IL-17 induces NSCLC cell migration and invasion by elevating MMP19 gene transcription and expression through the interaction of p300-dependent STAT3-K631 acetylation and its Y705-phosphorylation

The cancer cell metastasis is a major death reason for patients with non-small cell lung cancer(NSCLC).Although researchers have disclosed that interleukin 17(IL-17)can increase matrix metalloproteinases(MMPs)induction causing NSCLC cell metastasis,the underlying mechanism remains unclear.In the study,we found that IL-17 receptor A(IL-17RA),p300,p-STAT3,Ack-STAT3,and MMP19 were up-regulated both in NSCLC tissues and NSCLC cells stimulated with IL-17.p300,STAT3 and MMP19 overexpression or knockdown could raise or reduce IL-17-induced p-STAT3,Ack-STAT3 and MMP19 level as well as the cell migration and invasion.Mechanism investigation revealed that STAT3 and p300 bound to the same region(−544 to−389 nt)of MMP19 promoter,and p300 could acetylate STAT3-K631 elevating STAT3 transcriptional activity,p-STAT3 or MMP19 expression and the cell mobility exposed to IL-17.Meanwhile,p300-mediated STAT3-K631 acetylation and its Y705-phosphorylation could interact,synergistically facilitating MMP19 gene transcription and enhancing cell migration and invasion.Besides,the animal experiments exhibited that the nude mice inoculated with NSCLC cells by silencing p300,STAT3 or MMP19 gene plus IL-17 treatment,the nodule number,and MMP19,Ack-STAT3,or p-STAT3 production in the lung metastatic nodules were all alleviated.Collectively,these outcomes uncover that IL-17-triggered NSCLC metastasis involves up-regulating MMP19 expression via the interaction of STAT3-K631 acetylation by p300 and its Y705-phosphorylation,which provides a new mechanistic insight and potential strategy for NSCLC metastasis and therapy.

Effects of early enteral nutrition on Th17/Treg cells and IL-23/IL-17 in septic patients

BACKGROUND The imbalance of Th17/Treg cells and the IL-23/IL-17 axis have been confirmed to be associated with sepsis and various inflammatory diseases. Early enteral nutrition (EEN) can modulate the inflammatory response, improve immune dysfunction, and prevent enterogenic infection in critically ill patients;however, the precise mechanisms remain unclear. Considering the important roles of Th17 and Treg lymphocytes in the development of inflammatory and infectious diseases, we hypothesized that EEN could improve the immune dysfunction in sepsis by maintaining a balanced Th17/Treg cell ratio and by regulating the IL- 23/IL-17 axis. AIM To investigate the effects of EEN on the Th17/Treg cell ratios and the IL-23/IL-17 axis in septic patients. METHODS In this prospective clinical trial, patients were randomly divided into an EEN or delayed enteral nutrition (DEN) group. Enteral feeding was started within 48 h in the EEN group, whereas enteral feeding was started on the 4th day in the DEN group. The Th17 and Treg cell percentages and the interleukin levels were tested on days 1, 3, and 7 after admission. The clinical severity and outcome variables were also recorded. RESULTS Fifty-three patients were enrolled in this trial from October 2017 to June 2018. The Th17 cell percentages, Th17/Treg cell ratios, IL-17, IL-23, and IL-6 levels of the EEN group were lower than those of the DEN group on the 7th day after admission (P < 0.05). The duration of mechanical ventilation and of the intensive care unit stay of the EEN group were shorter than those of the DEN group (P <0.05). However, no difference in the 28-d mortality was found between the two groups (P = 0.728). CONCLUSION EEN could regulate the imbalance of Th17/Treg cell ratios and suppress the IL- 23/IL-17 axis during sepsis. Moreover, EEN could reduce the clinical severity of sepsis but did not reduce the 28-d mortality of septic patients.

慢性阻塞性肺病大鼠外周血中Foxp^(3+)调节性T细胞及IL-17、IL-6水平的变化

目的探讨Foxp3+调节性T细胞及白介素-6、白介素-17在慢性阻塞性肺疾病(COPD)大鼠发病机制中的作用。方法采用60只健康的雄性Wistar大鼠随机分为两组:正常对照组(30只),COPD组(30只)。香烟熏法复制Wistar大鼠慢性阻塞性肺疾病的动物模型。流式细胞仪检测香烟暴露大鼠(COPD)和正常对照大鼠外周血Foxp3+调节性T细胞数量,酶联免疫法(ELISA)测定实验组及正常对照组血清IL-6和IL-17的水平。结果 1Foxp3+主要表达在CD4+CD25+T细胞上,COPD大鼠外周血中CD4+CD25+T细胞、CD4+Foxp3+T细胞与对照组相比百分率均明显降低(P≤0.05);2大鼠COPD模型组血清IL-6、IL-17浓度含量较正常对照组明显增高,差异有统计学意义(P<0.05);3COPD组中外周血IL-17与IL-6水平呈正相关。结论香烟暴露致使(COPD)大鼠气道炎症模型中Foxp3+Treg细胞及相关炎症因子水平异常变化,提示IL-17、IL-6与Foxp3+Treg细胞共同参与了气道炎症的发生与发展过程。

IL-17A抗体抑制变应性鼻炎鼠模型中IL-17A及RORγt并升高Foxp3 mRNA表达

目的:探讨IL-17A抗体对小鼠变应性鼻炎鼻黏膜中IL-17A、RORγt、Foxp3 mRNA水平的影响及意义。方法:以卵清蛋白致敏的Balb/c小鼠变应性鼻炎模型作为实验组,同期以生理盐水替代作为对照组,使用IL-17A抗体治疗作为治疗组。实时定量PCR方法检测三组小鼠鼻黏膜中IL-17A、RORγt、Foxp3mRNA的含量。结果:实验组中RORγt以及IL-17AmRNA水平均较对照组升高(P<0.05),治疗组中RORγt以及IL-17AmRNA的含量均低于实验组(P<0.05)。而实验组中Foxp3 mRNA水平较对照组下降(P<0.0 5),治疗组中Foxp3 mRNA的含量高于实验组(P<0.0 5)。结论:IL-17A抗体抑制变应性鼻炎鼠模型中IL-17A及RORγt mRNA水平并升高Foxp3 mRNA表达水平。

IL-17^＋Foxp3^＋T细胞的研究进展

调节性T细胞是一类表达CD4^＋、CD25^＋及转录因子Foxp3的T细胞亚群,其免疫抑制功能对免疫稳态的动态调节必不可少。Th17细胞是一种新的CD4^＋T细胞亚群,优先分泌IL-17（即IL-17A）,还能分泌IL-17F、IL-21、IL-22、IL-26等细胞因子,Th17细胞及其效应细胞因子在宿主抵御真菌和细胞外细菌方面发挥重要作用。

IL-17＋Foxp3＋T细胞的研究进展

近来有文献报道复杂的细胞因子环境下,调节性T细胞(regulatory T cells,Tregs)可以转变为表型和功能上酷似Th17的新的T细胞亚群,即白细胞介素(IL)-17+Foxp3+T细胞。IL-17^+Foxp3^+T细胞分泌IL-17并表达维甲酸受体相关孤儿受体γt(RORγt),在免疫系统中表现出双重特性。IL-17^+Foxp3^+T细胞的发现为理解Tregs和Th17的关系提供了新的思路。本文重点介绍了IL-17^+Foxp3^+T细胞的表型特征、分化来源和多效性功能,并进一步总结了炎症性疾病和肿瘤微环境中IL-17^+Foxp3^+T细胞的作用。

血清sTim-3、Gal-9及IL-17水平与冠心病患者冠脉狭窄程度的关系及对预后的预测价值

目的探讨血清可溶性T细胞免疫球蛋白黏蛋白-3(sTim-3)、半乳糖凝集素-9(Gal-9)、白介素-17(IL-17)水平与冠心病患者冠脉狭窄程度及预后的关系,以期为冠心病患者的治疗及预后预测提供参考。方法选取2020年4月—2022年4月于延安市人民医院就诊的182例冠心病患者,根据Gensini评分将患者分为轻度狭窄组(49例)、中度狭窄组(76例)和重度狭窄组(57例);根据纽约心脏病协会(NYHA)心功能分级将患者分为心功能Ⅰ级组(32例)、心功能Ⅱ级组(85例)和心功能Ⅲ级组(65例);根据预后情况分为预后不良组(43例)和预后良好组(139例)。比较不同冠脉狭窄程度、不同心功能分级及不同预后冠心病患者血清sTim-3、Gal-9、IL-17水平,分析这几个指标与冠脉狭窄程度的相关性及其对患者预后的预测价值。结果患者Gensini评分及血清sTim-3、Gal-9、IL-17水平随冠脉狭窄程度的加重而升高,且不同冠脉狭窄程度患者Gensini评分及血清sTim-3、Gal-9、IL-17水平比较,差异有统计学意义(P<0.05)。患者血清sTim-3、Gal-9、IL-17水平随心功能分级的增加而升高,且不同心功能分级患者血清sTim-3、Gal-9、IL-17水平比较,差异有统计学意义(P<0.05)。Pearson相关分析结果显示,冠心病患者血清sTim-3、Gal-9、IL-17水平与Gensini评分均呈正相关(P<0.05)。预后良好组患者血清sTim-3、Gal-9、IL-17水平均低于预后不良组,差异均有统计学意义(P<0.05)。ROC曲线分析结果显示,血清sTim-3、Gal-9、IL-17水平以及三者联合预测冠心病患者预后的曲线下面积(AUC)分别为0.834、0.781、0.789、0.852(P<0.05)。结论冠心病患者血清sTim-3、Gal-9、IL-17水平随冠脉狭窄程度加重以及心功能分级增加而升高,且在冠心病预后不良患者中呈高表达水平,对冠心病患者预后具有一定预测价值,三者联合检测的预测价值更高。

HPV DNA负荷量、辅助性T细胞17、FoxP3+调节性T细胞及炎症因子与高危型HPV感染的相关性分析

目的探讨人乳头瘤病毒(HPV)DNA、辅助性T细胞17(Th17)、叉头状转录因子3阳性(FoxP3^(+))调节性T细胞(Treg)及宫颈灌洗液中炎症因子与高危型HPV感染的相关性,为高危型HPV感染防治提供新思路。方法将2020年9月至2021年2月该院收治的高危型HPV感染患者100例纳入研究作为研究组,然后将其中50例高危型HPV16/18阳性患者作为HPV16/18组,50例其他12种高危型阳性病例作为其他亚型组。另外,选取同期体检的健康女性作为对照组(n=50)。比较研究组和对照组HPV DNA负荷量、Th17、FoxP3^(+)Treg及宫颈灌洗液中炎症因子[白细胞介素(IL)-10、肿瘤坏死因子α(TNF-α)、IL-17A]水平,比较HPV16/18组和其他亚型组各项指标水平,比较不同宫颈病变患者各项指标水平。分析各项指标与高危型HPV感染、宫颈病变的相关性,以及对宫颈病变患者高危型HPV感染的诊断价值。结果研究组HPV DNA负荷量、Th17、FoxP3^(+)Treg及宫颈灌洗液中IL-10、TNF-α、IL-17A水平均高于对照组(P<0.05),HPV16/18组各项指标水平均高于其他亚型组(P<0.05)。不同宫颈病变患者各项指标水平比较,差异有统计学意义(P<0.05)。HPV DNA负荷量、FoxP3^(+)Treg、Th17及宫颈灌洗液中IL-10、TNF-α、IL-17A水平与高危型HPV感染分型(HPV16/18=1,其他亚型=2)均呈负相关(P<0.05),与宫颈病变(宫颈上皮内瘤变Ⅰ级=1,宫颈上皮内瘤变Ⅱ级=2,宫颈上皮内瘤变Ⅲ级=3,宫颈癌=4)均呈正相关(P<0.05)。各项指标联合诊断高危型HPV16/18阳性的曲线下面积(AUC)为0.911(95%CI:0.837~0.959),大于各指标单独诊断。结论高危型HPV感染患者HPV DNA负荷量、Th17、FoxP3^(+)Treg及炎症因子水平异常,与高危型HPV感染分型、宫颈病变密切相关,而且HPV DNA负荷量与Th17、FoxP3^(+)Treg、炎症因子相关,各项指标联合检测可为临床防治高危型HPV持续感染提供可靠依据。

0.1%纳米银溶液对慢性根尖周炎大鼠RORγT、IL-17及Foxp3表达的影响

目的:探讨0.1%纳米银溶液用于根管封药对慢性根尖周炎大鼠RORγT、IL-17、Foxp3表达的影响。方法:对40只雄性大鼠双侧下颌第一磨牙开髓建立根尖周炎模型,术后2周处死4只大鼠用RVG+HE判断造模成功,将剩余36只大鼠随机均分为空白对照组(K组),氢氧化钙组(Q组)及0.1%纳米银溶液组(N组)。于用药后7、14、21 d各组随机处死4只大鼠,免疫组织化学检测Foxp3表达,qRT-PCR检测RORγT、IL-17的表达。结果:免疫组化显示3组于用药7、14、21 d时Foxp3表达呈逐渐上升趋势,N、Q组均高于K组(P<0.05)。qRT-PCR显示,N组RORγT、IL-17表达在14、21 d均低于K组(P<0.05),21 d时,N较Q组IL-17表达无差异(P>0.05),RORγT表达有差异(P<0.05)。结论:0.1%纳米银溶液用于根管封药效果好,可降低RORγT、IL-17水平,促进Foxp3的表达。

Correlation of expression of STAT3,VEGF and differentiation of Th17 cells in psoriasis vulgaris of guinea pig

Objective:To investigate the role of T help 17 cells(Th17)and STAT3-VEGF pathway in pathogenesis of psoriasis.Methods:A total of 50 cases of psoriasis guinea pigs and 20 normal guinea pigs were selected.The ratio of Th17/IL-17 cell in peripheral blood were detected by flow cytometric analysis;STAT3 and VEGF concentrations were measured hy immunohistochemistry and Western blot.Results:The expression of Th17 in peripheral blood were significantly increased in psoriasis[(1.76±0.88)%]compared with controls[(0.48±0.27)%](P<0.05).Th17 related cytokine STAT3 and VEGF were significantly increased in psoriasis compared with controls(P<0.05),and were positively correlated the expression of Th17.Conclusions:The expressions of Th17,STAT3 and VEGF are elevated in psoriasis,which suggests Th17 cells have a potential role in the pathogenesis of psoriasis by STAT3-VEGF pathway.

Analysis of CD4^+CD25^+ Regulatory T Cells and Foxp3 mRNA in the Peripheral Blood of Patients with Asthma

The changes of CD4^＋CD25^＋ regulatory T cells （CD4^＋CD25^＋ Treg） and Foxp3 mRNA in peripheral blood mononuclear cells （PBMCs） from patients with asthma were investigated in order to elucidate the possible roles of CD4^＋CD25^＋ Treg in the development of asthma. The peripheral blood samples were collected from 29 healthy controls （normal control group） and 78 patients with asthma which included 30 patients in exacerbation group, 25 patients in persistent group, and 23 patients in remission group. By using flow cytometry and RT-PCR, the CD4^＋CD25^＋ Treg ratio and Foxp3 mRNA in PBMCs were detected. The CD4^＋CD25^＋ Treg ratio and Foxp3 mRNA in PBMCs of exacerbation and persistent groups were lower than that of remission and normal control groups （P〈0.05）. Although the CD4^＋CD25^＋ Treg ratio and Foxp3 mRNA of remission group were also lower than that of normal control group, there was no significant difference between them （P〉0.05）. As compared with persistent group, exacerbation group had lower CD4^＋CD25^＋ Treg ratio and Foxp3 mRNA （P〈0.05）. It was indicated that the decrease of CD4^＋CD25^＋ Treg ratio and its function in PBMCs may be responsible for pathogenesis of asthma.

Reduction of natural regulatory T cells in thymomas accompanying myasthenia gravis and its possible association with Foxp3 and thymic stromal lymphopoietin

Objective: To investigate the expression of both thymic regulatory T cells (CD4+CD25+Foxp3+cells, Treg) and thymic stromal lymphopoietin (TSLP) in thymomas accompanying myasthenia gravis. Methods: We used immunohistochemistry and real-time reverse trancription polymerase chain reaction (real-time RT-PCR) techniques to determine Foxp3+ Treg counts and the expression levels of Foxp3 mRNA and TSLP mRNA in thymomas of 23 MG patients and thymuses of 4 healthy controls. Results: The CD4+ Foxp3+ nTreg (natural regulatory T cells) counts in thymomas were significantly lower than those in normal thymuses (P<0.01), and the expression levels of Foxp3 mRNA and TSLP mRNA were also lower in thymomas(P<0.01). Among the thymoma types, type B1 thymoma had the highest Foxp3+ nTreg count and standard values of Foxp3 mRNA and TSLP mRNA. There was a strong positive correlation between the mRNA transcriptional levels of Foxp3 and TSLP. Conclusion: The insufficient expression of Foxp3 in thymoma, which may be caused by decreased transcription of TSLP, may result in the reduction of Tregs and cause autoimmune disorders.

Differentially expressed genes and signalling pathways are involved in mouse osteoblast-like MC3T3-E1 cells exposed to 17-β estradiol

Oestrogen is essential for maintaining bone mass, and it has been demonstrated to induce osteoblast proliferation and bone formation.In this study, complementary DNA（cDNA） microarrays were used to identify and study the expression of novel genes that may be involved in MC3T3-E1 cells’ response to 17-b estradiol. MC3T3-E1 cells were inoculated in minimum essential media alpha（a-MEM）cell culture supplemented with 17-b estradiol at different concentrations and for different time periods. MC3T3-E1 cells treated with1028mol？L2117-b estradiol for 5 days exhibited the highest proliferation and alkaline phosphatase（ALP） activity; thus, this group was chosen for microarray analysis. The harvested RNA was used for microarray hybridisation and subsequent real-time reverse transcription polymerase chain reaction（RT-PCR） to validate the expression levels for selected genes. The microarray results were analysed using both functional and pathway analysis. In this study, microarray analysis detected 5 403 differentially expressed genes,of which 1 996 genes were upregulated and 3 407 genes were downregulated, 1 553 different functional classifications were identified by gene ontology（GO） analysis and 53 different pathways were involved based on pathway analysis. Among the differentially expressed genes, a portion not previously reported to be associated with the osteoblast response to oestrogen was identified. These findings clearly demonstrate that the expression of genes related to osteoblast proliferation, cell differentiation, collagens and transforming growth factor beta（TGF-b）-related cytokines increases, while the expression of genes related to apoptosis and osteoclast differentiation decreases, following the exposure of MC3T3-E1 cells to a-MEM supplemented with 17-b estradiol. Microarray analysis with functional gene classification is critical for a complete understanding of complementary intracellular processes. This microarray analysis provides large-scale gene expression data that require further confirmatory studies.

慢性乙肝患者肝内IL-23/IL-17信号途径与Treg细胞之间的相互作用

目的探讨慢性乙肝患者体内FOXP3蛋白与炎症因子IL-23/IL-17信号途径之间的相互关系。方法采用qPCR、流式细胞术分析了39例慢性乙肝急性发作(ACLF)、56例慢性乙肝(CHB)及32例健康对照(HC)肝组织及外周血中的FOXP3及IL-23/IL-17信号途径相关炎症因子的表达,免疫组化及荧光共聚焦检测FOXP3在肝组织内的表达,定量PCR检测血清HBV DNA水平,酶联免疫吸附试验检测HBsAg浓度,全自动生化仪检测血清谷丙转氨酶(ALT)水平。结果 CHB患者外周血及肝组织内FOXP3 mRNA表达明显升高,肝内的FOXP3表达水平与患者血液中的HBV DNA水平及乙肝表面抗原(HB-sAg)浓度明显呈正相关。与健康对照组相比,慢性乙肝患者肝脏内的IL-23/IL-17信号通路相关的促炎因子也明显升高,且与FOXP3的表达水平升高呈正相关。结论 HBV感染的肝脏内IL-23/IL-17信号通路的活化并没有有效地被宿主的免疫机制如Treg细胞所抑制。因此,IL-23/IL-17信号通路可能是维持HBV病毒持续性感染的一个机制。

泡球蚴感染小鼠肝组织T-bet、GATA-3、ROR-γt和IL-17 mRNA表达水平的研究

目的:研究泡球蚴感染小鼠肝组织T-bet、GATA-3、ROR-γt和IL-17mRNA表达水平的变化特点,初步探讨Th17细胞与Th1/Th2在棘球蚴感染中的相互关系。方法:通过腹腔接种多房棘球绦虫的幼虫原头蚴混悬液制备泡球蚴感染小鼠模型,持续观察10个月,分别在感染2天、8天、1月、3月、6月和10月处死小鼠,收集肝脏组织。采用逆转录聚合酶链反应(RT-PCR)方法检测各组小鼠肝脏组织T-bet、GATA-3、ROR-γt和IL-17 mRNA的表达水平。结果:T-bet mRNA的表达水平在感染早期开始增高,1月时达高峰后又开始缓缓下降至感染前水平;GATA-3 mRNA的表达水平在感染1月时明显升高,随后一直保持较高的表达水平;ROR-γt mRNA的表达水平在感染早期开始增高,感染1月时达到高峰,随后缓慢下降。IL-17 mRNA表达水平自感染早期开始持续性增高,到感染3月时达高峰,之后又逐渐降低至感染前水平。各组小鼠肝组织ROR-γt mRNA表达水平与T-bet mRNA表达水平呈正相关(r=0.67),IL-17 mRNA表达水平与ROR-γt mRNA表达水平有相关性(r=0.35),P值均<0.05。结论:泡球蚴感染宿主早期,主要以Th1型和Th17型细胞免疫为主,感染中后期出现Th1向Th2转化,机体呈现出以Th2细胞为主的优势应答。

原发性肾病综合征患儿外周血Th17与CD4^+ CD25^+ Foxp3^+调节性T细胞的水平

目的：观察原发性肾病综合征（PNS）患儿外周血Th17细胞和CD4^＋CD25^＋Foxp^3＋调节性T细胞（Treg）及其相关因子的水平和功能，初步探讨其在儿童PNS发病中的作用。方法：PNS患儿分为单纯型肾病（SNS）20例、肾炎性肾病（NNS）15例；同时以20例健康体检儿童作为对照组。采用流式细胞术（FCM）检测外周血单个核细胞（PBMC）中Th17细胞和Treg细胞比例；Real—time PCR法检测PBMC中RORC、IL-23p19和Foxp3 mRNA的表达；ELISA分别检测血清中IL-1β、IL-6、TGF-β1的水平。结果：SNS、NNS组患儿Th17细胞及RORC、IL-23p19mRNA、IL-1β、IL-6的水平均明显高于正常组（P〈0．05），且NNS组患儿以上指标明显高于SNS组（P〈0．05）；SNS、NNS组患儿Treg细胞、Foxp3 mRNA和TGF-β1水平的表达明显低于正常组（P〈0．05），其中两组疾病患儿血清TGF—β1的水平无统计学意义（P〉0．05），其余指标NNS患儿均明显低于SNS组（P〈0．05）。结论：Th17／Treg细胞功能失衡可能在儿童PNS的发病中起到重要作用，其失调程度可能与PNS的临床表现、对激素的敏感性、病理类型、及预后有关。

苦参碱调节IL-6/STAT3/NF-κB信号通路对炎症性肠病大鼠Th17/Treg平衡的影响

目的探讨苦参碱(MT)调节白细胞介素-6(IL-6)/信号转导和转录激活因子3(STAT3)/核转录因子κB(NF-κB)通路对炎症性肠病大鼠辅助性T细胞17(Th17)/调节性T细胞(Treg)平衡的影响。方法按照随机数字表法将SD大鼠分为空白对照组(CK组)、Model组,低、中、高剂量MT组(MT-L组,50 mg/kg;MT-M组,100 mg/kg;MT-H组,200 mg/kg),美沙拉嗪组(MSLM组,0.42 g/kg)、MT-H+rIL-6(IL-6激活剂)组(200 mg/kg+0.05 mg/kg),每组18只。除CK组外,其余组大鼠均采用50 g/L三硝基苯磺酸(20 mg/kg)缓冲液与500 mL/L乙醇按照1∶1比例混匀后灌肠以构建炎症性肠病大鼠模型,建模成功后,分别进行给药处理,每天1次,持续7周。分别在给药1、3、5、7周时对大鼠进行体质量的测量;比较各组大鼠结肠长度的变化;HE染色检测大鼠结肠组织病理损伤;ELISA法检测大鼠血清中肿瘤坏死因子-α(TNF-α)、白细胞介素(IL)-17(IL-17)、IL-10水平;流式细胞术检测大鼠外周血中Th17、Treg细胞比例;Western blotting检测大鼠结肠组织中维甲酸相关孤核受体γt(RORγt)、叉头框蛋白P3(Foxp3)、IL-6、p-STAT3、p-NF-κB p65蛋白表达。结果与CK组比较,Model组大鼠结肠组织病理损伤严重,体质量(3、5、7周时)、IL-10水平、Treg细胞比例、Foxp3蛋白表达降低,结肠变短,TNF-α、IL-17水平、Th17细胞比例、Th17/Treg比值、RORγt、IL-6、p-STAT3、p-NF-κB p65蛋白表达增高(P<0.05);与Model组比较,MT-L组、MT-M组、MT-H组、MSLM组对应指标变化趋势与上述相反(P<0.05);rIL-6减弱了高剂量MT对炎症性肠病大鼠Th17/Treg平衡的促进作用。结论MT可能通过抑制IL-6/STAT3/NF-κB信号通路,促进炎症性肠病大鼠Th17/Treg平衡。

Immunotherapy of rat glioma without accumulation of CD4^+CD25^+FOXP3^+ regulatory T cells

Immunotherapy may be used for the treatment of glioblastoma multiforme; however, the induced immune response is inadequate when either T cells or dendritic cells are used alone. In this study we established a novel vaccine procedure in rats, using dendritic cells pulsed with C6 tumor cell lysates in combination with adoptive transfer of T lymphocytes from syngenic donors. On day 21 after tumor inoculation, all the rats were sacrificed, the brains were harvested for calculation of glioma volume, cytolytic T lymphocyte responses were measured by cytotoxic assay, and the frequency of regulatory T lymphocytes （CD4＋CD25~FOXP3~） in the peripheral blood was investigated by flow cytometric analysis. The survival rate of rats bearing C6 glioma was observed. Results showed that the co-immunization strategy had significant anti-tumor potential against the pre-established C6 glioma, and induced a strong cytolytic T lymphocyte response in rats. The frequency of peripheral blood CD4＊CD25＊FOXP3＊ regulatory T lymphocytes was significantly decreased following the combination therapy, and the rats survived for a longer period. Experimental findings indicate that the combined immunotherapy of glioma cell lysate-pulsed dendritic cell vaccination following adoptive transfer of T cells can effectively inhibit the growth of gliomas in rats, boost anti-tumor immunity and produce a sustained immune response while avoiding the accumulation of CD4＋CD25＋FOXP3＋ regulatory r lymphocytes.

Th9、Th17、CD4^+CD25^+FoxP3^+调节性T细胞在慢性淋巴细胞白血病患者外周血中的变化

目的探讨慢性淋巴细胞白血病患者外周血中Th9、Thl7和CD4^+CD25^+FoxP3^+调节性T细胞及相关细胞因子的表达水平及意义。方法采用流式细胞仪检测36例CLL患者和45例健康对照组外周血Th9、Th17和Treg细胞的表达率,采用酶联免疫吸附方法(ELISA)检测外周血中细胞因子IL-9、IL-17、TGF-β的表达水平。结果 1)CLL组患者外周血Th9、Th17细胞比例及细胞因子IL-9、IL-17浓度均明显高于健康对照组[(1.29±0.35)%vs(0.74±0.23)%,P<0.05;(2.15±0.46)%vs(1.08±0.37)%,P<0.05;(24.25±7.05)%vs(16.35±5.42)%,P<0.05;(11.47±3.99)%vs(7.36±2.85%),P<0.05]。CD4^+CD25^+FoxP3^+细胞比例及细胞因子TGF-β浓度明显低于对照组[(4.76±0.89)%vs(7.26±1.95)%,P<0.05;(3.82±1.39)%vs(6.82±2.01)%,P<0.05]。2)相关性分析:患者Th9、Th17细胞比例及IL-9、IL-17浓度与淋巴细胞计数负相关(γ_s=-0.374,P=0.034;γ_s=-0.382,P=0.032;γ_s=-0.445,P=0.011;γ_s=-0.413,P=0.024);患者Treg细胞比例及TGF-β浓度与淋巴细胞计数正相关(γ_s=0.416,P=0.023;γ_s=0.403,P=0.028)。结论慢性淋巴细胞白血病患者Th9、Th17与CD4^+CD25^+FoxP3^+调节性T细胞比例失衡,血清IL-9、IL-17和TGF-β水平变化,这些原因可能参与慢性淋巴细胞白血病的免疫发病过程。

转录因子Foxp3和IL-10、TGF-β在小鼠衰老过程中的变化规律

目的:研究衰老进程中各年龄段小鼠外周单个核细胞Foxp3 mRNA表达差异及其与血清白细胞介素-10(IL-10)、转化生长因子-β(TGF-β)的相关性。方法:选取SPF级健康雄性BALB/c小鼠24只,随机等分为2月龄组(青年)、6月龄组(中年)和15月龄组(老年),每组8只。采用流式细胞术检测分析小鼠脾细胞中CD4+CD25+Foxp3+细胞水平,采用SYBR GreenⅠ实时荧光定量RT-PCR法检测外周PBMC中Foxp3 mRNA的表达,同时利用酶联免疫吸附测定法(ELISA)检测血清中IL-10和TGF-β水平,并分析三者之间及其与免疫衰老之间的相关性。结果:流式细胞术检测提示15月龄组脾细胞中CD4+CD25+Foxp3+细胞比例较2月龄组明显增高(P<0.01),15月龄组外周PBMC中Foxp3 mRNA的表达水平显著高于2月龄组(P<0.01),15月龄组血清中IL-10和TGF-β的含量显著高于2月龄组(P<0.01);15月龄组血清中IL-10和TGF-β含量与外周PBMC中Foxp3 mRNA的表达水平无相关性。结论:小鼠衰老过程中,可能存在有CD4+CD25+Foxp3+Treg细胞分化增强的可能。在机体衰老进程中Foxp3 mRNA表达上调,抑制性细胞因子IL-10和TGF-β的生成增多,提示Foxp3、IL-10和TGF-β的表达提高可能参与了免疫衰老的发生机制。应用SYBR GreenⅠ实时荧光定量RT-PCR技术检测Foxp3 mRNA表达水平简便、经济、可行。