Cytokine flow cytometry differentiates the clinical status of multiple sclerosis (MS) patients

Abstract

In this study we have examined intracellular cytokines in peripheral blood mononuclear cells (PBMC) of MS patients by flow cytometry (cytokine flow cytometry). MS progressive patients showed an increased number of cells producing interferon-gamma (IFN-gamma) after activation with phorbol 12-myristate 13-acetate and ionomycin, compared with patients with clinically inactive forms (P < 0001) and with healthy controls (P = 0001). These cells belonged to the CD4+ and CD8+ subsets in similar proportions. Clinically inactive patients showed a lower level of cells producing IL-2 than controls (P = 0.03) and active MS patients (P = 0.03). Most IL-2-producing cells were CD4+ lymphocytes, although a small part of the IL-2 was also produced by CD8+ cells. The percentage of cells producing simultaneously IL-2 and IFN-gamma was increased in active MS and they were mainly CD4+ lymphocytes. No differences in the production of IL-4 were observed between groups. However, we found an increased IL-10 production in clinically active MS patients (P = 0.03). Treatment with IFN-beta of active MS patients showed lower levels of cytokines when compared with untreated MS patients. This methodological approach could help in the follow up and therapeutic monitoring of MS patients.

Orphan nuclear receptor NR4A2 expressed in T cells from multiple sclerosis mediates production of inflammatory cytokines

“…However, it was recently demonstrated that peripheral T cells from MS and healthy subjects significantly differ in surface phenotype or gene expression profiling (17,23,28). Using cDNA microarray, we have identified NR4A2 as a gene most significantly up-regulated in the peripheral T cells of MS (17).…”

Cytokine production and helper T cell subsets in Vogt-Koyanagi-Harada’s disease

“…4,26 It is therefore suggested that Th cells from patients with VKH disease are apt to deviate toward the Th1 type, and activated Th cells in the inflammatory foci produce Th1 cytokines, thereby promoting inflammation. In addition, RA, 24,25 multiple sclerosis (MS) 27 and Behçet's disease 9 are also considered to be Th1-prone diseases. In contrast to these diseases, allergic diseases 28 and systemic lupus erythematosus (SLE) 29,30 are considered to be Th2-prone diseases.…”

“…Cytokine treatments have been tried in some autoimmune diseases. 27,31 Inflammation of VKH disease may benefit from Th1 cytokine suppression or Th2 cytokine induction. However, artificial control of Th1/Th2 balance must be regulated cautiously, because reduction of excessive Th1 or Th2 cytokine production may precipitate the displacement of equilibrium toward another cytokine-prone condition.…”

Upregulation of Th1 Polarization by <i>Taraxacum officinale</i> in Normal and Immune Suppressed Mice

“…All the test drugs were added at a concentration 10 g/ml and incubated for 4 h at 37C. After adding permeabilizing agent, phycoerythrin (PE)-labelled antimouse IL-2, IFN-gamma and IL-4 were added to the samples in separate falcon tubes and again incubated for 30 min, resuspended in PBS (pH 7.4) and acquired directly on the flow cytometer (BD LSR; Becton Dickinson) 21 .…”

“…CA was added at concentrations of 0.25, 0.5, 1 and 2 g/ml and incubated for 4 h at 37C. PE-labelled antimouse IL-2, IFN-gamma and IL-4 were added to the samples, resuspended in PBS (pH 7.4) and acquired directly on the flow cytometer (BD LSR; Becton Dickinson) 21 .…”