Abstract
The mode of entry and intracellular fate of epimastigotes and trypomastigotes of Trypanosoma cruzi in cultured cells was studied. Electron microscopic observations indicated the uptake by phagocytosis of both forms into mouse peritoneal macrophages and of trypomastigotes and transition forms into other cultured cell types. In each instance the organisms were initially surrounded by a plasma membrane-derived phagosome. Trypsin and chymotrypsin treatment of the macrophages completely abolished attachment and ingestion of both forms, indicating that protease-sensitive structures on the macrophage plasma membrane mediate ingestion. The macrophage Fc or C3b receptors were not essential for uptake of T. cruzi in the conditions used. Cytochalasin B inhibited ingestion but not the attachment of both forms by macrophages. Epimastigotes were not taken up by HeLa, L cells, and calf embryo fibroblasts. In macrophages, epimastigotes were killed and digested within phagolysosomes. In contrast, trypomastigotes and transition forms escaped from the phagocytic vacuole and then multiplied in the cytoplasmic matrix. Amastigotes released from infected cells exhibited properties similar to those of trypomastigotes and were able to enter all cell types studied and multiply intracellularly.
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- Akiyama H. J., McQuillen N. K. Interaction and transformation of Leishmania donovani within in vitro cultured cells. An electron microscopical study. Am J Trop Med Hyg. 1972 Nov;21(6):873–879. doi: 10.4269/ajtmh.1972.21.873. [DOI] [PubMed] [Google Scholar]
- Alexander J. Effect of the antiphagocytic agent cytochalasin B on macrophage invasion by Leishmania mexicana promastigotes and Trypanosoma cruzi epimastigotes. J Protozool. 1975 May;22(2):237–240. doi: 10.1111/j.1550-7408.1975.tb05858.x. [DOI] [PubMed] [Google Scholar]
- Allison A. C., Davies P., De Petris S. Role of contractile microfilaments in macrophage movement and endocytosis. Nat New Biol. 1971 Aug 4;232(31):153–155. doi: 10.1038/newbio232153a0. [DOI] [PubMed] [Google Scholar]
- Axline S. G., Reaven E. P. Inhibition of phagocytosis and plasma membrane mobility of the cultivated macrophage by cytochalasin B. Role of subplasmalemmal microfilaments. J Cell Biol. 1974 Sep;62(3):647–659. doi: 10.1083/jcb.62.3.647. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Behbehani M. K. Multiplication of Trypanosoma (Schizotrypanum) cruzi in mouse peritoneal macrophages. Trans R Soc Trop Med Hyg. 1971;65(1):15–15. [PubMed] [Google Scholar]
- Bianco C., Griffin F. M., Jr, Silverstein S. C. Studies of the macrophage complement receptor. Alteration of receptor function upon macrophage activation. J Exp Med. 1975 Jun 1;141(6):1278–1290. doi: 10.1084/jem.141.6.1278. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Blanden R. V. Modification of macrophage function. J Reticuloendothel Soc. 1968 Jun;5(3):179–202. [PubMed] [Google Scholar]
- COHN Z. A., BENSON B. THE DIFFERENTIATION OF MONONUCLEAR PHAGOCYTES. MORPHOLOGY, CYTOCHEMISTRY, AND BIOCHEMISTRY. J Exp Med. 1965 Jan 1;121:153–170. doi: 10.1084/jem.121.1.153. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Dales S. Early events in cell-animal virus interactions. Bacteriol Rev. 1973 Jun;37(2):103–135. doi: 10.1128/br.37.2.103-135.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Dvorak J. A., Hyde T. P. Trypanosoma cruzi: interaction with vertebrate cells in vitro. 1. Individual interactions at the cellular and subcellular levels. Exp Parasitol. 1973 Oct;34(2):268–283. doi: 10.1016/0014-4894(73)90087-8. [DOI] [PubMed] [Google Scholar]
- Dvorak J. A., Schmunis G. A. Trypanosoma cruzi: interaction with mouse peritoneal macrophages. Exp Parasitol. 1972 Oct;32(2):289–300. doi: 10.1016/0014-4894(72)90036-7. [DOI] [PubMed] [Google Scholar]
- Edelson P. J., Zwiebel R., Cohn Z. A. The pinocytic rate of activated macrophages. J Exp Med. 1975 Nov 1;142(5):1150–1164. doi: 10.1084/jem.142.5.1150. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Griffin F. M., Jr, Bianco C., Silverstein S. C. Characterization of the macrophage receptro for complement and demonstration of its functional independence from the receptor for the Fc portion of immunoglobulin G. J Exp Med. 1975 Jun 1;141(6):1269–1277. doi: 10.1084/jem.141.6.1269. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hirsch J. G., Fedorko M. E. Ultrastructure of human leukocytes after simultaneous fixation with glutaraldehyde and osmium tetroxide and "postfixation" in uranyl acetate. J Cell Biol. 1968 Sep;38(3):615–627. doi: 10.1083/jcb.38.3.615. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hoff R. Killing in vitro of Trypanosoma cruzi by macrophages from mice immunized with T. cruzi or BCG, and absence of cross-immunity on challege in vivo. J Exp Med. 1975 Aug 1;142(2):299–311. doi: 10.1084/jem.142.2.299. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Holland P., Holland N. H., Cohn Z. A. The selective inhibition of macrophage phagocytic receptors by anti-membrane antibodies. J Exp Med. 1972 Mar 1;135(3):458–475. doi: 10.1084/jem.135.3.458. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hyde T. P., Dvorak J. A. Trypanosoma cruzi: interaction with vertebrate cells in vitro. 2. Quantitative analysis of the penetration phase. Exp Parasitol. 1973 Oct;34(2):284–294. doi: 10.1016/0014-4894(73)90088-x. [DOI] [PubMed] [Google Scholar]
- Jones T. C., Hirsch J. G. The interaction between Toxoplasma gondii and mammalian cells. II. The absence of lysosomal fusion with phagocytic vacuoles containing living parasites. J Exp Med. 1972 Nov 1;136(5):1173–1194. doi: 10.1084/jem.136.5.1173. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Jones T. C., Yeh S., Hirsch J. G. The interaction between Toxoplasma gondii and mammalian cells. I. Mechanism of entry and intracellular fate of the parasite. J Exp Med. 1972 Nov 1;136(5):1157–1172. doi: 10.1084/jem.136.5.1157. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kierszenbaum F., Knecht E., Budzko D. B., Pizzimenti M. C. Phagocytosis: a defense mechanism against infection with Trypanosoma cruzi. J Immunol. 1974 May;112(5):1839–1844. [PubMed] [Google Scholar]
- 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]
- Ladda R., Aikawa M., Sprinz H. Penetration of erythrocytes by merozoites of mammalian and avian malarial parasites. J Parasitol. 1969 Jun;55(3):633–644. [PubMed] [Google Scholar]
- Lay W. H., Nussenzweig V. Receptors for complement of leukocytes. J Exp Med. 1968 Nov 1;128(5):991–1009. doi: 10.1084/jem.128.5.991. [DOI] [PMC free article] [PubMed] [Google Scholar]
- MACKANESS G. B. THE IMMUNOLOGICAL BASIS OF ACQUIRED CELLULAR RESISTANCE. J Exp Med. 1964 Jul 1;120:105–120. doi: 10.1084/jem.120.1.105. [DOI] [PMC free article] [PubMed] [Google Scholar]
- MEYER H., DE OLIVEIRA MUSACCHIO M., DE ANDRADE MENDONCA I. Electron microscopic study of Trypanosoma cruzi in thin sections of infected tissue cultures and of blood-agar forms. Parasitology. 1958 May;48(1-2):1–8. doi: 10.1017/s0031182000021028. [DOI] [PubMed] [Google Scholar]
- Milder R. V., Kloetzel J., Deane M. P. Observation of the interaction of peritoneal macrophages with Trypanosoma cruzi. I. Initial phase of the relationship with blood stream and culture forms in vitro. Rev Inst Med Trop Sao Paulo. 1973 Nov-Dec;15(6):386–392. [PubMed] [Google Scholar]
- Nogueira N., Bianco C., Cohn Z. Studies on the selective lysis and purification of Trypanosoma cruzi. J Exp Med. 1975 Jul 1;142(1):224–229. doi: 10.1084/jem.142.1.224. [DOI] [PMC free article] [PubMed] [Google Scholar]
- North R. J. Cellular kinetics associated with the development of acquired cellular resistance. J Exp Med. 1969 Aug 1;130(2):299–314. doi: 10.1084/jem.130.2.299. [DOI] [PMC free article] [PubMed] [Google Scholar]
- PIZZI T., RUBIO M. Aspectos celulares de la immunidad en la enfermedad de Chagas. Bol Chil Parasitol. 1955 Jan-Mar;10(1):4–9. [PubMed] [Google Scholar]
- PIZZI T., RUBIO M., KNIERIM F. Immunologäa de la enfermedad de Chagas. Bol Chil Parasitol. 1954 Apr-Jun;9(2):35–47. [PubMed] [Google Scholar]
- Rabinovitch M. Attachment of modified erythrocytes to phagocytic cells in absence of serum. Proc Soc Exp Biol Med. 1967 Feb;124(2):396–399. doi: 10.3181/00379727-124-31749. [DOI] [PubMed] [Google Scholar]
- Rodriguez E., Marinkelle C. J. Trypanosoma cruzi: development in tissue culture. Exp Parasitol. 1970 Feb;27(1):78–87. doi: 10.1016/s0014-4894(70)80012-1. [DOI] [PubMed] [Google Scholar]
- SANABRIA A. ULTRASTRUCTURE OF TRYPANOSOMA CRUZI IN MOUSE MYOCARDIUM. I. TRYPANOSOME FORM. Exp Parasitol. 1963 Aug;14:81–91. doi: 10.1016/0014-4894(63)90013-4. [DOI] [PubMed] [Google Scholar]
- SANABRIA A. ULTRASTRUCTURE OF TRYPANOSOMA CRUZI IN MOUSE MYOCARDIUM. II. CRITHIDIAL AND LEISHMANIAL FORMS. Exp Parasitol. 1964 Apr;15:125–137. doi: 10.1016/0014-4894(64)90013-x. [DOI] [PubMed] [Google Scholar]
- Sanabria A. Ultrastructure of Trypanosoma cruzi in mouse liver. Exp Parasitol. 1971 Oct;30(2):187–198. doi: 10.1016/0014-4894(71)90083-x. [DOI] [PubMed] [Google Scholar]
- Scorza C., Scorza J. V. The role of inflammatory macrophages in experimental acute chagasic myocarditis. J Reticuloendothel Soc. 1972 Jun;11(6):604–616. [PubMed] [Google Scholar]
- Silverstein S. Macrophages and viral immunity. Semin Hematol. 1970 Apr;7(2):185–214. [PubMed] [Google Scholar]
- Sooksri V., Inoki S. Electron microscopic studies on penetration and development of Trypanosoma cruzi in HeLa cells. Biken J. 1972 Sep;15(3):179–191. [PubMed] [Google Scholar]
- TOBIE E. J., VON BRAND T., MEHLMAN B. Cultural and physiological observations on Trypanosoma rhodesiense and Trypanosoma gambiense. J Parasitol. 1950 Feb;36(1):48–54. [PubMed] [Google Scholar]
- 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]
- Unkeless J. C., Gordon S., Reich E. Secretion of plasminogen activator by stimulated macrophages. J Exp Med. 1974 Apr 1;139(4):834–850. doi: 10.1084/jem.139.4.834. [DOI] [PMC free article] [PubMed] [Google Scholar]