For treating infection in BALB/c mice, we used thalidomide in conjunction

For treating infection in BALB/c mice, we used thalidomide in conjunction with glucantime. mm, respectively). Cytokine study showed that the effect of thalidomide was not K02288 tyrosianse inhibitor dependent on IL-12; however, it up-regulated IFN- and down-regulated IL-10 production. Conclusively, thalidomide seems promising as a conjunctive therapy with antimony in murine model of visceral leishmaniasis. infection (Fu et al., 2002). The effects of combination therapy with thalidomide and leishmaniacidal drug, glucantime, on the course of disease induced by infection in BALB/c mice are unknown. It has been proposed that visceral leishmaniasis induced in BALB/c mice by the otherwise “dermatotropic” may be a better model of human visceral leishmaniasis (Handman et K02288 tyrosianse inhibitor al., 2001). We chose this model to investigate the effect of thalidomide on the course of visceral leishmaniasis induced by in BALB/c mice. Patients suffering from visceral leishmaniasis come to clinic centers when the disease completely develops. Therefore, in the present study, the infected mice were treated 2 weeks after challenge to see whether thalidomide would affect the course of already established disease. The treatments were performed for 12 days, and the progression of disease was monitored based on measuring foot pad swellings. MATERIALS AND K02288 tyrosianse inhibitor METHODS Parasites promastigotes, MHROM/IR/75/ER, were grown in Schneider’s medium supplemented with 10% heat-inactivated FBS, 292 g/ml L-glutamine, 4.5 mg/ml glucose, 100 g/ml streptomycin and 100 IU/ml penicilin at 23-25 as previously described (Alimohammadian et al., 2002). The parasites were kept within a virulent condition by regular passing in prone BALB/c Rabbit Polyclonal to PTTG mice. Fixed stage promastigotes had been centrifuged and harvested at 3,000 rpm for 10 min at 4. The pellet was cleaned three times in PBS (8 mM Na2HPO4, 1.75 mM KH2PO4. 0.25 mM KCl, 0.137 mM NaCl). Mice Feminine BALB/c mice (4-6 weeks outdated) had been obtained from the pet Breeding Stock Service of Pasteur Institute of Iran, Karaj, Iran. The mice had been split into six groupings (8 mice per group). The combined groups included Group C; control mice non-treated and non-infected, Group P; contaminated but non-treated, Goup OP; contaminated treated with sessami essential oil (carrier of thalidomide), Group TP; contaminated treated with in sessami essential oil thalidomide, Group GP; contaminated treated with glucantime, Group TGP; contaminated treated with glucantime and thalidomide. Infections and disease advancement The contaminated sets of mice received 5 103 infective fixed stage promastigotes in the hind feet pad subcutaneously (s.c). Every whole week after problem the feet pad swelling was measured utilizing a caliper. The info are portrayed in products (1 device = 10-2 cm of the web increase in feet pad thickness). Medications administration Mice treated with glucantime (Marial, France) had been received 100 mg/kg/time i.p. Mice treated with thalidomide (a sort present from Laphal Laboratories, France) had been received 30 mg/kg/time orally developed in sessami essential oil. K02288 tyrosianse inhibitor The medication administration was began 14 days after challenge. This era of time will do for establishment from the infections. Therefore, administration of medications was continuing for 12 times. Cell lifestyle condition for cytokine assay The draining lymph nodes (popliteal lymph nodes) had been taken out 4 and 7 weeks after problem and total draining lymph node cells had been isolated and cleaned three times with sterile PBS. The cells had been resuspended in RPMI supplemented with 2 mM/L-glutamine, 100 U/ml penicillin, 1 mg/ml streptomycin (all from Sigma) and 10% individual heat-inactivated fetal leg serum. Cells, 2 106 in amount in 1 ml of lifestyle medium had been plated on flat-bottomed 24-well plates with or without Phorbol Myristate Acetate (PMA, 50 ng/ml tissues culture moderate)/Ionomycin (1 M). The supernatants had been taken out after 24 hr of incubation and kept at -20. We examined cytokine creation by particular IL-10, IL-12 and IFN- enzyme-linked immunosorbent assay (ELISA) (Bender Med Systems, Austria) using a awareness of 10 pg/ml. Statistical evaluation Statistical significance between groupings was analyzed by student’s 0.05 were considered significant statistically. RESULTS Development of disease Fig. 1 shows the disease progression in different groups of mice. At the end of drug administration (4th week), there was no difference between groups of infected mice P and OP in terms of foot.