Interleukin-1 (IL-1), interleukin-2(IL-2) and interleukin-6(IL-6) activities and tumor necrosis factor (TNF) contents in plasma from patients with different sites of cancers as well as controls using bioassay technique were studied. The results showed that the levels of IL-1,IL-2,IL-6 s from patients with different sites of cancer were decreased remarkably in comparision with controls and the contents of TNF from patients with different sites of cancers increased significantly. But the difference between different sites of cancer was not statistically significant. The data suggest that the variations in the contents of TNF and the levels of interleukins may be related to the development of these caner patients.
Objective To investigate expression level of interleukin-27 (IL-27)and its impact on virus replication in lung after infected with respiratory syncytial virus (RSV). Methods In vivo,7-week aged female C57 mice were infected with RSV and sacrificed on day 0,0.5,1,2,4,6,8 (n=5).The left lung was extracted for RNA,and then real-time PCR was used to detect the mRNA levels of IL-27p28,IL-27EBI3, RSV-M protein and interferon-β (IFN-β).The right lung was used for HE staining.In vitro, human lung epithelial cells (A549)were infected with RSV,and stimulated with different concentrations of recombinant human interleukin 27 (rhIL-27)in doses of 0,1ng/mL,10ng/mL,and 50ng/mL.After 24 hours,the mRNA expression of interferon induced transmembrane protein 3 (IFITM3),IFN-β,and RSV-M protein were determined by real-time PCR.IFITM3 and STAT1/pSTAT1 protein expression levels were detected by Western blot. Results The viral load in the lungs of mice peaked on 4 day post infection (DPI),then gradually decreased,and almost returned to normal on 8 DPI.IFN-β increased transiently 12 hours after infection,and then quickly returned to normal baseline.IL-27 p28/EBI3 levels peaked on 1 DPI,then gradually decreased to normal on 4 DPI.Stimulating RSV-infected A549 cells with rhIL-27 of 10ng/mL and 50ng/mL significantly inhibited viral replication,enhanced IFTIM3 mRNA expression,and induced STAT1 phosphorylation.However,rhIL-27 stimulation did not affect IFN-β mRNA expression. Conclusions The IL-27 mRNA expression increases after RSV infection.The inhibition of IL-27 on RSV replication might be related to up-regulation of STAT1 phosphorylation and IFITM3 protein expression.
ObjectiveTo investigate the role of interleukin-25 (IL-25) and its receptor during allergen challenge test in allergic asthmatics as well as its underlining mechanism.MethodsFifteen allergic asthmatic patients with dual response in allergen challenge test were enrolled and blood samples were collected before and after challenge test. The expression levels of IL-25 receptor on the surface of eosinophils, plasma and intracellular IL-25 levels were measured by flow cytometry and enzyme-linked immunosorbent assay. Besides, the function of eosinophils from these patients was evaluated through the expression of type 2 cytokines, degranulation and chemotaxis after stimulation with IL-25.ResultsUpon allergen challenge, the expression of IL-17RB on the surface of eosinophils were increased from (7 426±2 824)/106 white blood cells to (19 446±5 593)/106 white blood cells (P<0.001). The expression of IL-17RA/RB on eosinophils were significantly increased from (4 508±1 360)/106 white blood cells to (9 025±3 166)/106 white blood cells (P<0.001). The plasma level of IL-25 increased from (650±45) pg/ml to (851±43) pg/ml (7 hours after allergen challenge) and (813±56) pg/ml (24 hours after allergen challenge) (P<0.001). The intracellular IL-25 expression of eosinophils was also upregulated from (10 398±1 909)/106 white blood cells to (147 684±46 222)/106 white blood cells (P<0.05). In vitro study, IL-25 (1 ng/ml) stimulated eosinophils for 2 hours promoted its expression of peroxidase [(12.5±4.2) ng/ml compared to control (1.26±0.4) ng/ml, P<0.05). The intracellular expression of IL-5 and IL-13 in eosinophils were also increased after stimulated by IL-25. IL-25 (1 pg/ml) stimulation compared to control could increase eosinophil migration in eotaxin [(36±3) vs. (69±5), P<0.05).ConclusionIL-25 and its receptor play a critical role in eosinophilic aggregation, activation and mobilization during allergic inflammation in allergic asthmatics.
Objective To study the regulative effect of angelica sinensis on cellular immune function in perioperative patients with obstructive jaundice. Methods Fourteen patients with obstructive jaundice were injected with angelica before and after operation for 14 days. The activity of IL-2 and the expression of IL-2R in lymphocytes in peripheral blood were measured, respectively. Results The activity of IL-2 and the expression of IL-2R decreased significantly in patients with obstructive jaundice (P<0.01). The activity of IL-2 and the expression of IL-2R in peripheral blood lymphocyte increased significantly before and after operations (after treatment using angelica) (P<0.01), though there was a little decrease after operation but they were still higher than that befor using angelica.Conclusion It maybe useful to use angelica to improve the cellular immune function in patients with obstructive jaundice.
ObjectiveTo investigate the effect of interleukin (IL)-23 receptor (IL-23R) overexpression on the balance of T helper 17 (Th17 cells)/regulatory T cells (Treg cells) in experimental autoimmune uveitis (EAU) mice. MethodsTwelve 8-week-old female C57BL/6J mice were randomly divided into LV-Ctrl group and LV-IL-23R group, with 6 mice in each group. Two groups of mice were injected with LV-Ctrl and LV-IL-23Rlentiviruses through the tail vein, respectively; 7 days after injection, the EAU mouse model was established by active immunization with vitamin A-binding protein 1-20 between photoreceptors. Starting from 13 days after immunization, the fundus of the mice was observed by indirect ophthalmoscopy every 2 days and clinical scores were performed; 30 days after immunization, hematoxylin-eosin staining was used to observe the histopathological changes of mouse retina. The levels of IL-17 in serum of the two groups of mice were detected by enzyme-linked immunosorbent assay; the proportion of Th17 cells and Treg cells was detected by flow cytometry. The relative mRNA expression of IL-23R, IL-17, retinoic acid-related orphan receptor γt (RORγt), IL-10 and forkhead transcripyion factor p3 (Foxp3) were detected by real-time quantitative polymerase chain reaction. Comparisons between groups were performed using repeated measures analysis of variance, independent samples Mann-Whitney U test, and independent samples t test. ResultsCompared with the LV-Ctrlgroup, the retinal inflammatory reaction of the LV-IL-23R group was more severe. At 13 days after immunization, there was no significant difference in fundus inflammation scores between LV-IL-23R group and LV-Ctrl group (t=-2.001, P=0.058); 15-29 days after immunization. The fundus inflammation scores of LV-IL-23Rgroup were higher than those of LV-Ctrl group, and the difference was statistically significant (t=-4.429, -6.578, -7.768, -10.183, -6.325, -7.304, -4.841, -6.872; P<0.001). Histopathological examination showed that the infiltration of inflammatory cells in the fundus increased, the retinal structure was damaged more seriously, and the histopathological score was significantly increased, and the difference was statistically significant (t=-4.339, P=0.001). Compared with the LV-Ctrl group, the relative expression of IL-23RmRNA in the spleen of the LV-IL-23R group was significantly increased, and the difference was statistically significant (Z=2.087, P=0.037). The relative expression of IL-17 and RORγt mRNA increased, while the relative expression of IL-10 and Foxp3 mRNA decreased, and the differences were statistically significant (t=-6.313,-5.922, 4.844, 7.572; P=0.003, 0.004, 0.008, 0.002). Compared with the LV-Ctrl group, the level of IL-17 in the serum of the mice in the LV-IL-23R group was significantly increased, and the difference was statistically significant (t=-5.423, P=0.002); the proportion of Th17 cells in the spleen and lymph nodes was significantly increased, whereas, the proportion of Treg cells was significantly reduced, and the difference was statistically significant (t=-4.290, 3.700; P=0.002, 0.006). ConclusionIL-23R overexpression can promote Th17/Treg imbalance in EAU mice, and aggravate the clinical and pathological manifestations of EAU.
To study the role of endotoxin in acute hemorrhagic necrotizing pancreatitis (AHNP), the change of endotoxin were studied in rats AHNP models by injection of 5% sodium taurocholate 1 ml/kg into pancreatic duct, and the effects of recombinant interleukin-2 (IL-2) in the treatment of AHNP were observed in this experiment. The results indicated that endotoxin involved the aggravation of AHNP and was associated with the increase of serum phospholipas-2 (PLA2), and these mediators were positively correlated with severe degrees of pancreatic damage. The results also suggeste that IL-2 might inhibit the overexpression of endotoxin and PLA2 and mitigate the pancreatic injury and decrease the 72h-mortality rate of AHNP from 66.7% to 26.7% (P<0.01). Endotoxin might play a major role in the pathogenesis of AHNP and IL-2 might have a potential role in the treatment of AHNP.
Effect of radical operation on expression of interleukin-2(IL-2)mRNA and production of IL-2 were markedly reduced preoperatively and four weeks after operation,expression of IL-2 mRNA significantly enhanced,but it was still lower than that in the normal group.Production of IL-2 nearly reached normal level,When PBL was activated by phytohemagglutinin(PHA),expresseion of IL-2 mRNA and production of IL-2 were much higher than that in non-activated PBL.These results suggested that expression of IL-2 were much higher than that in non-activated PBL.These results suggested that expression of IL-2 mRNA and production of IL-2 are dificient in gastric cancer patients,and radical surgery will help them to recover and they can also be improved through activation with PHA.
Objective To investigate the effects of cimetidine on the red cell immune function and interleukin-2(IL-2) in rats with obstructive jaundice. Methods Sixty SD rats were divided into bile duct ligation(BDL) group, cimetidine therapy (BDLC) group and sham operation(SO) group respectively. The red cell immue function and serum IL-2 level were determined with the red cell yeast-rosttes test and radioimmunoassay respectively. Results The red blood cell C3b receptor rosette rate(RBC-C3bRR), the red blood cell immune complex rosette rate(RICR), the red blood cell C3b receptor rosette-forming excited rate(RFER) and serum IL-2 level were significantly lower in BDL group as compared with SO group, the red blood cell C3b receptor rosette-forming inhibitory rate(RFIR) in BDL group was higher than that of SO group. After 7 days’ cimetidine therapy RBCC3bRR, RICR, RFER and IL-2 became higher than those of BDL group, but RFIR was lower than that of BDL group. Conclusion Supplemental cimetidine can significantly enhance the impaired red cell immune function and IL-2 production in rats with obstructive jaundice.
ObjectiveTo detect expressions of interleukin-17 (IL-17) and interleukin-27 (IL-27) proteins in liver tissue of hepatic alveolar echinococcosis. MethodsThe edge liver tissues (from the lesion edge 0.5 cm) and the normal liver tissues (from the lesion edge 5 cm) of 20 patients with hepatic alveolar echinococcosis in the Affiliated Hospital of Qinghai University were collected and stored at-80℃ freezer. The immunohistochemical method was used to detect the expressions of IL-17 and IL-27 proteins in these two tissues. ResultThe positive rates of IL-17 and IL-27 protein expressions in the edge liver tissues were significantly higher than those in the normal liver tissues[IL-17:80.0% (16/20) versus 10.0% (2/20), χ2=12.36, P < 0.01; IL-27:85.0% (17/20) versus 20.0% (4/20), χ2=12.36, P < 0.01]. ConclusionHigh expressions of IL-17 and IL-27 protein in edge liver tissue might participate in progress of hepatic alveolar echinococcosis.
ObjectiveTo observe whether interleukin-27 (IL-27) intervention could diminish allergic airway inflammation of mouse asthma induced by ovalbumin (OVA) and to investigate the related molecular mechanisms. MethodsSixty female C57/6J mice were randomly divided into six groups, a control group, an asthma group, two IL-27 prevention groups and two IL-27 treatment groups. Based on being sensitized and challenged with OVA in the asthma model, two kinds of IL-27 intervention asthma models were set up, one of which was low-dose multiple prevention model, the other was high-dose few times treatment model. HE stain and inflammation score were done for the lungs. CD4+ T cells were purified from mice spleen and cultured under Th2 medium with/without IL-27. Interleukin-4 (IL-4) was measured by ELISA. CD4+ T cells were cultured under different stringent Th2 medium and stimulated by IL-27. The level of total signal transducer and activator of transcription-1 (STAT1) protein and phos-STAT1 were tested by Western blot. ResultsIn low-dose multiple prevention group, IL-27 inhibited inflammation around bronchial and vascular obviously, the inflammation score was lower than the asthma group (P < 0.05), while the treatment group had no obvious statistical significance (P > 0.05). IL-27 repressed Th2 differentiation of naïve CD4+ T cells which was independent of interferon-γand IL-10. This effect was via STAT1 signaling pathway. CD4+ T cells from asthma mice or cultured under high-IL-4 inducing medium were found impairment of STAT1 phosphorylation. ConclusionsIL-27 could inhibit Th2 differentiation of naïve CD4+ T cells, but not in already committed Th2-CD4+ T cells. The inhibition effect of IL-27 for airway inflammation is obvious in prevention group, while the treatment group shows obviously resistance to inhibitory effect of IL-27. Already committed Th2-CD4+ T cells existed in asthma airway might be the reason for IL-27 resistance.