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.
Regulatory B cells (Bregs) are a subset of B cells with immunomodulatory effects. The study of Bregs began with a variety of animal models of immune diseases. Studies in patients with autoimmune diseases have further clarified that Bregs are a group of immune cells that secrete inhibitory cytokines such as interleukin-10. Abnormal functions and numbers of Bregs have been found in a variety of autoimmune diseases. The study of the negative immune regulatory network involving Bregs is expected to provide new therapeutic ideas for diseases such as immune diseases, cancer, infection and inflammation. Starting from the discovery and immune regulation mechanism of Bregs, this paper focuses on its regulatory mechanism and clinical research value in the occurrence and development of autoimmune diseases, tumors, infectious diseases and inflammation.
Objective To review the regulation of liver regeneration factors. Methods The literatures about liver regeneration related regulators in recent years were reviewed. Results With further advancement of researches on regulators of liver regeneration in recent years, there were more therapies for treatment of liver-related diseases. Regulators play important roles in the process of liver regeneration, as one of which, cell growth factor plays an essential role in liver cell proliferation, such as the proper expression of TNF-α and IL-6 promoting liver cell proliferation, HGF, TGF-α, EGF, ALR, FS, and others motivating liver cell proliferation, while TGF-β and IL-1 physically terminating liver cell proliferation. Conclusion By strengthening the studies on liver regeneration regulators, new methods may appear for treating liver-related diseases.
Objective To establish a method to isolate the CD4+CD25+ regulatory T cells (Tregs) and to identify the purity and function of these cells. Methods The peripheral blood (8 mL) were collected from the great saphenous vein of 10 rhesus monkeys (4 females and 6 males, aged 4-5 years, and weighing 5-8 kg). The mononuclear cells were isolated with density gradient centrifugation. CD4+ T cells were separated by the Magnetic cell sorting (MACS) negative selection and MACS positive selection. The cell yield rate, the cell viability, and the cell purity were compared between MACS negative selection and MACS positive selection. In CD4+ MACS negative selection, the anti-biotin MicroBeads and biotin-antibody cocktai in CD4+CD25+ Tregs isolation kit non-human primate were used, and in MACS positive selection, the anti-APC MicroBeads in CD4+CD25+ Tregs isolation kit non-human primate and CD4-APC were used. The CD4+ T cells separated by positive selection were selected to obtain CD4+CD25 Tregs with CD25 MicroBeads. The purity, activity, the FoxP3 level, and the suppressive function to concanavalin A (ConA) activated autologous CD4+CD24- effective T cells (Teffs) of CD4+CD25+ Tregs were detected by flow cytometry. Results After CD4+ T cells were separated by MACS negative selection and MACS positive selection, the cell viabilities were all up to 95%, showing no significant difference (P gt; 0.05). The cell yield rate and purity of CD4+ T cells by positive selection were significantly higher than those of CD4+ T cells by negative selection (P lt; 0.05). CD4+CD25+ Tregs can be successfully isolated by MACS double positive selection. The classifying purity was 76.2% ± 8.6%; the cell activity was 93.3% ± 4.7%; and the level of FoxP3 was 74.2% ± 6.9%. The CD4+CD25+ Tregs had suppressive effect on ConA activated autologous CD4+CD25- Teffs. Conclusion MACS double positive selection can be used to isolate high-purity CD4+CD25+ Tregs from the peripheral blood of rhesus monkeys and the process does not influence the activity of CD4+CD25+ Tregs.
ObjectiveThrough the analysis of quantitative and functional changes in peripheral blood CD4+ CD25+FOXP3+ regulatory T cells (Treg) of early HCC patients before and after operation, to discuss the operation effect on the immune function from the aspect of immune suppression. MethodsExtracted the lymphocytes of peripheral blood in HCC patients before and after operation (case group, n=15) and normal people (control group, n=5 cases), and analyze the number and function of Treg by flow cytometer after extracellular (CD4, CD25) and intracellular (FOXP3) staining. ResultsCD4+CD25+ T cells and CD25+FOXP3+ T cells in preoperative peripheral blood in case group were significantly higher than those in control group (12.43±2.57)% vs. (5.56±1.02)%, (5.14±1.4)% vs. (2.18±0.83)%, Plt;0.05). These two cells decreased at 1 week after operation. 〔(10.56±2.13)%, (4.28±1.08)%〕, but there was not statistically significant (Pgt;0.05), they decreased significantly at 2 weeks after operation 〔(7.30±0.89)%, (3.43±0.83)%， Plt;0.05〕. CD8+ T cells and CD4+CD25- T cells in preoperative peripheral blood in case group were significantly lower than those in control group 〔（23.42±1.80）% vs. （29.22±2.26）%， （36.14±1.12）% vs. （43.69±2.78）%， Plt;0.05〕， These two cells decreased significantly at 2 weeks after operation 〔（27.15±1.71）%， （40.30±2.00）%〕. The analysis on the Treg and AFP correlation found that they have low correlation (r=048, Plt;0.05 ). ConclusionsThe hepatectomy can improve the immune response of HCC patient. Treg may have a certain auxiliary significance in the diagnosis, treatment and prognosis of patients with hepatocellular carcinoma.
Objective To investigate the efficacy of continuous blood purification ( CBP) in the treatment of severe sepsis, and explore the related immune regulatory mechanisms. Methods Forty-eight patients with severe sepsis were randomly divided into a control group ( n =23) and a CBP group ( n =25) .CD4 + CD25 + regulatory T cells ( Treg% ) in peripheral blood and APACHEⅡ score were measured dynamically before treatment and 12, 24, 36, 48, 60, 72 hours after treatment. Meanwhile the length of ICUstay, duration of mechanical ventilation, and 28 day mortality were determined. Results Compared with the control group, the length of ICU stay, ventilator time, incidence of multiple organ failure, and mortality decreased significantly in the CBP group ( P lt; 0. 05) . And CBP also decreased Treg% and APACHEⅡ score significantly. There was a positive correlation between Treg% and APACHEⅡ score ( r =0. 804, P lt;0. 01) .Conclusion Early CBP treatment can reduce Treg%, improve cellular immunity and improve the prognosis of sepsis.
Legionella, one of the causative pathogens of atypical pneumonia, firstly outbroked during the period of American Legion’s convention in 1976, Philadelphia. Legionella infection can accompany multisystem involvement. In addition to pulmonary lesions, it also accompanies extrapulmonary manifestations, including gastrointestinal symptoms (primarily diarrhea and hepatic dysfunction), neurological symptoms (primarily headache, disorientation and confusion of consciousness), urinary symptoms (primarily hematuria, proteinuria and acute kidney injury), rhabdomyolysis, as well as electrolyte disorder mainly characterized by hyponatremia and hypophosphatemia. This article reviews the extrapulmonary manifestations and its regulatory mechanism of Legionella infection.
Iron death is an alternative to normal cell death and is regulated by a variety of cellular metabolic pathways. Iron death has become a hot topic of research because it can cause damage to various organs and degenerative diseases in the body. Metabolism, signalling pathways, endoplasmic reticulum stress, and immune cells can all affect the occurrence of iron death, and the blood-retina destruction induced by iron death plays an important role in autoimmune uveitis. Exploring the components of the blood-retina regulatory mechanism of iron death in autoimmune uveitis can lead to the search for targeted drug targets, which can provide a new research idea for the subsequent study of the diagnosis and treatment of autoimmune uveitis.
Objective To investigate the role of T helper 17 ( Th17) cells and CD4 + CD25 + Foxp3+regulatory T cells ( Treg) in the pathogenesis of asthma in a mouse model. Methods Twenty-four BALB/ c mice were randomly divided into an asthma group and a normal control group, with 12 mice in each group.Asthma model was established by ovalbumin sensitization and aerosol challenge in the asthma group. Airway reactivity was measured by plethysmography. The total and differential cell counts in bronchoalveolar lavage fluid ( BALF) were measured. The ratio of Th17 and Treg cells to mononuclear cells in the spleens of mice were detected by flow cytometry. The levels of IL-17 and IL-10 in BALF and lung homogenates were measured by ELISA. Results The bronchial provocation test showed that the average lung resistance increased remarkably in the asthma group. In spleens of the asthmatic mice, the percentage of Th17 cells was significantly higher [ ( 5.68 ±1. 99)% vs ( 2.80 ±0. 82) %, P lt; 0. 01] , and the percentage of Treg cells was significantly lower [ (2.88 ±0. 46) % vs ( 6.10 ±2.44) % , Plt; 0. 01] , with the ratio of Th17 to Treg significantly increased( 1. 93 ±0. 41 vs 0. 50 ±0. 15,P lt;0. 01) . In BALF and lung homogenates of the asthma group, the level of IL-17 was significantly higher[ ( 22. 37 ±3. 00) pg/mL vs ( 11. 42 ±2. 15) pg/mL, ( 52. 93 ±5. 39) pg/mL vs ( 19. 38 ±2. 65) pg/mL, both Plt; 0. 01] , and the level of IL-10 was significantly lower[ ( 6. 05 ±1. 25) pg/mL vs ( 14. 23 ±2. 94) pg/mL, ( 9. 33 ±1. 79) pg/mL vs ( 21. 40 ±2. 44) pg/mL, both P lt; 0. 01] compared with the control group.Conclusion The imbalance of Th17/ Treg plays an important role in the pathogenesis of asthma.
ObjectiveTo investigate the proportion of peripheral blood CD4+CD25+ regulatory T cells (Tregs) in patients with pancreatic head carcinoma, the dynamic changes of these cells before and after pancreatoduodenectomy were also analyzed. MethodsThe proportions of peripheral blood CD4+CD25+ Tregs in patients with pancreatic head carcinoma and normal individuals were examined by using flow cytometric analysis. The CD4+/CD8+ ratio was also studied before and after operation. ResultsThe patients with pancreatic head carcinoma showed higher ratio of CD4+CD25+ and CD4+CD25high Tregs compared with normal control before operation (Plt;0.05). However, the percentage of these T cells reduced significantly after pancreatoduodenectomy, which was most obviously on the 3rd day after operation (Plt;0.01, Plt;0.05). After operation, CA199 level began to decrease, which was obvious on the fourteen day after operation. This tendency of CD4+CD25high Tregs changes was similar to that of CA199. The patients showed an decreased ratios of CD4+/CD8+ compared with normal controls, which further declined after operation, and reached the lowest point on the seventh day after operation (Plt;0.05). ConclusionsPancreatoduodenectomy may be helpful for the recovery of antitumor immunity. The perioperative period of patients with pancreatic head carcinoma may be a beneficial windowphase for immune intervention and Tregs may be served as target cells.