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。     

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。     

正常妊娠大鼠的Th2型免疫反应

采用流式细胞仪,^3H-TdR掺入和酶联免疫打点(enzyme-linked immunospot,ELISPOT)方法研究妊娠免疫学指标的改变。妊娠晚期大鼠脾脏单个核细胞表面分子主要组织相容性抗原Ⅱ(MHCⅡ)明显下调,外周血单个核细胞表达CD11c明显减少,共激活分子B7-1和B7-2未见改变;脾脏和外周血单个核细胞中Th2细胞因子IL-10、IL-4表达增多,TGFD阳性细胞数也明显增加,而Th1细胞因子IFNγ的产生未受抑制。此外,脾脏和外周血中单个核细胞的抗原特异性增殖未见改变,而腹腔淋巴结细胞的增殖明显升高。脾脏单个核细胞在妊娠晚期分泌较少的抗原特异性抗体。提示妊娠期性激素具有免疫调节作用,可能与怀孕时Th1细胞介导的自身免疫性疾病得到缓解有关。     

糖尿病患者外周血Th17/Treg细胞及相关细胞因子的表达变化

目的:探讨糖尿病患者外周血Th17细胞和Treg细胞数量以及相关细胞因子表达的变化。方法:以本院2013年1月至2015年12月收治的50例糖尿病患者为研究对象,其中1型糖尿病患者25例,2型糖尿病患者25例,同时以25例健康体检人群为正常对照,检测各组外周血Th17/Treg细胞数量以及血清IL-17、IL-10和TGF-β水平。结果:糖尿病患者外周血中Th17细胞比例均显著高于正常对照组(P0.05)而Treg细胞的比例均显著低于正常对照组(P0.05),且1型糖尿病患者Treg细胞比例显著低于2型糖尿病患者(P0.05)。糖尿病患者血清中IL-10和TGF-β水平均显著低于正常对照组(P0.05)而IL-17水平均显著高于正常对照组(P0.05),且1型糖尿病患者的IL-10水平显著低于2型糖尿病患者(P0.05)。结论:糖尿病患者体内Treg细胞数量及相关细胞因子低于正常而Th17细胞数量及相关细胞因子高于正常,这可能与患者体内的自身免疫失调有关。     

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。     

乳腺癌肺转移过程中Th1/Th2型细胞因子的动态变化

探究乳腺癌肺转移模型鼠肺组织内Th1/Th2型细胞因子的动态变化。利用皮下接种或尾静脉注射乳腺癌细胞系-4T1细胞的方式,建立乳腺癌肺转移模型。在4T1细胞接种后第7、14、21天分离模型鼠肺组织,通过HE染色、墨汁染色确认模型建立成功后,提取肺组织总RNA,采用Real-time RT-PCR方法,检测肺组织内IFN-γ、IL-4、IL-5、IL-13表达水平的变化。结果显示,尾静脉注射4T1细胞的BALB/c鼠,肺组织内Th1型细胞因子水平呈现先升高后逐渐下降的趋势;而皮下接种4T1细胞的BALB/c鼠,其肺组织内Th1型细胞因子水平一致处于较低水平。与Th1型细胞因子不同,肺组织内Th2型细胞因子水平无论在皮下接种鼠亦或是尾静脉注射鼠均表现为逐渐升高趋势。结果表明,乳腺癌肺转移过程中出现Th1/Th2型细胞因子失衡,并且由刚开始的Th1优势应答逐渐转变为Th2优势应答。     

妊娠期肝内胆汁淤积症患者外周血中维生素D受体 与Th1/Th2 型细胞因子的变化

目的:研究妊娠期肝内胆汁淤积症患者外周血中维生素D受体的表达与Th1/Th2型细胞因子干扰素-γ/白细胞介素-4(IFN-γ/IL-4)的变化关系,探讨ICP发病机制。方法:选取ICP患者31例(ICP组),孕周相匹配的正常孕妇31例(正常对照组)。采用酶联免疫吸附试验(ELISA法),检测两组孕妇血清中Th1型细胞因子(IFN-γ)和Th2型细胞因子(IL-4)的水平;采用实时荧光定量逆转录-多聚酶链反应(qRT-PCR),检测两组孕妇外周血单个核细胞维生素D受体(VDR)mRNA的表达水平,采用3-磷酸甘油醛脱氢酶(GAPDH)为内参,根据相对定量公式:2-△△CT分析VDR mRNA的表达水平。结果:(1)ICP组外周血清中IFN-γ的浓度[(230.93±36.04)pg/ml]明显高于正常对照组[(138.37±25.08)pg/ml],差异有统计学意义(P<0.01)。ICP组血清中IL-4浓度[(9.99±3.19)pg/ml]和正常对照组[(8.58±2.43)pg/ml]比较,差异无统计学意义(P>0.05)。ICP组IFN-γ/IL-4比值(24.56±6.91)高于正常对照组(17.13±4.84),差异有统计学意义(P<0.05)。(2)ICP组外周血单个核细胞维生素D受体mRNA的表达明显低于正常对照组(P<0.01),正常对照组VDR的表达定义为1.0,ICP组的表达量为0.4。(3)ICP组外周血中VDR的表达水平与IFN-γ浓度呈明显负相关(r=-0.833,P<0.01),与IL-4浓度无明显相关(r=-0.109,P>0.05),与IFN-γ/IL-4比值呈负相关,但相关性不强(r=-0.356,P=0.049<0.05)。结论:ICP患者外周血Th1/Th2型细胞因子平衡由Th2向Th1偏移,可能与ICP孕妇外周血单个核细胞VDR的表达减少有关。     

Th17及Th1相关细胞因子与支气管哮喘的关系研究

目的:分析外周血Th17、Th1及相关细胞因子表达水平和支气管哮喘(bronchial asthma,BA)发生、发展的相关性研究。方法:回顾选取我院收治的BA病例57份,称作BA组,另选取呼吸系统正常的病例55例为对照组,检测两组入选者的外周血IL-2、TNF-α、Th17、IL-6、Th1指标表达差异,并进行多因素回归分析。结果:BA组Th17(0.62±1.67)%、Th1(1.45±0.48)%及Th1/Th17(2.33±1.28)均显著低于对照组(P均0.05);BA组TNF-α(27.46±8.12)pg/mL、IL-6(11.69±2.14)pg/mL表达量显著高于对照组,IL-2(2.58±3.89)pg/mL、IFN-γ(3.74±6.15)pg/mL含量均显著低于对照组(P均0.05);经Logistic回归分析,TNF-α、IL-6、IFN-γ、Th1/Th17、IL-2均和BA有密切相关性(P均0.05)。结论:IL-2、IFN-γ、Th1/Th17、TNF-α、IL-6表达水平均与BA有密切关联,可能是参与BA发病的主要原因,及早进行Th17、Th1及相关细胞因子检查有助于明确病情。     

异源反应性NK细胞对骨髓移植小鼠脾脏Th1/Th2细胞亚群的影响

目的:观察异源反应性自然杀伤细胞(NK细胞)对骨髓移植小鼠脾脏辅助性T细胞Th1/Th2亚群的影响。方法:以BALB/c和CB6F1小鼠为受体,在γ射线照射后通过尾静脉输入C57BL/6J小鼠的骨髓细胞和脾脏单个核细胞,建立移植物抗宿主病模型;然后输入供体的NK细胞,检测受体小鼠脾脏中Th1/Th2淋巴细胞亚群和外周血中γ干扰素(IFN-γ)、白细胞介素10(IL-10)的变化。结果:与单纯输入骨髓细胞和单个核细胞组小鼠相比,输入异源反应性NK细胞后,BALB/c和CB6F1小鼠脾脏中Th1细胞比例均下降,Th2细胞比例均上升,CB6F1小鼠脾脏中Th2虽有升高但没有统计学意义;外周血中IL-10水平显著升高,IFN-γ的水平显著下降。结论:异源反应性NK细胞可能通过降低脾脏中Th1细胞亚群比例和升高Th2细胞亚群比例减轻移植物抗宿主病。     

变应性鼻炎患者外周血中Tr1/Th3细胞的检测和分析

目的:检测变应性鼻炎(Allergic rhinitis,AR)患者和健康对照者外周血中IL-10+CD4+T细胞、TGF-β+CD4+T细胞(分别代表Tr1细胞和Th3细胞的特性)的比例,并探讨其在AR发病中的意义,为AR的治疗提供临床参考。方法:分离19例对粉尘螨过敏的AR患者和19例健康对照者外周血单个核细胞(PBMCs),采用流式细胞术分别检测外周血中IL-10+CD4+T细胞、TGF-β+CD4+T细胞的比例。结果:同健康对照者相比,AR患者外周血中IL-10+CD4+T细胞的比例显著降低[(1.66±0.48)%vs.(3.80.92)%,t=-9.08,P0.01)],AR患者外周血中TGF-β+CD4+T细胞的比例降低[(1.92±0.54)%vs.(4.76±1.12)%,t=-9.94,P0.01)]。结论:外周血中IL-10+CD4+T(Tr1)细胞比例的降低可能是AR发病的一个重要因素,提高AR患者外周血中分泌IL-10的Tr1细胞的比例可能在AR的治疗中具有重要意义。外周血中TGF-β1+CD4+T(Th3)细胞的比例显著降低,可能是AR发病的一个重要因素。但TGF-β1与AR关系的研究较少,特别是外周血中TGF-β1水平与AR的关系研究较少,需进一步研究。     

原发性肝癌患者Th1/Th2 亚群研究*

目的:探讨原发性肝癌患者血清中淋巴细胞亚群中Th1/Th2的变化,为分析肝癌的发生发展状症和临床治疗提供免疫学指标。方法:应用放射免疫分析及酶联免疫分析法(ELISA),测定46例肝癌患者,及43例正常对照组进行比较。以IL-2、INF-γ和TNF-α水平代表Th1型细胞因子,以IL-4,IL-6、IL-8、IL-10的水平代表Th2型细胞因子。结果:肝癌患者IL-2、TNF-γ、IL-6的水平明显低正常对照组,P<0.01。IL-4、IL-8、IL-10、TNF-α的水平明显高于正常对照组,P<0.01。结论:肝癌患者体内存在Th1/Th2细胞因子失衡,其中Th1亚群功能抑制,Th2亚群功能亢进,其与肿瘤在宿主体内生长密切相关。通过纠正这些免疫失调将成为肝癌治疗的重要手段。     

21例侵袭性肺曲霉病患者Th以及Treg细胞的检测

目的：分析侵袭性肺曲霉病患者辅助性T细胞（Th）以及调节性T细胞（Treg）在外周血中单个核细胞中的表达情况及其临床相关性,探讨Th和Treg细胞介导的免疫反应在侵袭性肺曲霉病中的作用。方法分离21例侵袭性肺曲霉病患者及19例健康人外周血的单个核细胞,采用流式细胞术分析Th1、Th2、Th17、Treg细胞群的表达情况,Real-timePCR方法检测相关转录因子T-bet、GATA-3、RORγt以及Foxp3的表达,ELISA法检测血清中相关细胞因子IFN-γ、IL-4、IL-17以及TGF-β的表达。结果与健康人对照组相比,侵袭性肺曲霉病患者Th1细胞以及Treg细胞占CD4＋T细胞的比例较之对照组明显降低;Th1、Th17、Treg细胞相关转录因子T-bet、RORγt、Foxp3以及相关细胞因子IFN-γ、IL-17A、TGF-β与对照组相比表达明显降低。结论IPA患者的Th1、Th17以及Treg细胞所介导的免疫反应受抑制。     

Proinflammatory cytokines in bovine colostrum potentiate the mitogenic response of peripheral blood mononuclear cells from newborn calves through IL-2 and CD25 expression

Abstract: Bovine colostrum contains high concentrations of cytokines, and colostral cytokines are considered to be an important factor in stimulation of maturation of the immune system in newborns. In this study, 5 proinflammatory cytokines (IL-1beta, IL-6, TNF-alpha, IFN-gamma and IL-1 receptor antagonist, IL-1ra) present in colostrum were tested for their potential to enhance mitogenic response and to elicit expression of IL-2 mRNA and CD25 in peripheral blood mononuclear cells (PBMC) from newborn calves before being fed colostrum. PBMC were pretreated with each recombinant bovine cytokine for 2 hr before stimulation with concanavalin A (ConA). Pretreatment of PBMC from newborn calves with IL-1beta, TNF-alpha or IFN-gamma significantly enhanced the ConA response, whereas IL-1ra inhibited the response. The degree of enhancement or inhibition of mitogenic response by these cytokines was more pronounced in PBMC from newborn calves than in those from adult cows. Although IL-2 mRNA expression in ConA-stimulated PBMC from newborn calves was weaker than that in those from adult cows of ConA-stimulated controls, the expression levels became comparable after pretreatment with IL-1beta, TNF-alpha or IFN-gamma. The CD25 expression in PBMC from newborn calves was also enhanced by pretreatment with IL-1beta, TNF-beta and IFN-gamma. These results suggest that pretreatment of neonatal PBMC with IL-1beta, TNF-alpha or IFN-gamma promotes mitogenic response to ConA through up-regulating the production of IL-2 and the expression of the mature IL-2 receptor.     

Kinetics of cytokines and chemokines gene expression distinguishes Paracoccidioides brasiliensis infection from disease

The human infection with Paracoccidioides brasiliensis may result in three major outcomes: the paracoccidioidomycosis-infection (PI), the adult form (AF) and the juvenile form (JF) of the disease. The aim of this study was to compare the immunological response among these groups. The gene expression of multiple cytokines, including IL-4, IL-5, IL-6, IL-10, IFN-gamma, TNF-alpha and TGF-beta1, and chemokines, CXCL8, CXCL9 and CXCL10 was evaluated by RT-PCR in peripheral blood mononuclear cells unstimulated or following phytohemagglutinin stimulation for 3, 6, 12, 24 and 48 h. PI individuals expressed earlier and higher levels of mRNA of IFN-gamma, TNF-alpha, CXCL9 and CXCL10 when compared to JF patients. In relation to AF patients, the PI group presented similar levels of CXCL10 and IFN-gamma and higher levels of CXCL9. On the other hand, mRNA expression of Th2 cytokines (IL-4, IL-10, IL-5 and TGF-beta1) was higher and earlier in JF and AF groups, when compared to PI individuals. At some time intervals it was possible to differentiate JF from AF, mainly in relation to IL-4 and TGF-beta1 mRNA, expressed in higher levels in the JF patients. The distinct patterns of cytokines and chemokines expression support their important role in determining the different outcomes observed in this disease.     

宫颈癌组织中Th1/Th2类细胞因子的漂移研究

目的:观察宫颈癌组织中Th1/Th2类细胞因子的漂移情况。方法:选以IL-2和IFN-γ代表Th1类细胞因子,IL-4和IL-6代表Th2类细胞因子,通过逆转录聚合酶链反应(RT-PCR)检测25例宫颈癌癌组织中Th1/Th2类细胞因子mRNA的表达。结果:IIIB期宫颈癌组织中,Th1型细胞因子的表达显著低于IB期、IIA期、IIB期,Th2型细胞因子的表达显著高于IB期、IIA期、IIB期,差异均有统计学意义(P0.05)。Ⅰ期和Ⅱ期宫颈癌以Th1型细胞因子表达为主。25例宫颈癌组织中,13例呈典型的Th1类细胞因子的强势表达,7例为Th2型,5例为Th0型,随着宫颈癌分期的增高,由Th1向Th2漂移(P0.05)。结论:IB期、IIA期、IIB期宫颈癌患者组织中细胞因子呈Th1状态,IIIA期呈Th0状态,IIIB期呈Th2状态,随着宫颈癌分期的增高,由Th1向Th2漂移。     

Constitutive expression of CIITA directs CD4 T cells to produce Th2 cytokines in the thymus

We generated mice expressing a human type III CIITA transgene (CIITA Tg) under control of the CD4 promoter to study the role of CIITA in CD4 T cell biology. The transgene is expressed in peripheral CD4 and CD8 T cells, as well as in thymocytes. When CD4 T cells were differentiated towards the Th2 lineage, both control and CIITA Tg Th2 cells expressed similar levels of Th2 cytokines. Th1 cells from control and CIITA Tg mice cells produced comparable levels of IFN-gamma. CIITA Tg Th1 cells also expressed IL-4, IL-5, and IL-13 in the absence of Stat6. There was an approximate 10-fold increase in the number of peripheral na?ve CD4 T cells and NK1.1- thymocytes producing IL-4 from CIITA Tg mice compared to control mice. Finally, Th1 cells from irradiated control mice reconstituted with CIITA Tg bone marrow displayed the same cytokine production profiles as Th1 cells from CIITA Tg mice. Together, our data demonstrate that CIITA expression pre-disposes CD4 T cells to produce Th2 type cytokines. Moreover, phenotypic similarities between Th1 cells expressing the CIITA transgene and CIITA deficient Th1 cells suggest that the role of CIITA in cytokine regulation is complex and may reflect both direct and indirect mechanisms of T cell development and differentiation.