Role of CD8+ CD25+ Foxp3+ regulatory T cells in multiple sclerosis

Ann Neurol. 2010 May;67(5):625-38. doi: 10.1002/ana.21944.


Objective: The objective of this study was to investigate the role of CD8+ CD25+ FoxP3+ cells during the course of multiple sclerosis (MS).

Methods: Peripheral blood and cerebrospinal fluid (CSF) CD8+ T-cell clones (TCCs) recognizing autoreactive CD4+ T cells were isolated from 20 MS patients during exacerbations, 15 patients in remission, 15 healthy subjects, and 10 patients with other inflammatory neurological diseases. Characteristics of noncytotoxic CD8+ CD25+ regulatory T cells were studied. Cell phenotype was evaluated using flow cytometry. Cytokine production and phospho-signal transducer and activator of transcription 3 (STAT3) concentration were determined using enzyme-linked immunosorbent assay. To assess 2,3-dioxygenase (IDO) activity on dendritic cells (DCs), kynurenine concentration was measured by high-performance liquid chromatography.

Results: Inhibition of CD4+ self-reactive T-cell proliferation, and of interferon-gamma and interleukin (IL)-17 secretion, was observed after adding CD8+ CD25+ FoxP3+ cells to cultures. Suppression was abrogated by silencing FoxP3 using small interfering RNA. Cells were CD122+, CTLA-4+, GITR+, CCR7+, and CD62L+, producing IL-10 and transforming growth factor-beta. CD8+ CD25+ FoxP3+ cells downregulated costimulatory molecule expression on dendritic cells through a STAT3-mediated pathway, resulting in less efficient antigen presentation, and induced IDO expression on DCs through STAT3 and cytotoxic T-lymphocyte antigen 4-dependent mechanisms. CD8+ regulatory TCC cloning frequency studied in blood and CSF was suppressed to a greater degree during exacerbations than during remission or in controls. Likewise, in CSF of MS patients during acute exacerbations, lower levels of CD8+ CD25+ FoxP3+ T cells were detected using flow cytometry.

Interpretation: CD8+ CD25+ FoxP3+ cells are novel regulatory cells exerting significant influence over self-reactive CD4+ T-cell regulation during the course of MS. Induction of these cells may provide new therapeutic alternatives for MS by eliminating or inhibiting self-reactive T cells.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • CD4 Antigens / pharmacology
  • CD8 Antigens / metabolism*
  • Cell Proliferation
  • Chromatography, High Pressure Liquid / methods
  • Dendritic Cells / metabolism
  • Enzyme-Linked Immunosorbent Assay / methods
  • Female
  • Flow Cytometry / methods
  • Forkhead Transcription Factors / metabolism*
  • Gene Expression Regulation / drug effects
  • Humans
  • Indoleamine-Pyrrole 2,3,-Dioxygenase / metabolism
  • Interferon-gamma / metabolism
  • Interleukin-10 / metabolism
  • Interleukin-2 Receptor alpha Subunit / metabolism*
  • Logistic Models
  • Male
  • Multiple Sclerosis / pathology*
  • RNA, Small Interfering / pharmacology
  • STAT3 Transcription Factor / metabolism
  • T-Lymphocytes, Regulatory / drug effects
  • T-Lymphocytes, Regulatory / physiology*


  • CD4 Antigens
  • CD8 Antigens
  • FOXP3 protein, human
  • Forkhead Transcription Factors
  • Indoleamine-Pyrrole 2,3,-Dioxygenase
  • Interleukin-2 Receptor alpha Subunit
  • RNA, Small Interfering
  • STAT3 Transcription Factor
  • STAT3 protein, human
  • Interleukin-10
  • Interferon-gamma