Protocol detail

Quantifying EBV-specific T cell Responses

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EBV-specific T cell responses were analyzed using an IFN-γ enzyme-linked immunospot (ELISpot) assay as previously described (10 (link), 54 (link)). Briefly, splenocytes were depleted of human CD19⁺ cells using anti-CD19 microbeads (Miltenyi Biotec). The CD19-depleted fraction was stimulated with autologous LCLs at a ratio of 1:4 for 18 h, and incubation with R10 and phorbol myristate acetate (PMA)-ionomycin served as negative and positive controls, respectively. Each condition was performed in duplicates. Spots were counted on an ELISpot reader system (ELR02; Autoimmun Diagnostika GmbH).

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Murer A., Rühl J., Zbinden A., Capaul R., Hammerschmidt W., Chijioke O, & Münz C. (2019). MicroRNAs of Epstein-Barr Virus Attenuate T-Cell-Mediated Immune Control In Vivo. mBio, 10(1), e01941-18.

Publication 2019

anti-CD19 microbeads ELR02

Cells Enzyme Enzyme linked immunospot assay Human Ifn γ Ionomycin Lcls Microbeads Phorbol myristate acetate Spots T cell

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Variable analysis

independent variables

Stimulation with autologous LCLs at a ratio of 1:4

dependent variables

EBV-specific T cell responses analyzed using an IFN-γ enzyme-linked immunospot (ELISpot) assay

control variables

Incubation with R10 (negative control)
Incubation with phorbol myristate acetate (PMA)-ionomycin (positive control)

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