Multiparameter Flow Cytometry Characterization

After stimulation, cells were stained for surface and intracellular markers as
performed previously [23 (link), 24 (link)]. The following antibodies and dyes were
used; Aqua LIVE/DEAD (Invitrogen), anti-CD3 BUV396 (UCHT1; BD Biosciences),
anti-CD19 BV510 (SJ25C1; BioLegend), anti-CD14 BV510 (M5E2; BioLegend), anti-CD4
BUV805 (SK3; BD Biosciences), anti-CD8 BV786 (RPA-T8; BD Biosciences), anti-CCR7
PE/Cy7 (G043H7; BioLegend), anti-CD45RA PE/TR (MEM-56; Invitrogen), anti-KLRG1
eFlour710/PerCP (13F12F2; eBioscience), anti-IFN-γ AF700 (B27;
BioLegend), anti-IL-2 APC/Cy7 (MQ1-17H12; BioLegend), anti-TNF-α Pacific
Blue (MAb11; BioLegend), anti-Perforin-1 PE (B-D48; Cell Sciences),
anti-Granzyme B PE/Cy5.5 (GB11; Invitrogen), anti-T-bet BV605 (4B10; BioLegend),
and anti-Eomesodermin (EOMES) eFlour660 (WD1928; eBioscience). Flow cytometric
analysis was performed on a LSRFortessa (BD Biosciences) and analyzed by Flowjo
v10.6.1. Statistics analysis was done by SPICE V6 [25 (link)] and GraphPad Prism 8.

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Jin W., Fang M., Sayin I., Smith C., Hunter J.L., Richardson B., Golden J.B., Haley C., Schmader K.E., Betts M.R., Tyring S.K., Cameron C.M., Cameron M.J, & Canaday D.H. (2023). Differential CD4+ T-Cell Cytokine and Cytotoxic Responses Between Reactivation and Latent Phases of Herpes Zoster Infection. Pathogens and Immunity, 7(2), 171-188.

Publication 2022

Aqua LIVE/DEAD anti-CD14 BV510 anti-CD8 BV786 (RPA-T8; anti-CCR7PE/Cy7 anti-IFN-γ AF700 LSRFortessa

Anti cd3 Anti t Antibodies Cell Cy5 5 Dyes Granzyme Ifn γ Intracellular Perforin 1 Prism Spice Tnf α

Corresponding Organization :

Other organizations : Case Western Reserve University, Geriatric Research Education and Clinical Center, Ohio University, Center for Clinical Studies, Durham VA Medical Center, Duke University Hospital, Duke Medical Center, University of Pennsylvania

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Variable analysis

independent variables

Stimulation of cells

dependent variables

Expression of surface and intracellular markers
Immune cell phenotypes and functions

control variables

Staining procedures performed as in previous studies [23, 24]

controls

Positive controls: Not explicitly mentioned
Negative controls: Not explicitly mentioned

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