AIM: To investigated the interaction between toll-like receptor 4 (TLR4)-activated hepatoma cells and macrophages in the induction of tumor-immune suppression mediated by CD4+CD25high family of transcription factor P3...AIM: To investigated the interaction between toll-like receptor 4 (TLR4)-activated hepatoma cells and macrophages in the induction of tumor-immune suppression mediated by CD4+CD25high family of transcription factor P3 (FOXP3) regulatory T cells (Tregs). METHODS: The proportion of FOXP3+ Tregs was identified in peripheral blood and tumor tissues of 60 hepatocellular carcinoma (HCC) patients. TLR4 expression was examined in tumor tissues and cell lines. The correlation was examined between FOXP3+ Tregs in peripheral blood and TLR4 expression of HCC tissues. Following activation of TLR4 in H22 murine hepatoma cells pre-incubated with lipopolysaccharide (LPS) and co-cultured with macrophage cell line RAW246.7, the synthesis of cytokines tumor necrosis factor-α, CCL22, and interleukin (IL)-10 by the two cell lines was detected and analyzed. RESULTS: FOXP3+ Tregs were enriched in tumor sites, and circulating FOXP3+ Tregs were increased in HCC patients in correlation with multiple tumor foci and up-regulated TLR4 expression in HCC tissues. Semi-quantitative analysis indicated that TLR4 was over-expressed in HCC compared with the matched normal tissues. Cell cultivation experiments indicated that the mRNAs of IL-10 and CCL22 were significantly up-regulated in the RAW246.7 cell line when co-cultured with LPS preincubated H22 cells. CONCLUSION: In hepatoma cell lines, TLR4 may indirectly facilitate the recruitment of Tregs to the tumor site and promote intrahepatic metastasis through its interaction with macrophages.展开更多
As a part of a basic research project on Xeno-transplantion. we have been engaged in the derivation of embryonic stein cell lines from Chinese mini swine. Here, we reported for the first time the establishment of two ...As a part of a basic research project on Xeno-transplantion. we have been engaged in the derivation of embryonic stein cell lines from Chinese mini swine. Here, we reported for the first time the establishment of two porcine EG cell lines (BPEG1 and BPEG2) from primordial germ cells of genital ridges of a 28 and a 27 d embryos respectively. Their pluripotent nature has been identified by colony morphology, marker characterization as well as by in vitro and in vivo differentiation. These porcine EG cells are potentially useful for further basic studies.展开更多
AIM:To assess BGC823 gastric cancer(GC) cell metastasis after knockdown of liver-intestine cadherin(CDH17) and the therapeutic value of CDH17-RNAilentivirus in vivo.METHODS:We evaluated primary tumor growth and assess...AIM:To assess BGC823 gastric cancer(GC) cell metastasis after knockdown of liver-intestine cadherin(CDH17) and the therapeutic value of CDH17-RNAilentivirus in vivo.METHODS:We evaluated primary tumor growth and assessed local infiltration and systemic tumor dissemination using an orthotopic implantation technique.The therapeutic value of CDH17 knockdown was examined by intratumoral administration of CDH17-RNA interference(RNAi)-lentivirus in an established GC tumor xenograft mouse model.Furthermore,a comparative proteomic approach was utilized to identify differentially expressed proteins in BGC823 and lenti-CDH17-miRneg cells following CDH17 knockdown.RESULTS:Metastases in the liver and lung appeared earlier and more frequently in animals with tumors derived from BGC823 or lenti-CDH17-miR-neg cells than in tumors derived from lenti-CDH17-miR-B cells.Average tumor weight and volume in the CDH17-RNAi-lentivirus-treated group were significantly lower than those in the control group(tumor volume:0.89 ± 0.04 cm 3 vs 1.16 ± 0.06 cm 3,P < 0.05;tumor weight:1.15 ± 0.58 g vs 2.09 ± 0.08 g,P < 0.05).Fifteen differentially expressed proteins were identified after CDH17 silencing in BGC823 cells,including a variety of cytoskeletal and chaperone proteins as well as proteins involved in metabolism,immunity/defense,cell proliferation and differentiation,cell cycle,and signal transduction.CONCLUSION:Our data establish a foundation for future studies of the comprehensive protein expression patterns and effects of CDH17 in GC.展开更多
Objective To investigate the effect of Coriolus versicolor polysaccharide-B （CVPs-B） on the biological characteristics of human esophageal carcinoma cell line Ecal09 in vitro. Methods The cells of experimental group...Objective To investigate the effect of Coriolus versicolor polysaccharide-B （CVPs-B） on the biological characteristics of human esophageal carcinoma cell line Ecal09 in vitro. Methods The cells of experimental group （EG） were cultured in DMEM with 10% FCS and 150μg/mL CVPs-B, the cells of control group （CG） were cultured in DMEM with 10% FCS without CVPs-B. MTT reduction assay was performed to detect the effect of CVPs-B on the proliferation of Ecal09 cells after the compound was administrated in varying concentrations. The living conditions of the Ecal09 cells were determined using trypan blue exclusion. Then, cell growth curves were drawn. Flow cytometry was performed to detect the effect of CVPs-B on the apoptosis and cell cycle of Ecal09. Results In comparison with the CG, a marked decrease in the proliferation of Eca09 cells was observed in the EG, after incubation with CVPs-B. The survival rate of Eca09 cells decreased as the time of CVPs-B incubation prolonged. Comparing the cell cycles and apoptotic rates between the two groups, the proportions of cells in the G0/G1, S, and G2/M phases in the EG were found to be （68.4±3.7）%, （13.9±2.1）%, and （17.7±1.4）%, respectively, after 24 h incubation with CVPs-B. The cells had an apoptotic rate of （9.7±0.7）%. On the other hand, the proportions of the G0/G1, S, and G2/M cells of the CG were found to be （53.9±3.6）%, （26.6±2.8）%, and （19.5±2.3）%, respectively, with an apoptotic rate of （5.7±1.4）%. In comparison with the CG cells, significant cell growth in the G0/G1 phase was observed in the EG （P〈0.05）. Furthermore, a significant decrease in the number of cells in the S phase was observed （P〈0.05） in the EG. Conclusions CVPs-B can inhibit proliferation and enhance apoptosis of Ecal09 cells and may be useful in the treatment of esophageal carcinoma.展开更多
TGF-β and Foxp3 expressions are crucial for the induction and functional activity of CD4＋Foxp3＋ regulatory T （iTreg） cells. Here, we demonstrate that although TGF-β-primed CD8＋ cells display much lower Foxp3 ex...TGF-β and Foxp3 expressions are crucial for the induction and functional activity of CD4＋Foxp3＋ regulatory T （iTreg） cells. Here, we demonstrate that although TGF-β-primed CD8＋ cells display much lower Foxp3 expression, their suppressive capacity is equivalent to that of CD4＋ iTreg cells, and both Foxp3- and Foxp3＋ CD8＋ subsets have suppressive activities in vitro and in vivo. CD8＋Foxp3- iTreg cells produce little IFN-γ but almost no IL-2, and display a typical anergic phenotype. Among phenotypic markers expressed in CD8＋Foxp3- cells, we identify CD103 expression particularly crucial for the generation and function of this subset. Moreover, IL-IO and TGF-β signals rather than cytotoxicity mediate the suppressive effect of this novel Treg population. Therefore, TGF-β can induce both CD8＋Foxp3- and CD8＋Foxp3＋ iTreg subsets, which may represent the unique immunoregulatory means to treat autoimmune and inflammatory diseases.展开更多
OBJECTIVE CD4^＋CD25^＋ T regulatory （Treg） cells are a population of T cells which suppress an overactive immune system. CCR4 is a chemokine receptor involved in the recruitment of lymphocytes. Nasopharyngeal carci...OBJECTIVE CD4^＋CD25^＋ T regulatory （Treg） cells are a population of T cells which suppress an overactive immune system. CCR4 is a chemokine receptor involved in the recruitment of lymphocytes. Nasopharyngeal carcinoma （NPC） is resistant to immunosurveillance, owing to the increased number of tumor-infiltrating Treg cells which are recruited to the tumor bv CCR4.展开更多
The paucity of naturally occurring CD4^＋FoxP3^＋ regulatory T cells （Tregs） and difficulties in the identification and isolation of such cells are major obstacles in the study of human Tregs. The availability of hu...The paucity of naturally occurring CD4^＋FoxP3^＋ regulatory T cells （Tregs） and difficulties in the identification and isolation of such cells are major obstacles in the study of human Tregs. The availability of human Treg-like cell lines is likely to facilitate a better understanding of the molecular basis of Treg function and aid the discovery of pharmacological reagents to regulate Treg activity in a disease state.展开更多
Objective:To analyze the effect of spleen on the Treg cells during pregnancy. Methods: The mononuclear cells were separated from the peripheral, spleen and uterus or placenta blood. Flow cytometry was employed to anal...Objective:To analyze the effect of spleen on the Treg cells during pregnancy. Methods: The mononuclear cells were separated from the peripheral, spleen and uterus or placenta blood. Flow cytometry was employed to analyze the percent of Treg cells in total T cells in different stages of pregnancy. Immunohistochemical staining was used to make sure the distribution of Treg in spleen in different stages of pregnancy. Results: The results of immunohistochemical staining showed that compared with spleen Treg cells in normal unpregnant mice, spleen Treg cells on day 7 and 14 of pregnancy significantly increased. After splenectomy, peripheral blood and placenta Treg cells on day 7 of pregnancy markedly decreased as compared with the normal pregnancy(P<0.01). And the cells on day 14of pregnancy were markedly recovered as compared with the normal pregnancy. Conclusion: Our study indicated that the spleen and its Treg cells might play important roles in transient tolerance during pregnancy.展开更多
动脉粥样硬化(atherosclerosis,AS)是动脉系统的慢性炎症性疾病,免疫细胞和炎症介质对其发生、发展均有重要作用。高迁移率族蛋白1(high mobility group box 1 protein,HMGB1)这一新发现的晚期炎症介质,具有促血管损伤后再狭窄和AS等多...动脉粥样硬化(atherosclerosis,AS)是动脉系统的慢性炎症性疾病,免疫细胞和炎症介质对其发生、发展均有重要作用。高迁移率族蛋白1(high mobility group box 1 protein,HMGB1)这一新发现的晚期炎症介质,具有促血管损伤后再狭窄和AS等多种病理生理效应。Treg细胞负向调节免疫应答,有效抑制机体炎症反应。AS患者Treg细胞的数量减少或功能下降,提示Treg细胞与AS病变相关。Toll样受体(Toll like receptors,TLRs)是诱导机体炎症反应重要的模式识别受体(pattern recogination receptors,PRRs),现认为HMGB1是其家族中TLR2、4的内源性配体,而Treg细胞表达这两种受体。HMGB1与Treg细胞上的TLR2、4结合,可能直接影响其抑制作用,导致机体持续慢性炎症反应,参与AS的形成和发展。展开更多
最近，来自比利时鲁文盖斯堡大学的Jan L．Ceuppens课题组研究了Treg细胞在共刺激因子信号通路阻断的情况下如何减缓免疫排斥效应的分子机制，发现结合MR1以及低浓度的CTLA-4Ig能够起到最佳的免疫抑制效果，相关结果发表在最近一期的《J...最近，来自比利时鲁文盖斯堡大学的Jan L．Ceuppens课题组研究了Treg细胞在共刺激因子信号通路阻断的情况下如何减缓免疫排斥效应的分子机制，发现结合MR1以及低浓度的CTLA-4Ig能够起到最佳的免疫抑制效果，相关结果发表在最近一期的《Journal of Immunology》杂志上。展开更多
最近，来自migham and Women医院的研究者们在对多发性硬化疾病进行研究的过程中发现了治疗癌症的新线索。在最新一期的《Science Immunology》杂志上。由Howard Weiner博士领导的研究团队发现了一种利用抗体精确打击Treg细胞来促进癌...最近，来自migham and Women医院的研究者们在对多发性硬化疾病进行研究的过程中发现了治疗癌症的新线索。在最新一期的《Science Immunology》杂志上。由Howard Weiner博士领导的研究团队发现了一种利用抗体精确打击Treg细胞来促进癌症免疫反应的方法。展开更多
文摘AIM: To investigated the interaction between toll-like receptor 4 (TLR4)-activated hepatoma cells and macrophages in the induction of tumor-immune suppression mediated by CD4+CD25high family of transcription factor P3 (FOXP3) regulatory T cells (Tregs). METHODS: The proportion of FOXP3+ Tregs was identified in peripheral blood and tumor tissues of 60 hepatocellular carcinoma (HCC) patients. TLR4 expression was examined in tumor tissues and cell lines. The correlation was examined between FOXP3+ Tregs in peripheral blood and TLR4 expression of HCC tissues. Following activation of TLR4 in H22 murine hepatoma cells pre-incubated with lipopolysaccharide (LPS) and co-cultured with macrophage cell line RAW246.7, the synthesis of cytokines tumor necrosis factor-α, CCL22, and interleukin (IL)-10 by the two cell lines was detected and analyzed. RESULTS: FOXP3+ Tregs were enriched in tumor sites, and circulating FOXP3+ Tregs were increased in HCC patients in correlation with multiple tumor foci and up-regulated TLR4 expression in HCC tissues. Semi-quantitative analysis indicated that TLR4 was over-expressed in HCC compared with the matched normal tissues. Cell cultivation experiments indicated that the mRNAs of IL-10 and CCL22 were significantly up-regulated in the RAW246.7 cell line when co-cultured with LPS preincubated H22 cells. CONCLUSION: In hepatoma cell lines, TLR4 may indirectly facilitate the recruitment of Tregs to the tumor site and promote intrahepatic metastasis through its interaction with macrophages.
基金an introductory part of a project supported by National Science Foundation of China(No.39993430).
文摘As a part of a basic research project on Xeno-transplantion. we have been engaged in the derivation of embryonic stein cell lines from Chinese mini swine. Here, we reported for the first time the establishment of two porcine EG cell lines (BPEG1 and BPEG2) from primordial germ cells of genital ridges of a 28 and a 27 d embryos respectively. Their pluripotent nature has been identified by colony morphology, marker characterization as well as by in vitro and in vivo differentiation. These porcine EG cells are potentially useful for further basic studies.
基金Supported by The National Natural Science Foundation of China,No.30871147
文摘AIM:To assess BGC823 gastric cancer(GC) cell metastasis after knockdown of liver-intestine cadherin(CDH17) and the therapeutic value of CDH17-RNAilentivirus in vivo.METHODS:We evaluated primary tumor growth and assessed local infiltration and systemic tumor dissemination using an orthotopic implantation technique.The therapeutic value of CDH17 knockdown was examined by intratumoral administration of CDH17-RNA interference(RNAi)-lentivirus in an established GC tumor xenograft mouse model.Furthermore,a comparative proteomic approach was utilized to identify differentially expressed proteins in BGC823 and lenti-CDH17-miRneg cells following CDH17 knockdown.RESULTS:Metastases in the liver and lung appeared earlier and more frequently in animals with tumors derived from BGC823 or lenti-CDH17-miR-neg cells than in tumors derived from lenti-CDH17-miR-B cells.Average tumor weight and volume in the CDH17-RNAi-lentivirus-treated group were significantly lower than those in the control group(tumor volume:0.89 ± 0.04 cm 3 vs 1.16 ± 0.06 cm 3,P < 0.05;tumor weight:1.15 ± 0.58 g vs 2.09 ± 0.08 g,P < 0.05).Fifteen differentially expressed proteins were identified after CDH17 silencing in BGC823 cells,including a variety of cytoskeletal and chaperone proteins as well as proteins involved in metabolism,immunity/defense,cell proliferation and differentiation,cell cycle,and signal transduction.CONCLUSION:Our data establish a foundation for future studies of the comprehensive protein expression patterns and effects of CDH17 in GC.
基金supported by the Guangdong Provincial Sci-Tech Planning(No.2010B030700051)
文摘Objective To investigate the effect of Coriolus versicolor polysaccharide-B （CVPs-B） on the biological characteristics of human esophageal carcinoma cell line Ecal09 in vitro. Methods The cells of experimental group （EG） were cultured in DMEM with 10% FCS and 150μg/mL CVPs-B, the cells of control group （CG） were cultured in DMEM with 10% FCS without CVPs-B. MTT reduction assay was performed to detect the effect of CVPs-B on the proliferation of Ecal09 cells after the compound was administrated in varying concentrations. The living conditions of the Ecal09 cells were determined using trypan blue exclusion. Then, cell growth curves were drawn. Flow cytometry was performed to detect the effect of CVPs-B on the apoptosis and cell cycle of Ecal09. Results In comparison with the CG, a marked decrease in the proliferation of Eca09 cells was observed in the EG, after incubation with CVPs-B. The survival rate of Eca09 cells decreased as the time of CVPs-B incubation prolonged. Comparing the cell cycles and apoptotic rates between the two groups, the proportions of cells in the G0/G1, S, and G2/M phases in the EG were found to be （68.4±3.7）%, （13.9±2.1）%, and （17.7±1.4）%, respectively, after 24 h incubation with CVPs-B. The cells had an apoptotic rate of （9.7±0.7）%. On the other hand, the proportions of the G0/G1, S, and G2/M cells of the CG were found to be （53.9±3.6）%, （26.6±2.8）%, and （19.5±2.3）%, respectively, with an apoptotic rate of （5.7±1.4）%. In comparison with the CG cells, significant cell growth in the G0/G1 phase was observed in the EG （P〈0.05）. Furthermore, a significant decrease in the number of cells in the S phase was observed （P〈0.05） in the EG. Conclusions CVPs-B can inhibit proliferation and enhance apoptosis of Ecal09 cells and may be useful in the treatment of esophageal carcinoma.
文摘TGF-β and Foxp3 expressions are crucial for the induction and functional activity of CD4＋Foxp3＋ regulatory T （iTreg） cells. Here, we demonstrate that although TGF-β-primed CD8＋ cells display much lower Foxp3 expression, their suppressive capacity is equivalent to that of CD4＋ iTreg cells, and both Foxp3- and Foxp3＋ CD8＋ subsets have suppressive activities in vitro and in vivo. CD8＋Foxp3- iTreg cells produce little IFN-γ but almost no IL-2, and display a typical anergic phenotype. Among phenotypic markers expressed in CD8＋Foxp3- cells, we identify CD103 expression particularly crucial for the generation and function of this subset. Moreover, IL-IO and TGF-β signals rather than cytotoxicity mediate the suppressive effect of this novel Treg population. Therefore, TGF-β can induce both CD8＋Foxp3- and CD8＋Foxp3＋ iTreg subsets, which may represent the unique immunoregulatory means to treat autoimmune and inflammatory diseases.
基金the National Natural Science Foundation of China
文摘OBJECTIVE CD4^＋CD25^＋ T regulatory （Treg） cells are a population of T cells which suppress an overactive immune system. CCR4 is a chemokine receptor involved in the recruitment of lymphocytes. Nasopharyngeal carcinoma （NPC） is resistant to immunosurveillance, owing to the increased number of tumor-infiltrating Treg cells which are recruited to the tumor bv CCR4.
文摘The paucity of naturally occurring CD4^＋FoxP3^＋ regulatory T cells （Tregs） and difficulties in the identification and isolation of such cells are major obstacles in the study of human Tregs. The availability of human Treg-like cell lines is likely to facilitate a better understanding of the molecular basis of Treg function and aid the discovery of pharmacological reagents to regulate Treg activity in a disease state.
基金Supported by National Natural Science Foundation of China(NO.81070536)
文摘Objective:To analyze the effect of spleen on the Treg cells during pregnancy. Methods: The mononuclear cells were separated from the peripheral, spleen and uterus or placenta blood. Flow cytometry was employed to analyze the percent of Treg cells in total T cells in different stages of pregnancy. Immunohistochemical staining was used to make sure the distribution of Treg in spleen in different stages of pregnancy. Results: The results of immunohistochemical staining showed that compared with spleen Treg cells in normal unpregnant mice, spleen Treg cells on day 7 and 14 of pregnancy significantly increased. After splenectomy, peripheral blood and placenta Treg cells on day 7 of pregnancy markedly decreased as compared with the normal pregnancy(P<0.01). And the cells on day 14of pregnancy were markedly recovered as compared with the normal pregnancy. Conclusion: Our study indicated that the spleen and its Treg cells might play important roles in transient tolerance during pregnancy.
文摘动脉粥样硬化(atherosclerosis,AS)是动脉系统的慢性炎症性疾病,免疫细胞和炎症介质对其发生、发展均有重要作用。高迁移率族蛋白1(high mobility group box 1 protein,HMGB1)这一新发现的晚期炎症介质,具有促血管损伤后再狭窄和AS等多种病理生理效应。Treg细胞负向调节免疫应答,有效抑制机体炎症反应。AS患者Treg细胞的数量减少或功能下降,提示Treg细胞与AS病变相关。Toll样受体(Toll like receptors,TLRs)是诱导机体炎症反应重要的模式识别受体(pattern recogination receptors,PRRs),现认为HMGB1是其家族中TLR2、4的内源性配体,而Treg细胞表达这两种受体。HMGB1与Treg细胞上的TLR2、4结合,可能直接影响其抑制作用,导致机体持续慢性炎症反应,参与AS的形成和发展。
文摘炎症是肥胖和2型糖尿病之间的关键联系，过量的能量摄入可诱导内脏脂肪组织（visceral adipose tissue，VAT）的慢性炎症反应，最终引起血脂异常、胰岛素抵抗等代谢异常，甚至发展为2型糖尿病等代谢性疾病。前期研究发现，正常体重小鼠VAT 中存在一类调节性 T 细胞（regulatory T cell，Treg cell），参与调控局部及系统性的炎症和代谢，其转录组与淋巴组织来源Treg细胞截然不同。然而，在不同生理或病理条件下，VAT Treg细胞的变化及调控机制尚不清楚。
