| Research Abstract |
In this project, attempt have been made to establish CD4-positive lymphocytes which are resistant to the infection by human immunodeficiency virus (HIV) based on the fact that CD4-positive cells which are killed by HIV in AIDS play a major role in elimination of viruses.
First, we have investigated the effect of introduction of the plasmid expressing antisense RNA against Tat and Rev derived by a strong metallothioneine promoter. A human CD4-positive T cell line, MOLT4, was transfected with HIV-LTR-CAT and Tat-expression plasmids. Trans-activation of HIV-LTR by Tat was significantly blocked by co-transfection with the plasmid expressing antisense RNA against Tat and Rev as observed by transient CAT assays. We then selected MOLT4 cells stably transfected with the Tat-antisense RNA plasmid for further biological studies. However, the substantial evidence has not yet been obtained since viabilities of these cell clones were not good enough to demonstrate the actual remistence to HIV infection.
In addition to the above experiments, construction of a retrovirus vector which is able to endow cells with ability to produce antisense RNA to the HIV Tat (and Rev) is now underway. Since this retrovirus vector contains neomycin-resistance gene as well, selection and establishment of those cells producing the antisense RNA has now become much easier. We are attempting to test eagerly if these cells are resistant to HIV infection.
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