α cd45 apc h7

T-cell phenotyping was performed on fresh PBMNC or follow-up samples stored in liquid nitrogen. For the analysis of GrB+ T-cells, the cells were first stained for cell surface markers; α-CD45 APC-H7 (BD 641417), α-CD3 APC (BD 345767), α-CD4 PerCP (BD 345770), and α-CD8 PE-Cy7 (BD 335822), and then fixed and permeabilized with Fix/Perm (BD 554714), and intracellular GrB was stained (Alexa Fluor 700, BD 561016)). The memory cell subsets of CD4+ and CD8+ T-cells were studied by the following panel: α-CD45-APC-H7, α-CD3-PeCy7 (BD 557851), α-CD4-PerCP, α-CD45RA AlexaFluor700 (BD 560673) and α-CCR7-PE (R&D Systems FAB197P). 50´000 CD45+ cells were acquired with FACSAria (BD) and analyzed with FlowJo.