Cd5 apc l17f12

High sensitivity multiparameter flow cytometric immunophenotyping was performed with a FACS Canto 10-color instrument (BD Biosciences, San Jose, CA) using the following antibodies: CD45-V500C (2D1, BD Biosciences), CD3-PC7 (UCHT1, Beckman-Coulter, Miami, FL), CD2-BV421 (RPA-2.10, BD Horizon, San Jose, CA), CD5-APC (L17F12, BD Biosciences), CD7-BB515 (M-T701, BD Horizon), CD4-APC-A700 (13B8.2, Beckman-Coulter), CD8-APC-H7 (SK1, BD Biosciences), CD56-PE (N901, Beckman-Coulter), CD26-PerCP-Cy5.5 (BA5B, BioLegend, San Diego, CA) and CD279 (PD-1)-BV605 (EH12.2H7, BioLegend). At least 200,000 cells were acquired and at least 20 events were required for a positive result. The data was analyzed using custom software (“Woodlist,” gift of B.L. Wood, University of Washington, Seattle, WA). Abnormal T cell populations were identified by visual assessment based on aberrant antigen expression. Median fluorescent intensity (MFI) of markers was evaluated. Abnormal populations were considered “dim” or “bright” for a given marker if the fluorescent intensity was ≥1/3 log different from that of corresponding internal normal CD4+ T cells. In select cases, clonality of T cell populations was assessed with a Beta Mark TCR Vβ Repertoire Kit (Beckman-Coulter) using a modified, single-tube method as described previously (25 (link)).