Cell and viral regulatory elements enhance the expression and function of a human immunodeficiency virus inhibitory gene.
Regulated expression of recombinant genes in CD4+ cells is an important objective for gene therapy of AIDS, as these cells represent the principal target for viral replication of human immunodeficiency virus (HIV). We report here that specific combinations of CD4 cell-specific and viral regulatory elements can enhance expression of an antiviral gene product. Different viral regulatory elements were incorporated into a previously reported CD4 locus control region to increase the expression of reporter genes in T and monocytic cell lines. The CD4-specific regulatory elements were included to enhance expression in CD4 cells, and viral regulatory regions, including the cytomegalovirus immediate-early (CMV IE) upstream enhancer, which contains the kappa B and Ap1 regulatory elements and a Tat-responsive element of the HIV type 1 long terminal repeat, were used to increase gene expression and modulate its activity in response to viral infection. In transient transfection assays, this vector was 100- to 1,000-fold more active than the original CD4 regulatory elements alone. Expression of an inhibitory form of the Rev protein, Rev M10, was more effective than previously described vectors and protected against productive viral replication in CD4+ peripheral blood mononuclear cells. The combination of CD4 lineage-specific and viral regulatory elements will facilitate the development of more effective antiviral genetic strategies for AIDS.
Ranga
U
U
Department of Internal Medicine, University of Michigan Medical Center, Ann Arbor 48109-0650, USA.
Woffendin
C
C
Yang
Z Y
ZY
Xu
L
L
Verma
S
S
Littman
D R
DR
Nabel
G J
GJ
D015704:CD4 Antigens / Q000235:genetics; D002460:Cell Line; D003587:Cytomegalovirus; D004742:Enhancer Elements, Genetic; D005786:Gene Expression Regulation; D015694:Gene Products, rev / Q000235:genetics*; D015696:Gene Products, tat; D005822:Genetic Vectors; D016325:HIV Long Terminal Repeat; D015497:HIV-1 / Q000235:genetics*; D006367:HeLa Cells; D006801:Humans; D010442:Peptide Chain Initiation, Translational; D011401:Promoter Regions, Genetic; D012045:Regulatory Sequences, Nucleic Acid; D054321:rev Gene Products, Human Immunodeficiency Virus; D054322:tat Gene Products, Human Immunodeficiency Virus
D015704:CD4 Antigens; D015694:Gene Products, rev; D015696:Gene Products, tat; D054321:rev Gene Products, Human Immunodeficiency Virus; D054322:tat Gene Products, Human Immunodeficiency Virus