1. Lorestan University Of Medical Sciences, Khorramabad, Iran 2. Lorestan University Of Medical Sciences, Khorramabad, Iran 3. Shahid Beheshti University Of Medical Sciences,Tehran, Iran
Leishmaniasis is a hygienic issue which the number of affected people to all kinds is surging in the world. until now there are no affordable vaccines which can be used. at economical view vaccine should be easy accessed and simple produced with no complications and can be done without advanced equipment. in cutaneous leishmaniasis in result of th1 activation, il12 will be secreted which activates macrophages to secrete interferon γ (ifnγ). ifnγ activates macrophages to eliminate phagocyted parasites and also it could activate cytotoxic responses similar to no induction and tnf α secretion in macrophages so that they kill parasites. in result of ifnγ increasing in cutaneous leishmaniasis leisure is capable of immunologic response escalation will be efficacious in parasite inhibition. the aim of this research was to provide promastigote of leishmania tarentolae (l. tarentolae) transgene including of ifnγ gene.
Ifnγ gene sequence has been determined and synthetized in expression plasmid. recombinant plasmid was confirmed by pcr and restriction enzyme. then l. tarentolae was transfected with the recombinant plasmid by electroporation and confirmed by pcr. in order to evaluate ifnγ expression in promastigote of leishmania tarentolae, rt-pcr and elisa were used.
Recombinant plasmid was well designed and approved. attachment of the gene to promastigote genome was confirmed by restriction enzyme. then expression and production of ifnγ confirmed.
Expression system of l. tarentolae was able to produce and secrete ifnγ. culture simplicity, being cheaper and the ability of glycosylation in this system are the advantages. also it can be used to for leishmanisation and help in the treatment of patients with cutaneous leishmaniasis lesion after complementary studies.