Document Type : Original Article
Authors
- Fatemeh Roodbari 1, 2
- Farzaneh Sabahi 1
- Mohamad Nabi Sarbolouki 3
- Farzaneh Barkhordari 4
- Ahmad Adeli 4
- Amel Jamedar 1, 5
- Fereidoun Mahboudi 4
1 Department of Virology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran
2 Department of Cellular and Molecular Biology, Faculty of Basic Sciences, University of Mazandaran, Babolsar
3 Institute of Biochemistry and Biophysics, Tehran University of Basic Sciences
4 Biotechnology Research Center, Pasteur Institute of Iran, Tehran
5 Department of Microbiology and Virology, Faculty of Medicine, Mashad University of Medical Sciences, Mashad, Iran
Abstract
Background: Development of an effective vaccine is highly needed in order to restrict the AIDS pandemic. DNA vaccines initiate both arms of immunity without the potential of causing disease. HIV-1 p24 and gp41 (gag and env) proteins play important roles in viral pathogenesis and are effective candidates for immune induction and vaccine design.
Objective: In this study, new DNA vaccine candidates constructed from HIV-1 fused p24- gp41 or gp41 alone were evaluated in Balb/c mice for induction of cellular and humoral immune responses.
Methods: Recombinant plasmids, pcDNA3.1/Hygro expression vector containing immunogenic sequences of fused p24-gp41 or gp41alone were produced. Dendrosome used as a system for carrying vectors in laboratory animals, and an IL-12 containing vector (pCAGGS-IL-12) was co-immunized with the p24-gp41 vector as a genetic adjuvant. Induction of effective immune responses against the designed vectors as DNA vaccine candidates in Balb/c mice was evaluated. Levels of total antibodies, IgG isotypes (IgG2a and IgG1); IFN-γ and IL-4 were measured by ELISA. MTT assay was used to evaluate lymphoproliferation.
Results: The results confirmed that the immunogenic epitopes of both p24 and gp41 genes are highly effective inducers of immune responses, and administration of fused p24-gp41 alone or along with IL-12 resulted in further enhancement of immune responses. Group 4 that received fused fragments (p24-gp41) along with an IL-12 expressing vector demonstrated a significantly higher Stimulation Index (SI) and IFN- production (p<0.0001) with a significant increase in IgG2a/IgG1 ratio, indicating the stimulation of CMI towards Th1. Although gp41 containing vector (group 6) also showed significant increases in both proliferation and IFN- production, the responses were persistently lower than that of p24-gp41 containing vectors. Total antibody production was highest in group 6 as expected.
Conclusion: Dendrosome proved to be an efficient carrier of recombinant plasmids constructed in this study. Further studies are necessary to evaluate these constructs as HIV vaccine candidates.
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