Because of this close association between chemotherapy and cell-mediated immunity, treatment for L. donovani infection has been thought to be more amenable to combined therapy, that is, immunochemotherapy [16]. Therefore, we tested immunochemotherapy to determine the safety, immunogenicity and probable curative potential of 78 kDa antigen in combination with a newly tested drug cisplatin in mice infected with L. donovani. The current MK-1775 research buy study is expected to assist in the evaluation of immunochemotherapy as a better alternative antileishmanial therapy. Promastigotes of L. donovani, strain MHOM/IN/80/Dd8, were grown at 22°C in NNN medium

supplemented with MEM (pH 7·2), 200U of streptomycin, 200U of benzyl penicillin and 40 μg of gentamycin per mL and subcultured in

the same medium after every 48–72 h. Inbred BALB/c mice of either sex weighing 20–25 g were used for the present study. During the start of the experiment, the mice weigh around 20–25 g, but by the time, infection was given and treatment was completed weight increased to 25–30 g. These animals were obtained from Institute of Microbial Technology, Chandigarh, India, and then maintained in the Central Animal House, Panjab University, Chandigarh. All the mice were kept in appropriate cages and fed with water and food ad libitum throughout the study period. The ethical clearance for conducting various experiments on BALB/c mice was taken from Institutional Animal Ethics Committee (IAEC) of the Panjab check details University, Chandigarh. Cis-diamminedichloroplatinum (II) dichloride (CP) was purchased from Sigma-Aldrich Co. (St. Louis, MO, USA) in the pure form, and then it was dissolved in distilled water to get the

requisite concentration of 0·5 mg/kg body wt [14]. The 78 kDa antigen of L. donovani was identified and eluted as described by Nagill and Kaur [6]. The 78 kDa antigen alone (without any adjuvant) was also used as a vaccine candidate for immunization. 78 kDa + MPL-A vaccine was prepared by the addition pentoxifylline of 144 μL solution of MPL-A (conc. 10 mg/mL) to 360 μg of 78 kDa antigen. Subcutaneous route was used for immunization of mice in all the groups [6]. Mice were infected intracardially with 107 promastigotes/0·1 mL [14]. Animals were divided into different groups, and each group consisted of eighteen mice. Animals of Group 1 (Chemotherapy) received intraperitoneal injection of cisplatin at a dose of 0·5 mg/kg body wt. continuously for 5 days in two cycles with an interval of 14 days between each cycle, while Group 2 (cisplatin + 78 kDa) and Group 3 (cisplatin + 78 kDa + MPL-A) received immunochemotherapy, respectively.