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. 2014 Nov 27;3(4):1116–1130. doi: 10.3390/cells3041116

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© 2014 by the authors; licensee MDPI, Basel, Switzerland.

This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution license (http://creativecommons.org/licenses/by/4.0/).

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Linearity of PBMC responses to CA, analyzed by IFN-γ/IL-17A/IL-22 triple FluoroSpot. PBMC from three donors were incubated with stimuli or medium alone and analyzed for spots representing cells secreting single-, double- and triple combinations of IFN-γ, IL-17A and IL-22. Quadruplicate wells with 200,000, 300,000, 400,000 or 500,000 PBMC/well were analyzed. The graphs to the left show the mean total number of spots/well and standard deviation obtained for each cell concentration. The charts to the right show the proportion of single-, double- and triple spots for each cytokine/cytokine combination per total number of spots.