Expression and identification of immunological activities of the HIV-gp120N-human interferon gamma fusion protein.
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.
Feng
Ting-Ting
TT
State Key Laboratory for Infectious Disease Diagnosis and Treatment, The First Affiliated Hospital, College of Medical Sciences, Zhejiang University, Hangzhou, China.
Zhao
Gang
G
Yao
Hang Ping
HP
Wang
Qiao Ping
QP
Wu
Nan-Ping
NP
D000276:Adjuvants, Immunologic; D000818:Animals; D049109:Cell Proliferation; D016207:Cytokines / Q000378:metabolism; D017931:DNA Primers / Q000737:chemistry; D004797:Enzyme-Linked Immunosorbent Assay; D015699:HIV Envelope Protein gp120 / Q000276:immunology*; D006801:Humans; D007114:Immunization; D007371:Interferon-gamma / Q000276:immunology*; D008213:Lymphocyte Activation / Q000276:immunology; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D010957:Plasmids; D011993:Recombinant Fusion Proteins / Q000276:immunology*; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology; D018417:Th1 Cells / Q000276:immunology; D018418:Th2 Cells / Q000276:immunology
D000276:Adjuvants, Immunologic; D016207:Cytokines; D017931:DNA Primers; D015699:HIV Envelope Protein gp120; D011993:Recombinant Fusion Proteins; D007371:Interferon-gamma