Leptosome-entrapped leptospiral antigens conferred significant higher levels of protection than those entrapped with PC-liposomes in a hamster model

We prepared novel liposomes from total polar lipids of non-pathogenic Leptospira biflexa serovar Potac (designated leptosomes) and evaluated their vaccine delivery/adjuvant potential with novel protective antigens (Lp0607, Lp1118 and Lp1454) of L. interrogans serovar Pomona in a hamster model. The immune response induced by three individual antigens and protective efficacy were evaluated and compared to those induced by same antigens entrapped with PC-liposomes and E. coli lipid liposomes (escheriosomes). Four-week-old hamsters were immunized subcutaneously twice at a 3-week interval, bled at various time points to evaluate antibody response and sacrificed to isolate splenocytes for lymphocyte proliferation and cytokine profiles in response to recall antigen. For the challenge test, 10× MLD50 (modified lethal dose 50%) of virulent L. interrogans serovar Pomona were administered intraperitoneally. Our results demonstrate that leptosome are better adjuvant than PC-liposomes as revealed by enhanced long term antibody response, lymphocyte proliferation and significant enhancement of both Th1 (IFN-γ) and Th2 (IL-4 and IL-10) cytokines. Additionally, leptosomes and escheriosomes induced significantly higher level of memory responses than PC-liposome did. Moreover, the novel leptosomal vaccine induced significantly higher levels of protection than those prepared with PC-liposomes as revealed by enhanced survival, reduced histopathological lesions in vital organs and reduced leptospiral load in kidneys. Taken together, the results of the present study clearly reveal that both leptosomes and escheriosomes have emerged as promising delivery vehicles/adjuvants that can be widely exploited with newly discovered antigens in future leptospira vaccines.