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Objective: To evaluate soluble interleukin-2 receptor alpha chain (sIL-2Rα, sCD25) in serum for the determination of rheumatoid arthritis (RA) activity., Methods: Peripheral blood was collected from 108 patients with RA, 39 patients with osteoarthritis (OA) and 50 healthy control subjects, and synovial fluids were from 40 patients with RA. The sera from the patients with RA, the disease control group (osteoarthritis), the healthy control group, and the synovial fluids of the RA patients were detected by enzyme-linked immunosorbent assay (ELISA).The clinical manifestations and laboratory parameters of the patients with RA were recorded and the correlation with the serum sCD25 level was analyzed., Results: The serum sCD25 concentration in RA group was (2 886±1 333) ng/L, the serum sCD25 concentration in OA group was (2 090±718) ng/L, and the serum sCD25 concentration in healthy group was (1 768±753) ng/L. The serum sCD25 level in the patients with RA was significantly higher than that in the disease controls and healthy controls (P<0.001). Sensitivity of serum sCD25 in the diagnosis of RA was 66.1% and specificity was 83.0%;serum sCD25 levels and erythrocyte sedimentation rate (r=0.321, P=0.001), C-reactive protein (r=0.446, P<0.001), DAS28 score (r=0.324, P<0.001), joint tenderness count (r=0.203, P=0.024), D-dimer levels (r=0.383, P<0.001), age (r=0.24, P=0.007), IgG (r=0.207, P=0.028), HRF-IgG (r=0.345, P=0.034) showed a significant positive correlation, and disease duration (r=-0.206, P=0.021) showed a negative correlation with sCD25;In patients with rheumatoid arthritis, the positive rates of serum ESR, CRP, and sCD25 were 14.3% (2 cases), 14.3% (2 cases), and 71.4% (10 cases) in the low disease activity group. The positive rates of serum ESR, CRP and sCD25 in the moderate disease activity group were 94.2% (49 cases), 82.7% (43 cases), and 86.5% (45 cases). The positive rates of serum ESR, CRP, and sCD25 in the high disease activity group were 100% (42 cases), 95.2% (40 cases), and 90.5% (38 cases);36 cases of ESR and/or CRP were negative (about 33.3%) in 108 patients, serum sCD5 levels of 17 cases in these 36 cases (about 47.2%)increased, of which 14 cases (about 82.4%) had a DAS28 score higher than 3.2., Conclusion: The serum sCD25 has a high specificity for diagnosis of RA and a poor sensitivity. The serum level is closely related to the activity of RA, indicating that sCD25 may be involved in the inflammatory process of RA and may become a new inflammatory marker of RA. It is more meaningful for detection of serum sCD25 when RA is active, but ESR and/or CRP is negative.

Objective: To study the levels of CD4+CD25+CD127- and CD3+CD4-CD8- regulatory T (Treg) cells in peripheral blood of children with idiopathic thrombocytopenic purpura (ITP)., Methods: The flow cytometry was used to detect the expression of CD4+CD25+CD127- and CD3+CD4-CD8- Treg cells in peripheral blood of 33 children with ITP and 21 healthy children., Results: The expression levels of CD4+CD25+CD127-［(2.7±1.7)% vs (4.8±1.6)%; P<0.01］and CD3+CD4-CD8-［(5.2±3.1)% vs (8.1±3.5)%; P<0.01］Treg cells in children with ITP were significantly lower than in the controls., Conclusions: The expression levels of CD4+CD25+CD127- and CD3+CD4-CD8- Treg cells decrease in children with ITP, suggesting that CD4+CD25+CD127- and CD3+CD4-CD8- Treg cells might play a role in the pathogenesis of ITP.

Objective: To investigate the expression of CD4+CD25+ regulatory T cell (Treg) in labial gland (SG) of the patients with Sjögren's syndrome (SS)., Methods: Specimens of labial gland were obtained from 25 primary SS patients and 6 healthy controls during biopsy to be made into serial sections. Immunohistochemistry was used to examine the expression of CD25, CD4, CD8, CD68 and Foxp3., Results: Infiltration of a great amount of mononuclear cells and atrophy of part of the labial glands were seen in the specimens from SS patients. The percentage of CD4+ T cells in the infiltrating leucocytes of the SS patients was (44.2 +/- 20.5)%, significantly higher than that of the controls [(9.2 +/- 6.0)%, P < 0.01]. CD25 and Foxp3 were completely absent in the specimens from both the controls and SS patients., Conclusion: CD4+CD25+ regulatory T cell lacks in the labial gland of SS patients. CD4+CD25+ regulatory T cells play a key role in the progression of SS.

As a functionally and phenotypically distinctive T cell subpopulation, CD4+CD25+ regulatory T cells are anergic and retain their ability to suppress antigen-driven response of CD4+CD25- cells in a contact-dependent manner or through a way of secreting immunosuppressive cytokines such as IL-10 and TGF-beta. Graft-versus-host disease (GVHD) is one of the major complications following allogeneic hematopoietic stem cell transplantation (HSCT). Recently, some researches on the relationship between donor CD4+CD25+ regulatory T cells and GVHD severity produced two contradictory conclusions: one is CD4+CD25+ regulatory T cells that can prevent GVHD efficiently; the other is that GVHD is associated with the increased numbers of peripheral blood CD4+CD25+ regulatory T cells. The answer to this question will provide a new idea for clinic therapy of GVHD. In this review some new research progresses in the related area, such as the CD4+CD25+ regulatory T cells, the phenotype, characteristics, immunoregulatory mechanisms of CD4+CD25+ regulatory T cells, as well as the relation of CD4+CD25+ with GVHD were presented.

This study was aimed to analyze the proportion of T cell subsets, CD4(+) CD25(high) regulating T cells (Tr) in peripheral blood of B-NHL patients and their change regularity, and to investigate the immunosuppression mechanism and influence of chemotherapy on immunosuppression function of B-NHL patients. The peripheral blood was collected from 42 patients with B-NHL, 36 patients with B-NHL who achieved partial remission (PR) or complete remission (CR) after 4 - 6 cycles of chemotherapy and 15 healthy controls. By using monoclonal antibodies, the blood samples were evaluated with the flow cytometry for lymphocyte subsets and Tr cells. The results showed that the proportion of CD3(+) and CD4(+) T cells, and the ratio of CD4/CD8 in patients with B-NHL group was significantly less than those in the healthy controls, i.e. (68.33 +/- 15.27)% versus (72.06 +/- 9.26)%; (34.47 +/- 12.84)% versus (42.45 +/- 9.2)%; 1.36 +/- 0.26 versus 1.92 +/- 0.20, but the prevalence of the CD4(+) CD25(high) Tr cells was significantly higher than those in the healthy group [(4.10 +/- 1.21)% versus (2.04 +/- 1.03)%, P < 0.001]. The ratio of CD4/CD8 in chemotherapy group was lower than that in control, but the proportion of CD4(+) CD25(high) Treg cells in chemotherapy group was higher than those before chemo-/radio-therapy and the control. It is concluded that the relative increase of CD4(+) CD25(high) Tr cells in peripheral blood of B-NHL patients may be related to immunosuppression and tumor progression.

Objective: To investigate the role of CD4(+)CD25(+) regulatory T cells (Tr cells) in the pathogenesis of asthma in children., Methods: Peripheral blood samples were collected from 20 pediatric patients with asthma, 10 male and 10 female, aged 7 (3-12), and 20 healthy children, 10 male and 10 female, aged 6.5 (2-11). Lymphocytes were isolated. Flow cytometry was used to examine the percentages of CD4(+)CD25(+) regulatory T cells, IL-10 secreting CD4(+)CD25(+) regulatory T cells (CD4(+)CD25(+)-IL-10), and transforming growth factor (TGF)-beta secreting CD4(+)CD25(+) regulatory T cells (CD4(+)CD25(+)-TGF-beta). RT-PCR and real-time PCR were used to detect the mRNA expression of suppressor of cytokine signal 1 (SOSC1) and Foxp3., Results: The percentages of CD4(+)CD25(+) regulatory T cells of the asthma children was 6.51% +/- 1.94%, significantly lower than that of the healthy children (11.96% +/- 2.30%, P < 0.01); the percentage of CD4(+)CD25(+)-IL-10 of the asthma children was 1.46% +/- 0.35%, significantly lower than that of the healthy children (5.65% +/- 1.70%, P < 0.01); and the percentage of CD4(+)CD25(+)-TGF-beta of the asthma children was 1.24% +/- 0.21%, significantly lower than that t of the healthy children (4.23% +/- 1.65%, P < 0.01). The Foxp3 mRNA expression of the asthma children was 0.12 +/- 0.05, significantly lower than that of the healthy children (1.71 +/- 0.58, P < 0.01); and the SOCS1 mRNA expression of the asthma children was 0.38 +/- 0.19, significantly lower than that of the healthy children (1.51 +/- 0.41, P < 0.01)., Conclusion: The decrease of CD4(+)CD25(+) regulatory T cells may be involved in the pathogenesis of asthma. The decreased mRNA expression of Foxp3 and SOCS1 may be associated with the aberrant development of CD4(+)CD25(+) regulatory T cells.

Objective: To investigate the influence of dendritic cells (DCs) on the prevalence and function of regulatory T cells (Tregs) in cytokine induced killer (CIK) cells., Methods: The blood samples of 20 patients with solid tumors were collected. The peripheral mononuclear cells (PBMCs) were isolated. CIK cells were added into the culture fluid without CD(4)(+)CD(25)(+)T cells (CIK-Treg(del) cells) and the culture fluid of regular PBMCs respectively, and the proliferation and cytotoxicity of the CIK cells were detected by BrdU method and with the cells of human lung carcinoma, breast carcinoma, colon carcinoma, and lymphoma as target cells respectively. CD(4)(+)CD(25)(+)T cells were added into another culture fluid of CIK-Treg(del) cells at the proportions of 20:1, 10:1, and 5:1 respectively, then the proliferation and cytotoxicity of the CIK cells were detected as described above. Flow cytometry was used to detect the surface markers of CIK cells. To identify the influence of DCs on the anti-tumor activity of CIK, PBMCs were isolated from the patients with solid tumor to culture the DCs and CIK cells. Dendritic cells were harvested on day 7 and co-cultured with the CIK cells (DC+CIK cells). The frequency of Tregs in CIK was determined by flow cytometry. The cytotoxicity was examined by LDH assay. The levels of TGF-beta, IL-10, IFN-gamma, IL-2, and IL-6 were analyzed by ELISA., Results: The rates of the main effector cells in CIK cells (CD(3)(+)CD(56)(+) cells) were 17% +/- 5% and 28% +/- 5% in the regular CIK cells and CIK-Treg(del) cells respectively. LDH method showed that the cytotoxicity towards tumor cells of the CIK-Treg(del) cells The rates of the main effector cells in CIK cells (CD(3)(+)CD(56)(+) cells) were 17% +/- 5% and 28% +/- 5% in the regular CIK cells and CIK-Treg(del) cells respectively. LDH method showed that the cytotoxicity towards tumor cells of the CIK-Treg(del) cells was higher than that of the regular CIK cells (P < 0.05), however, after the addition of selected cells, the cytotoxicity of the CIK-Treg(del) cells decreased. Flow cytometry showed that the proportions of CD(4)(+)CD(25)(+) Treg cells in the CIK cells and DC-CIK cells were 13% +/- 5% and 10% +/- 4% respectively (t = 3.977, P = 0.001). After the DC induction the cytotoxicity of CIK cells was significantly higher than that of the regular CIK cells. ELISA showed that after DC induction the levels of TGF-beta and IL-10 of the DC+CIK group were significantly lower than those of the regular CIK cells (t = 2.136, P = 0.046; and t = 2.965, P = 0.008), and the level of IFN-gamma was significantly higher in the DC+CIK group (t = 2.220, P = 0.039)., Conclusion: CD(4)(+)CD(25)(+) regulatory T cells inhibit the anti-tumor activity of CIK cells. The interaction between CIK cells and DCs is sufficient for the blockage of the properties of regulatory T cells. CIK cells have the desirable properties for immunotherapy approaches, especially after co-culture with DCs.

The purpose was to study the effect of mesenchymal stem cell (MSC) on immune function, and to explore the new strategy to prevent graft versus host disease (GVHD) and host versus graft reaction (HVGR) in allogeneic bone marrow transplantation. MSCs from human bone marrow cells were isolated and cultured. The purity of MSCs were identified with the spindle-fibroblastic morphological characterization by microphotography, and the phenotypes were tested by flow cytometry. MSCs were planted in 6-well plates (8 x 10(4)/well for group A, 4 x 10(4)/well for group B and 2 x 10(4)/well for group C), and cocultured for 7 days with T cell isolated from peripheral blood. Peripheral blood T cells non-cocultured with MSC acted as the control group. CD3, CD4, CD8, and CD25 expressed on T cells were analyzed by flow cytometry after coculture with MSCs for 0, 24, 72 hours and 7 days respectively. The results showed that CD4(+)CD25(+) T cells and CD8(+) T cells of allogeneic T lymphocytes cocultured with bone marrow MSCs (group A and group B) increased obviously as compared with control group (T lymphocytes uncocultured with MSCs), and there were no difference between groups A and B. CD3, CD4, CD8 and CD25 expressed on T cells had no significant difference between experimental groups and control group. The result suggested that CD4(+)CD25(+)-regulatory T cells and CD8(+) T cells were increased apparently when cocultured with allogeneic MSCs. It is concluded that MSCs pretreatment may be useful in the prevention of GVHD and HVGR in allogeneic bone marrow transplantation and provides a new strategy to induce transplantation tolerance.