15353-j-veluchamy

70 | Chapter 2 Supplementary figure 8: Paired comparison analysis of CD56dim CD16+ subset using modifed NK cell gating strategy (A) The gating procedure to analyse NK cell phenotypes from PBMC population is shown. Lymphocytes negative for exclusion channel antigens and positive for CD45 were plotted for CD45+CD56+ cells. CD56+ NK cells were sub-divided into NK bright and NK dim populations based on CD56 expression and further the CD56dimCD16+ NK cells (NK effectors) were defined by gating CD45+CD3-CD56+ NK cells against CD16. (B) CD16 levels were compared between non activated fresh and cryopreserved NK cells (C) CD16 levels were compared between cytokine activated fresh and cryopreserved NK cells. In, B and C the following conditions: i) NK only ii) NK + A431 and iii) NK + A431 + CET were compared between fresh and cryopreserved NK cells for changes in CD56dim CD16+ marker expression. Data points represent the mean of triplicate values from independent experiments from 12 PBMC donors (6 donors: BD LSRFortessa + 6 donors: MACSQuant). Statistical analysis was done using Wilcoxon test and only statistically significant differences are mentioned (***p<0.001).