Objective To investigate the role of T cell factor-4 (TCF-4) in the carcinogenesis of colorectal cancer. Methods Relevant references about TCF-4 and the carcinogenesis of colorectal cancer, which were published recently domestic and abroad, were collected and reviewed. Results For TCF-4 gene, multiple isoforms are generated by way of alternative splicing, which encode different proteins. TCF-4 protein is sequence-specific DNA binding protein and is incapable of activating or repressing transcription independently, but it can interact with distinct partners to lead to different effects through multiple domains. Conclusion TCF-4 might be viewed as nuclear vehicles targeting other auxiliary proteins to a specific set of promoters and functions as molecular switch during the carcinogenesis of colorectal cancer.
Objective To investigate the percentage of CD4+CD25+ Treg in peripheral blood of patients with severe multiple trauma and systemic inflammatory response syndrome（SIRS） and its effects on cellular immunity and secondary infection.Metheds Peripheral blood of 23 patients with severe multiple trauma was collected in 24 h after SIRS was diagnosed，and flow cytometry was used to determine the percentage of CD4+CD25+ Treg and CD4/CD8 ratio.Simultaneously，in order to explore the cell proliferation，silver staining was used to determine Ag-NORs of leukomonocyte in peripheral blood represented by IS%.In order to investigate the infection in patients，sputum and secretion sample were collected for bacteriological examination on 1 and 5 day after SIRS was established.Forty healthy volunteers were enrolled as control.Results Compared with the control，the percentage of CD4+CD25+ Treg was significant higher[（14.21±3.43）% vs（9.53±3.22），Plt;0.01] and the ratio of CD4/CD8 and IS% were significant lower in patients with severe multiple trauma[（5.94±0.66）% vs（6.74±0.95）%，（1.22±0.25）% vs（1.72±0.36）%，respectively，both Plt;0.01].In those patients（n=14） who developed secondary infection，Treg% was significant higher [（18.69±4.21）% vs（12.58±2.49）%，Plt;0.01]，while IS% and CD4/CD8 were significant lower [（5.79±0.68）% vs（6.15±0.57）%，（1.15±0.25）% vs（1.39±0.25）%，both Plt;0.01].compared to the patients without secondary infection Conlusion CD4+CD25+ Treg is valuable to estimate the cellular immunity and predict secondary infection in patients with severe multiple trauma.
Objective To explore the role of CD4+CD25+ Treg cells in chronic pulmonary infection caused by Pseudomonas aeruginosa(PA).Methods Sixty SD rats were randomly divided into a PA group and a control group(n=30 in each group).Chronic lung infection model was established by implantation of silicone tube precoated with PA into the main bronchus.Twenty-eight days later Treg cells in peripheral blood were measured by fluorescence-activated cell sorting（FACS）.Levels of IL-10 and TGF-β in serum were assayed by ELISA.The expression of Foxp3 mRNA in spleen was measured by RT-PCR.Pathological changes of lung tissue were studed by HE staining.Results Treg/CD4+ T cells in the PA group were significantly more than those in the control group[（19.79±6.45）% vs （5.15±0.47）%，Plt;0.05].The levels of IL-10 and TGF-β were (231.52±54.48)pg/mL and （121.05±7.98）pg/mL in the PA group respectively，which were significantly higher than those in the control group[（35.43±23.56）pg/mL and （36.02±8.94）pg/mL].The expression of Foxp3 mRNA in the PA group was significantly higher compared with the control group(0.80±0.044 vs 0.25±0.054，Plt;0.05）.HE staining revealed that PA caused a intensive inflammatory reaction with lymphocytes infiltration.Conclusion CD4+CD25+ Treg cell is up-regulated and plays an important role in chronic lung infection caused by Pseudomonas aeruginosa.
目的 检测系统性红斑狼疮（SLE）患者CD4+CD25+调节性T细胞（Treg）上Ⅰ型干扰素受体（IFNAR）的分布格局，了解Ⅰ型干扰素对SLE患者CD4+CD25+Treg产生直接影响的作用靶点。 方法 选取2010年9月－2011年10月间20例初次确诊的SLE患者（SLE组）和20例健康女性（对照组），分离SLE患者和对照组的外周血单个核细胞，采用流式细胞术测定CD4+CD25+Treg上IFNAR的表达。 结果 ① IFNAR1、IFNAR2在Treg和CD4+CD25? T细胞表面均有表达；两组Treg表面IFNAR1和IFNAR2的表达水平均高于CD4+CD25? T细胞。② 与对照组相比，SLE组Treg表面IFNAR1表达的平均荧光强度明显增高（P=0.001）。③ SLE组Treg表面IFNAR1表达平均荧光强度与疾病活动指数评分呈正相关（rs=0.505，P=0.023）。 结论 SLE患者CD4+CD25+Treg表面相对高表达IFNAR1且与疾病活动性相关，提示Ⅰ型干扰素以Treg上IFNAR为靶点在SLE发病机制中可能发挥重要作用，为SLE等自身免疫性疾病治疗寻找新的干预手段提供了理论基础。
ObjectiveTo evaluate the diagnostic value of T-SPOT.TB assay in patients with hematological disorders and tuberculosis. MethodsA total of 82 patients with hematological disorders and suspected tuberculosis diagnosed between March 2012 and April 2013 received T-SPOT. TB assay in the peripheral blood mononuclear cells. ResultsThe positive detection rate of T-SPOT.TB assay for patients with hematological disorders and tuberculosis was 59.09% (13/22), which was higher than the positive detection rate of anti-TB antibody test[13.64% (3/22)]. The sensibility and specificity of T-SPOT.TB assay for patients with hematological disorders and tuberculosis was 59.09% (13/22) and 68.33% (41/60), respectively. ConclusionT-SPOT.TB assay is of great value on diagnosis of tuberculosis for patients with hematological disorders and suspected tuberculosis. The diagnostic value of T-SPOT.TB assay is more important for tuberculosis infected patients; it can be used as an accessorial diagnostic method for patients with hematological disorder and suspected tuberculosis.
Objective To investigate the therapeutic effects of transplanting allogeneic marrow mesenchymal stem cells (MSCs) via subarachnoid space on spinal cord injury(SCI) and the T cell subpopulation. Methods Density gradient centrifugation was used to isolate and expand MSCs from bone marrow of 10 six-week-old SD rats. The SCI model was produced by weightbeating from 60 eight-week-old female SD rats. Forty survival SCI rats,which BBB scores were zero, were divided randomly into 2 groups:experimental group(group A) and control group(group B). In addition, 20 normal eightweekold SD ratswere used as blank group (group C). In group A, 1 ml cells suspention containing MSCs(the 6th generation, 2×106/ml) was injected via subarachnoid space. Ingroup B, equal volume of L-DMEM was injected in the same way. The BBB score was obtained after 1st,2nd and 3rd weeks of injection. At the same time,T cell subpopulation was detected by flow cytometry. Results The BBB score in group A was better than that in group B, but fewer than that in group C in the 3rd week. CD4+T cells in group A were less than those in groups B and C in the 1st, 2nd, and 3rd weeks. CD8+T cells in group A were less than those in groups B and C in the 2nd and 3rd weeks. The ratio of CD4+/CD8+T cells in group A was less than those in groups B and C in the 1st week. Above differences showed statistically significant difference(P＜0.05). However, there were no statistically significant differences in the ratio of CD4+/CD8+T cells between group A and groups B, C in the 2nd and 3rd weeks (P＞0.05). Conclusion The above results suggest that allogeneic MSCs transplantation via subarachnoid space is beneficial to SCI to some extend, do not result in rejection in vivo. Furthermore, it can lead to immunosuppression in short time. So, it provides clues to apply MSCs to treat SCI and other diseases.
The pathogenesis of Vogt-Koyanagi Harada disease (VKH) has not yet been fully defined. Current studies mainly suggest that VKH is actually an autoimmune disease, especially related to the immune response mediated by various signal transduction pathways involved in the function of T cells. In recent years, the influence of the balance imbalance of various T cell subsets in cellular immunity on the pathogenesis of VKH has been a hot research direction. Currently, T helper cell 17/T regulatory cells, balance is the focus of clinical research, meanwhile, new discoveries and potential clinical treatment schemes have been made for related cellular pathways, particularly the Janus kinase/signal transducers and activators of transcription pathway and NF-kappa B pathway. The exploration of B cells in the pathogenesis of VKH has also achieved initial results through the successful application of various targeted drugs. In the future, further screening and localization of genes or proteins that are abnormally regulated or expressed in VKH, for which early comprehensive and in-depth exploration will be helpful, thus improve the efficacy of clinical treatment programs and develop new therapeutic targets.
ObjectiveTo investigate the levels of regulatory T cells (Treg) and FoxP3 gene in patients with gastric cancer before and after operation. MethodsTwenty patients with definite diagnosis of gastric cancer and 15 healthy volunteers were selected. The levels of Treg and T cell subsets in peripheral blood were determined by detecting of CD4 and CD25 with immunefluorescence stain and flow cytometry, the expressions of FoxP3 mRNA in these Treg were detected by RTPCR technique. The expression of FoxP3 protein in the gastric cancer tissue was measured by immunohistochemistry assay. ResultsThe percentage of Treg cells in total CD4+ T isolated from the patients with gastric cancer was higher than that of healthy volunteers 〔(19.39±5.58)% versus (9.91±3.23)%, Plt;0.01〕, and it markedly decreased after operation 〔（13.50±5.93）% versus (19.39±5.58)%, Plt;0.05〕. The FoxP3 mRNA expression in the patients with gastric cancer was also higher than that of healthy volunteers (0.86±0.03 versus 0.64±0.02, Plt;0.01), and decreased after operation (0.73±0.04 versus 0.86±0.03, Plt;0.05). The percentage of CD4+T cell in mononucleocytes of peripheral blood of patients with gastric cancer was significantly lower than that of healthy volunteers (Plt;0.01), but the difference was not significant between before and after operation. FoxP3 protein expressed in cytoplasm of 13 patients with gastric cancer, in which bly positive in 2 cases, middle positive in 6 cases, weakly positive in 5 cases. FoxP3 protein didn’t express in cytoplasm of 7 patients with gastric cancer. ConclusionsTreg may have a significant effect on the onset and development of gastric cancer through immunosuppressive effect. Tumor tissue is an important initiating agent on Treg proliferation.
【Abstract】ObjectiveTo find a stable and efficient method for ex vivo concentration of CD4+CD25+ regulatory T cells in rats. MethodsCD4+CD25+ regulatory T cells were separated from the rat splenic cells in two steps by magnetic cell sorting (MACS) system. CD4+ T cells were first negatively sorted by cocktail antibodies and antiIgG magnetic microbeads. And then CD4+CD 25+T cells were positively sorted by antiCD25 PE and antiPE magnetic microbeads. The purity and the cell survival rate of the sorted cells were measured by flow cytometry and trypan blue dyeing respectively, and the immunosuppressive action of CD4+CD25+ T cells on the proliferation of CD4+CD25- T cells was also assessed by in vitro cell proliferation assay. ResultsThe purity of negatively sorted CD4+ T cells were (83.6±2.5)% (79%~87%), and the purity of positively sorted CD4+CD25+ T cells was (90.2±1.8)% (86%~93%) with the survival rate of (92.8±3.4)% (92%~95%). These concentrated cells significantly suppressed the proliferation of CD4+CD25- T cells in mixed lymphocyte culture （CD25+/CD25- versus CD25-, P＜0.01）. ConclusionWe created a twostep procedure of magnetic cell sorting system for CD4+CD25+ regulatory T cells sorting, which insures the cells to be satisfactorily purified and well functioned.
ObjectiveTo approach the role of CD4+CD25+ regulatory T cells in the maintenance of immunotolerance in mouse liver allograft. MethodsThe mouse orthotopic liver transplantation was performed. After the liver transplantation immunotolerance induction, antiCD25 monoclonal antibody (PC61) was injected into the recipients with a delayed timing to remove the CD4+CD25+ T cells. The percentage of CD4+CD25+ T cells and the expression of forkhead/winged helix transcription factor (Foxp3) in the recipients were examined. Furthermore, the survival time of the recipient was observed. ResultsC3H/HeJ recipients receiving DBA/2 hepatic allografts survived over 70 d as in the syngeneic liver transplantation (C3H/HeJ recipients receiving C3H/HeJ hepatic grafts). With various protocols of the delayed PC61 treatment, the CD4+CD25+ T cell was completely disappeared as observed. However, the removal of CD4+CD25+ regulatory T cells after the induction of transplantation immunotolerance did not affect the survival of hepatic allografts. ConclusionCD4+CD25+ regulatory T cells are not essential for the maintenance of spontaneous mouse liver transplantation immunotolerance.