Zonal Difference and Prognostic Significance of Foxp3 Regulatory T Cell Infiltration in Breast Cancer / 한국유방암학회지

PURPOSE: Forkhead box P3 (Foxp3) is known as the most specific marker for regulatory T lymphocytes, which play an important role in immune tolerance to disturb antitumor immunity. The present study aimed to investigate the prognostic significance of Foxp3 regulatory T lymphocyte (Foxp3 Treg) infiltration in breast cancer. METHODS: Immunohistochemical studies with Foxp3, CD4, and CD8 were performed on representative full tissue sections from 143 patients with invasive ductal carcinoma, not otherwise specified. Foxp3 Treg infiltration and the ratios between Foxp3 Treg and CD4 or CD8 T cells were separately analyzed for the tumor bed and tumor periphery to evaluate their association with different clinicopathological parameters and patients' outcome. RESULTS: The tumor periphery was considerably more densely infiltrated by Foxp3 Treg, CD4, and CD8 T cells than the tumor bed. Unfavorable clinicopathological parameters (a Ki-67 labeling index of > or =14%, a worse histologic grade, a worse nuclear grade, hormone receptor negativity, human epidermal growth factor receptor 2 positivity, and tumor recurrence) were associated with increased Foxp3 Treg infiltration and a high ratio between Foxp3 Treg and CD4/CD8 T cells. In the tumor periphery, as Foxp3 Treg infiltration and the Foxp3 Treg/CD8 ratio increased, patients' 5-year disease-free survival rate decreased. CONCLUSION: The infiltration densities of Foxp3 Treg, CD4, and CD8 T cells were markedly different between the tumor bed and periphery. Besides the absolute count of Foxp3 Treg, the ratio between Foxp3 Treg and effector T cells was a significant prognostic factor in breast cancer.

Study on the prevention and therapeutic effects of Faecalibacterium prausnitzii on colitis of experimental rats / 中华消化杂志

Objective To explore the therapeutic effects and the mechanisms of Faecalibacterium prausnitzii (Fp) on trinitro-benzene-sulfonic acid-induced colitis.Methods Sixty Sprague-Dawley rats were divided into healthy control group, colitis model control group, Fp pretreated group,Fp supernatant pretreated group,Fp treated group and Fp supernatant treated group.Disease activity index (DAI),histological injury of colonic tissue,the content of butyrate in feces,forkhead box protein 3 (Foxp3) regulatory T cells (Treg) in peripheral blood and spleen and the level of interlenkin (IL)-17 and IL-6 in serum were evaluated.All the data were statistical analyzed by single factor analysis of variance. Results Compared with colitis model control group, DAI significantly lowered and histological injury obviously improved in Fp pretreated group, Fp supernatant pretreated group,Fp treated group and Fp supernatant treated group.The effects of Fp pretreated group were better than those of Fp treated group and Fp supernatant pretreated group were better than Fp supernatant treated group.The concentration of butyrate in Fp pretreated group,Fp supernatant pretreated group,Fp treated group and Fp supernatant treated group was (3091.08 ±485.50) × 106 mol/L,(1714.64 ± 351.25) × 10(-8) mol/L,(2064.75 ± 295.04) × 10-6 mol/L and (1089.13±321.23) × 10-6 mol/L respectively,there was significant difference between Fp pretreated group and other groups (F=49.796,P＜0.01).The peripheral blood level of Foxp3+ Treg in Fp supernatant pretreated group was highest.The spleen level of Foxp3+ Treg in Fp pretreated group and Fp supernatant pretreated group were significantly higher than that of other groups.The serum level of IL-17 and IL-6 in Fp pretreated group,Fp supernatant pretreated group,Fp treated group and Fp supernatant treated group was significantly lower than that of colitis model group.Conclustons Fp plays a role in promoting the repair of intestinal inflammatory reaction in colitis model rats.The mechanism may be related with butyrate producing,the peripheral blood and spleen level of Foxp3+ Treg up-regulating,suppressing the secretion of proinflammatory cytokine IL-17 and IL-6.Rebuilding the balance of Treg/Th17 to reduce local intestinal inflammation.

The Regulation of FOXP3 Expression by the Treatment of TGF-beta and the Modification of DNA Methylation in Lung Cancer Cell Lines / 결핵및호흡기질환

BACKGROUND: Transcription factor FOXP3 characterizes the thymically derived regulatory T cells. FOXP3 is expressed by cancer cell itself and FOXP3 expression was induced by TGF-beta treatment in pancreatic cancer cell line. However, the expression of FOXP3 expression is not well known in patients with lung cancer. This study was conducted to investigate the expression of FOXP3 in patients with lung cancer and to investigate the regulation of FOXP3 expression by the treatment of TGF-beta and DNA methyltransferase inhibitor in lung cancer cell lines. METHODS: FOXP3 expression in the tissue of patients with resected non-small cell lung cancer (NSCLC) was evaluated by immunohistochemistry. The regulation of FOXP3 expression was investigated by Western blot and RT-PCR after lung cancer cell lines were stimulated with TGF-beta1 and TGF-beta2. The regulation of FOXP3 expression was also investigated by RT-PCR and flow cytometry after lung cancer cell lines were treated with DNA methyltransferase inhibitor (5-AZA-dC). RESULTS: FOXP3 expression was confirmed in 27% of patients with NSCLC. In NCI-H460 cell line, TGF-beta2 decreased FOXP3 mRNA and protein expressions. In A549 cell line, both TGF-beta1 and TGF-beta2 decreased FOXP3 mRNA and protein expressions. 5-AZA-dC increased FOXP3 mRNA expression in NCI-H460 and A549 cell lines. Moreover, 5-AZA-dC increased intracellular FOXP3 protein expression in A549 cell lines. CONCLUSION: It was shown that FOXP3 is expressed by cancer cell itself in patients with NSCLC. Treatment of TGF-beta2 and DNA methyltransferase inhibitor seems to be associated with the regulation of FOXP3 expression in lung cancer cell lines.

Higher infiltration by Th17 cells compared with regulatory T cells is associated with severe acute T-cell-mediated graft rejection

The aim of this study was to evaluate whether the Th17 and Treg cell infiltration into allograft tissue is associated with the severity of allograft dysfunction and tissue injury in acute T cell-mediated rejection (ATCMR). Seventy-one allograft tissues with biopsy-proven ATCMR were included. The biopsy specimens were immunostained for FOXP3 and IL-17. The allograft function was assessed at biopsy by measuring serum creatinine (Scr) concentration, and by applying the modified diet in renal disease (MDRD) formula, which provides the estimated glomerular filtration rate (eGFR). The severity of allograft tissue injury was assessed by calculating tissue injury scores using the Banff classification. The average numbers of infiltrating Treg and Th17 cells were 11.6 +/- 12.2 cells/mm2 and 5.6 +/- 8.0 cells/mm2, respectively. The average Treg/Th17 ratio was 5.6 +/- 8.2. The Treg/Th17 ratio was significantly associated with allograft function (Scr and MDRD eGFR) and with the severity of interstitial injury and tubular injury (P < 0.05, all parameters). In separate analyses of the number of infiltrating Treg and Th17 cells, Th17 cell infiltration was significantly associated with allograft function and the severity of tissue injury. By contrast, Treg cell infiltration was not significantly associated with allograft dysfunction or the severity of tissue injury. The results of this study show that higher infiltration of Th17 cell compared with Treg cell is significantly associated with the severity of allograft dysfunction and tissue injury.

Significance of Foxp3 Positive Regulatory T Cell and Tumor Infiltrating T Lymphocyte in Triple Negative Breast Cancer

BACKGROUND: Triple negative breast cancer (TNBC) is defined as a lack of the expression of estrogen receptor, progesterone receptor and human epidermal growth factor receptor 2 in breast cancer. Many TNBCs show a profound infiltration of tumor infiltrating lymphocytes (TILs). It is still uncertain whether these TILs are protumoral or antitumoral. Regulatory T cells (Tregs) play a role in inducing immune tolerance to antigens, and they may be selectively recruited by cancer cells. This study was conducted to evaluate the significance of TILs with an emphasis on forkhead box p3 (Foxp3), which is a marker for CD25+CD4+ Treg in TNBC. METHODS: We investigated the Foxp3, CD8 and CD4 expressions in 100 cases of TNBC by immunohistochemistry and using a tissue microarray. The Foxp3 expression was divided as the high and low infiltration groups (cut-off value=20). RESULTS: The high infiltration group was correlated with higher histologic and nuclear grades. However, Foxp3+ Tregs were decreased in the T3 and T4 TNBCs as compared to that of the T1 and T2 TNBCs. No significant differences were found for the nodal status, lymphovascular invasion, stage, recurrence and overall survival. CONCLUSIONS: High Foxp3+ Treg infiltration in TNBC is correlated with the nuclear and histologic grades, but there was no relation to recurrence and overall survival.

The Prognostic Significance of the Tumor-Infiltrating FoxP3-Positive Regulatory T Cells in Gastric Carcinoma

BACKGROUND: Regulatory T cells (Tregs) are known to be key regulators of immune responses in patients with autoimmune disease and infection and also for attenuating antitumor immunity by the host. It has been reported that high numbers of tumor-infiltrating Tregs might be associated with poor clinical outcomes for several malignant tumors. Therefore, this study aimed to examine the impact of tumor-infiltrating Tregs on the prognosis of gastric carcinoma patients. METHODS: The immunohistochemical staining for anti-fork head Box P3 (FoxP3) antibody was performed by using a 3 mm core from the tumor specimens of each of the 173 gastric cancer patients for constructing a tissue microarray. FoxP3-positive Tregs were quantified by calculating the numbers of positive cells per 5 high-power fields on light microscopy. Thereafter, the 173 patients were subdivided into the low Tregs group ( 3/5 HPF, n = 132). RESULTS: The high Tregs group was significantly associated with a higher stage, more invasion depth and lymph node metastasis (p = 0.009, p = 0.036, p = 0.006, respectively). The high Tregs group showed significantly poorer overall survival and event-free survival (p = 0.004, p = 0.017, respectively) on the univariate analysis. The Tregs group and the tumor, node and metastasis stage were also independent prognostic factors that were significantly associated with overall survival (p = 0.025, p < 0.001, respectively) by multivariate analysis. CONCLUSIONS: Our results indicated that a high number of tumor-infiltrating FoxP3-positive Tregs could be an indicator of poor long term survival for gastric carcinoma patients.

Studies on the relationship between pulmonary function changes with Foxp3,TGF-β1/Smads signal transduction pathway in adjuvant arthritis rats / 中国免疫学杂志

Objective:To observe the changes of lung index,lung function,regulatory T cells and Foxp3,TGF-β1,Smad3,Smad7 protein expression in adjuvant arthritis(AA) rats,and to investigate the possible mechanism of Foxp3 and TGF-β/Smads signal transduction pathway in reducing lung function.Methods: 24 Wistar rats were randomly divided into normal control group and model group,12 rats in each group.The rats were injected with 0.1 ml Freunds'complete adjuvant to induce inflammation at the part of their rightback foot in model group.On the 48~(th) day post-inflammatory,the swelling degree of toe and arthritis index(AI) were observed.Lung index of the rats was calculated,and pulmonary function of the rats was detected.The percentage of regulatory T cells was determined,and pathological changes of the lungs was observed by HE staining.Foxp3,TGF-β1,Smad3,Smad7 protein expression in the lungs of the two groups of rats was examined by immunohistochemistry.Results:①Compared with those of the normal,the swelling degree of toes,AI,and the lung index were increased.The average peak expiratory flow in 1 second (FEV1/FVC),alveolitis points,TGF-β1 and Smad3 protein expression was significantly higher in the AA model group rats(P＜0.01).Forced vital capacity (FVC),25% of vital capacity of the peak expiratory flow (FEF25),50% of vital capacity of the peak expiratory flow (FEF50),75% of vital capacity of the peak expiratory flow (FEF75),maximum mid-expiratory flow (MMF),forced maximal expiratory flow (PEF),pulmonary dynamic compliance (Cldyn),the percentage of CD4~+ T cell,CD25~+ T cell,CD4~+ CD25+~ T cell,Foxp3 and Smad7 protein expression was obviously lower in the AA model group rats(P＜0.01).②FEF50,Cldyn and MMF were negative correlated with the swelling degree of toes,pulmonary coefficient and TGF-β1.FEF50,FEF75,MMF and FEV1/FVC were positiyely correlated with the arthritis index,pulmonary coefficient and Foxp3(P＜0.05 or P＜0.01).Conclusion:The decline in lung function in AA rats.Tip-induced may be sustained after the inflammation,and the development of cause inflammation,to chronic inflammatory response leading to lung injury.Simultaneously,the decline in lung function and Foxp3.Smad3,Smad7,TGF-β1 protein expression was associated.Description transcription factor Foxp3 and TGF-β1/Smads signal transducation pathway may participate in the mechanism.

The role of CD4+CD25+Foxp3 regulatory T cells in patients with chronic hepatitis B / 中华传染病杂志

Objective To investigate the role of CD4+CD25+Foxp3 regulatory T cells in chronicity of hepatitis B and viral clearance of hepatitis B virus(HBV).Methods Nineteen patients with chronic active hepatitis B(CAH).21 HBV carriers(AsC)and 12 patients with resolved HBV infection and 1 5 healthy controls were enrolled.The frequency and phenotype of peripheral CD4+CD25+Foxp3+ T cells were detected by flow cytometry.CD4+CD25+T cells were sorted by magnetic-activated cell sorting(MACS)assay.Level of Foxp3 mRNA in CD4+CD25+T cells was examined by real time polymerase chain reaction(PCR)assay.The data were analyzed by one-way ANoVA or nonparametric statistics.Results Both frequencies of CD4+CD25+Foxp3+T cells and levels of Foxp3 mRNA in CD4+CD25+T ceils in patients with CAH or AsC were significantly higher than those in healthy controls Or resolved HBV infection(F=6.8,F=3.72,respectively;both P<0.05).Accumulation of Foxp3+T cells in liver tissue of CAH patients was higher than that of healthy controls,while that in AsC was lower than CAH.The frequency of CD4+CD25+Foxp3+T cells of hepatitis B e antigen(HBeAg)positive patients(including CAH and AsC)was significantly higher than that of HBeAg negative patients(t=2.3,P<0.05),and that of antFHBe negative patients were significantly higher than anti-HBe positive patients(t=2.4,P<0.05).Furthermore,the frequency of CD4+CD25+Foxp3 regulatory T cells was positively correlated with serum HBV DNA level of patients with chronic hepatitis B(r=0.56,P<0.01).Conclusion The findings have important implication in the understanding of the role of CD4'CD25'regulatory T cells in chronicity and viral clearance in HBV infection.

Progress on the regulatory T cells and immune tolerance in organ transplantation / 解放军医学杂志

CD4+CD25+FoxP3+Treg may regulate the immune responses and induce tolerance to alloantigen in vivo in organ transplantation. A high expression of CD4+CD25+FoxP3+Treg may be a possible indicator of acute allograft rejection. The patients with renal transplantation showing a high expression of CD4+CD25+FoxP3+Treg might be more prone to the development of acute allograft rejection,and the detection of its expression may contribute to predict allograft acceptance or rejection. It seems plausible that the imbalance between effector T cells and Treg might reflect an immune state. Additional experimentation and clinical studies are needed to investigate the long-term impact of development and function of Treg cells on immunosuppression in allogeneic renal transplantation.