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. 2014 Jan 16;63(2):632–644. doi: 10.2337/db13-0981

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© 2014 by the American Diabetes Association.

Readers may use this article as long as the work is properly cited, the use is educational and not for profit, and the work is not altered. See http://creativecommons.org/licenses/by-nc-nd/3.0/ for details.

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AW-recipient mice show higher autoreactive/proinflammatory T cells in the pancreatic microenvironment compared with NW recipients. Four-week-old NOD mice were continued on AW or switched to NW for 4 weeks and killed at 8 weeks of age, and single suspensions from spleen and PnLN were used for immunological analysis. A: PnLN and spleen cells were labeled with CFSE, incubated with immunodominant β-cell antigen for 4 days, and examined for CFSE dilution among CD4⁺ and CD8⁺ cells by FACS. Representative FACS analysis graphs with percentage values (left panel) and mean ± SD of percentage of CD4 and CD8 cells with CFSE dilution (4 mice/group tested independently) (right panel) are shown. B: PnLN cells were also activated ex vivo with phorbol myristic acid and ionomycin in the presence of brefeldin A for 4 h and stained for surface CD4 and CD8, followed by intracellular IFN-γ and IL-17. Representative FACS analysis graphs with percentage values (left panel) and mean ± SD of percentage values of IFN-γ– and IL-17–positive cells among CD4 and CD8 populations from 4 mice/group tested independently (right panel) are shown.