Clustered epitopes within the Gag-env HIV-1 fusion proteins DNA vaccine enhance immune responses in mice
Abstract number: 1733_566
Roodbari F., Sabahi F., Sarbolouki M.N., Barkhordary F., Mahdavi M.A., Sarami Forooshani R., Bamdad T., Adeli A., Mahboudi F.
Objectives: The global HIV epidemic continues to expand and exceeding previous predictions. An effective vaccine represents the best hope to curtail the HIV epidemic. DNA vaccines induce conformational-dependent humoral and cellular response and mimic live vaccines without their pathogenic potential. The importance of CD8+ CTL responses in controlling HIV and SIV viraemia has led to a series of vaccine candidates that effectively induce these responses. It is now widely believed that an HIV vaccine strategy must stimulate both a strong humoral (antibody) as well as cell-mediated (CTL) immune response. Neutralising antibodies reduce viral burden by latching onto and destroying HIV circulating in body fluids. CTLs eliminate HIV-infected cells, thereby inhibiting viral replication. Both types of immune response are invoked by the presence of antigens, such as HIV envelope glycoproteins, in the body.
The P24 and gp41 play many of important roles in host-virus-interaction and pathogenesis. These proteins are considered as attractive vaccine candidate in which their immungenecity and immunomodulatory effects have been confirmed.
Methods: In this study, an expression vector (PCDNA3.1 hygro) containing P24-gp41 immunogenic sequences under the control of IE HCMV promoter was designed. The expression of the recombinant peptides was analyzed in an eukaryotic systems, COS-7 and Hela cells. Immunofluorescence and western blotting confirmed the presence of expressed proteins The cited P24-gp41 fusion gene that is able to express in a proper folding form. This fusion gene was studied as DNA vaccine in mice(BALB/c) for evaluation and generation of effective immune responses. For immunising, we used dendrosome, a novel family of vehicles for transfection and therapy. Some groups of mice were immunised with our construct and pCAGGS-IL-12 as co-injection. Humoral response and IFN-g and IL-2 was detected by ELISA. Lymphoprolifration assay was detected by MTT.
Results: ELISA and MTT assays confirmed, the cited P24-gp41 fusion gene that is able to enhance immune responses in mice.
Conclusions: Our construct is a good candidate for DNA vaccine against HIV-1.OF course, it needs much more research for clearing some things that they are unclear.
|Session name:||European Society of Clinical Microbiology and Infectious Diseases|
|Location:||ICC, Munich, Germany|
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