Flow Cytometry Analysis of T Cell Phenotypes

Flow cytometry was used to analyze the phenotypes of freshly isolated cells. T cell phenotype was determined using the following fluorochrome-conjugated monoclonal antibodies from BD Biosciences, San Jose, CA: anti-CD4 FITC (clone RPA-T4, RRID: AB_2562052), and anti-CD8AF700 (clone RPA-T8, RRID: AB_396953) (15 (link)). The samples were exposed to monoclonal antibodies for 15 min at room temperature in the dark, washed, and events were monitored using a Beckman Coulter Navios instrument (Beckman, San Jose, CA, USA). T helper and cytotoxic lymphocytes were gated by positive surface staining for CD4 and CD8 and were expressed as a percentage of gated lymphocytes (16 (link)).