Expression and identification of immunological activities of the HIV-gp120N-human interferon gamma fusion protein

Anat Rec (Hoboken). 2009 Mar;292(3):381-6. doi: 10.1002/ar.20853.

Abstract

The human immunodeficiency virus type 1 (HIV-1) envelope glycoprotein gp120 is a vaccine immunogen that has been studied extensively. To enhance the immune response of cells against HIV-1 gp120, we tested the coexpression of gp120N with interferon-gamma (IFN-gamma) as an immune adjuvant. Two recombinant prokaryotic plasmids were constructed: the pET44b-HIV-1-gp120N plasmid construct carried the HIV-1 gp120N gene (pET44-gp120N), whereas the pET44b-HIV-1-gp120N-IFN-gamma plasmid construct carried a fusion gp120N-IFN-gamma gene (pET44b-gp120N-IFN-gamma). Target protein expression was achieved in E. coli BL21 (DE3) cells by chemical induction. To test the immunological activity of the proteins, mice were injected with a control, gp120N, or the fusion gp120N-IFN-gamma protein. The serum and spleen cells of the mice were collected for immunological detection. Results showed that specific T lymphocyte proliferation and the expression of the Th1-type cytokines (IL-2 and IFN-gamma) were higher in the gp120N-IFN-gamma group than the other two groups (P < 0.05). No difference was observed in the expression levels of the Th2-type cytokines (IL-4 and IL-10; P > 0.05). These results suggest that IFN-gamma plays a prominent role as an immune adjuvant when coexpressed with HIV-1 gp120N. IFN-gamma enhances the specific cell immune response of mice against HIV-1 gp120.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adjuvants, Immunologic
  • Animals
  • Cell Proliferation
  • Cytokines / metabolism
  • DNA Primers / chemistry
  • Enzyme-Linked Immunosorbent Assay
  • HIV Envelope Protein gp120 / immunology*
  • Humans
  • Immunization
  • Interferon-gamma / immunology*
  • Lymphocyte Activation / immunology
  • Male
  • Mice
  • Mice, Inbred BALB C
  • Plasmids
  • Recombinant Fusion Proteins / immunology*
  • T-Lymphocytes, Cytotoxic / immunology
  • Th1 Cells / immunology
  • Th2 Cells / immunology

Substances

  • Adjuvants, Immunologic
  • Cytokines
  • DNA Primers
  • HIV Envelope Protein gp120
  • Recombinant Fusion Proteins
  • Interferon-gamma