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. 2017 Aug 21;8:987. doi: 10.3389/fimmu.2017.00987

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Copyright © 2017 Cabral, Hanley, Gerlach, O’Leary, Cunningham, Ritter, Ceredig, Joshi and Griffin.

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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Manipulation of murine Treg surface N-glycosylation impairs suppressive potency. (A) Naïve and (B) memory CD4⁺ Tconv and (C) naïve and (D) memory CD8⁺ Tconv were cocultured in the presence of CD4⁻CD8⁻ antigen presenting cells and no enzyme or PNGase F-treated Treg populations at Treg: responder T-cell ratios of 0:1, 1:2, 1:4, 1:6, 1:8, and 1:16 for 4 days with anti-CD3 stimulation. Suppressive function was quantified based on responder T-cell division index (DI) after 4 days and presented as the calculated percent suppression [%Suppression (DI)]. (E,F) Day 4 frequencies of viable Treg were expressed as percentages of the total viable cells (Viable Treg frequency) in the cocultures with (E) naïve and (F) memory CD4⁺ responder T-cells. Data represent mean ± SD (n = 3 technical replicates). Results were confirmed in three independent experiments. Statistical analysis was performed by permutation test with an unpaired design comparing the coculture with the different Treg populations to the no Treg condition (*p value ≤ 0.1).