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. 1990 Feb 1;171(2):401–413. doi: 10.1084/jem.171.2.401

The exit of Trypanosoma cruzi from the phagosome is inhibited by raising the pH of acidic compartments

PMCID: PMC2187728  PMID: 2406362

Abstract

The protozoan parasite Trypanosoma cruzi can infect many distinct mammalian cell types. The parasites enter cells through the formation of phagocytic vacuoles, but later are found free in the cytosol, where they multiply as amastigotes. Using transmission electron microscopy we found that within 2 h after infection 70% of the parasites, including examples of both mammalian forms (trypomastigotes and amastigotes), were inside partially disrupted vacuoles or free in the cytosol. We demonstrated that the pH of vacuoles containing recently interiorized parasites is acidic, through immunocytochemical localization of the acidotropic compound DAMP (18) in their interior. Increasing the vacuolar pH with chloroquine, ammonium chloride, methylamine, or monensin significantly inhibited the escape of the parasites into the cytosol. These results are compatible with the hypothesis that an acid- active hemolysin of T. cruzi (15) might be involved in the escape mechanism.

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Selected References

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  1. Anderson R. G., Falck J. R., Goldstein J. L., Brown M. S. Visualization of acidic organelles in intact cells by electron microscopy. Proc Natl Acad Sci U S A. 1984 Aug;81(15):4838–4842. doi: 10.1073/pnas.81.15.4838. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Anderson R. G., Pathak R. K. Vesicles and cisternae in the trans Golgi apparatus of human fibroblasts are acidic compartments. Cell. 1985 Mar;40(3):635–643. doi: 10.1016/0092-8674(85)90212-0. [DOI] [PubMed] [Google Scholar]
  3. Andrews N. W., Hong K. S., Robbins E. S., Nussenzweig V. Stage-specific surface antigens expressed during the morphogenesis of vertebrate forms of Trypanosoma cruzi. Exp Parasitol. 1987 Dec;64(3):474–484. doi: 10.1016/0014-4894(87)90062-2. [DOI] [PubMed] [Google Scholar]
  4. Andrews N. W., Robbins E. S., Ley V., Hong K. S., Nussenzweig V. Developmentally regulated, phospholipase C-mediated release of the major surface glycoprotein of amastigotes of Trypanosoma cruzi. J Exp Med. 1988 Feb 1;167(2):300–314. doi: 10.1084/jem.167.2.300. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Andrews N. W., Whitlow M. B. Secretion by Trypanosoma cruzi of a hemolysin active at low pH. Mol Biochem Parasitol. 1989 Mar 15;33(3):249–256. doi: 10.1016/0166-6851(89)90086-8. [DOI] [PubMed] [Google Scholar]
  6. Brener Z. Biology of Trypanosoma cruzi. Annu Rev Microbiol. 1973;27:347–382. doi: 10.1146/annurev.mi.27.100173.002023. [DOI] [PubMed] [Google Scholar]
  7. Diment S., Leech M. S., Stahl P. D. Generation of macrophage variants with 5-azacytidine: selection for mannose receptor expression. J Leukoc Biol. 1987 Nov;42(5):485–490. doi: 10.1002/jlb.42.5.485. [DOI] [PubMed] [Google Scholar]
  8. Gonzalez A., Prediger E., Huecas M. E., Nogueira N., Lizardi P. M. Minichromosomal repetitive DNA in Trypanosoma cruzi: its use in a high-sensitivity parasite detection assay. Proc Natl Acad Sci U S A. 1984 Jun;81(11):3356–3360. doi: 10.1073/pnas.81.11.3356. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Heuser J. Changes in lysosome shape and distribution correlated with changes in cytoplasmic pH. J Cell Biol. 1989 Mar;108(3):855–864. doi: 10.1083/jcb.108.3.855. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Horwitz M. A., Maxfield F. R. Legionella pneumophila inhibits acidification of its phagosome in human monocytes. J Cell Biol. 1984 Dec;99(6):1936–1943. doi: 10.1083/jcb.99.6.1936. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Kress Y., Bloom B. R., Wittner M., Rowen A., Tanowitz H. Resistance of Trypanosoma cruzi to killing by macrophages. Nature. 1975 Oct 2;257(5525):394–396. doi: 10.1038/257394a0. [DOI] [PubMed] [Google Scholar]
  12. Ley V., Andrews N. W., Robbins E. S., Nussenzweig V. Amastigotes of Trypanosoma cruzi sustain an infective cycle in mammalian cells. J Exp Med. 1988 Aug 1;168(2):649–659. doi: 10.1084/jem.168.2.649. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Martínez-Silva R., López V. A., Colón J. I., Chiriboga J. Isolation of Trypanosoma cruzi from blood of acutely and chronically infected mice in tissue culture. Am J Trop Med Hyg. 1969 Nov;18(6):878–884. doi: 10.4269/ajtmh.1969.18.878. [DOI] [PubMed] [Google Scholar]
  14. Maxfield F. R. Weak bases and ionophores rapidly and reversibly raise the pH of endocytic vesicles in cultured mouse fibroblasts. J Cell Biol. 1982 Nov;95(2 Pt 1):676–681. doi: 10.1083/jcb.95.2.676. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Meirelles M. N., de Araujo-Jorge T. C., Miranda C. F., de Souza W., Barbosa H. S. Interaction of Trypanosoma cruzi with heart muscle cells: ultrastructural and cytochemical analysis of endocytic vacuole formation and effect upon myogenesis in vitro. Eur J Cell Biol. 1986 Aug;41(2):198–206. [PubMed] [Google Scholar]
  16. Milder R., Kloetzel J. The development of Trypanosoma cruzi in macrophages in vitro. Interaction with lysosomes and host cell fate. Parasitology. 1980 Feb;80(1):139–145. doi: 10.1017/s0031182000000597. [DOI] [PubMed] [Google Scholar]
  17. Murphy R. F., Powers S., Cantor C. R. Endosome pH measured in single cells by dual fluorescence flow cytometry: rapid acidification of insulin to pH 6. J Cell Biol. 1984 May;98(5):1757–1762. doi: 10.1083/jcb.98.5.1757. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Nogueira N., Cohn Z. Trypanosoma cruzi: mechanism of entry and intracellular fate in mammalian cells. J Exp Med. 1976 Jun 1;143(6):1402–1420. doi: 10.1084/jem.143.6.1402. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Pipkin A. C., Sr Transmission of Trypanosoma cruzi by arthropod vectors: anterior versus posterior route infection. Int Rev Trop Med. 1969;3:1–47. [PubMed] [Google Scholar]
  20. Poole B., Ohkuma S. Effect of weak bases on the intralysosomal pH in mouse peritoneal macrophages. J Cell Biol. 1981 Sep;90(3):665–669. doi: 10.1083/jcb.90.3.665. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Tanasugarn L., McNeil P., Reynolds G. T., Taylor D. L. Microspectrofluorometry by digital image processing: measurement of cytoplasmic pH. J Cell Biol. 1984 Feb;98(2):717–724. doi: 10.1083/jcb.98.2.717. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Tanowitz H., Wittner M., Kress Y., Bloom B. Studies of in vitro infection by Trypanosoma cruzi. I. Ultrastructural studies on the invasion of macrophages and L-cells. Am J Trop Med Hyg. 1975 Jan;24(1):25–33. doi: 10.4269/ajtmh.1975.24.25. [DOI] [PubMed] [Google Scholar]
  23. Teixeira M. M., Yoshida N. Stage-specific surface antigens of metacyclic trypomastigotes of Trypanosoma cruzi identified by monoclonal antibodies. Mol Biochem Parasitol. 1986 Mar;18(3):271–282. doi: 10.1016/0166-6851(86)90085-x. [DOI] [PubMed] [Google Scholar]
  24. Vale R. D. Intracellular transport using microtubule-based motors. Annu Rev Cell Biol. 1987;3:347–378. doi: 10.1146/annurev.cb.03.110187.002023. [DOI] [PubMed] [Google Scholar]
  25. de Carvalho T. M., de Souza W. Early events related with the behaviour of Trypanosoma cruzi within an endocytic vacuole in mouse peritoneal macrophages. Cell Struct Funct. 1989 Aug;14(4):383–392. doi: 10.1247/csf.14.383. [DOI] [PubMed] [Google Scholar]

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