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 September 2, 2008
Citation Information: J Clin Invest. 2008;118(10):3411-3419. https://doi.org/10.1172/JCI35365.
View: Text | PDF
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

×

Figure 2

Suppressive activity and proliferation of the top 2% of sorted CD4+CD25high cells from MS patients and healthy controls.

Options: View larger image (or click on image) Download as PowerPoint
Suppressive activity and proliferation of the top 2% of sorted CD4+CD25h...
(A) CD4+ lymphocytes obtained from the peripheral blood of MS patients and healthy controls were stained with Pe-Cy7–conjugated anti-CD3, FITC-conjugated anti-CD8, Alexa Fluor 647–conjugated anti-CD25, and PE-conjugated anti-CD127. Sorting was performed on the CD4+CD25highCD127low and CD4+CD25highCD127high populations. Sorting was also performed on the CD4+CD25high population with 2 different gates. In the example provided for 1 MS patient, CD4+CD25+ T cells appear in orange, CD4+CD25+CD127low T cells appear in green, and CD127high cells appear in violet. The presence of CD127high activated cells can be observed in the CD4+CD25high sorted T cells when the gate is not stringent enough (4% gating stringency, area above the red line), while in the case of a 2% gating stringency (area above the blue line), only a few CD127high cells remain within the CD4+CD25high T cell subset. FSC, forward scatter. (B) CD4+CD25– responder cells were stimulated with anti-CD3 antibody (0.1 μg/ml) in coculture with the top 2% of CD4+CD25high sorted cells. Data are the mean of duplicate wells. Regulatory properties of CD4+CD25high cells are comparable in both HIs (n = 10) and patients (n = 12). (C) Comparison of the proliferation of the top 2% of sorted CD4+CD25high T cells in MS patients and HIs relative to the proliferation of the CD4+CD25– T cell subset. The proliferation of CD4+CD25high T cells was minimal and almost null in both patients and controls. Data are mean ± SD. (D) Intracellular FOXP3 staining was performed on the top 2% of sorted CD4+CD25high T cells in 10 patients and 9 HIs. In B and D, the mean values for each group are indicated by horizontal lines.