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Construction and characterization of a recombinant fowlpox virus containing HIV-1 multi-epitope-p24 chimeric gene in mice

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Abstract

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-P7.5×20) 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-γ and IL-2) of spleen lymphocytes stimulated by H-2d-restricted CTL peptide were observed in immunized mice. We concluded that the rFPV may induce HIV-1 specific immunity especially cellular immunity in mice.

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Authors and Affiliations

  1. The 11th Institute, Academy of Military Medical Sciences, Changchun, 130062, China

    Zhang LiShu & Jin NingYi

  2. College of Agricultural and Biological Engineer, Tianjin University, Tianjin, 300072, China

    Song YingJin

  3. College of Life Science, Jinan University, Guangzhou, 510632, China

    Wang Hong

  4. Department of Biochemistry and Molecular Biology, Oklahoma State University, Stillwater, 74078, USA

    Ma HeWen

  5. Institute of Vascular Medicine, Third Affiliated Hospital of Peking University, Beijing, 100083, China

    Li ZiJian

  6. School of Life Science, East China Normal University, Shanghai, 200026, China

    Jiang WenZheng

Authors
  1. Zhang LiShu
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  2. Jin NingYi
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  3. Song YingJin
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  4. Wang Hong
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  5. Ma HeWen
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  6. Li ZiJian
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  7. Jiang WenZheng
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Correspondence to Jin NingYi.

Additional information

Supported by the China “863” Program (Grant No. 2003AA219051)

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Zhang, L., Jin, N., Song, Y. et al. Construction and characterization of a recombinant fowlpox virus containing HIV-1 multi-epitope-p24 chimeric gene in mice. SCI CHINA SER C 50, 212–220 (2007). https://doi.org/10.1007/s11427-007-0017-1

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  • DOI: https://doi.org/10.1007/s11427-007-0017-1

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