Effect of promoters on cellular immune response induced by recombinant fowlpox virus expressing multi-epitope polypeptides from HIV-1.
HIV stimulates strong immune CD8(+) cytotoxic T lymphocytes (CTL) response in infected people, despite causing an immunodeficiency. It has been demonstrated that this response could be very important for the control of the virus. We have shown previously that a recombinant fowlpox virus (rFWPV), expressing the multi-epitope polypeptide (MEP) from HIV-1 TAB9, induces strong and protective Th1 and CTL responses in Balb/c mice. Here, we have studied the CTL response against MEPs TAB9 and CR3 after immunizing with rFWPVs, where these genes are under the control of a strong synthetic early/late promoter or the 7.5 kDa promoter from vaccinia virus. TAB9 expression was increased by more than 9-fold using the strong promoter, which was translated into a two times increase in CTL response. The overall expression of CR3 was already ten times higher when compared with TAB9 with the 7.5 kDa promoter, but the use of a stronger promoter showed no effect either on the expression or CTL response. Moreover, rFWPV expressing TAB9 induced a stronger CTL response than those expressing CR3, measured as the number of interferon- gamma -secreting splenocytes, in spite of its lower antigen expression levels. These results suggest that the capacity of a stronger promoter to increase the MEP expression and/or CTL response against their epitopes is highly dependent on the nature of the polypeptide used.
Vázquez-Blomquist
Dania
D
División de Vacunas, Centro de Ingeniería Genética y Biotecnología, Apdo 6162, Ciudad Habana 10600, Cuba. dania.vazquez@cigb.edu.cu
González
Syduet
S
Duarte
Carlos A
CA
D000818:Animals; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology*; D002230:Carbidopa / Q000276:immunology*; D002460:Cell Line; D002645:Chickens; D004338:Drug Combinations; D000939:Epitopes / Q000235:genetics / Q000276:immunology*; D018984:Epitopes, T-Lymphocyte / Q000235:genetics / Q000276:immunology*; D005260:Female; D005587:Fowlpox virus / Q000235:genetics / Q000276:immunology*; D015967:Gene Expression Regulation, Viral; D015488:HIV Antigens / Q000235:genetics / Q000276:immunology; D007111:Immunity, Cellular / Q000276:immunology; D007980:Levodopa / Q000235:genetics / Q000276:immunology*; D051379:Mice; D010455:Peptides / Q000235:genetics / Q000276:immunology; D011401:Promoter Regions, Genetic / Q000276:immunology; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology; D011995:Recombination, Genetic; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology; D014616:Vaccinia virus / Q000276:immunology
D004338:Drug Combinations; D000939:Epitopes; D018984:Epitopes, T-Lymphocyte; D015488:HIV Antigens; D010455:Peptides; D011993:Recombinant Fusion Proteins; C417682:TAB9; C009265:carbidopa, levodopa drug combination; D007980:Levodopa; D002230:Carbidopa