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  • After in vitro stimulation, surface staining was performed and the frequency of CD4+ T cells and intracellular expression of CD40L, IFN- γ and TNF was assessed by flow cytometry using the following antibodies: PerCP/Cy5.5 anti-human CD3, PE/Cy7 anti-human CD4 and FITC anti-human CD154, PE anti-human IFN- γ and APC anti-human TNF (all from Biolegend); the fixable viability stain 780 (e-Biosciences) was used to discriminate dead cells.
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