Development of live vaccine used transgenic Leishmania parasite

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Leishmaniasis is often a zoonosis in which dog is known to be one of the most important animal reservoirs. For control of such zoonotic deseases, development of a vaccine directed against the natural reservoir host is desirable. In this study, we successfully produced, by molecular engineering, Leishmania amazonensis promastigotes expressing biologically active canine interleukin-8 (cIL-8). The PCR products of cIL-8 cDNA were inserted into the L.amazonensis expression vector (p6.5) which was designed to contain a tunicanycin (TM) resistance gene for selection. The constructed expression vector (p6.5-cIL-8) was transfected into promastigotes of a L.amazonensis clone (C_1/12) by electroporation. Transfectants were selected in culture madium with TM.By PCR of extrachromosomal DNAs from the transfectants using cIL-8 specific primers, a predicted product of a 330 bp long was amplified. Western blot analysis of the trnsfectants demonstrated a peptide of about 8.8kDa in molecular size, as expected of cIL-8. The culture supernatant of the transfectant showed a chemotactic activity with dog and mouse polymorphonuclear cells (PMN).