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. 2015 Nov 25;6:598. doi: 10.3389/fimmu.2015.00598

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Copyright © 2015 Ameres, Liang, Wiesner, Mautner and Moosmann.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Enrichment of IE-1-specific CD4 T cells. (A) After 6–8 weeks of restimulation with the autologous IE-1 mini-LCL, T-cell cultures were immunomagnetically separated into CD4⁺ and CD4⁻ fractions, and the purity of these fractions was analyzed by flow cytometry. Results for two exemplary donors are shown. (B) PBMCs ex vivo and the separated CD4-enriched (CD4⁺) and CD4-depleted (CD8⁺) fractions of IE-1 mini-LCL-stimulated T-cell cultures were tested for their reactivity to IE-1 peptides in an IFN-γ ELISPOT assay. A peptide library covering the entire IE-1 protein sequence from HCMV strain AD169 plus some sequence variants from strains Toledo and TB40 was used for the stimulation of specific T cells in this assay. PBMCs were stimulated with peptide only, CD4⁺ and CD8⁺ fractions were stimulated with CD40-activated B cells loaded with peptides. Results are shown for two representative donors.