Enolase has been reported as vaccine candidate in Plasmodium sp and other microorganisms such as Candida albicans, Chlamydia pneumonia and Streptococcus sp. There have been reports regarding the presence of anti-enolase antibodies among malaria patients and prophylactic potential of recombinant P. falciparum enolase in mice against a challenge with a lethal strain of P. yoelii suggested that enolase is a potential Estradiol Benzoate protective antigen. Since there has been no report of enolase as well as aldolase as 
vaccine candidates in case of Leishmania, it was pertinent to assess their immunogenic and prophylactic potential against VL- a fatal infection. Therefore, in the present study we undertook molecular cloning and characterization of enolase as well as aldolase, in vitro cellular responses of both proteins using lymphocytes/PBMCs of cured Leishmania hamsters/patients and evaluation of prophylactic efficacy against L. donovani infection in golden hamster, since this is the perfect experimental model for VL as it closely mimics the situation as found in human VL bioinformatic approach for predicting T-cell epitopes in both the proteins. Similarly, FBA has also been found to be protective in Onchocerca volvulus, Fasciola hepatica and Schistosoma mansoni. As these proteins have not been evaluated as vaccine candidates in case of VL, it was pertinent to assess these proteins for their immunogenicity and prophylactic potential against VL. For this, we first cloned, overexpressed and purified the proteins to homogeneity. The antibody generated against rLdEno and rLdAld in rabbit were observed to be specific to Leishmania and Atropine sulfate detected single band against whole cell lysate and SLD of Leishmania promastigotes. Both the proteins were purified in native conditions and were enzymatically active as determined by studying their kinetic parameters which were similar to that observed earlier. In-silico sequence and structural comparison of LdAld and LdEno with human counterpart exposed key differences in the active site of both glycolytic enzymes. These differences may provide platform to design specific inhibitors. A major factor contributing to healing in leishmaniasis is the development of strong cell mediated immune response. The measures of cell-mediated immunity are Leishmania specific lymphoproliferation and the stimulation of T-cells to produce macrophage activating factor, including IFN-c which in turn activate macrophages to kill the intracellular parasites. It has been reported earlier that upon stimulation with SLD and its subfractions, a T-cell response develops in cells from exposed or individuals infected with Leishmania and cured using anti-leishmanials. Further, it is well established that recovery from Leishmania infection, relies on induction of Th1 response with production of IFN-c, IL-12 and enhanced expression of nitric oxide synthase. In the absence of cytokine reagents for hamsters, nitric oxide assay was used to indirectly estimate the IFN-c response, as NO is up regulated by IFN-c. Therefore, when assessed for their immunogenicity in vitro, rLdAld gave significantly higher LTT response than rLdEno while the NO response against cured hamsters in comparison to normal and infected ones was almost 2�C3 times enhanced than that observed in case of SLD and was better in case of rLdEno than rLdAld. The generation of NO supports the up-regulation of iNOS by Th1 cellassociated cytokines. The characterization of the cellular immune response was first performed in cured Leishmania infected hamsters and then the responses of both the proteins were validated in endemic non immune donors and in immune patients of VL that were cured with amphotericin B.