Document Type : Original Article
Authors
- Sayeed Bayanolhagh 1
- Mahtab Alinezhad 2
- Kooroosh Kamali 3
- Maryam Foroughi 4
- Hamid Reza Khorram Khorshid 5
- Minoo Mohraz 4
- Fereidoun Mahboudi 6
- Ali Akbar Pourfathollah 1
1 Department of Immunology, Faculty of Medical Sciences, Tarbiat Modares University
2 Research and Development Complex, Pasteur Institute of Iran
3 Department of Epidemiology and Biostatics, Tehran University of Medical Sciences
4 Iranian Research Center for HIV/AIDS, Tehran University of Medical Sciences
5 Genetic Research Centre, University of Social Welfare and Rehabilitation Sciences
6 Biotechnology Research Center, Pasteur Institute of Iran, Tehran, Iran
Abstract
Background: Numerous evidences indicate that in some HIV-1 positive patients, the humoral and cellular immune responses are induced against HIV-1 proteins and this is inversely related to the progress of infection.
Objective: The aim of this study was the evaluation of the Adenovectors containing HIV genes in induction of immune responses in mice.
Methods: The HIV-1 genes including gag p24, rev, nef and exon-1 of tat were amplified from HIV-1 RNA (clade-A). The cDNA of each gene was cloned into a transfer vector. The transfer vector was then co-transformed into E. coli strain BJ5183 together with pAdenovector ΔE1/E3. The recombinant adenoviral construct was transfected into QBI-293A cells. Recombinant viruses were purified and titrated on 293 cell plates. Expression of transgenes was evaluated using western blotting. Then 1012 viral particles were injected into 15 groups of 5 mice and all patterns of combination of these 4 HIV-1 genes were evaluated. After 2 weeks, humoral and cellular immune responses were evaluated using ELISA, cell proliferation and ELISpot (IL-2, IL-4 and IFN-γ) assays, consecutively.
Results: It was demonstrated that each gene was expressed. The response targets were mostly toward Th1, though several Th2 responses were also observed. Single injection in our study induced a good cellular response but the humoral responses were not as strong as the cellular ones.
Conclusion: Considering and comparing all results and evaluating the various possible interactions revealed that simultaneous injection of tat and gag has enhanced the humoral and cellular responses.
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