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The induction of Trypanosoma cruzi trypomastigote to amastigote transformation by low pH

Published online by Cambridge University Press:  06 April 2009

S. Tomlinson
Affiliation:
Michael Heidelberger Division of Immunology, Department of Pathology
F. Vandekerckhove
Affiliation:
Michael Heidelberger Division of Immunology, Department of Pathology
U. Frevert
Affiliation:
Department of Parasitology, New York University Medical Center, 550 First Avenue, New York, NY 10016, USA
V. Nussenzweig
Affiliation:
Michael Heidelberger Division of Immunology, Department of Pathology

Summary

Following cell invasion, Trypanosoma cruzi trypomastigotes transform into amastigotes, which are the mammalian replicative forms of the parasite. Although amastigotes represent a critical stage in the life-cycle of T. cruzi, little is known of the factors controlling trypomastigote to amastigote transformation. Kanbera et al. (1990) observed that exposure of trypomastigotes to acidic pH induced their transformation into rounded forms resembling amastigotes. We confirm their observation and, using two strains of T. cruzi, establish that these transformants are ultrastructurally and biochemically indistinguishable from natural amastigotes. Incubation of trypomastigotes in medium at pH 5·0 for 2 h was sufficient to trigger their transformation into forms resembling amastigotes. Electron microscopical analysis confirmed that the kinetoplast structure, and general morphological features of the acid-induced, extracellular amastigotes were indistinguishable from those of intracellular-derived amastigotes. The extracellular transformation was accompanied by the acquisition of the stage-specific surface antigen of the naturally transformed amastigotes (Ssp-4), and loss of a stagespecific trypomastigote antigen (Ssp-3). Trypomastigotes incubated at neutral pH did not transform into amastigotes, and did not acquire the Ssp-4 epitope or lose the Ssp-3 epitope. Finally, acid-induced amastigotes subsequently incorporated [3H]thymidine into their DNA, indicating that the important replicative property of intracellular amastigotes is also exhibited by these in vitro transformants. This effect of low pH appears to be of physiological relevance, and acid-induced extracellular transformation appears to represent a valid experimental technique for studies of the molecular mechanisms involved in the differentiation process.

Type
Research Article
Copyright
Copyright © Cambridge University Press 1995

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