Human Immunodeficiency Virus (HIV) Genes in Human Monocytes
Abstract
To investigate the expression of Human Immunodeficiency Virus (HIV) genes in human monocytes, a DNA transfection system was developed and characterized using cultured primary monocytes. Monocytes that were cultured 6-7 days in an adherent monolayer were efficiently recovered and transfected by electroporation with an expression vector containing the E. coli lacZ gene under control of the cytomegalovirus immediate --- early promotor. Successful transfection was detected by expression of Beta-galactosidase activity and by histochemical staining for Beta-galactosidase in cells that were allowed to readhere to plastic following transfection. Over 30% of the surviving adherent monocytes expressed the transfected Beta-galactosidase gene. In the same manner, monocytes were transfected with HIV provirus clones pIIIB and pIIIB/PB. The provirus pIIIB/PB differes from pIIIB only in that it contains a small sequence from the 'env' gene of a macrophage tropic HIV-1. Virus derived from pIIIB will not replicate in monocytes whereas virus derived from pIIIB/PB will. Monocytes transfected with either provirus DNA expressed high levels of p24 antigen within day of transfection, and cell-free supernatants contained virus that was infectious for T cells. HIV Genes in human monocytes.
Document Details
- Document Type
- Technical Report
- Publication Date
- Jan 01, 1993
- Accession Number
- ADA274527
Entities
People
- Jerry P. Weir
- Monte S. Meltzer
Organizations
- Walter Reed Army Institute of Research