BACKGROUND: Liver failure in chronic hepatitis B (CHB) patients is a severe, life-threatening condition. Intestinal endotoxemia plays a significant role in the progress to liver failure. High mobility group box-1 (HMG...BACKGROUND: Liver failure in chronic hepatitis B (CHB) patients is a severe, life-threatening condition. Intestinal endotoxemia plays a significant role in the progress to liver failure. High mobility group box-1 (HMGB1) protein is involved in the process of endotoxemia. Regulatory T (Treg) cells maintain immune tolerance and contribute to the immunological hyporesponsiveness against HBV infection. However, the roles of HMGB1 and Treg cells in the pathogenesis of liver failure in CHB patients, and whether HMGB1 affects the immune activity of Treg cells are poorly known at present, and so were explored in this study. METHODS: The levels of HMGB1 expression were detected by ELISA, real-time RT-PCR, and Western blotting, and the percentage of CD4(+)CD25(+)CD127(low) Treg cells among CD4(+) cells was detected by flow cytometry in liver failure patients with chronic HBV infection, CHB patients, and healthy controls. Then, CD4(+)CD25(+)CD127(low) Treg cells isolated from the peripheral blood mononuclear cells from CHB patients were stimulated with HMGB1 at different concentrations or at various intervals. The effect of HMGB1 on the immune activity of Treg cells was assessed by a suppression assay of the allogeneic mixed lymphocyte response. The levels of forkhead box P3 (Foxp3) expression in Treg cells treated with HMGB1 were detected by RT-PCR and Western blotting. RESULTS: A higher level of HMGB1 expression and a lower percentage of Treg cells within the population of CIA(+) cells were found in liver failure patients than in CHB patients (82.6+/-20.1 mu g/L vs. 34.2+/-13.7 mu g/L; 4.55+/-1.34% vs. 9.52+/-3.89%, respectively). The immune activity of Treg cells was significantly weakened and the levels of Foxp3 expression were reduced in a dose- or time-dependent manner when Treg cells were stimulated with HMGB1 in vitro. CONCLUSIONS: The high level of HMGB1 and the low percentage of Treg cells play an important role in the pathogenesis of liver failure in patients with chronic HBV infection. Moreover, HMGB1 can weaken the immune activity of Treg cells. It is suggested that effectively inhibiting HMGB1 expression could be a feasible way to treat liver failure by suppressing endotoxemia and enhancing Treg cell activity.展开更多
Objective To investigate the expression of Snail in bladder urothelial carcinoma and evaluate its relationship with E-cadherin and a subset of T cell groups. Methods Immunohistochemical method was used to detect the e...Objective To investigate the expression of Snail in bladder urothelial carcinoma and evaluate its relationship with E-cadherin and a subset of T cell groups. Methods Immunohistochemical method was used to detect the expression of Snail and E-cadherin proteins in tissue展开更多
Objoctive： There is heterogeneity in the prognosis of gastric cancers staged according to the tumornodes-metastasis （TNM） system. This study evaluated the prognostic potential of an immune score system to supplemen...Objoctive： There is heterogeneity in the prognosis of gastric cancers staged according to the tumornodes-metastasis （TNM） system. This study evaluated the prognostic potential of an immune score system to supplement the TNM staging system. Mothodsg An immunohistochemical analysis was conducted to assess the density of T cells, B cells, and myeloid-derived suppressor cells （MDSCs） in cancer tissues from 100 stage IIIA gastric cancer patients; the expression of the high-mobility group protein B1 （HMGB1） was also evaluated in cancer cells. The relationship between the overall survival （OS）, disease-free survival （DFS）, and immunological parameters was analyzed.Results： An immune score system was compiled based on the prognostic role of the density ofT cells, B cells, MDSCs, and the expression of HMGB1 in cancer tissues. The median 5-year survival of this group of patient was 32%. However, the 5-year survival rates of 80.0%, 51.7%, 0%, 5.8%, and 0% varied among the patients with an immune score of 4 to those with an immune score of 0 based on the immune score system, respectively. Similarly, differences in DFS rates were observed among the immune score subgroups. Concluslons： An immune score system could effectively identify the prognostic heterogeneity within stage IliA gastric cancer patients, implying that this immune score system may potentially supplement the TNM staging system, and help in identifying a more homogeneous group of patients who on the basis of prognosis can undergo adjuvant therapy.展开更多
目的探讨儿童过敏性鼻炎(allergic rhinitis,AR)和支气管哮喘(bronchial asthma,BA)患儿外周血高迁移率族蛋白B1(high mobility group box 1,HMGB1)水平与辅助性T细胞17(T helper 17 cell,Th17)和调节性T细胞(regulatory T cell,Treg)...目的探讨儿童过敏性鼻炎(allergic rhinitis,AR)和支气管哮喘(bronchial asthma,BA)患儿外周血高迁移率族蛋白B1(high mobility group box 1,HMGB1)水平与辅助性T细胞17(T helper 17 cell,Th17)和调节性T细胞(regulatory T cell,Treg)的比例(Th17/Treg)的相关性。方法采用前瞻性研究方法,选取2023年3月至12月在解放军总医院第七医学中心门急诊就诊患儿及健康体检者共113例作为研究对象,按照临床症状分成健康对照组(n=25)、过敏性鼻炎组(AR组,n=28)、支气管哮喘组(BA组,n=31)和两者并有组(AR+BA组,n=29),比较4组肺功能、免疫球蛋白E(immunoglobulin E,IgE)、白细胞介素(interleukin,IL)-17、HMGB1水平和Th17/Treg比例。统计学方法采用t检验、方差分析和Pearson相关性分析。结果各组的第1秒用力呼气容积(forced expiratory volume in the first second,FEV1)的结果显示,健康对照组高于AR组、BA组和AR+BA组[(96.4±2.5)%、(90.7±4.9)%、(88.6±4.1)%、(82.1±5.5)%,F=47.193,P值均<0.001],显示AR和BA患儿肺功能可能变差。AR组、BA组和AR+BA组HMGB1水平高于健康对照组[(30.4±12.4)mg/L、(47.2±17.8)mg/L、(51.4±21.8)mg/L、(4.3±2.1)mg/L,F=48.896,P值均<0.001]。AR组、BA组和AR+BA组Th17/Treg比例高于健康对照组(0.68±0.14、0.71±0.14、1.36±0.40、0.30±0.07,F=105.547,P值均<0.001)。Pearson相关性分析结果显示,HMGB1与Th17(r=0.521,P<0.001)、Th17/Treg(r=0.419,P<0.05)、IgE(r=0.502,P<0.05)、IL-17(r=0.472,P<0.05)呈正相关,与FEV1(r=-0.645,P<0.001)呈负相关。结论在儿童AR合并BA的发病过程中,HMGB1水平及Th17细胞百分比增加,而Treg细胞百分比降低,Th17/Treg比例存在失衡的情形,HMGB1水平与Th17细胞百分比、Th17/Treg比例、IgE、IL-17及FEV1存在一定的相关性,HMGB1和Th17/Treg的动态平衡可能通过影响细胞因子的表达而起到一定的调节作用。展开更多
目的构建经MHCⅡ通路的屋尘螨1类变应原Der p 1的T细胞表位肽疫苗重组载体。方法分别合成TAT、Ih C和含编码Der p 1的3段T细胞表位的融合核苷酸序列,用特异性引物PCR扩增相应的基因片段,分别用相应的双酶切后,用T4 DNA连接酶连接形成TAT...目的构建经MHCⅡ通路的屋尘螨1类变应原Der p 1的T细胞表位肽疫苗重组载体。方法分别合成TAT、Ih C和含编码Der p 1的3段T细胞表位的融合核苷酸序列,用特异性引物PCR扩增相应的基因片段,分别用相应的双酶切后,用T4 DNA连接酶连接形成TAT-Ih C-Der p 1-3T融合基因,并插入至原核表达载体p ET-28a(+)中,构建重组原核表达载体p ET-28a(+)-TATIh C-Der p 1-3T,Bam HⅠ和XhoⅠ进行双酶切和测序鉴定。重组载体转化大肠杆菌E.coli BL21(DE3)菌株,IPTG诱导后,经SDS-PAGE电泳分析和Western blot验证,纯化TAT-Ih C-Der p 1-3T蛋白后进行Ig E结合试验。结果双酶切和测序结果表明,成功构建了p ET-28a-TAT-Ih C-Der p 1-3T重组原核表达载体;SDS-PAGE电泳分析显示TAT-Ih C-Der p 1-3T可诱导表达;Western blot检测表明该融合蛋白纯化成功;Ig E结合试验表明TAT-Ih C-Der p 1-3T结合屋尘螨过敏病人血清Ig E的能力强于Der p 1变应原(P<0.001)。结论成功构建了可表达经MHC通路的编码Der p 1的3段T细胞表位的重组p ET-28a-TAT-Ih C-Der p 1-3T载体,纯化的TAT-Ih C-Der p 1-3T具有较强的Ig E结合能力,从而为后续经MHC通路的特异性免疫治疗奠定基础。展开更多
基金supported by a grant from the National Natural Science Foundation of China (No. 81071342)
文摘BACKGROUND: Liver failure in chronic hepatitis B (CHB) patients is a severe, life-threatening condition. Intestinal endotoxemia plays a significant role in the progress to liver failure. High mobility group box-1 (HMGB1) protein is involved in the process of endotoxemia. Regulatory T (Treg) cells maintain immune tolerance and contribute to the immunological hyporesponsiveness against HBV infection. However, the roles of HMGB1 and Treg cells in the pathogenesis of liver failure in CHB patients, and whether HMGB1 affects the immune activity of Treg cells are poorly known at present, and so were explored in this study. METHODS: The levels of HMGB1 expression were detected by ELISA, real-time RT-PCR, and Western blotting, and the percentage of CD4(+)CD25(+)CD127(low) Treg cells among CD4(+) cells was detected by flow cytometry in liver failure patients with chronic HBV infection, CHB patients, and healthy controls. Then, CD4(+)CD25(+)CD127(low) Treg cells isolated from the peripheral blood mononuclear cells from CHB patients were stimulated with HMGB1 at different concentrations or at various intervals. The effect of HMGB1 on the immune activity of Treg cells was assessed by a suppression assay of the allogeneic mixed lymphocyte response. The levels of forkhead box P3 (Foxp3) expression in Treg cells treated with HMGB1 were detected by RT-PCR and Western blotting. RESULTS: A higher level of HMGB1 expression and a lower percentage of Treg cells within the population of CIA(+) cells were found in liver failure patients than in CHB patients (82.6+/-20.1 mu g/L vs. 34.2+/-13.7 mu g/L; 4.55+/-1.34% vs. 9.52+/-3.89%, respectively). The immune activity of Treg cells was significantly weakened and the levels of Foxp3 expression were reduced in a dose- or time-dependent manner when Treg cells were stimulated with HMGB1 in vitro. CONCLUSIONS: The high level of HMGB1 and the low percentage of Treg cells play an important role in the pathogenesis of liver failure in patients with chronic HBV infection. Moreover, HMGB1 can weaken the immune activity of Treg cells. It is suggested that effectively inhibiting HMGB1 expression could be a feasible way to treat liver failure by suppressing endotoxemia and enhancing Treg cell activity.
文摘Objective To investigate the expression of Snail in bladder urothelial carcinoma and evaluate its relationship with E-cadherin and a subset of T cell groups. Methods Immunohistochemical method was used to detect the expression of Snail and E-cadherin proteins in tissue
基金support from the National Nature Science Foundation of China ( Grant No.81272341, 81401156)Research Program of Guangzhou Municipal Health Bureau Foundation of China (Grant No.20141A011085, 20141A011088)The PhD Start-up Fund Guangzhou Medical University (Grant No.2013C49)
文摘Objoctive： There is heterogeneity in the prognosis of gastric cancers staged according to the tumornodes-metastasis （TNM） system. This study evaluated the prognostic potential of an immune score system to supplement the TNM staging system. Mothodsg An immunohistochemical analysis was conducted to assess the density of T cells, B cells, and myeloid-derived suppressor cells （MDSCs） in cancer tissues from 100 stage IIIA gastric cancer patients; the expression of the high-mobility group protein B1 （HMGB1） was also evaluated in cancer cells. The relationship between the overall survival （OS）, disease-free survival （DFS）, and immunological parameters was analyzed.Results： An immune score system was compiled based on the prognostic role of the density ofT cells, B cells, MDSCs, and the expression of HMGB1 in cancer tissues. The median 5-year survival of this group of patient was 32%. However, the 5-year survival rates of 80.0%, 51.7%, 0%, 5.8%, and 0% varied among the patients with an immune score of 4 to those with an immune score of 0 based on the immune score system, respectively. Similarly, differences in DFS rates were observed among the immune score subgroups. Concluslons： An immune score system could effectively identify the prognostic heterogeneity within stage IliA gastric cancer patients, implying that this immune score system may potentially supplement the TNM staging system, and help in identifying a more homogeneous group of patients who on the basis of prognosis can undergo adjuvant therapy.
文摘目的探讨儿童过敏性鼻炎(allergic rhinitis,AR)和支气管哮喘(bronchial asthma,BA)患儿外周血高迁移率族蛋白B1(high mobility group box 1,HMGB1)水平与辅助性T细胞17(T helper 17 cell,Th17)和调节性T细胞(regulatory T cell,Treg)的比例(Th17/Treg)的相关性。方法采用前瞻性研究方法,选取2023年3月至12月在解放军总医院第七医学中心门急诊就诊患儿及健康体检者共113例作为研究对象,按照临床症状分成健康对照组(n=25)、过敏性鼻炎组(AR组,n=28)、支气管哮喘组(BA组,n=31)和两者并有组(AR+BA组,n=29),比较4组肺功能、免疫球蛋白E(immunoglobulin E,IgE)、白细胞介素(interleukin,IL)-17、HMGB1水平和Th17/Treg比例。统计学方法采用t检验、方差分析和Pearson相关性分析。结果各组的第1秒用力呼气容积(forced expiratory volume in the first second,FEV1)的结果显示,健康对照组高于AR组、BA组和AR+BA组[(96.4±2.5)%、(90.7±4.9)%、(88.6±4.1)%、(82.1±5.5)%,F=47.193,P值均<0.001],显示AR和BA患儿肺功能可能变差。AR组、BA组和AR+BA组HMGB1水平高于健康对照组[(30.4±12.4)mg/L、(47.2±17.8)mg/L、(51.4±21.8)mg/L、(4.3±2.1)mg/L,F=48.896,P值均<0.001]。AR组、BA组和AR+BA组Th17/Treg比例高于健康对照组(0.68±0.14、0.71±0.14、1.36±0.40、0.30±0.07,F=105.547,P值均<0.001)。Pearson相关性分析结果显示,HMGB1与Th17(r=0.521,P<0.001)、Th17/Treg(r=0.419,P<0.05)、IgE(r=0.502,P<0.05)、IL-17(r=0.472,P<0.05)呈正相关,与FEV1(r=-0.645,P<0.001)呈负相关。结论在儿童AR合并BA的发病过程中,HMGB1水平及Th17细胞百分比增加,而Treg细胞百分比降低,Th17/Treg比例存在失衡的情形,HMGB1水平与Th17细胞百分比、Th17/Treg比例、IgE、IL-17及FEV1存在一定的相关性,HMGB1和Th17/Treg的动态平衡可能通过影响细胞因子的表达而起到一定的调节作用。
文摘目的构建经MHCⅡ通路的屋尘螨1类变应原Der p 1的T细胞表位肽疫苗重组载体。方法分别合成TAT、Ih C和含编码Der p 1的3段T细胞表位的融合核苷酸序列,用特异性引物PCR扩增相应的基因片段,分别用相应的双酶切后,用T4 DNA连接酶连接形成TAT-Ih C-Der p 1-3T融合基因,并插入至原核表达载体p ET-28a(+)中,构建重组原核表达载体p ET-28a(+)-TATIh C-Der p 1-3T,Bam HⅠ和XhoⅠ进行双酶切和测序鉴定。重组载体转化大肠杆菌E.coli BL21(DE3)菌株,IPTG诱导后,经SDS-PAGE电泳分析和Western blot验证,纯化TAT-Ih C-Der p 1-3T蛋白后进行Ig E结合试验。结果双酶切和测序结果表明,成功构建了p ET-28a-TAT-Ih C-Der p 1-3T重组原核表达载体;SDS-PAGE电泳分析显示TAT-Ih C-Der p 1-3T可诱导表达;Western blot检测表明该融合蛋白纯化成功;Ig E结合试验表明TAT-Ih C-Der p 1-3T结合屋尘螨过敏病人血清Ig E的能力强于Der p 1变应原(P<0.001)。结论成功构建了可表达经MHC通路的编码Der p 1的3段T细胞表位的重组p ET-28a-TAT-Ih C-Der p 1-3T载体,纯化的TAT-Ih C-Der p 1-3T具有较强的Ig E结合能力,从而为后续经MHC通路的特异性免疫治疗奠定基础。
