恶性淋巴瘤患者TH 1/TH 2细胞因子表达水平的研究

目的 探讨恶性淋巴瘤患者血清中TH1/TH2细胞因子变化及其临床意义,为肿瘤的免疫治疗提供实验依据.方法 用流式细胞小球微阵列术(cytometric bead array,CBA)检测92例恶性淋巴瘤患者及70例健康人群血清中γ干扰素(IFN-γ)、肿瘤坏死因子-α仪(TNF-α)、白细胞介素(IL-2、IL-4、IL-5、IL-10)表达水平.结果 92例恶性淋巴瘤患者血清中TH1型细胞因子的水平分别为:IFN-γ(34.26±33.4g)pg/ml、TNF-α(8.17±10.09)pg/ml、IL-2(3.74 4±1.72)pg/ml;TH2型细胞因子的水平分别为:IL-10(6.28±8.56)pg/ml、IL-5(3.53±3.20)pg/ml、IL-4(6.22±7.13)pg/ml.除TNF-α表达水平降低外,其余5项均明显高于健康体检组,差异有统计学意义(P＜0.01).TH1细胞因子IL-2与TH2细胞因子IL-4的比值明显下降(0.78±O.44),与健康体检组(1.09±0.45)比较差异有统计学意义(P＜0.01).IL-10与疾病的进展相关,Ⅲ/Ⅳ期恶性淋巴瘤患者的表达水平为(9.58±13.96)pg/ml,Ⅰ/Ⅱ期的表达水平为(4.77±3.50)pg/ml,二者比较差异有统计学意义(P＜0.01).IFN-γ在大于60岁的恶性淋巴瘤患者中表达水平明显降低,与其他年龄段恶性淋巴瘤患者比较差异有统计学意义(P ＜0.05).结论 恶性淋巴瘤患者血清中TH1/TH2细胞因子平衡失调,检测TH1/TH2细胞因子可作为评价淋巴瘤临床进展及预后指标.TH1/TH2平衡向TH2方向漂移,这可能是肿瘤细胞发生免疫逃逸,从而导致肿瘤的发生或者转移的原因之一.     

IL—12对慢性乙型肝炎患者TH2／TH2分化的影响

目的 观察IL-12对慢性乙肝患者TH1/TH2类细胞分化的影响。方法 分离50例慢性乙型肝炎患者PBMC,分别与植物血凝素（PHA,100ug/mL）、HBcAg(1ug/mL）、HBeAg(1ug/mL）单独或联合IL-12（10ng/mL）体外培养48h,ELISA法检测培养上清液 IL-2、IFN-γ、IL-4、IL-10水平。20例健康人群做对照。结果 IL-12对健康人群PBMC产生TH1/TH2类细胞因子无显著影响,但对慢性乙型肝炎患者则显著增强PBMC产生IFN-γ,且慢性中度患者最为突出。IL-12与HBeAg联合诱导,不但显著增强慢性乙型肝炎患者PBMC产生IL-12和IFN-γ,还抑制IL-4和IL-10的产生。结论 IL-12可增强慢性乙型肝炎患者IFN-γ优势表达,可促进HBeAg诱导的TH2型优势表达向TH1型优势表达转换。     

黄芪在树突状细胞水平对哮喘TH 1/TH 2平衡的调节作用

目的 探讨黄芪在树突状细胞(DC)水平对过敏性哮喘TH/TH2平衡的调节作用.方法 用rhGM-CSF和rhIL-4诱导培养外周血来源的DC并予鉴定,ELISA法检测其分泌的细胞因子IL-12、IL-10以及与自身T细胞反应后,RT-PCR检测T-bet和GATA-3 mRNA含量,流式细胞术检测T细胞分泌的胞内细胞因子IL-4和IFN-γ水平.结果 哮喘患儿外周血DC分泌IL-10高于对照组(P＜0.05);黄芪干预后DC分泌IL-10降低,与哮喘组比较差异有统计学意义(P＜0.05).哮喘患儿外周血DC分泌IL-12低于对照组(P＜0.05);黄芪干预后DC分泌IL-12增加,但与哮喘组比较差异无统计学意义.混合培养第7天哮喘组T细胞内IL-4水平显著高于正常对照组(P＜0.01);而IFN-γ水平则显著低于正常对照组(P＜0.05);哮喘组IL-4/IFN-γ比值高于正常对照组(P＜0.01).黄芪干预后T细胞内IL-4水平与哮喘组比较差异无统计学意义,而IFN-γ水平增加,与哮喘组比较差异有统计学意义(P＜0.05),IL-4/IFN-γ比值降低,与哮喘组比较差异有统计学意义(P＜0.01).哮喘组T-bet mRNA的表达强度明显低于正常对照组(P＜0.01);而哮喘组GATA-3 mRNA的表达强度则明显高于正常对照组(P＜0.05);哮喘组GATA-3/T-bet比值高于正常对照组(P＜0.05).黄芪干预后T细胞GATA-3 mRNA的表达强度与哮喘组比较差异无统计学意义,而T-bet mRNA水平增加,与哮喘组比较差异有统计学意义(P＜0.05),GATA-3/T-bet比值降低,与哮喘组比较差异有统计学意义(P＜0.01).结论 哮喘患儿DC功能缺陷,产生IL-12减少、IL-10增加导致TH2优势分化,从而使TH1/TH2平衡向TH2倾斜,合成IFN-γ减少,进而造成气道慢性炎症、气道高反应性而致哮喘发作.黄芪对DC的调节主要通过降低IL-10的分泌水平,从而降低其抑制TH0细胞向TH 1分化的功能,即间接抑制了TH0细胞向TH2的分化.     

抗CD86单克隆抗体对哮喘小鼠TH1/TH2细胞因子的影响

目的观察抗CD86单克隆抗体对卵蛋白干粉(OVA)致敏并刺激小鼠TH1和TH2细胞因子比例的变化,为防治哮喘提供实验依据.方法经OVA致敏的雌性Balb/c于激发前腹腔注射抗CD86单克隆抗体及同型对照IgG2α 50 μg,末次激发48 h后收集支气管肺泡灌洗液(BALF),以酶联免疫吸附法(ELISA)测定BALF中白细胞介素-4(IL-4)、白细胞介素-5(IL-5)和干扰素-γ(INF-γ)水平.结果以OVA致敏并激发的哮喘对照组小鼠BALF中IL-4、IL-5水平升高[分别为(64.23±3.91)pg/mL、(379.84±73.02)pg/mL],与PBS对照组[分别为(22±2)pg/mL、(245.75±50.01)pg/mL]比较,差异有显著性(P＜0.05);OVA致敏并激发的哮喘对照组小鼠BALF中INF-γ水平降低[(98.73±16.41)pg/mL],与PBS对照组[(218.35±48.63)pg/mL]比较,差异有显著性(P＜0.05).抗CD86单克隆抗体组小鼠BALF中IL-4、IL-5水平降低[分别为(35.78±4.88)pg/mL、(222.98±58.68)pg/mL],而INF-γ水平升高[(206.92±47.8)pg/mL],与哮喘对照组及同型对照IgG2α组(98.73±16.41)pg/mL、(102.32±15.49)pg/mL比较差异有显著性(P＜0.05).结论抗CD86单克隆抗体通过阻断共刺激信号,纠正TH1/TH2失衡,从而达到抗气道炎症的作用.     

慢性丙型肝炎患者抗病毒治疗前PBMC细胞因子的研究

目的 研究准备抗病毒治疗的慢性丙型肝炎患者的免疫特点.方法 将30例慢性丙型肝炎患者和10例正常对照外周血单个核细胞(PBMC)体外培养72 h后,用ELISA法检测培养上清中细胞因子IL-2、IL-4、IL-10、IL-12、IFN-γ和TNF-α的浓度.结果 (1)慢性丙型肝炎患者PBMC培养上清中IFN-γ、IL-10和TNF-α的水平明显升高(P＜0.05),没有检测到IL-2、IL-4、IL-12的基础分泌.(2)不同病情患者间的细胞因子的分泌水平差异无统计学意义(P＞0.05).结论 慢性丙型肝炎患者体内TH2型细胞因子的分泌占优势.提示通过调整TH1/TH2失衡可能达到抗病毒治疗目的.     

白细胞介素4及γ干扰素与哮喘发病关系的研究

选择12名正常儿童和27名过敏性哮喘患儿,对其PBMC在体外培养时加入丝裂原PMA和PHA刺激后产生的IL-4和IFN-γ用BA-ELISA方法进行测定。结果正常人和哮喘患儿PBMC经刺激后,上清液中IL-4的浓度(?)±s分别为86.25±36.97pg/ml和212.70±92.86pg/ml。经统计学处理t=4.53,P<0.01有显著差异,表明过敏性哮喘患儿PBMC体外合成IL-4明显高于正常儿童,上清液中IFN-γ的浓度(?)±s分别为     

BCG30主要分泌蛋白对尘螨变应性患者TH1/TH2平衡调节作用的观察

目的　研究结核杆菌相对分子质量 (Mr)为 30× 10 3的主要分泌蛋白 (BCG30 ,也称Ag85B)对尘螨变应原过敏所致的哮喘等变应性患者外周血单个核细胞 (PBMC)分泌TH1、TH2细胞因子的调控作用。方法　常规分离 2 7例尘螨过敏的变应性患者 (A) ,其中 18例哮喘患者 (A1)、9例变应性鼻炎或皮炎患者 (A2 )和 13例正常人 (B)的PBMC ,在离体与螨变应原、BCG30单独或复合共培养 ,ELISA法测定培养上清IL 5、IFN γ的水平。结果　A、A1、A2、B 4组无刺激状态下的IL 5、IFN γ水平无差异 (P >0 .0 5 )。尘螨变应原刺激可显著促进A、A1、A2 3组PBMC分泌IL 5 ,P均 <0 .0 5 ,同时 ,A组的IFN γ水平自身比较亦明显增高 (73.5 8pg ml± 30 .2 3pg mlvs 30 .71pg ml± 18.87pg ml,P <0 .0 5 )。将尘螨变应原与BCG30和PBMC进行复合培养 ,并与单独尘螨变应原刺激组比较 ,其IFN γ水平在A组 (92 .89pg ml± 2 9.0 7pg mlvs 73.5 8pg ml± 30 .2 3pg ml)和A1组 (87.6 0pg ml± 36 .4 5pg mlvs 5 8.75pg ml± 7.84pg ml)均较后者为高 (P均 <0 .0 5 ) ,且A组的IFN γ IL 5的比值较单独尘螨变应原刺激组显著增高 (5 .0 3± 1.36vs 4 .2 0± 1.6 4 ,P <0 .0 5 ) ,而A1组其比值亦有增高趋势 (P =0 .0 7)。正常人中各刺激组间所有指标均     

PI3K信号途径在哮喘患者外周血单个核细胞中的表达及意义

目的初步了解磷脂酰肌醇3激酶(PI3K)在急性发作期哮喘患者外周血单个核细胞(PBMC)中的活化情况,并观察其特异性抑制剂(Wortmannin)对Th1、Th2型细胞因子IFN-γ和IL-4表达的影响,探讨PI3K信号途径在哮喘T细胞免疫紊乱中可能的作用机制.方法以20例急性发作期哮喘患者作为实验组,15例健康人为对照组.PBMC提取采用Ficoll密度梯度离心法,半定量RT-PCR法测定PBMC中PI3KmRNA的表达,ELISA法测定不同浓度组Wortmannin处理的PBMC培养上清液中IFN-γ、IL-4水平.结果与对照组相比,急性发作期哮喘患者PI3KmRNA表达增高(P＜0.05).哮喘患者PBMC培养上清液IFN-γ水平低下、IL-4水平升高,与对照组比较差异均有统计学意义(P＜0.05和P＜0.01);同时加入不同浓度Wortmannin共培养后,实验发现均浓度依赖性地抑制了哮喘组及对照组IL-4的产生(P＜0.05或P＜0.01),而同组不同浓度梯度组间比较IFN-γ产生升高,但差异无统计学意义.结论哮喘患者急性发作时可能存在PI3K信号途径的过度活化,其过度活化对Th1型细胞因子IFN-γ影响不明显,有可能主要通过介导Th2型细胞因子IL-4的产生而参与Th1/Th2的失衡.     

TH1/TH2细胞因子与习惯性流产的关系研究

目的通过测定习惯性流产患者（RSA）及正常妊娠妇女外周血血清TH1/TH2细胞因子的含量,探讨TH1/TH2细胞因子与习惯性流产发生的关系。方法采用酶联免疫吸附法检测30例RSA患者,72例正常妊娠妇女血清IL-2、IL-4、IL-10、IFN-γ、TNF-α及TNF-β水平并比较两组之间的差异。结果RSA患者IL-2、IL-4、IL-10、IFN-γ、TNF-α血清水平显著高于正常妊娠妇女（P〈0.05）,TNF-β血清水平显著低于正常妊娠妇女（P〈0.05）。结论血清IL-2、IL-4、IL-10、IFN-γ、TNF-α水平可能在习惯性流产发生中起重要作用。     

体外阻断CD134对LN患者外周血单个核细胞TH1/TH2细胞因子表达的影响

目的 探讨CD134／CD134L共刺激分子对TH1，TH2细胞因子表达的影响及在狼疮性肾炎（LN）发病机制中的可能作用。方法活动期LN患者10例，分别采取外周静脉血，用密度梯度离心法制备新鲜PBMC，分成6组：（1）对照培养组；（2）单纯刺激组；（3）抗CD134组；（4）抗Ⅱ，4组；（5）rhCD134：Fc组；（6）地塞米松（Dex）组。另选取10例健康体检者作为健康对照组。采用ELISA法分别测定培养液上清中IFN-γ、IL-4、IL-10表达水平。结果（1）在抗CD3ε单抗／rIL-2刺激前，活动期LN患者PBMC培养液上清分泌的IFN-γ、IL-4和IL-10水平都明显高于健康对照组；（2）在抗CD3ε单抗／rIL-2刺激下，活动期LN患者PBMC分泌的IFN-γ，4和IL-10又较刺激前明显增加；（3）抗IL-4单抗、抗CDl34单抗均可使经抗CD3ε单抗／rIL-2刺激的LN患者PBMC分泌IL-4、IL-10水平显著下降，导致IFN-γ水平显著增高，免疫应答朝TH1方向偏离；（4）Dex能显著抑制抗CD3ε单抗／rIL-2刺激的原代培养PBMC中IL-4、IFN-γ的分泌水平，但对IL-10的产生并没有明显的抑制或促进作用；（5）rhCD134：Fc能显著降低抗CD3ε单抗／rIL-2刺激的原代培养PBMC中IFN-γ及IL-4、IL-10的分泌水平，对TH1和TH2细胞因子的产生都有显著抑制作用。结论糖皮质激素的抗炎机制具有多重性，对TH1、TH2细胞因子的作用并不均衡；抗CD134单抗对减轻LN患者PBMC的异常活化有一定作用；CD134-IgG融合蛋白与抗CD134单抗相比，抑制作用更明显，对急性期LN有更好的治疗作用。     

Biology of human TH1 and TH2 cells

Evidence is accumulating to suggest the existence of polarized human T-cell responses, reminiscent of T_H1 and T_H2 subsets described for mouse T cells. Human T_H1 cells preferentially develop during infections by intracellular bacteria and trigger phagocyte-mediated host defense, whereas T_H2 cells, which predominate during helminthic infestations and in response to common environmental allergens, are responsible for phagocyte-independent host response. Human T_H1 and T_H2 cells exhibit not only different functional properties but probably also distinct surface markers; T_H2, but not T_H1, clones express membrane CD30 and release the soluble form of CD30, a member of the TNF receptor superfamily. The cytokine profile of natural immunity evoked by different offending agents in the context of different host genetic backgrounds appears to be the most critical factor in determining the phenotype of the subsequent specific response. IL-12 and IFN- and produced by macrophages and NK cells favor the development of T_H1 cells, whereas the early production of IL-4 by a stillunidentified cell type favors the development of T_H2 cells. Clearly, polarized human T_H1 and T_H2 responses not only play different roles in protection, they can also promote different immunopathological reactions. Strong and persistent T_H1 responses seen to be involved in organ-specific autoimmunity, contact dermatitis, and some chronic inflammatory disorders of unknown etiology. In contrast, polarized T_H2 responses favor a reduced protection against the majority of infectious agents (including HIV) and, in genetically predisposed hosts, are responsible for triggering of allergic atopic disorders.     

Genetic polymorphism and TH1/TH2 orientation

BACKGROUND: It is likely that besides developmental and environmental factors, genetic factors also play an important role in Th1/Th2 orientation and susceptibility to related disorders. Thus, for each genetic factor involved one would expect an opposite pattern of susceptibility towards Th1- and Th2-associated diseases. METHODS: We report a comparative analysis of the pattern of association of four genetic polymorphisms with bronchial asthma (Th2 disease) and Crohn's disease (CD; Th1 disease). The study population included 291 Roman children with bronchial asthma and 72 adult Romans with CD, and haptoglobin, adenosine deaminase (ADA), acid phosphatase locus 1 (ACP1) and MN phenotypes were determined. RESULTS: Compared with controls from the same population, the pattern of phenotype association observed in bronchial asthma is exactly opposite to that observed in CD. The analysis of pairwise gametic type distribution for ACP1, ADA and MN polymorphisms has shown that the pattern of differences between bronchial asthma and controls is opposite to that observed between CD and controls. CONCLUSIONS: The pattern of differences between bronchial asthma versus CD is compatible with the hypothesis that some of the genetic systems considered contribute to Th1/Th2 orientation.     

第9、10、11肋间神经和肋下神经的血供

本文观察了25具成人专供研究用的尸体,对50侧第9、10、11肋间神经和肋下神经的血供进行了研究,对该四对神经的营养动脉数目、外径、长度、入神经干部位和来源动脉进行观察测量。发现营养动脉外径较细,长度亦较短,不宜作为肋间神经的血管蒂进行吻合,而肋间神经营养动脉的来源动脉——肋间后动脉,其外径较粗,并有足够长度,作为血管蒂进行吻合较为理想。     

哮喘患儿激素治疗后TH1/TH2的变化

目的观察哮喘患儿血清中白细胞介素IL12、IL13和IgE水平的变化及糖皮质激素对其的影响。方法采用ELISA法分别检测哮喘患儿急性期及口服强的松5～7天后、健康对照组血清中IL12、IL13和IgE的水平。结果哮喘组患儿IL12水平急性发作期较治疗后和健康对照组低,差异均有显著性（P〈0.01）。IL13水平急性发作期较治疗后和健康对照组高,差异均有显著性（P〈0.01）,IgE水平急性发作期较治疗后和健康对照组高,差异均有显著性（P〈0.01）。直线相关分析表明,血清中IL12与IL13负相关,与IgE呈负相关,IL13与IgE呈正相关。结论哮喘患者血清中IL13、IgE水平升高,IL12水平下降,3者的变化相关,糖皮质激素可使IL13水平下降,IL12水平升高。     

Coevolution of TH1, TH2, and TH17 responses during repeated pulmonary exposure to Aspergillus fumigatus conidia

Aspergillus fumigatus, a ubiquitous airborne fungus, can cause invasive infection in immunocompromised individuals but also triggers allergic bronchopulmonary aspergillosis in a subset of otherwise healthy individuals repeatedly exposed to the organism. This study addresses a critical gap in our understanding of the immunoregulation in response to repeated exposure to A. fumigatus conidia. C57BL/6 mice were challenged intranasally with A. fumigatus conidia weekly, and leukocyte composition, activation, and cytokine production were examined after two, four, and eight challenges. Approximately 99% of A. fumigatus conidia were cleared within 24 h after inoculation, and repeated exposure to A. fumigatus conidia did not result in hyphal growth or accumulation of conidia with time. After 2 challenges, there was an early influx of neutrophils and regulatory T (T(reg)) cells into the lungs but minimal inflammation. Repeated exposure promoted sustained expansion of the draining lymph nodes, while the influx of eosinophils and other myeloid cells into the lungs peaked after four exposures and then decreased despite continued A. fumigatus challenges. Goblet cell metaplasia and low-level fibrosis were evident during the response. Repeated exposure to A. fumigatus conidia induced T cell activation in the lungs and the codevelopment by four exposures of T(H)1, T(H)2, and T(H)17 responses in the lungs, which were maintained through eight exposures. Changes in CD4 T cell polarization or T(reg) numbers did not account for the reduction in myeloid cell numbers later in the response, suggesting a non-T-cell regulatory pathway involved in dampening inflammation during repeated exposure to A. fumigatus conidia.