Designing and engineering of DNA-vaccine construction encoding multiple CTL-epitopes of major HIV-1 antigens.
A synthetic T cell immunogen (TCI) has been designed as a candidate DNA-based vaccine against Human immunodeficiency virus (HIV)-1 using cytotoxic T lymphocytes (CD8(+) CTL) and T-helper lymphocytes (CD4(+) Th) epitopes retrieved from the Los Alamos HIV Molecular Immunology Database. The protein 392 amino acids in length contains about eighty CTL-epitopes, many of which are overlapping and are totally restricted by ten different HLA class I molecules. To be able to detect CTL responses induced by a DNA vaccine in experimental animals, additional epitopes, restricted by mouse and Macaque rhesus major histocompatibility complex (MHC) class I molecules, were included in the target immunogen. The gene encoding the TCI protein was assembled, cloned into vector plasmids and expressed in a prokaryotic and a eukaryotic system. The presence of HIV-1 protein fragments in the immunogen structure was ascertained by ELISA and immunoblotting using panels of HIV-1-positive sera and monoclonal antibodies to p24. It has been demonstrated that DNA vaccine can induce both specific T cell responses (CTL and blast transformation) and specific antibodies in mice immunized with pcDNA-TCI.
Bazhan
Sergei I
SI
The State Research Center of Virology and Biotechnology Vector, 630559 Koltsovo, Novosibirsk Region, Russia. bazhan@vector.nsc.ru
Belavin
Pavel A
PA
Seregin
Sergei V
SV
Danilyuk
Nadezhda K
NK
Babkina
Irina N
IN
Karpenko
Larisa I
LI
Nekrasova
Nadezhda A
NA
Lebedev
Leonid R
LR
Ignatyev
George M
GM
Agafonov
Alexander P
AP
Poryvaeva
Vera A
VA
Aborneva
Irina V
IV
Ilyichev
Alexander A
AA
D016915:AIDS Vaccines / Q000235:genetics* / Q000276:immunology*; D000818:Animals; D000918:Antibody Specificity; D001483:Base Sequence; D002455:Cell Division; D004279:DNA, Viral / Q000235:genetics / Q000276:immunology; D015195:Drug Design; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000235:genetics* / Q000276:immunology*; D004926:Escherichia coli / Q000235:genetics; D005818:Genetic Engineering; D015483:HIV Antibodies / Q000032:analysis / Q000096:biosynthesis; D015488:HIV Antigens / Q000235:genetics* / Q000276:immunology*; D015497:HIV-1 / Q000235:genetics* / Q000276:immunology*; D006801:Humans; D007120:Immunochemistry; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D010957:Plasmids / Q000276:immunology; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology*; D019444:Vaccines, DNA / Q000276:immunology
D016915:AIDS Vaccines; D004279:DNA, Viral; D000939:Epitopes; D015483:HIV Antibodies; D015488:HIV Antigens; D019444:Vaccines, DNA