Korean J Physiol Pharmacol.  2015 Mar;19(2):83-88. 10.4196/kjpp.2015.19.2.83.

Dynamic Frequency of Blood CD4+CD25+ Regulatory T Cells in Rats with Collagen-induced Arthritis

  • 1Department of Pharmacy, Shanghai Ninth People's Hospital, Shanghai Jiaotong University School of Medicine, Shanghai 200011, China.
  • 2School of Pharmacy, Anhui Medical University, Hefei 230032, Anhui, China. aydlijun@sohu.com
  • 3Bone Research Program, ANZAC Research Institute, University of Sydney, NSW 2139, Sydney, Australia.


CD4+CD25+ regulatory T cells (CD4+CD25+ Tregs) have been shown to play a regulatory or suppressive role in the immune response and are possibly relevant to the pathogenesis of autoimmune diseases. In the present study, we attempted to investigate the frequency of CD4+CD25+ Tregs in peripheral blood (PB) of collagen-induced arthritis (CIA) rats during the development of arthritis, to determine whether their frequency is involved in the immunoregulation of this disease. The results showed that normal rats had similar frequencies of CD4+CD25+ Tregs in PB during the experiment time, expressed as a percentage of CD4+CD25+Foxp3+ T cells among the CD4+ T lymphocyte population. In contrast, the frequency of CD4+CD25+Foxp3+ T cells in CIA rats was found to change during the development of arthritis. In CIA rats, there is a significant negative correlation between the frequency of CD4+CD25+Foxp3+ T cells and paw swelling (r=-0.786, p< 0.01). The relationship between the frequency of CD4+CD25+Foxp3+ T and immune activation was not found in normal rats. During the time course, the frequency of CD4+CD25+Foxp3+ T was lower in CIA rats than in normal ones. The data suggest that the frequency of PB CD4+CD25+ Tregs may be a promising marker for arthritis activity.


Arthritis; CD25; Correlation; Foxp3; Frequency

MeSH Terms

Arthritis, Experimental*
Autoimmune Diseases
T-Lymphocytes, Regulatory*


  • Fig. 1 Paw swelling of CIA rats. The paw swelling was expressed as an increase in mean bi-hind paw volume by subtracting that at day 0. (A) Paw swelling during arthritis progress in CIA rats. (B) Photo of the hind paw of a normal rat on day 17. (C) Photo of the hind paw of a CIA rat on day 17. Values are expressed as the means±SEM for ten animals in each group. Compared with normal group, ##p<0.01.

  • Fig. 2 Gating strategy used to determine the proportion of CD4+CD25+Foxp3+ Tregs. Erythrocytes were lysed with NH4Cl lysing solution. Cells were stained with FITC-conjugated anti-CD4 antibody and PE-conjugated anti-CD25 antibody. After permeabilization, cells were stained with PE-Cy5-conjugated anti-Foxp3 antibody. The labeled cells were detected on a FACS Calibur. Lymphocytes were gated using an FSC vs. SSC plot (A), followed by a CD4+ gate in a CD4 (FL-1) vs. CD25 (FL-2) plot (B). Then, in CD4+ T cells, Tregs were discriminated according to their CD25 (FL-2) and Foxp3 (FL-3) expression, by which CD4+CD25+Foxp3+ cells are considered as Tregs (C).

  • Fig. 3 Proportion of peripheral blood CD4+CD25+ Tregs during the arthritis progress. (A) Flow cytometry of peripheral blood CD4+CD25+ Tregs of normal and CIA rats at different time points. (B) Proportion of peripheral blood CD4+CD25+ Tregs during arthritis progress. Values are expressed as the means±SEM for five animals in each group. Compared with normal group, #p<0.05, ##p<0.01.

  • Fig. 4 Correlation between the frequency of CD4+CD25+Foxp3+ Tregs and paw swelling. Linear regression was used to analyze the correlation between the frequency of peripheral blood CD4+CD25+ Foxp3+ T cells and paw swelling during arthritis progress. (A) The frequency of CD4+CD25+ Tregs was not correlated with paw swelling in normal rats. (B) The frequency of CD4+CD25+ Tregs was negatively correlated with paw swelling with statistical significance in CIA rats (r=-0.786).


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