Flow cytometric quantification of HIV-1 Tat protein in tat-transfected Jurkat T cell lines

The transactivator Tat protein represents a pivotal factor for the replicat ion of human immunodeficiency virus type 1 (HIV-1). in this report, we desc ribe a flow cytometry procedure designed to quantify the intracellular cont ent of Tat protein in Jurkat CD4(+) T lymphoblastoid cell lines, stably tra nsfected with plasmids expressing full-length Tat protein. Various expressi on vectors were compared for their effectiveness to yield Tat protein in Ju rkat cells, and several technical parameters were analyzed to optimize the assay. This method offers a quick and efficient approach to select stably t ransfected cell. lines expressing different levels of specific protein. (C) 1998 Elsevier Science B.V. All rights reserved.