Objective:To elucidate immunoprophylactic potential of recombinant Wuchereria bancrofti(W.bancrofti)cuticular collagen(COL-4)in BALB/c mice and filarial clinical samples.Methods:col-4 gene was PCR amplified from W.ban...Objective:To elucidate immunoprophylactic potential of recombinant Wuchereria bancrofti(W.bancrofti)cuticular collagen(COL-4)in BALB/c mice and filarial clinical samples.Methods:col-4 gene was PCR amplified from W.bancrofti L3 cDNA library and cloned in pRSET B vector.Recombinant COL-4 was over expressed in salt inducible system and was purified by nickel afiinity chromatography.Humoral and cellular responses were measured by EUSA and peripheral blood mononuclear cells(PBMC)of various filarial clinical samples respectively using purified recombinant COL-4 antigen.Then the protective immune responses of COL-4 immunized BALB/c mice were characterized.Results:Sequence analysis of COL-4 with human host proteins reveals lack of homology.The recombinant COL-4 was found to be at 15 kDa fusion protein.The affinity purified COL-4 showed significant reactivity with putatively immune sere and in a similar fashion it demonstrated marked proliferation in PBMC samples.Immunization studies in experimental filarial host(mice)elicited significant liters with protective antibody isotype profile(IgM and IgG).Cellular immune responses were also significant in terms of splenocytes proliferation assay on mice samples.Conclusions:Our immunological findings in experimental host suggest Th2mediated immune response.Hence,we propose that W.bancrofti COL-4 could be an efficacious vaccine candidate against lymphatic filariasis.展开更多
基金supported by Department of Biotechnology(DBT),(Grant No:BT/01/COE/06/13)New Delhi.Government of India
文摘Objective:To elucidate immunoprophylactic potential of recombinant Wuchereria bancrofti(W.bancrofti)cuticular collagen(COL-4)in BALB/c mice and filarial clinical samples.Methods:col-4 gene was PCR amplified from W.bancrofti L3 cDNA library and cloned in pRSET B vector.Recombinant COL-4 was over expressed in salt inducible system and was purified by nickel afiinity chromatography.Humoral and cellular responses were measured by EUSA and peripheral blood mononuclear cells(PBMC)of various filarial clinical samples respectively using purified recombinant COL-4 antigen.Then the protective immune responses of COL-4 immunized BALB/c mice were characterized.Results:Sequence analysis of COL-4 with human host proteins reveals lack of homology.The recombinant COL-4 was found to be at 15 kDa fusion protein.The affinity purified COL-4 showed significant reactivity with putatively immune sere and in a similar fashion it demonstrated marked proliferation in PBMC samples.Immunization studies in experimental filarial host(mice)elicited significant liters with protective antibody isotype profile(IgM and IgG).Cellular immune responses were also significant in terms of splenocytes proliferation assay on mice samples.Conclusions:Our immunological findings in experimental host suggest Th2mediated immune response.Hence,we propose that W.bancrofti COL-4 could be an efficacious vaccine candidate against lymphatic filariasis.
