Patients with relapsing-remitting multiple sclerosis have normal Treg function when cells expressing IL-7 receptor α-chain are excluded from the analysis
Laure Michel, … , Jean-Paul Soulillou, David-Axel Laplaud
Laure Michel, … , Jean-Paul Soulillou, David-Axel Laplaud
Published October 1, 2008; First published September 2, 2008
Citation Information: J Clin Invest. 2008;118(10):3411-3419. https://doi.org/10.1172/JCI35365.
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Research Article Autoimmunity

Patients with relapsing-remitting multiple sclerosis have normal Treg function when cells expressing IL-7 receptor α-chain are excluded from the analysis

Abstract

Multiple sclerosis (MS) is a chronic inflammatory disease that results in demyelination in the central nervous system, and a defect in the regulatory function of CD4+CD25high T cells has been implicated in the pathogenesis of the disease. Here, we reanalyzed the function of this T cell subset in patients with MS, but we depleted cells expressing IL-7 receptor α-chain (CD127), a marker recently described as present on activated T cells but not Tregs. Similar to other studies, we observed a marked defect in the suppressive function of unseparated CD4+CD25high T cells isolated from MS patients. However, when CD127high cells were removed from the CD4+CD25high population, patient and control cells inhibited T cell proliferation and cytokine production equally. Likewise, when the CD25 gate used to sort the cells was stringent enough to eliminate CD127high cells, CD4+CD25high T cells from patients with MS and healthy individuals had similar regulatory function. Additional analysis indicated that the CD127high cells within the CD4+CD25high T cell population from patients with MS appeared more proliferative and secreted more IFN-γ and IL-2 than the same cells from healthy individuals. Taken together, we conclude that CD4+CD25highCD127low Tregs from MS patients and healthy individuals exhibit similar suppressive functions. The decreased inhibitory function of unfractioned CD4+CD25high cells previously observed might be due to abnormal activation of CD127high T cells in patients with MS.

Authors

Laure Michel, Laureline Berthelot, Ségolène Pettré, Sandrine Wiertlewski, Fabienne Lefrère, Cécile Braudeau, Sophie Brouard, Jean-Paul Soulillou, David-Axel Laplaud

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Figure 1

Frequency, proliferation, and suppressive activity of the top 4% of sorted CD4+CD25high T cells from MS patients and HIs.

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Frequency, proliferation, and suppressive activity of the top 4% of sort...
(A) Comparison of the percentage of CD4+CD25high T cells from MS patients and controls. The cut-off for high-staining CD25 was placed at 6 × 103 of mean fluorescence intensity. No statistical difference can be shown between the groups (patients, n = 21; HI, n = 17). (B) Regulatory properties of CD4+CD25high cells were examined in 11 untreated patients with RR-MS and 11 healthy controls. Cocultures of CD4+CD25– and CD4+CD25high cells were performed at a 1:1 ratio and under anti-CD3 stimulation. Proliferation was measured by incorporation of 3H-thymidine after 5 days of incubation. The percentage of suppression of responding cell proliferation (CD4+CD25–) by CD4+CD25high cells was determined as 1 – (proliferation of coculture / proliferation of responder population alone) × 100, where proliferation is expressed as cpm. CD4+CD25high T cells from MS patients exhibited less suppressive activity when the gate for sorting was positioned as shown in Supplemental Figure 1 (P < 0.05, Mann-Whitney U test). Mean values in A and B are indicated by horizontal lines. (C) Comparison of the proliferation of CD4+CD25high T cells in MS patients and HIs relative to the proliferation of the CD4+CD25– T cell subset. Proliferation of CD4+CD25high cells was not significantly different between patients (n = 11) and HIs (n = 11). When the top 4% of CD4+CD25high T cells were sorted, the cells were not fully anergic, suggesting the presence of activated T cells. Data represent mean ± SD.