Leishmania (Viannia) braziliensis: Influence of successive in vitro cultivation on the expression of promastigote proteinases

Rebello, K. M. et al. (2010) Leishmania (Viannia) braziliensis: Influence of successive in vitro cultivation on the expression of promastigote proteinases. Experimental Parasitology, 126(4), pp. 570-576. (doi: 10.1016/j.exppara.2010.06.009) (PMID:20553928)

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Abstract

Cysteine proteinases are an important virulence factor in Leishmania parasites. In this study we analyzed the cysteine proteinase expression of infective Leishmania (Viannia) braziliensis promastigotes, examining the expression induced by successive in vitro passages in culture. We observed that this parasite presents a decrease in its virulence over BALB/c macrophages, after successive passages in culture, but still they present proteinase activity, being capable of hydrolyzing the substrate pGlu-Phe-Leu-p Nitroanilide at pH 7.0. This proteinase activity also decreases in the course of the successive passages. Additionally, the decrease in the amount of CPB proteins following successive passages of promastigotes was verified by immunoblotting assays, using an anti-CPB antiserum. Real-time PCR assays were performed to assess the relative cpb expression when compared to a housekeeping gene in promastigote cDNA preparations from the first, fourth and seventh passages. Interestingly, the data indicate a relative increase in cpb gene transcripts as the promastigotes were maintained under in vitro culture: 2.2 times higher for fourth and 2.7 times higher for seventh passages when compared to the first passage. Thus, the information gathered here shows that the expression of cysteine proteinases is modified during in vitro cultivation of L. (V.) braziliensis promastigotes.

Item Type:Articles
Status:Published
Refereed:Yes
Glasgow Author(s) Enlighten ID:Otto, Dr Thomas
Authors: Rebello, K. M., Britto, C., Pereira, B. A. S., Pita-Pereira, D. d., Moraes, M. O., Ferreira, A. B. R., Cysne-Finkelstein, L., Otto, T. D., Côrtes, L. M. d. C., da-Silva, G. G., and Alves, C. R.
College/School:College of Medical Veterinary and Life Sciences > Institute of Infection Immunity and Inflammation
Journal Name:Experimental Parasitology
Publisher:Elsevier
ISSN:1090-2449
ISSN (Online):0014-4894
Published Online:08 June 2010

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