Immunophenotyping of T Cells in HIV

Staining of cells for different immune markers has been described previously [2 (link), 43 (link)]. Briefly, lymphocytes isolated from the blood samples obtained from HIV-infected or normal patients were stained for cell surface markers using specific antibodies: CD3-Cy7, CD4-Tx red, CD8-APC (Beckman Coulter), CD38 PE, HLA-DR FITC, CCR5 PE (BD Pharmingen) and CaspACE FITC-VAD-FMK (Promega). Stained cells were fixed using the Cytofix reagent (Beckman Coulter) and run on a 10 color Beckman Coulter Gallios Flow Cytometer. At least 20,000 events for each sample were acquired. Data was analyzed using the FlowJo software (Tree Star). Cells were first gated on the CD3+ population and CD4+ and CD8+ T cell subsets determined.