Biological and genetic characterization of two Colombian clones of Trypanosoma cruzi groups I and II
Keywords:
Trypanosoma cruzi, Chagas disease, variability (Genetics), tropism
Abstract
Introduction. Genetic differences between T. cruzi I and T. cruzi II may determine differences in their tissue tropism in the vertebrate host and may also be responsible for the differences in clinical manifestations of Chagas disease.Objective. Two Colombian clones of the T. cruzi groups I and II were characterized biologically and genetically in a murine model.
Materials and methods. Strains Cas15 and AF1 belonging to the T. cruzi groups I and II were cloned in semisolid medium. A clone of each strain and a mix of both were used to infect mice; the mice were subsequently sacrificed at selected post-infection intervals. In order to identify the parasite presence in blood and organs, two methods were used (a) microhematocrit and (b) polymerase chain reaction with primers for satellite DNA and the intergenic region of miniexon.
Results. The T. cruzi I clone was more infectious, with a preferential tropism observed in heart, rectum and skeletal muscle, whereas clone T. cruzi II exhibited a preferential tropism for spleen and liver. During the infection with the clone mixture a predominance of the T. cruzi I clone over clone II in blood as well as in organs was observed.
Conclusion. The results corroborate that the genetic differences between the T. cruzi groups correlate with their tissue tropism, and can play an essential role in explaining the clinical manifestations of Chagas disease observed in Colombia.
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References
1. Andrade LO, Machado CR, Chiari E, Pena SD, Macedo AM. Differential tissue distribution of diverse clones of Trypanosoma cruzi in infected mice. Mol Biochem Parasitol 1999;100:269-75.
2. Moncayo A. Chagas disease: current epidemiological trends after the interruption of vectorial and transfusional transmission in the Southern Cone countries. Mem Inst Oswaldo Cruz 2003;98:577-91.
3. Lenzi HL, Oliveira DN, Lima MT, Gatas CR. Trypanosoma cruzi: paninfectivity of CL strain during murine acute infection. Exp Parasitol 1996;84:16-27.
4. Prata A. Clinical and epidemiological aspects of Chagas disease. Lancet Infect Dis 2001;1:92-100.
5. Vago AR, Macedo AM, Oliveira RP, Andrade LO, Chiari E, Galvao LM, et al. Kinetoplast DNA signatures of Trypanosoma cruzi strains obtained directly from infected tissues. Am J Pathol 1996;149:2153-9.
6. Vago AR, Andrade LO, Leite AA, d'Avila Reis D, Macedo AM, Adad SJ, et al. Genetic characterization of Trypanosoma cruzi directly from tissues of patients with chronic Chagas disease: differential distribution of genetic types into diverse organs. Am J Pathol 2000;156:1805-9.
7. Macedo AM, Machado CR, Oliveira RP, Pena SD. Trypanosoma cruzi: genetic structure of populations and relevant of genetic variability to the pathogenesis of Chagas disease. Mem Inst Oswaldo Cruz 2004;99:1-12.
8. Oliveira RP, Broude NF, Macedo AM, Cantor CR, Smith CL, Pena SD. Probing the genetic population structure of Trypanosoma cruzi with polymorphic microsatellites. Proc Natl Acad Sci USA 1998;95:3776-80.
9. Andrade LO, Machado CR, Chiari E, Pena SD, Macedo AM. Trypanosoma cruzi: role of host genetic background in the differential tissue distribution of parasite clonal populations. Exp Parasitol 2002;100:269-75.
10. Macedo AM, Martins MS, Chiari E, Pena SD. DNA fingerprinting of Trypanosoma cruzi: a new tool for characterization of strains and clones. Mol Biochem Parasitol 1992;55:147-53.
11. Tibayrenc M, Neubauer K, Barnabé C, Guerrini F, Skarecky D, Ayala FJ. Genetic characterization of six parasitic protozoa: parity betweem random-primer DNA typing and multilocus enzyme electrophoresis. Proc Natl Acad Sci USA 1993;90:1335-9.
12. Souto RP, Fernandes O, Macedo AM, Campbell DA, Zingales B. DNA markers define two major phylogenetic lineages of Trypanosoma cruzi. Mol Biochem Parasitol 1996;83:141-52.
13. Fernandes O, Souto RP, Castro JA, Pereira JB, Fernandes NC, Junqueira AC, et al. Brazilian isolates of Trypanosoma cruzi from humans and triatomines classified into two lineages using mini-exon and ribosomal RNA sequences. Am J Trop Med Hyg 1998;58:807-11.
14. Fernandes O, Mangia RH, Lisboa CV, Pinho AP, Morel CM, Zingales B, et al. The complexity of the sylvatic cycle of Trypanosoma cruzi in Rio de Janeiro state (Brazil) revealed by the non-transcribed spacer of the mini-exon gene. Parasitology 1999;118:161-6.
15. Miles MA, Feliciangeli MD, de Arias AR. American trypanosomiasis (Chagas disease) and the role of molecular epidemiology in guiding control strategies. BMJ 2003;326:1444-8.
16. Añez N, Crisante G, da Silva FM, Rojas A, Carrasco H, Umezawa E, et al. Predominance of lineage I among Trypanosoma cruzi isolates from Venezuelan patients with different clinical profiles of acute Chagas disease. Trop Med Int Health 2004;9:1319-26.
17. Magalhães-Santos IF, Souza MM, Lima CS, Andrade S. Infection of Calomys callosus (Rodentia Cricetidae) with strains of different Trypanosoma cruzi biodemes: Pathogenicity, histotropism, and fibrosis induction. Mem Inst Oswaldo Cruz 2004;99:407-13.
18. Mejía AM, Triana O. Análisis por LSSP-PCR de la variabilidad genética de Trypanosoma cruzi en sangre y órganos de ratones. Biomédica 2005;25:76-86.
19. Freitas JM, Lages-Silva E, Crema E, Pena SD, Macedo AM. Real time PCR strategy for the identification of major lineages of Trypanosoma cruzi directly in chronically infected human tissues. Int J
Parasitol 2005;35:411-7.
20. Bernabé C, Tibayrenc M. Trypanosoma cruzi: longterm sub-cultures in two different culture media do not confirm the existence of highly versatile multilocus genotypes. Int J Parasitol 2004;34:779-84.
21. Goldberg SS, Chiari E. Growth and isolation of single colonies of Trypanosoma cruzi on solid medium. J Parasitol 1980;66:677-9.
22. Camargo EP. Growth and differentiation in Trypanosoma cruzi. Origin of metacyclic trypanosomes in liquid media. Rev Inst Med Trop São Paulo 1964;12:93-100.
23. Miller SA, Dykes DD, Polesky HF. A simple salting out procedure for extracting DNA from human nucleated cells. Nucleic Acids Res 1988;16:1215.
24. Sturm N, Degrave W, Morel C, Simpson L. Sensitive detection and schizodeme classification of Trypanosoma cruzi cells by amplification of kinetoplast minicircle DNA sequences. Use in diagnosis of Chagas disease. Mol Biochem Parasitol 1989;33:205-14.
25. Pena SD, Barreto G, Vago AR, De Marco L, Reinach F, Dias Neto E, et al. Sequence-specific gene signatures can be obtained by PCR with single specific primers at low stringency. Proc Natl Acad Sci USA 1994;91:1946-9.
26. Campos RF, Goncalves MS, dos Reis EA, dos Reis MG, Andrade SG. Comparative analysis by polymerase chain reaction amplified minicircles of kinetoplast DNA of a stable strain of Trypanosoma cruzi
from Sao Felipe, Bahia, its clones and subclones: possibility of predominance of a principal clone in this area. Mem Inst Oswaldo Cruz 1999;94:23-9.
27. Walsh PS, Metzger DA, Higuchi R. Chelex® 100 as a medium for simple extraction of DNA for PCR-based typing from forensic material. Biotechniques 1991;10:506-13.
28. Sambrook J, Fritsch EF, Maniatis T. Molecular cloning. A laboratory manual. Second edition. Cold Spring: Harbor Laboratory Press; 1989.
29. Virreira M, Torrico F, Truyens C, Alonso-Vega C, Solano M, Carlier Y, et al. Comparison of polymerase chain reaction methods for reliable and easy detection of congenital Trypanosoma cruzi infection. Am J Trop Med Hyg 2003;68:574-82.
30. Franco DJ, Vago AR, Chiari E, Meira FC, Galvao LM, Machado CR. Trypanosoma cruzi: mixture of two populations can modify virulence and tissue tropism in rat. Exp Parasitol 2003;104:54-61.
31. Campos RM, Andrade SG. Characterization of subpopulations (clones and subclones) of the 21 SF strain of Trypanosoma cruzi after long lasting maintenance in the laboratory. Mem Inst Oswaldo Cruz
1996;91:795-800.
32. Di Noia JM, Buscaglia CA, De Marchi CR, Almeida IC, Frasch CC. A Trypanosoma cruzi small surface molecule provides the first immunological evidence that Chagas disease is due to a single parasite lineage. J Exp Med 2002;195:401-13.
33. Rios JF. Estudio del tropismo tisular de dos cepas colombianas de Trypanosoma cruzi (Tesis). Medellín: Universidad de Antioquia; 1995.
34. Camandaroba EL, Campos RF, Magalhaes JB, Andrade SG. Clonal structure of Trypanosoma cruzi Colombian strain (biodeme type III): biological, isoenzymic and histopathological analysis of seven
isolated clones. Rev Soc Bras Med Trop 2001;34:151-7.
35. Cummings KL, Tarleton RL. Rapid quantitation of Trypanosoma cruzi in host tissue by real-time PCR. Mol Biochem Parasitol 2003;129:53-9.
36. Macedo AM, Pena SD. Genetic variability of Trypanosoma cruzi: implications for the pathogenesis of Chagas disease. Parasitol Today 1998;14:119-23.
2. Moncayo A. Chagas disease: current epidemiological trends after the interruption of vectorial and transfusional transmission in the Southern Cone countries. Mem Inst Oswaldo Cruz 2003;98:577-91.
3. Lenzi HL, Oliveira DN, Lima MT, Gatas CR. Trypanosoma cruzi: paninfectivity of CL strain during murine acute infection. Exp Parasitol 1996;84:16-27.
4. Prata A. Clinical and epidemiological aspects of Chagas disease. Lancet Infect Dis 2001;1:92-100.
5. Vago AR, Macedo AM, Oliveira RP, Andrade LO, Chiari E, Galvao LM, et al. Kinetoplast DNA signatures of Trypanosoma cruzi strains obtained directly from infected tissues. Am J Pathol 1996;149:2153-9.
6. Vago AR, Andrade LO, Leite AA, d'Avila Reis D, Macedo AM, Adad SJ, et al. Genetic characterization of Trypanosoma cruzi directly from tissues of patients with chronic Chagas disease: differential distribution of genetic types into diverse organs. Am J Pathol 2000;156:1805-9.
7. Macedo AM, Machado CR, Oliveira RP, Pena SD. Trypanosoma cruzi: genetic structure of populations and relevant of genetic variability to the pathogenesis of Chagas disease. Mem Inst Oswaldo Cruz 2004;99:1-12.
8. Oliveira RP, Broude NF, Macedo AM, Cantor CR, Smith CL, Pena SD. Probing the genetic population structure of Trypanosoma cruzi with polymorphic microsatellites. Proc Natl Acad Sci USA 1998;95:3776-80.
9. Andrade LO, Machado CR, Chiari E, Pena SD, Macedo AM. Trypanosoma cruzi: role of host genetic background in the differential tissue distribution of parasite clonal populations. Exp Parasitol 2002;100:269-75.
10. Macedo AM, Martins MS, Chiari E, Pena SD. DNA fingerprinting of Trypanosoma cruzi: a new tool for characterization of strains and clones. Mol Biochem Parasitol 1992;55:147-53.
11. Tibayrenc M, Neubauer K, Barnabé C, Guerrini F, Skarecky D, Ayala FJ. Genetic characterization of six parasitic protozoa: parity betweem random-primer DNA typing and multilocus enzyme electrophoresis. Proc Natl Acad Sci USA 1993;90:1335-9.
12. Souto RP, Fernandes O, Macedo AM, Campbell DA, Zingales B. DNA markers define two major phylogenetic lineages of Trypanosoma cruzi. Mol Biochem Parasitol 1996;83:141-52.
13. Fernandes O, Souto RP, Castro JA, Pereira JB, Fernandes NC, Junqueira AC, et al. Brazilian isolates of Trypanosoma cruzi from humans and triatomines classified into two lineages using mini-exon and ribosomal RNA sequences. Am J Trop Med Hyg 1998;58:807-11.
14. Fernandes O, Mangia RH, Lisboa CV, Pinho AP, Morel CM, Zingales B, et al. The complexity of the sylvatic cycle of Trypanosoma cruzi in Rio de Janeiro state (Brazil) revealed by the non-transcribed spacer of the mini-exon gene. Parasitology 1999;118:161-6.
15. Miles MA, Feliciangeli MD, de Arias AR. American trypanosomiasis (Chagas disease) and the role of molecular epidemiology in guiding control strategies. BMJ 2003;326:1444-8.
16. Añez N, Crisante G, da Silva FM, Rojas A, Carrasco H, Umezawa E, et al. Predominance of lineage I among Trypanosoma cruzi isolates from Venezuelan patients with different clinical profiles of acute Chagas disease. Trop Med Int Health 2004;9:1319-26.
17. Magalhães-Santos IF, Souza MM, Lima CS, Andrade S. Infection of Calomys callosus (Rodentia Cricetidae) with strains of different Trypanosoma cruzi biodemes: Pathogenicity, histotropism, and fibrosis induction. Mem Inst Oswaldo Cruz 2004;99:407-13.
18. Mejía AM, Triana O. Análisis por LSSP-PCR de la variabilidad genética de Trypanosoma cruzi en sangre y órganos de ratones. Biomédica 2005;25:76-86.
19. Freitas JM, Lages-Silva E, Crema E, Pena SD, Macedo AM. Real time PCR strategy for the identification of major lineages of Trypanosoma cruzi directly in chronically infected human tissues. Int J
Parasitol 2005;35:411-7.
20. Bernabé C, Tibayrenc M. Trypanosoma cruzi: longterm sub-cultures in two different culture media do not confirm the existence of highly versatile multilocus genotypes. Int J Parasitol 2004;34:779-84.
21. Goldberg SS, Chiari E. Growth and isolation of single colonies of Trypanosoma cruzi on solid medium. J Parasitol 1980;66:677-9.
22. Camargo EP. Growth and differentiation in Trypanosoma cruzi. Origin of metacyclic trypanosomes in liquid media. Rev Inst Med Trop São Paulo 1964;12:93-100.
23. Miller SA, Dykes DD, Polesky HF. A simple salting out procedure for extracting DNA from human nucleated cells. Nucleic Acids Res 1988;16:1215.
24. Sturm N, Degrave W, Morel C, Simpson L. Sensitive detection and schizodeme classification of Trypanosoma cruzi cells by amplification of kinetoplast minicircle DNA sequences. Use in diagnosis of Chagas disease. Mol Biochem Parasitol 1989;33:205-14.
25. Pena SD, Barreto G, Vago AR, De Marco L, Reinach F, Dias Neto E, et al. Sequence-specific gene signatures can be obtained by PCR with single specific primers at low stringency. Proc Natl Acad Sci USA 1994;91:1946-9.
26. Campos RF, Goncalves MS, dos Reis EA, dos Reis MG, Andrade SG. Comparative analysis by polymerase chain reaction amplified minicircles of kinetoplast DNA of a stable strain of Trypanosoma cruzi
from Sao Felipe, Bahia, its clones and subclones: possibility of predominance of a principal clone in this area. Mem Inst Oswaldo Cruz 1999;94:23-9.
27. Walsh PS, Metzger DA, Higuchi R. Chelex® 100 as a medium for simple extraction of DNA for PCR-based typing from forensic material. Biotechniques 1991;10:506-13.
28. Sambrook J, Fritsch EF, Maniatis T. Molecular cloning. A laboratory manual. Second edition. Cold Spring: Harbor Laboratory Press; 1989.
29. Virreira M, Torrico F, Truyens C, Alonso-Vega C, Solano M, Carlier Y, et al. Comparison of polymerase chain reaction methods for reliable and easy detection of congenital Trypanosoma cruzi infection. Am J Trop Med Hyg 2003;68:574-82.
30. Franco DJ, Vago AR, Chiari E, Meira FC, Galvao LM, Machado CR. Trypanosoma cruzi: mixture of two populations can modify virulence and tissue tropism in rat. Exp Parasitol 2003;104:54-61.
31. Campos RM, Andrade SG. Characterization of subpopulations (clones and subclones) of the 21 SF strain of Trypanosoma cruzi after long lasting maintenance in the laboratory. Mem Inst Oswaldo Cruz
1996;91:795-800.
32. Di Noia JM, Buscaglia CA, De Marchi CR, Almeida IC, Frasch CC. A Trypanosoma cruzi small surface molecule provides the first immunological evidence that Chagas disease is due to a single parasite lineage. J Exp Med 2002;195:401-13.
33. Rios JF. Estudio del tropismo tisular de dos cepas colombianas de Trypanosoma cruzi (Tesis). Medellín: Universidad de Antioquia; 1995.
34. Camandaroba EL, Campos RF, Magalhaes JB, Andrade SG. Clonal structure of Trypanosoma cruzi Colombian strain (biodeme type III): biological, isoenzymic and histopathological analysis of seven
isolated clones. Rev Soc Bras Med Trop 2001;34:151-7.
35. Cummings KL, Tarleton RL. Rapid quantitation of Trypanosoma cruzi in host tissue by real-time PCR. Mol Biochem Parasitol 2003;129:53-9.
36. Macedo AM, Pena SD. Genetic variability of Trypanosoma cruzi: implications for the pathogenesis of Chagas disease. Parasitol Today 1998;14:119-23.
How to Cite
1.
Botero LA, Mejía AM, Triana O. Biological and genetic characterization of two Colombian clones of Trypanosoma cruzi groups I and II. biomedica [Internet]. 2007 Jan. 1 [cited 2024 May 19];27(1esp):64-7. Available from: https://revistabiomedica.org/index.php/biomedica/article/view/249
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