350-4354, 1979.
15. VILLALTA, F. & KIERSZENBAUM, F. Cross-reactivity of vectorborne metacyclic forms of Trypanosoma cruzi with mammalian and culture stages. J. Protozool., 30: 329-331, 1983.
16. YOSHIDA, N. Trypanosoma cruzi: Recognition of trypomastigote surface antigens by lytic antisera from mice resistant to acute infection. Exp. Parasit. (In press).
17. YOSHIDA, N. Surface antigens of metacyclic trypomastigotes of Trypanosoma cruzi Infect Immun., 40: 836-839, 1983.
18. YOSHIDA, N.; POTOCNJAK, P.; NUSSENZWEIG, V. & NUSSENZWEIG, R. S. Biosynthesis of Pb44, the protective antigen of sporozoites of Plasmodium berghei. J. exp. Med., 154: 1225-1236, 1981.
19. YOSHIDA, N.; TEIXEIRA, M. M. G.; CHRISTO, C B. & SBRAVATE, C. A. Resistance of mice immunized with killed culture trypomastigotes against infection by insect-derived trypomastigotes of Trypanosoma cruzi Rev. Inst. Med. trop. S. Paulo, 36: 329-333, 1984.

Publication Dates

Publication in this collection
16 Oct 2012
Date of issue
Apr 1986

History

Received
08 Apr 1985

This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.

[1] 1. BRENER, Z. & CHIARI, E. Variações morfológicas observadas em diferentes amostras de Trypanosoma cruzi Rev. Inst. Med. trop. S. Paulo, 5: 220-224, 1963.

[2] 2. CAMARGO, E. P. Growth and differentiation in Trypanosoma cruzi I. Origin of metacyclic trypanosomes in liquid media. Rev. Inst. Med. trop. S. Paulo, 6: 93-100, 1964.

[3] 3. CAMARGO, E. P.; MATTEI, D. M.; BARBIERI, C. L. & MOREL, C. M. Electrophoretic analysis of endonuclease-generated fragments of k-DNA, of esterase isoenzymes and surface proteins as aids for species identification of insect trypanosomatids. J. Protozool., 29: 251-258, 1982.

[4] 4. HANSON, W. L. Immune response and mechanisms of resistance in Trypanosoma cruzi Pan Amer. Hlth. Org. Scient. Publ., 347: 22-34, 1977.

[5] 5. KESSLER, S. W. Rapid isolation of antigens from cells with staphylococcal protein A antibody absorbent: parameters of the interaction of antibody antigen complexes with protein A. J. Immunol., 115: 1617-1624, 1975.

[6] 6. KIERSZENBAUM, F. & LIMA, M. F. Susceptibility of insectborne, metacyclic forms of Trypanosoma cruzi to antibody-mediated mechanisms of destruction. Amer. J. trop. Med. Hyg., 32: 1236-1241, 1983.

[7] 7. KOHLER, G. & MILSTEIN, C. Continuous cultures of fused celts secreting antibody of predefined specificity. Nature (London), 856: 495-497, 1975.

[8] 8 KRETTLI, A. U. & BRENER, Z. Protective effects of specific antibodies in Trypanosoma cruzi infection. J. Immunol., 116: 755-760, 1976.

[9] 9. KRETTLI, A. U. & BRENER, Z. Resistance against Trypanosoma cruzi associated to anti-living trypomastigote antibodies. J. Immunol., 128: 2009-2012, 1982.

[10] 10. LAEMMLI, U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature (London), 227: 680-685, 1970.

[11] 11. MARKWELL, M. A. K. & FOX, C. F. Surface-specific iodination of membrane proteins of viruses and eukaryotic cells using 1,3,4,6-tetraehloro-3a,6a-diphenylglycoluril. Biochemistry. 17: 4807-4817, 1978.

[12] 12. NOGUEIRA, N.; UNKELESS, J. & COHN, Z. Specific glycoprotein antigens on the surface of insect and mammalian stages of Trypanosoma cruzi Proc. nat. Acad. Sci. (Wash.), 79: 1259-1263, 1982.

[13] 13. POTOCNJAK, P.; YOSHIDA, N.; NUSSENZWEIG, R. S. & NUSSENZWEIG, V. - Monovalent fragments (Fab) of monoclonal antibodies to a sporozoite surface antigen (Pb44) protect mice against malarial infection. J. exp. Med., 151: 1504-1513, 1980.

[14] 14. TOWBIN, H.; STAEHELIN, T. & GORDON, J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: Procedures and some applications. Proc. natl. Acad. Scl. (Wash.), 76: 4350-4354, 1979.

[15] 15. VILLALTA, F. & KIERSZENBAUM, F. Cross-reactivity of vectorborne metacyclic forms of Trypanosoma cruzi with mammalian and culture stages. J. Protozool., 30: 329-331, 1983.

[16] 16. YOSHIDA, N. Trypanosoma cruzi: Recognition of trypomastigote surface antigens by lytic antisera from mice resistant to acute infection. Exp. Parasit. (In press).

[17] 17. YOSHIDA, N. Surface antigens of metacyclic trypomastigotes of Trypanosoma cruzi Infect Immun., 40: 836-839, 1983.

[18] 18. YOSHIDA, N.; POTOCNJAK, P.; NUSSENZWEIG, V. & NUSSENZWEIG, R. S. Biosynthesis of Pb44, the protective antigen of sporozoites of Plasmodium berghei. J. exp. Med., 154: 1225-1236, 1981.

[19] 19. YOSHIDA, N.; TEIXEIRA, M. M. G.; CHRISTO, C B. & SBRAVATE, C. A. Resistance of mice immunized with killed culture trypomastigotes against infection by insect-derived trypomastigotes of Trypanosoma cruzi Rev. Inst. Med. trop. S. Paulo, 36: 329-333, 1984.

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Brasil

Antigen characterization of vector-borne and cultured metacyclic trypomastigotes of Trypanosoma cruzi

Caracterização antigênica de tripomastigotas metacíclicos de Trypanosoma cruzi provenientes do vetor e de cultura

Abstracts

Publication Dates

History