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. 2014 Jul 2;20:956–969.

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Copyright © 2014 Molecular Vision.

This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited, used for non-commercial purposes, and is not altered or transformed.

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Cytokine-secretion assay with selected clones stimulated with individual proteins. After induction of protein expression by Isopropyl β-D-thiogalactoside (IPTG) and opsonization with complement, the bacteria were added to autologous Epstein-Barr virus-transformed B cells (EBV-B cells) and incubated overnight. CD4 T cells (5,000/well) were added to EBV-B cells (30,000/well). The amount of tumor necrosis factor (TNF)-α, interferon (IFN)-γ, interleukin (IL)-5, and IL-17 in the supernatant of overnight co-cultures was estimated with a multicytokine bioplex kit. As the positive control, anti-MAGE-3 T cell clone R12C9 was used. The screening of the clones was performed in triplicates. Error bars represent SD.