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Vol 47(2013) N 2 p. 228-235;
M.R. Aghasadeghi1, R. Zabihollahi1*, S.M. Sadat1, M. Salehi2, S.H. Ashtiani3, R. Namazi2, N. Kashanizadeh4, K. Azadmanesh1

Production and Evaluation of Immunologic Characteristics of mzNL4-3, a Non-Infectious HIV-1 Clone with a Large Deletion in the Pol-Sequence

1Hepatitis and AIDS Department, Pasteur Institute of Iran, Tehran, Iran
2Department of pharmaceutical biotechnology, School of Pharmacy and Pharmaceutical Sciences, Isfahan , Iran
3Institute of Biochemistry and Biophysics, University of Tehran, Tehran, Iran
4Gynecology Department, Baghiata-allah Hospital, University of Baghiata-allah, Iran

*Rezvan_z_m@yahoo.com
Received - 2012-06-20; Accepted - 2012-08-09

Inactivation of integrase (IN) and reverse transcriptase (RT) can revoke the replication of HIV virions, and non-infectious HIV particles are desirable virus-like particle (VLP) vaccine candidates. Here, we produced inactive HIV-1 particles fit for vaccine and virological purposes by introducing a mutation into the pol sequence. Proviral DNA (pNL4-3) was cut at two points in the pol region using the BalI restriction enzyme and then religated. HEK 293T cells were transfected with the resultant plasmid (pmzNL4-3) to produce mutated virions. To confirm a production of VLPs and evaluate their biological activity the p24 load and syncytium formation (MT2 cells) were analyzed. The assay indicated that mzNL4-3 virions were assembled and contained functional envelope glycoproteins (ENV). In addition, mzNL4-3 virions were able to infect neither the MT2 nor HEK 293T cells. Furthermore, the immunogenicity of VLPs was investigated in a mouse model. According to the data on vaccinated mice, the titer of ENV-specific antibodies rose rapidly after a boosting injection. Moreover, lymphoid cells extracted from these mice proliferated after exposure to the antigen. The mzNL4-3 virus particles possessed immunogenic antigens of HIV and can effectively trigger humoral and CD4 immune responses. Non-infectious mzNL4-3 virions may also be used in biomedical experiments for improving the biological safety conditions. Moreover, the mzNL4-3 seems to be a promising candidate for further HIV-1 vaccine investigations.

human immunodeficiency virus, virus-like particles, reverse transcriptase, integrase, envelope glycoproteins, HIV-1 vaccine



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