Construction and characterization of a recombinant fowlpox virus containing HIV-1 multi-epitope-p24 chimeric gene in mice.
The epidemic of HIV/AIDS is sweeping across the world. It is of great importance to figure out new ways to curb this disease. Epitope-based vaccine is one of these solutions. In this study, a chimeric gene was obtained by combination of a designed HIV-1 multi-epitope gene (MEG) and HIV-1 p24 gene. A recombinant plasmid pUTA2-MEGp24 was then constructed by inserting MEGp24 gene into the downstream of the promoter (ATI-P7.5x20) of fowlpox virus (FPV) transfer vector pUTA2. The recombinant plasmid and wild-type FPV 282E4 strain were then co-transfected into CEF cells and homologous recombination occurred. A recombinant virus expressing HIV-1 protein MEGp24 was screened by genome PCR and Western blot assay. Large scale preparation and purification of the recombinant fowlpox virus (rFPV) were then carried out. BALB/c mice were immunized intramuscularly with the rFPV for three times on day 0, 14 and 42. Mice were executed and sampled one week after the third inoculation. Anti-HIV-1 antibody in serum and Th1 cytokines in the supernatant of cultured spleen cells were assayed by ELISA. The count of T lymphocyte subsets and the CTL activity of spleen lymphocytes were analyzed by flow cytometry and lactate dehydrogenase (LDH) release assay, respectively. The results showed that HIV-1 specific antibody in serum and increased T lymphocyte subsets (CD4(+) T, CD8(+) T) were detected in the immunization group. CTL target-killing activity and higher secretion of Th1 cytokines (IFN-gamma and IL-2) of spleen lymphocytes stimulated by H-2(d)-restricted CTL peptide were observed in immunized mice. We concluded that the rFPV may induce HIV-1 specific immunity especially cellular immunity in mice.
Zhang
LiShu
L
The 11th Institute, Academy of Military Medical Sciences, Changchun 130062, China.
Jin
NingYi
N
Song
YingJin
Y
Wang
Hong
H
Ma
HeWen
H
Li
ZiJian
Z
Jiang
WenZheng
W
D000818:Animals; D000914:Antibodies, Viral / Q000097:blood; D018791:CD4 Lymphocyte Count; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology; D017931:DNA Primers; D005587:Fowlpox virus / Q000235:genetics*; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D016655:HIV Core Protein p24 / Q000235:genetics*; D015497:HIV-1 / Q000235:genetics* / Q000276:immunology; D018655:Lymphocyte Count; D051379:Mice; D050506:Mutant Chimeric Proteins / Q000235:genetics; D016133:Polymerase Chain Reaction; D011995:Recombination, Genetic; D013154:Spleen / Q000276:immunology; D013601:T-Lymphocytes / Q000276:immunology; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology
D000914:Antibodies, Viral; D017931:DNA Primers; D016655:HIV Core Protein p24; D050506:Mutant Chimeric Proteins