HIV promoter activity in primary antigen-specific human T lymphocytes.
Human retroviruses, such as the HIV, infects human T cells, and efficient HIV replication occurs primarily in activated T cells rather than resting cells. Increased HIV production is likely caused by the activation of the retroviral promoter, and the HIV promoter may be regulated by intracellular signals induced during immune stimulation. To examine the regulation of retroviral promoter activity in normal, Ag-specific primary T lymphoblasts, a heterogeneous population of primary human T cells was transfected with either the HIV promoter or a promoter from a different retrovirus, Rous sarcoma virus (RSV) by protoplast fusion technique. Transfected T cells responded normally to Ag or mitogen stimulation, and activation of these T cells increased both the HIV and RSV promoter activity. Promoter activity was assessed by using transient expression assays after the T cells were restimulated with Ag, mitogen, or IL-2. In situ hybridization of transfected human T cells showed that 68 to 95% of activated lymphocytes expressed CAT mRNA directed by HIV or RSV. Thus, protoplast transfection of primary T cells was efficient in that the majority of cells expressed CAT message. By deletion of different regions of the HIV promoter, the enhancer region was identified as necessary for effective HIV promoter activity. In addition these deletion studies identified a region that negatively affects HIV promoter activity in primary T cells. Cotransfection of the HIV promoter with the HIV transactivator protein, tat, increases HIV promoter activity in both resting and activated primary human T cells only when the tat target sequences were present.
Horvat
R T
RT
Department of Microbiology, University of Kansas, Lawrence 66045.
Wood
C
C
D015500:Chloramphenicol O-Acetyltransferase / Q000378:metabolism; D000939:Epitopes / Q000276:immunology*; D006678:HIV / Q000235:genetics*; D006801:Humans; D008213:Lymphocyte Activation; D009693:Nucleic Acid Hybridization; D011401:Promoter Regions, Genetic; D012091:Repetitive Sequences, Nucleic Acid; D013601:T-Lymphocytes / Q000201:enzymology / Q000276:immunology / Q000382:microbiology*; D014162:Transfection
D000939:Epitopes; D015500:Chloramphenicol O-Acetyltransferase