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Tolerogenic dendritic cells (tolDCs) facilitate the suppression of autoimmune responses by differentiating regulatory T cells (Treg). The dysfunction of immunotolerance results in the development of autoimmune diseases, such as rheumatoid arthritis (RA). As multipotent progenitor cells, mesenchymal stem cells (MSCs), can regulate dendritic cells (DCs) to restore their immunosuppressive function and prevent disease development. However, the underlying mechanisms of MSCs in regulating DCs still need to be better defined. Simultaneously, the delivery system for MSCs also influences their function. Herein, MSCs are encapsulated in alginate hydrogel to improve cell survival and retention in situ, maximizing efficacy in vivo. The three-dimensional co-culture of encapsulated MSCs with DCs demonstrates that MSCs can inhibit the maturation of DCs and the secretion of pro-inflammatory cytokines. In the collagen-induced arthritis (CIA) mice model, alginate hydrogel encapsulated MSCs induce a significantly higher expression of CD39+CD73+ on MSCs. These enzymes hydrolyze ATP to adenosine and activate A2A/2B receptors on immature DCs, further promoting the phenotypic transformation of DCs to tolDCs and regulating naïve T cells to Tregs. Therefore, encapsulated MSCs obviously alleviate the inflammatory response and prevent CIA progression. This finding clarifies the mechanism of MSCs-DCs crosstalk in eliciting the immunosuppression effect and provides insights into hydrogel-promoted stem cell therapy for autoimmune diseases.
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@#[Abstract] Objective: To detect the expression of CD39 in head and neck squamous cell carcinoma (HNSCC) tisseus, and to analyze its correlation with patients’clinicopathological features and its prognostic significance. Methods: Tissue specimens and case data of 85 patients with HNSCC underwent surgery at Cancer Hospital of Tianjin from May 2012 to December 2013 were collected for this study. Gene chips were obtained from Oncomine database, and HNSCC cell lines SCC15, UM1, and Cal25 were selected for this study. Online analysis was performed to compare the differential expression of CD39 in buccal mucosa (BM) tissues and HNSCC tissues, Western blotting and Immunohistochemistry (IHC) were used to detect the protein expression of CD39 in HNSCC tissues. Spearman’ s correlation analysis was used to study the correlation between the expressions of CD39 and clinicopathological features of HNSCC patients. Both Kaplan-Meier curve analysis and Log rank test were used to analyze the association between the expression of CD39 in HNSCC tissues and the survival of patients, and Cox risk proportional regression model was used to evaluate the relationship between CD39 expression and the risk of relapse. Results: The transcription level of CD39 was obviously up-regulated in HNSCC tissues than in BM tissues (P<0.01), and CD39 expression was detected in HNSCC cell lines SCC15, UM1 and Cal25. Dexamethasone (DXM) could enhance the expression of CD39 in UM1 cells in dose-dependent manner. CD39 was highly expressed in 53 (62.4%) HNSCC patients, which was positively correlated with preoperative chemotherapy (r=0.234, P<0.05). The recurrence-free survival (RFS) of patients with high CD39 expression was significantly shortened (P<0.05), and high CD39 expression was an independent relapse risk factor (HR=2.328, 95%CI=1.091-4.967; P<0.05) for patients with HNSCC. Conclusion: CD39 is DXM-inducively and constitutively expressed in HNSCC. And over-expression of CD39 is an independent predictor of poor prognosis in HNSCC patients, indicating its important role in the progression of HNSCC.
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Objective To evaluate the changes of adenosine metabolism pathway related molecules and their contribution to inflammatory injury in primary biliary cholangitis (PBC).Methods The consecutive samples of 49 subjects with PBC from The First People's Hospital of Taicang and The Second People's Hospital of Changshu were recruited from October 2016 to October 2017,and 36 healthy controls were involved in this study.The expression of CD39 and CD73 on CD4+T cells and Foxp3 + regulatory T cells were assayed by flow cytometry and the concentration of adenosine triphosphate (ATP) in serum was analyzed by ultraperformance liquid chromatography-quadrupole time-of-flight mass spectrometry (UHPLC-Q-TOF/MS).The correlations between Tregs,ATP and liver function were analyzed,i.e.,alanine aminotransferase (ALT),aspartate aminotransferase (AST),gamma-glutamyltransferase (GGT),alkaline phosphatase (ALP) and Mayo scores.Results In the patients with PBC,low proportions of CD4+CD39+T cells were noted compared with healthy controls [(5.28 ± 1.92) ％ vs (11.0l ± 3.19) ％,t =10.25,P ＜ 0.01].The patients with PBC also had significantly low proportion of CD4+CD25 + Foxp3+ CD39+ T cells compared with healthy controls [(23.75 ± 9.48) ％ vs (54.68 ± 5.18) ％,t =13.79,P ＜0.01].No significant difference of the proportion of CD4+CD73+T or CD4+CD25+ Foxp3+CD39+T cells was found between PBC and control groups (t values were 2.235 and 1.083,P ＞ 0.05).The level of serum ATP was higher in the patients with PBC than that of healthy controls [(200.28 ± 79.41) μg/L vs (89.20 ± 33.76) μg/L,t =8.367,P ＜ 0.01].A significant correlation was demonstrated between the proportion of CD39 + Treg in total Treg cells and the levels of ATP (r =-0.413,P =0.003),GGT (r=-0.378,P=0.007) and Mayo score (r=-0.382,P=0.007).Conclusion The low proportion of CD39+ Treg cells may contribute to the down-regulation of ATP hydrolysis in the patients with PBC.No significant change of CD73 + Treg cells was found in PBC patients.
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Objective The objectives of this study were to investigate the clinical significance of CD39,CD73,double positive subgroups for CD39 and CD73,and other lymphocytes with clinicopathological parameters in the microenvironment of colorectal cancer.Methods Tumor infiltrating lymphocyte(TIL)was collected from 24 patients with colorectal cancer after radical resection.The expression of CD39+,CD73+ or CD39+ with CD73+ in T cells were measured by flow cytometry.The association between these subgroups and clinicopathologic parameters was analyzed.Results The CD73+ and CD39+ with CD73+ subgroups were associated with lymph node metastasis and poor degree of differentiation,and this mechanism was closely related to tumor-associated inflammation.Conclusion CD39+ with CD73+ colorectal tumor infiltration Treg has a more unique biological activity than other Treg group.This study provides a new idea and theoretical basis for predicting the prognosis of colorectal cancer.
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Objective To study the clinical significance of CD39 on regulatory T (Treg) cells in the peripheral blood (PB) of patients following liver transplantation and to evaluate the relationship between the levels of CD39+ Treg cells in the PB and acute rejection.Methods A prospective study was conducted to compare the CD39+ Treg cells from 76 liver transplant patients with those coming from 20 age-matched healthy individuals.The PB samples were collected within one year at different time points post-transplant.Blood samples and liver biopsies were collected at the time when acute rejection was diagnosed.The percentages of CD39 within the CD4+ CD25+ T cells were measured by using flow cytometry.The liver transplant patients were classified into two groups:the rejection group which consisted of 17 patients who an episode of acute rejection,and the non-rejection group consisted of the remaining 59 patients who had no acute rejection episodes.The percentages of CD39 within the CD4 + CD25 + T cells and the inhibition function of the CD39+ Treg cells were compared between the two liver transplant groups.Results The percentages of CD39 within the CD4+ CD25+ cells were significantly lower in the rejection group during acute rejection as compared to the non-rejection group (P ＜ 0.05).The percentages of CD39 within the CD4 + CD25 + cells were negatively correlated with the Rejection Activity Index (r =-0.86,P ＜ 0.05).The inhibition rate regarding the CD4+ CD25+ CD39+ Treg cells in patients with acute rejection was significantly lower than those without rejection (P ＜ 0.05).Conclusions The percentages of CD39 within the CD4+ CD25+ T cells were significantly lower in the rejection group during acute rejection and were negatively correlated with the RAI.The inhibition rate regarding the CD4+ CD25+ CD39+ Treg cells in patients with acute rejection was significantly lower than those without rejection.
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Objective To investigate expression and suppressive function of CD39 + regulatory T cells (Treg) in kidney transplant recipients.Method Thirty recipients of first kidney transplants were treated with tacrolimus,mycophenolate mofetil and prednisone.Within 28 days posttransplantation,there were 14 patients subject to acute rejection (AR group),and the rest 16 patients had no episodes of acute rejections (NR group).Twelve healthy volunteers served as healthy controls (HC group).We collected peripheral blood from the three groups and separated PBMC by density gradient centrifugation,and sorted Tresp,CD39-Treg and CD39+ Treg by flow cytometry.We next analyzed the ratio of CD39 + Treg/CD4+ T cells.ELISA was used to determine the suppressive ability of CD39-Treg and CD39+ Treg on secretion of IFN-γ and IL-17 by Tresp.Results The ratio of CD39 + Treg/CD4 + T cells in AR group was significantly reduced as compared with HC group and NR group (P＜0.05).In HC group and NR group,the secretion of IFN-γ and IL-17 by Tresp was suppressed significantly (P＜0.05) by CD39+ Treg.CD39Treg could suppress secretion of IFN-γ but not IL-17 production by Tresp.CD39+ Treg in AR group AR could suppress the secretion of IFN-γ significantly (P＜0.01),but not to IL-17 production.Conclusion CD39+ Treg have important immunoregulation function.The relative amount of CD39+ Treg was reduced and their regulatory function was impaired in patients with acute rejection.
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Objective To analyze the influence of ionizing radiation on the viability of the regulatory T cells and the relationship between the expression of CD39 and apoptosis related factors in mice Treg cells.Methods Mice were administered to whole body irradiation with 2 Gy of γ-rays,then lymphocytes were separated from thymus and spleen.The percentages of CD4 + T and CD4 + FOXP3 +CD25 + Treg lymphocytes,apoptosis related genes of Bax,Bcl-2 and CD39 were analyzed with FACS.Results Treg cells had a higher survival fraction than CD4 + FOXP3-Tcon cells after irradiation,and the time-responses of Treg cells in thymus and spleen were different.The apoptosis rate of Treg cells increased at 4 d after irradiation(t =-3.6,-7.4,P ＜0.05).The Bax/Bcl-2 ratio and the CD39 expression were increased in thymus Treg cells (t =-3.4,-6.6,P ＜ 0.05),but decreased in spleen Treg cells (t =2.4,2.6,P ＜0.05).Conclusions Thechanges of apoptosis and Bax/Bcl-2 are in consistent with the expression of CD39 in the irradiated Treg cells.