蛋氨酸脑啡肽单独或联合IL-2、IFN-γ对小鼠CD4+T细胞作用的研究

通过体内外MENK单独或联合IL-2、IFN-γ对C57BL/6小鼠CD4+T细胞数量,CD4+T细胞mRNA表达量以及细胞因子产生量的变化,来阐明MENK对CD4+T细胞的免疫效应.应用流式细胞术、酶联免疫吸附试验、RT-PCR方法检测C57 BL/6小鼠体内外单独应用MENK,或联合IL-2、IFN-γ后,CD4+T细胞数量,CD4+T细胞mRNA的表达量及上清中细胞因子含量.MENK单独或联合IL-2、IFN-γ,在体内能显著增加小鼠CD4+T细胞数量,CD4+T细胞mRNA表达量及细胞因子IL-2、IFN-γ含量;体外MENK单独应用可以显著增加小鼠CD4+T细胞数量,CD4+T细胞mRNA表达量及细胞因子IL-2、IFN-γ含量;而体外MENK联合IL-2或IFN-γ与MENK单独作用时相比无明显差异.MENK单独或联合IL-2、IFN-γ,在体内能上调CD4+T细胞的免疫效应.     

不同RF型类风湿性关节炎患者Th17相关细胞因子表达研究

目的:探讨类风湿因子阳性与阴性类风湿关节炎(rheumatoid arthritis,RA)患者外周血中辅助T细胞(Th17)及相关细胞因子白介素17(interleukin,IL-17),白介素6(interleukin,IL-6)表达的差异。方法:收集RA患者51例,根据RF测定分为RF+、RF-组,健康查体者(对照组)20例,采用流式细胞术检测受检者外周血单个核细胞(PBMC)的Th17细胞的百分率;以酶联免疫吸附法(ELISA)检测受检者血浆中IL-17,IL-6的水平。结果:RA患者CD4+IL-17+T细胞,IL-17、IL-6水平均高于对照组,RF因子阳性与阴性RA患者之间CD4+IL-17+T细胞,IL-17、IL-6表达水平均存在差异有统计学意义。结论:在RA中不同RF型免疫反应和炎症表达的不同,可能与Th17及相关细胞因子表达差异有关。     

EV71相关手足口病患儿T细胞及细胞因子检测及意义

目的:探讨EV71相关手足口病患儿外周血T细胞及细胞因子的水平及其意义.方法:将90例EV71感染手足口病患儿分为脑炎并发肺水肿组8例,单纯脑炎组30例,无并发症组52例.采用间接免疫荧光法检测外周血CD4+T及CD8+T细胞百分比,用双抗体夹心ABC-ELISA法检测血清IL-6、IL-10、TNF-α及IFN-γ含量.结果:并发症组CD4+及CD8+T淋巴细胞亚群的百分比相对无并发症组的降低,差异有统计学意义(F=9.304,P＜0.05;X2=26.075,P＜0.05),并发症组血清1L-10、IFN-γ和IL-6含量较无并发症组增高,差异有统计学意义(F=4.232,P＜0.05;X2=16.975,P＜0.05;X2=15.562,P＜0.05),并发症组TNF-α含量和无并发症组差异无统计学意义.结论:外周血T细胞及细胞因子均参与了EV71相关手足口病重症并发症的发生过程,在手足口病的临床诊疗中可予以参考,早期鉴别出现重症并发症的病例,及时给与相应的治疗措施,降低手足口病的病死率和致残率.     

95例SARS患者的Th2细胞因子反应及其临床意义

本研究共收集95例罹患严重急性呼吸综合征(SARS)的医护人员的急性期和恢复期血清,用ELISA方法测定血清中白细胞介素2(IL-2)、白细胞介素4(IL-4)、白细胞介素10(IL-10)、干扰素γ(IFN-γ)、肿瘤坏死因子β(TNF β)以及转化生长因子β(TGF-β)水平,用流式细胞仪测定急性期患者外周血CD4+和CD8+T细胞比例.结果显示,SARS患者病程初、中期血清IFN-γ无明显改变;IL-2在起病2月内无显著变化,但在病程3～5个月时降低;IL-4在健康对照和患者均未能测出;TNF-β在SARS感染的1周内和病程第2个月内下降.与上述细胞因子形成对比,患者IL-10以及TGF-β均在发病起即持续升高直至整个病程.这些细胞因子的变化均与激素的使用无关(P＞0.05).SARS患者急性期CD4+T细胞和CD8+T细胞急剧减少.可见,SARS相关冠状病毒感染可引起Th2细胞因子反应,可能损害患者的细胞免疫功能而促进体液免疫,IL-10和TGF-β可能在SARS的发病中起作用.     

细胞免疫功能及辅助型T细胞因子在口腔扁平苔藓中的作用

目的探讨细胞免疫功能及辅助型T细胞(Th)细胞因子在口腔扁平苔藓(OLP)中的作用。方法以35例糜烂型(糜烂组)及29例网纹型OLP(网纹组)患者为对象,选取同期20例健康人为对照组,流式细胞术检测外周血中T细胞、B细胞、NK细胞和调节性T细胞亚群表达百分率。ELISA法检测外周血中TNF-α和INF-γ(Th1型细胞因子),IL-4和IL-10(Th2型细胞因子),IL-17、IL-23(Th17型细胞因子)含量;分离外周血单个核细胞,qRT-PCR检测上述因子mRNA的表达。结果与健康组相比,OLP患者CD3~+、CD4~+T细胞亚群比例(F=3.211和3.565,P0.05)及CD4~+/CD8~+降低(F=3.430,P0.05),CD4~+Foxp3~+调节性T细胞亚群比例(F=3.370,P0.05)及外周血中TNF-α、INF-γ、IL-4、IL-10、IL-17、IL-23含量增高(F=0.923、0.820、1.043、1.132、0.745和0.802,P0.05)。与网纹组相比,糜烂组CD8~+T细胞亚群比例较低(t=2.450,P0.05);IL-4、IL-10蛋白(t=22.780和25.112,P0.05)及mRNA较低(t=3.781和6.710,P0.05),IL-17、IL-23蛋白(t=15.765和19.307,P0.05)及mRNA较高(t=4.022和8.569,P0.05)。结论口腔扁平苔藓患者存在细胞免疫紊乱及Th细胞因子异常,其中CD8~+及Th17型细胞与糜烂型OLP发病有关,Th2型细胞与网纹型OLP发病有关。     

分泌性中耳炎患者外周血Th1Th2细胞及T淋巴细胞亚群水平的表达及临床意义

目的:探讨分泌性中耳炎(SOM)患者外周血T辅助细胞1(Th1)、T辅助细胞2(Th2)细胞因子及T淋巴细胞亚群水平的表达及其临床意义。方法:选取我院于2015年1月至2018年1月期间收治的SOM患者135例记为SOM组,根据病程将患者分为急性组(病程14d,49例)、亚急性组(病程14-30d,53例)、慢性组(病程30d,33例)。另外选择同期于我院进行体检的100例健康者为对照组。分别对比SOM组和对照组受试者、不同病程SOM患者外周血Th1细胞因子[干扰素(INF-γ)、白细胞介素-2(IL-2)]、Th2细胞因子[白细胞介素-4(IL-4)、白细胞介素-10(IL-10)]以及T淋巴细胞亚群[CD3~+、CD4~+、CD8~+、CD4~+/CD8~+]水平。采用Pearson相关性分析INF-γ、IL-2、IL-4、IL-10与CD3~+、CD4~+、CD8~+、CD4~+/CD8~+的相关性。结果:SOM组患者外周血INF-γ、IL-2、IL-4、IL-10水平高于对照组(P0.05);急性组患者外周血INF-γ、IL-2、IL-4、IL-10水平低于亚急性组、慢性组,亚急性组患者外周血INF-γ、IL-2、IL-4、IL-10水平低于慢性组(P0.05)。SOM组患者外周血CD3~+、CD4~+、CD4~+/CD8~+水平低于对照组,CD8~+水平高于对照组(P0.05);急性组患者外周血CD3~+、CD4~+、CD4~+/CD8~+水平高于亚急性组、慢性组,CD8~+水平低于亚急性组、慢性组,亚急性组患者外周血CD3~+、CD4~+、CD4~+/CD8~+水平高于慢性组,CD8~+水平低于慢性组(P0.05)。Pearson相关性分析结果显示,SOM患者外周血INF-γ、IL-4与CD8~+呈正相关(P0.05),IL-4与CD3~+、CD4~+呈负相关(P0.05)。结论:SOM患者外周血Th1Th2细胞因子、T淋巴细胞亚群水平均表现异常,且其水平与疾病发生和发展存在一定联系,通过监测Th1Th2细胞因子、T淋巴细胞亚群有助于评估SOM患者病情。     

阴道微生态与宫颈HPV感染患者炎性因子和T细胞亚群的相关性分析CSCD

目的分析阴道微生态与宫颈感染人乳头瘤病毒(HPV)患者炎性因子、 T细胞亚群的相关性。方法 选取120例宫颈感染HPV患者作为观察组,另选取95例同期体检健康女性作为对照组,观察2组阴道菌群情况、T细胞亚群水平(CD4+、CD8+、CD4+/CD8+)、IFN-γ和IL-4等炎性因子水平。观察组根据患者阴道微生态情况又分为阴道微生态失衡组和阴道微生态正常组,比较两组T细胞亚群和炎性因子水平;ROC曲线评估T细胞亚群和炎性因子预测阴道微生态失调的价值;采用Spearman相关性分析分析阴道菌群失调与HPV感染患者T细胞亚群变化和炎性因子水平的关系。结果 观察组阴道微生态失衡率明显46.7%高于对照组6.3%(P<0.05);观察组CD4+T细胞、CD4+/CD8+、IFN-γ水平均较对照组低,CD8+T细胞、IL-4水平均较对照组高(P<0.05);阴道微生态失衡者CD4+T细胞、CD4+/CD8+和IFN-γ水平低于阴道微生态正常者,而CD8+T细胞、IL-4水平高于阴道微生态正常者(P<0.05);血清IFN-γ、IL-4、CD4+T细胞、CD8+T细胞、CD4+/CD8+联合预测HPV感染患者阴道微生态失衡的AUC为0.972,灵敏度、特异度分别为85.7%、98.4%;Spearman相关性分析显示,HPV感染患者血清IFN-γ、IL-4、CD4+T细胞、CD8+T细胞、CD4+/CD8+水平与阴道菌群失调呈正相关(P<0.05)。结论 HPV感染患者阴道微生态失衡,出现免疫功能紊乱和炎症反应,血清T细胞亚群和炎性因子与患者阴道微生态失衡有相关性,在一定程度上能预测HPV感染患者阴道微生态失衡的发生。     

黄芪多糖对免疫抑制模型小鼠Treg细胞及Th17细胞亚群的影响

本研究旨在探讨黄芪多糖对免疫抑制模型小鼠CD4+CD25+调节性T细胞(Treg细胞)和CD4+Th17细胞亚群功能的影响。通过小鼠腹腔注射环磷酰胺建立免疫抑制动物模型,应用黄芪多糖对免疫抑制小鼠进行治疗,采用流式细胞仪分析脾脏中CD4+T细胞、CD4+CD25+Treg细胞的比例,ELISA检测血清IL-17的水平。结果表明:与模型对照组比较,黄芪多糖高、中剂量明显降低小鼠脾脏CD4+T细胞数的比例(P0.05),同时黄芪多糖上调小鼠脾脏CD4+CD25+Treg细胞数的比例(P0.05),降低免疫抑制小鼠血清IL-17的水平,尤其1.0 g/(kg·d)剂量组的IL-17水平下降较为明显(P0.01)。提示黄芪多糖具有提高免疫抑制小鼠CD4+CD25+Treg细胞数的比例、降低CD4+T细胞的数量及抑制IL-17分泌活性的作用。     

95例SARS患者细胞免疫功能抑制的特点及其原因

用ELISA方法测定95例SARS患者急性期、恢复期血清细胞因子水平,用流式细胞仪检测该组患者急性期、恢复期淋巴细胞亚群,并分析细胞因子水平和淋巴细胞亚群变化的关系。结果发现,IL-10和TGF—β在观察期（急性期和恢复期）持续升高,急性期CD4^＋和CD8^＋T细胞显著减少,恢复期患者外周血CD8^＋记忆T细胞减少36．78％。IL-10和TGF-β升高与淋巴细胞变化具有统计学相关。结果提示,SARS病毒感染抑制宿主的细胞免疫功能,引起急性期CD4^＋和CD8^＋T细胞显著降低和恢复期的免疫记忆细胞减少。而IL-10和TGF—β过表达可能在SARS免疫病理中起重要作用。     

rhG-CSF 动员对供者CD4+ T 细胞免疫表型和功能影响

目的:研究重组人粒细胞集落刺激因子(rhG-CSF)动员对供者CD4+T细胞表面分子淋巴细胞功能相关抗原-1(LFA-1)、细胞间黏附分子-1(ICAM-1)、L-选择素(LAM-1)和人整合素-4(VLA-4)的表达及其介导的CD4+T细胞功能的影响,探讨外周血干细胞移植过程中CD4+T细胞免疫耐受机制。方法:使用三色荧光标记检测动员前及动员后第5天供者外周血LFA-1、ICAM-1、LAM-1和VLA-4的表达率,ELISA方法检测动员前后CD4+T细胞分泌IFN-γ和IL-4能力,免疫磁性分选法分离纯化CD4+T细胞,检测动员前后CD4+T细胞对基质细胞衍生因子-1α(SDF-1α)的迁移能力和对ICAM-1的黏附能力。结果:动员前后CD4+T细胞LFA-1(CD11a)和VLA-4(CD49d)表达率差异无统计学意义(P>0.01),动员前后CD4+T细胞LAM-1(CD62L)和ICAM-1(CD54)的表达率差异均有统计学意义,动员前显著高于动员后(P<0.01);动员前后CD4+T淋巴细胞向SDF-1α的迁移率差异无统计学意义(P>0.01);动员后CD4+T细胞对ICAM-1的黏附率降低(P<0.01);动员后IL-4和IFN-γ两个细胞因子在外周血血清的浓度均降低(P<0.01)。结论:rhG-CSF动员不影响CD4+T细胞LFA-1和VLA-4表达及CD4+T细胞迁移,但影响CD4+T细胞ICAM-1和LAM-1表达以及CD4+T细胞通过LFA-1对ICAM-1的黏附能力影响,并可能影响CD4+T细胞分泌细胞因子IL-4及IFN-γ的功能。     

Cutting edge: in the absence of TGF-β signaling in T cells, fewer CD103+ regulatory T cells develop, but exuberant IFN-γ production renders mice more susceptible to helminth infection

Multiple factors control susceptibility of C57BL/6 mice to infection with the helminth Heligmosomoides polygyrus, including TGF-β signaling, which inhibits immunity in vivo. However, mice expressing a T cell-specific dominant-negative TGF-β receptor II (TGF-βRII DN) show dampened Th2 immunity and diminished resistance to infection. Interestingly, H. polygyrus-infected TGF-βRII DN mice show greater frequencies of CD4(+)Foxp3(+)Helios(+) Tregs than infected wild-type mice, but levels of CD103 are greatly reduced on both these cells and on the CD4(+)Foxp3(+)Helios(-) population. Although Th9 and Th17 levels are comparable between infected TGF-βRII DN and wild-type mice, the former develop exaggerated CD4(+) and CD8(+) T cell IFN-γ responses. Increased susceptibility conferred by TGF-βRII DN expression was lost in IFN-γ-deficient mice, although they remained unable to completely clear infection. Hence, overexpression of IFN-γ negatively modulates immunity, and the presence of Helios(+) Tregs may maintain susceptibility on the C57BL/6 background.     

The role of inflammatory and anti-inflammatory cytokines in the pathogenesis of human tegumentary leishmaniasis

In tegumentary leishmaniasis caused by Leishmania braziliensis, there is evidence that increased production of IFN-γ, TNF-α and absence of IL-10 is associated with strong inflammatory reaction and with tissue destruction and development of the lesions observed in cutaneous leishmaniasis (CL) and mucosal leishmaniasis (ML). We evaluate the role of regulatory cytokines and cytokine antagonists in the downregulation of immune response in L. braziliensis infection. Peripheral blood mononuclear cells from CL and ML were stimulated with soluble Leishmania antigen in the presence or absence of regulatory cytokines (IL-10, IL-27 and TGF-β) or antagonists of cytokines (α-TNF-α and α-IFN-γ). Cytokines production (IL-10, IL-17, TNF-α and IFN-γ) was measured by ELISA. IL-10 and TGF-β downmodulate TNF-α and IL-17 production, whereas IL-27 had no effect in the production of TNF-α, IFN-γ and IL-17 in these patients. Neutralization of TNF-α decreased IFN-γ level and the neutralization of IFN-γ decreased TNF-α level and increased IL-10 production. This study demonstrate that IL-10 and TGF-β are cytokines that appear to be more involved in modulation of immune response in CL and ML patients. IL-10 might have a protective role, since the neutralization of IFN-γ decreases the production of TNF-α in an IL-10-dependent manner.     

Transforming growth factor-β and Th17 responses in resistance to primary murine schistosomiasis mansoni

Discovery of the T-helper (Th) 17 cell lineage and functions in immune responses of mouse and man prompted us to investigate the role of transforming growth factor-beta (TGF-β) and interleukin (IL)-17 in innate resistance to murine schistosomiasis mansoni. Schistosoma mansoni-infected BALB/c and C57BL/6 mice were administered with recombinant TGF-β or mouse monoclonal antibody to TGF-β to evaluate the impact of this cytokine on host immune responses against lung-stage schistosomula, and subsequent effects on adult worm parameters. Developing schistosomula elicited increase in peripheral blood mononuclear cells (PBMC) mRNA expression and/or plasma levels of IL-4, IL-17, and interferon-gamma (IFN-γ), cytokines known to antagonize each other, resulting in impaired Th1/Th2, and Th17 immune responses and parasite evasion. Mice treated with TGF-β showed elevated PBMC mRNA expression of IL-6, IL-17, TGF-β, and TNF-α mRNA and increased IL-23 and IL-17 or TGF-β plasma levels, associated with significantly (P < 0.02–<0.0001) lower S. mansoni adult worm burden compared to controls in both mouse strains, thus suggesting that TGF-β led to heightened Th17 responses that mediated resistance to the infection. Mice treated with antibody to TGF-β showed increase in PBMC mRNA expression and plasma levels of IL-4, IL-12p70, and IFN-γ, and significantly (P < 0.02 and <0.0001) reduced worm burden and liver worm egg counts than untreated mice, indicating that Th1/Th2 immune responses were potentiated, resulting in significant innate resistance to schistosomiasis. The implications of these observations for schistosome immune evasion and vaccination were discussed.     

Effect of short-term interferon-β treatment on cytokines in multiple sclerosis: significant modulation of IL-17 and IL-23

Therapeutic effect of interferon-β (IFN-β) treatment has been associated with modulation of the balance between Th1, Th17, Th2 and regulatory T (Treg) cells, whereas the impact of disease modifying drugs on Th9-immunity in multiple sclerosis (MS) has not been studied. To investigate the short-term effects of IFN-β treatment on cytokines in MS, we determined serum levels of IL-17, IL-23, IL-10, IL-4, IFN-γ, IL-9 and TGF-β in relapsing remitting MS patients before and 2 months after IFN-β treatment by ELISA. MS patients showed increased IL-17, IL-23 and IL-4 levels and decreased IL-9 levels as compared to healthy controls. IFN-β treatment only reduced IL-17 and IL-23 levels, whereas the levels of other cytokines remained unchanged. IFN-β treatment appears to exert its earliest therapeutic effect on Th17-immunity. The influence of IL-9 on MS pathogenesis needs to be further studied.     

A regulatory instead of an IL-17 T response predominates in Helicobacter pylori-associated gastritis in children

Th17 cells seem to have an important role in the efficacy of vaccines against Helicobacter pylori. Because children are a target group for human vaccination and Th17/T(reg) cells have intrinsically linked and antagonic commitments, we compared the gastric levels of Th17- and T(reg)-associated cytokines of children and adults. IL-6, IL-10 and TGF-β1 levels and Foxp3(+) cell numbers were higher, but IL-1β, IL-17A and IL-23 were lower in infected children than in infected adults. In conclusion T(reg) instead of Th17 cell response to H. pylori-infection predominates in children.     

Nitric oxide modulates TGF-beta-directive signals to suppress Foxp3+ regulatory T cell differentiation and potentiate Th1 development

TGF-β can induce Foxp3(+) inducible regulatory T cells (Treg) and also synergize with IL-6 and IL-4 to induce Th17 and Th9 cells. We now report that NO modulates TGF-β activity away from Treg but toward the Th1 lineage. NO potentiated Th1 differentiation in the presence of TGF-β in both IL-12-independent and -dependent fashions by augmenting IFN-γ-activated STAT-1 and T-bet. Differentiation into Treg, Th1, and Th17 lineages could be modulated by NO competing with other cofactors, such as IL-6 and retinoic acid. NO antagonized IL-6 to block TGF-β-directed Th17 differentiation, and together with IL-6, NO suppressed Treg development induced by TGF-β and retinoic acid. Furthermore, we show that physiologically produced NO from TNF and inducible NO synthase-producing dendritic cells can contribute to Th1 development predominating over Treg development through a synergistic activity induced when these cells cocluster with conventional dendritic cells presenting Ag to naive Th cells. This illustrates that NO is another cofactor allowing TGF-β to participate in development of multiple Th lineages and suggests a new mechanism by which NO, which is associated with protection against intracellular pathogens, might maintain effective Th1 immunity.     

Analysis of the expression of toll-like receptors 2 and 4 and cytokine production during experimental Leishmania chagasi infection

Toll-like receptors (TLRs) recognise pathogen-derived molecules and influence immunity to control parasite infections. This study aimed to evaluate the mRNA expression of TLRs 2 and 4, the expression and production of the cytokines interleukin (IL)-12, interferon (IFN)-γ, tumor necrosis factor (TNF)-α, IL-17, IL-10 and transforming growth factor (TGF)-β and the production of nitric oxide (NO) in the spleen of mice infected with Leishmania chagasi. It also aimed to evaluate any correlations between mRNA expression TLR2 and 4 and cytokines and NO production. Infection resulted in increased TLR2-4, IL-17, TNF-α and TGF-β mRNA expression during early infection, with decreased expression during late infection correlating with parasite load. IFN-γ and IL-12 mRNA expression decreased at the peak of parasitism. IL-10 mRNA expression increased throughout the entire time period analysed. Although TGF-β, TNF-α and IL-17 were highly produced during the initial phase of infection, IFN-γ and IL-12 exhibited high production during the final phase of infection. IL-10 and NO showed increased production throughout the evaluated time period. In the acute phase of infection, there was a positive correlation between TLR2-4, TNF-α, IL-17, NO, IL-10 and TGF-β expression and parasite load. During the chronic phase of infection, there was a positive correlation between TLR2-4, TNF-α, IL-17 and TGF-β expression and parasite load. Our data suggest that infection by L. chagasi resulted in modulation of TLRs 2 and 4 and cytokines.     

Astragalus polysaccharides enhance the humoral and cellular immune responses of hepatitis B surface antigen vaccination through inhibiting the expression of transforming growth factor β and the frequency of regulatory T cells

Astragalus polysaccharides (APS), extracted from the root of Astragalus membranaceus, a traditional Chinese medicinal herb, have extensive pharmacological and strong immunomodulatory effects. In this study, the potential adjuvant effect of APS on humoral and cellular immune responses to hepatitis B subunit vaccine was investigated. Coadministration of APS with recombinant hepatitis B surface antigen significantly increased antigen-specific antibody production, T-cell proliferation and CTL (cytotoxic T lymphocyte) activity. Production of interferon-γ (IFN-γ), interleukin-2 (IL-2) and IL-4 in CD4(+) T cells and of IFN-γ in CD8(+) T cells were dramatically increased. Furthermore, expression of the genes PFP, GraB, Fas L and Fas were up-regulated; interestingly, expression of transforming growth factor β (TGF-β) and the frequency of CD4(+)CD25(+)Foxp3(+) regulatory T cells (Treg cells) were down-regulated. Expression of Toll-like receptor 4 (TLR4) was significantly increased by administration of APS. Together, these results suggest that APS is a potent adjuvant for the hepatitis B subunit vaccine and can enhance both humoral and cellular immune responses via activating the TLR4 signaling pathway and inhibit the expression of TGF-β and frequency of Treg cells.