The purpose of this study is to establish reliable markers of AIDS infection, access viral burden and to identity mechanisms of HIV replication susceptible to intervention. Current FACS technology permits simultaneous acquisition and subsequent analysis of multiple parameters, including nuclear factors. Cell typing is readily achieved using labelled antibodies to well characterized surface markers. Internalization of HIV can be monitored with labelled antibodies to HIV gpl2O. Nuclear factors can be identified by labelled oligonucleotides. Cells exhibiting any select parameter can be enriched by FACS sorting and nucleic acids further identified with standard hybridization techniques. We have established fixation techniques and characterized useful HIV reagents by IFA and probe hybridization to Northern and Southern blots. Initial FACS analysis of chronically infected cell lines with these reagents show promising results. FACS and IFA analysis of the HIV replication cycle in T-cells is in progress. Work is scheduled to access HIV infection in patient PBL's.
