Expression of recombinant plasmid of HIV-1 gp120 and IFN-gamma in E. coli and its immunoregulatory effect / 浙江大学学报·医学版

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the regulating effect of interferon-gamma (IFN-gamma) gene on the immune response induced by human immunodeficiency virus (HIV) gp120 in mice.</p><p><b>METHODS</b>The recombinant prokaryotic plasmid pet44b with HIV-1gp120 N and pet44b with HIV-1gp120 N linking IFN-gamma were constructed and expressed in E. coli BL21 (DE3) through IPTG. The purified proteins gp120 N and gp120 N/IFN-gamma were used in the following experiments mice were injected with PBS i. s as control group 1, injected i. s with gp120 N as control group 2, and injected i. s with gp120 N/IFN-gamma as experiment group. The serum samples and spleen cells were collected 2 weeks after completion of immune program.T cells enlargement stimulated by gp120, CTL test and the detection of Th1 cytokines (SKs) IL-2, IFN-gamma and Th2 CKs IL-4, IL-10 were performed.</p><p><b>RESULT</b>SDS-PAGE and Western blot confirmed that gp120 N and gp120 N/IFN-gamma fusion proteins were successfully expressed. The immune experiments showed that the enlargement of specific T lymphocytes, the response of CTL and the expression of Th1 type CKs IL-2 and IL-gamma against gp120 were significantly different among the three groups (P < 0.05). And the response of gp120 N/IFN-gamma group was stronger than that of gp120 N group, the latter was stronger than that of PBS group (P < 0.05). The differences in expression of Th2 type CKs IL-4 and IL-10 were not significant among the three groups (P >0.05).</p><p><b>CONCLUSION</b>The cell specific immune responses of BALB/c mice to HIV-1CNgp120 antigen can be enhanced when HIV-1CNgp120 gene and IFN-gamma gene are used in coordination.</p>