Computational protocol: HCV Specific IL-21 Producing T Cells but Not IL-17A Producing T Cells Are Associated with HCV Viral Control in HIV/HCV Coinfection

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Protocol publication

[…] Crypopreserved PBMC were stained with fluorophore-conjugated monoclonal antibodies to human CD4 (Biolegend OKT-4), CD8 (Biolegend 53–6.7), IL-21R (BD Biosciences 17A12) or an IgG1 isotype control (ebioscience P3). Cells were washed and fixed with BD Cyotofix/Cytoperm fixation buffer (BD Biosciences). Cells were then washed with Perm/Wash buffer (BD Biosciences) and intracellular cytokine staining was performed using anti-IFN-γ (Biolegend B27). An aqua amine dye (Invitrogen) was used as a discriminating marker for live and dead cells. Background for intracellular cytokine staining was calculated based on Fluorescence-minus-one (FMO) and no-peptide negative controls. Data were acquired on an LSRII flow cytometer using BD FACSDiva software (both BD Biosciences) and analyzed by FlowJo (Tree Star Inc., San Carlos, CA). […]

Pipeline specifications

Software tools BD FACSDiva, FlowJo
Application Flow cytometry
Organisms Hepacivirus C, Human poliovirus 1 Mahoney
Diseases Hepatitis C, HIV Infections