Design of four-helix bundle protein as a candidate for HIV vaccine.
To be efficient, a synthetic vaccine should contain different T and B cell epitopes of human immunodeficiency virus (HIV) antigens, and the B epitope regions in the vaccine and in the HIV should be conformationally similar. We have suggested previously the construction of vaccines in the form of a protein with a predetermined tertiary structure, namely a four-alpha-helix bundle. Antigenic determinants of cellular and humoral immunity are blocks for the vaccine design. From experimentally studied HIV-1 T and B cell epitopes, we constructed a sequence of a four-helix protein TBI (T and B cell epitopes containing immunogen). The gene of the protein was synthesized and the protein was produced in C600 Escherichia coli cells under recA promoter from Proteus mirabelis. CD spectroscopy of the protein demonstrated that 30% of amino acid residues adopt an alpha-helical conformation. Mice immunized with TBI have shown both humoral and cellular immune responses to HIV-1. The obtained data show that the design of TBI was successful. The synthesized gene structure makes possible further reconstruction and improvement of the protein vaccine structure.
Eroshkin
A M
AM
Research Institute of Molecular Biology, NPO VECTOR, Koltsovo, Novosibirsk Region, Russia.
Karginova
E A
EA
Gileva
I P
IP
Lomakin
A S
AS
Lebedev
L R
LR
Kamyinina
T P
TP
Pereboev
A V
AV
Ignat'ev
G M
GM
D016915:AIDS Vaccines / Q000737:chemistry* / Q000276:immunology; D000595:Amino Acid Sequence; D000818:Animals; D000917:Antibody Formation; D001402:B-Lymphocytes / Q000276:immunology; D001483:Base Sequence; D015195:Drug Design; D000939:Epitopes / Q000737:chemistry; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D005813:Genes, Synthetic; D015497:HIV-1 / Q000276:immunology*; D007111:Immunity, Cellular; D007114:Immunization; D051379:Mice; D008969:Molecular Sequence Data; D011487:Protein Conformation; D017433:Protein Structure, Secondary; D011506:Proteins / Q000737:chemistry* / Q000235:genetics / Q000276:immunology; D011994:Recombinant Proteins; D013601:T-Lymphocytes / Q000276:immunology
D016915:AIDS Vaccines; D000939:Epitopes; D011506:Proteins; D011994:Recombinant Proteins; C085784:TBI protein, recombinant