Putative markers of infective life stages in Leishmania (Viannia) braziliensis.
Gene expression is known to vary significantly during the Leishmania life-cycle. Its monitoring might allow identification of molecular changes associated with the infective stages (metacyclics and amastigotes) and contribute to the understanding of the complex host-parasite relationships. So far, very few studies have been done on Leishmania (Viannia) braziliensis, one of the most pathogenic species. Such studies require, first of all, reference molecular markers. In the present work, we applied differential display analysis (DD analysis) in order to identify transcripts that might be (i) candidate markers of metacyclics and intracellular amastigotes of L. (V.) braziliensis or (ii) potential controls, i.e. constitutively expressed. In total, 48 DNA fragments gave reliable sequencing data, 29 of them being potential markers of infective stages and 12 potential controls. Eight sequences could be identified with reported genes. Validation of the results of DD analysis was done for 4 genes (2 differentially expressed and 2 controls) by quantitative real-time PCR. The infective insect stage-specific protein (meta 1) was more expressed in metacyclic-enriched preparations. The oligopeptidase b showed a higher expression in amastigotes. Two genes, glucose-6-phosphate dehydrogenase and a serine/threonine protein kinase, were found to be similarly expressed in the different biological samples.
Gamboa
D
D
Instituto de Medicina Tropical Alexander von Humboldt, Universidad Peruana Cayetano Heredia, A.P. 4314, Lima 100, Peru.
Van Eys
G
G
Victoir
K
K
Torres
K
K
Adaui
V
V
Arevalo
J
J
Dujardin
J-C
JC
D000818:Animals; D001483:Base Sequence; D015415:Biomarkers / Q000378:metabolism; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation; D017125:Genes, Protozoan / Q000235:genetics*; D006801:Humans; D007892:Leishmania braziliensis / Q000235:genetics* / Q000378:metabolism*; D016773:Leishmaniasis, Cutaneous / Q000469:parasitology*; D008969:Molecular Sequence Data; D015203:Reproducibility of Results; D020133:Reverse Transcriptase Polymerase Chain Reaction
D015415:Biomarkers