Panels used for flow cytometry.

Displayed are the combinations of conjugated monoclonal antibodies (mAb) against specific antigens within each panel. In addition, the clone for each specific mAb is shown. Each panel was used for flow cytometric (FCM) analysis of bone marrow, cord blood, peripheral blood, inguinal LN, liver LN, and spleen samples of human donors (all n = 5). Thawed MNC fractions of the human tissue samples were assessed on a NaviosTM 10-color flow cytometer and analyzed using Kaluza Software® 1.0 (Beckman coulter). Panel 1 was used to identify different NK cell developmental stages based on CD34, CD117, CD94 and CD56 expression profiles.10 Additionally, expression of early development markers CD133 and CD33, stimulatory co-receptor 2B4 (CD244), and C-type lectin NKG2A were analyzed to refine the definition of the different NK cell developmental stages. Panel 2 and 3 were used to analyze the NK cell receptor repertoire of CD45+CD56brightCD16+/−CD3− and CD45+CD56dimCD16+CD3− NK cells consisting of inhibitory and stimulatory receptors. Inhibitory receptors contain KIR (CD158a, CD158b, CD158e1) and NKG2A (CD159a). Stimulatory receptors contain NCR (CD335/336/337), NKG2C (CD159c), NKG2D (CD314), and 2B4 (CD244).