Flow Cytometry Analysis of PBMCs and LMNCs

PBMCs and LMNCs were analyzed on a modified 4-laser LSR Fortessa (BD Biosciences). Antibody capture beads (BD Bioscience) were used for single-stain compensation controls. Flow cytometry data were analyzed with FlowJo 8.8.7 (Treestar) and when applicable, fluorescence minus one (FMO) gating strategies were used to set the manual gates9 (link)37 (link)38 (link). A virus-specific response was considered positive when ≥0.05% of the cells produced IFN-γ after background reduction and if the response was larger than twice the negative background signal9 (link)37 (link)38 (link).

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Tauriainen J., Scharf L., Frederiksen J., Naji A., Ljunggren H.G., Sönnerborg A., Lund O., Reyes-Terán G., Hecht F.M., Deeks S.G., Betts M.R., Buggert M, & Karlsson A.C. (2017). Perturbed CD8+ T cell TIGIT/CD226/PVR axis despite early initiation of antiretroviral treatment in HIV infected individuals. Scientific Reports, 7, 40354.

Publication 2017

LSR Fortessa Antibody capture beads

Antibody Cells Flow cytometry Fluorescence Ifn γ Stain Virus

Corresponding Organization :

Other organizations : Karolinska University Hospital, Karolinska Institutet, Technical University of Denmark, University of Pennsylvania, San Francisco General Hospital, University of California, San Francisco

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Variable analysis

independent variables

None explicitly mentioned

dependent variables

Percentage of cells producing IFN-γ

control variables

None explicitly mentioned

controls

Antibody capture beads (BD Bioscience) were used for single-stain compensation controls.
Fluorescence minus one (FMO) gating strategies were used to set the manual gates.

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