[Expression of recombinant plasmid of HIV-1 gp120 and IFN-gamma in E. coli and its immunoregulatory effect]

Zhejiang Da Xue Xue Bao Yi Xue Ban. 2006 Nov;35(6):610-4. doi: 10.3785/j.issn.1008-9292.2006.06.007.
[Article in Chinese]

Abstract

Objective: To investigate the regulating effect of interferon-gamma (IFN-gamma) gene on the immune response induced by human immunodeficiency virus (HIV) gp120 in mice.

Methods: 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.

Result: 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).

Conclusion: 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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cytotoxicity, Immunologic / immunology
  • Escherichia coli / genetics*
  • HIV Envelope Protein gp120 / biosynthesis*
  • HIV Envelope Protein gp120 / genetics
  • HIV Envelope Protein gp120 / immunology
  • HIV-1 / genetics
  • HIV-1 / metabolism
  • Immunization / methods
  • Interferon-gamma / biosynthesis*
  • Interferon-gamma / genetics
  • Interferon-gamma / immunology
  • Male
  • Mice
  • Mice, Inbred BALB C
  • Plasmids / genetics
  • Prokaryotic Cells / metabolism
  • Recombinant Fusion Proteins / biosynthesis*
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / immunology
  • T-Lymphocytes, Cytotoxic / immunology

Substances

  • HIV Envelope Protein gp120
  • Recombinant Fusion Proteins
  • Interferon-gamma