外周血T细胞亚群及细胞因子对外源性哮喘IgE生成…

对３０例哮喘患者及３０例健康成年人的外周血，采用单克隆抗体（ＭｃＡｂ）间接免疫荧光法测定Ｔ细胞亚群；ＥＬＩＳＡ双抗体夹心法测定ＩｇＥ、ＩＬ－４；ＦＩ２细胞株，生物学方法测定ＩＬ－２；ＩＬ－６依赖细胞株７ＴＤ１，掺入法测定ＩＬ－６；用抗人ＣＤ２３的ＭｃＡｂ测定ＣＤ２３。为研究Ｔ细胞、细胞因子对哮喘ＩｇＥ生成调节机理及细胞因子在哮喘发病过程中的作用。结果显示：发作期ＩｇＥ、ＩＬ－２、ＩＬ－４、ＣＤ２３     

病毒性肝炎患者血清IL—6和T细胞亚中的关系

采用ＭＴＴ比色法和抗体致敏的红细胞花环试验（间接法）检测了９１例病毒性肝炎患者血清ＩＬ－６和Ｔ细胞亚群。结果显示，病毒性肝炎患者血清ＩＬ－６活性均明显高于正常对照组（Ｐ〈０．０１）；以重型肝炎为最高；且与肝细胞损害程度呈正比（Ｐ〈０．０１）；ＣＤ４＾＋细胞、ＣＤ４＾＋／ＣＤ８＾＋均明显降低，与血清ＩＬ－６呈负相关；ＣＤ８＾＋细胞明显增高，与血清ＩＬ－６呈正相关（Ｐ〈０．０１）。表明，血清ＩＬ－６与     

慢性肾炎患者白细胞介素－6及T细胞亚群变化的探讨

本文报告慢性肾炎患者白细胞介素－６（ＩＬ－６）的水平（７例）及Ｔ细胞亚群的变化（１９例）．Ｔ细胞亚群分析用ＣＤ３、ＣＤ４、ＣＤ８、Ｔγδ等单克隆抗体进行间接免疫荧光法测定；ＩＬ－６用ＩＬ－６依赖细胞Ｂ９细胞增殖的生物学测定法。７例患者ＩＬ－６的活性均显著增高（Ｐ＜０．０１）。１９例患者的Ｔ细胞亚群有异常改变：ＣＤ_４~＋细胞明显下降，ＣＤ_８~＋细胞在多数病例内增加，但可因慢性肾炎的类型而不同，ＣＤ_４~＋／ＣＤ_８~＋比值降低。１７例患者Ｔγδ细胞显著高于正常对照组。根据上述变化，本文对慢性肾炎发病的有关机理作了初步探讨。     

细胞因子与哮喘发病机理的关系

研究分设３组，哮喘发作组、哮喘缓解组和正常对照组各２０例，外周血ＩＦＮ－ｒ测定采用微量细胞病变抑制法。ＩＬ－４分泌细胞阳性率测定采用ＡＰＡＡＰ法。Ｃｏｎ－Ａ诱导Ｔｓ细胞活性测定采用Ｓｍｉｔｈ改良法。血清ＩｇＥ测定采用ＥＬＩＳＡ法。试验结果如下：（１）哮喘发作组外周血Ｔ淋巴细胞检测ＩＬ－４分泌细胞阳性率（７．６±３．５％）明显高于哮喘缓解组（３．８±２．０％），Ｐ＜０．０１，更高于正常对照组（１．８±０．５％），Ｐ＜０．０００１。在体外用ＰＨＡ诱导检测产生ＩＰＮ－Ｖ的能力，发现哮喘发作组（７３２．…     

病毒性肝炎患者血清IL-6和T细胞亚群的关系

采用ＭＴＴ比色法和抗体致敏的红细胞花环试验（间接法）　检测了９１例病毒性肝炎患者血清ＩＬ－６和Ｔ细胞亚群。　结果显示，病毒性肝炎患者血清ＩＬ－６活性均明显高于正常对照组（Ｐ＜０．０１），以重型肝炎为最高；且与肝细胞损害程度呈正比（Ｐ＜０．０１）；ＣＤ４~＋细胞、ＣＤ４＋／ＣＤ８~＋均明显降低，与血清ＩＬ－６呈负相关；ＣＯ８~＋细胞明显增高，与血清ＩＬ－６呈正相关（Ｐ＜０．０１）。表明，血清ＩＬ－６与机体细胞免疫功能密切相关，二者互为因果，共同参与病毒性肝炎的免疫病理损害，可作为判断病情和预后及免疫调节治疗的监测指标。     

MTEC 1分泌的趋化因子引起特定亚群胸腺细胞的定向迁移

分析胸腺髓质上皮样细胞系ＭＴＥＣ１分泌的化学趋化因子对胸腺细胞亚群的趋化作用。方法以抗体加补体杀伤结合免疫磁珠及ｐａｎｎｉｎｇ法，将小鼠胸腺细胞分离纯化，获得ＣＤ４＋ＣＤ８＋（ＤＰ），ＣＤ４－ＣＤ８－（ＤＮ），ＣＤ４＋ＣＤ８－（ＣＤ４ＳＰ）及ＣＤ４－ＣＤ８＋（ＣＤ８ＳＰ）四亚群细胞，用Ｂｏｙｄｅｎ小室分析ＭＴＥＣ１┐ＳＮ对四群胸腺细胞的趋化作用。结果ＭＴＥＣ１┐ＳＮ对ＤＰ及ＣＤ４ＳＰ胸腺细胞有趋化活性（ＣＩ＝６．６±１．０及６．１±１．８）；对ＣＤ８ＳＰ细胞有中度趋化活性（ＣＩ＝３．２±１．０）；对ＤＮ趋化活性微弱（ＣＩ＝１．３±０．６）。化学趋化因子ＭＣＰ┐１纯品对ＣＤ４ＳＰ胸腺细胞显示强趋化活性（ＣＩ＝５．６），对ＤＮ胸腺细胞则无可测出趋化活性。结论ＭＴＥＣ１分泌的化学趋化因子对ＤＰ，ＣＤ４ＳＰ及ＣＤ８ＳＰ胸腺细胞有显著趋化作用，对ＤＮ胸腺细胞几乎无趋化作用。提示此类化学趋化因子有趋使胸腺发育中后期阶段的细胞向胸腺髓质区迁移和定位的作用。     

癫痫患者免疫状态的探讨

本文观察了７０例癫痫（ＥＰ）患者的免疫状态，首次检测了Ｂ淋巴细胞亚群，ｓＩＬ－２Ｒ及ＴＮＦ，对各项免疫学指标进行了对比研究，发现ＩｇＡ，ＩｇＭ含量明显降低，ＣＩＣ含量明显增高（Ｐ＜０．０１），Ｃ３明显降低（Ｐ＜０．０５），显示患者体液免疫功能低下；淋巴细胞亚群测定发现ＣＤ３＾＋，ＣＤ４＾＋明显降低，ＣＤ８＾＋明显增高，ＣＤ４＾＋／ＣＤ８＾＋比值明显低于对照组（Ｐ＜０．０１），表明细胞免疫功能低下；     

九代清源口服液增强反复呼吸道感染患儿免疫功能的临床实验研究

目的：探讨九代清源口服液在增强反复呼吸道感染患儿免疫功能中的作用。方法：将１００例ＲＲＩ患儿随机分为２组,即常规治疗组和九代清源口服液辅助治疗组（简称九代组）,分别于治疗前、治疗后抽取静脉血检测Ｔ淋巴细胞亚群（ＳＰ法）、ＮＫ细胞活性（ＭＴＴ法）、ｓＩＬ－２Ｒ（ＥＬＩＳＡ法）、ＩｇＧ、ＩｇＡ、ＩｇＭ（透比浊法）。结果：ＲＢＩ患儿治疗前ＣＤ３＾＋、ＣＤ４＾＋、ＣＤ４＾＋／ＣＤ８＾＋、ＮＫ细胞活性及ＩｇＧ、ＩｇＡ显著降低,ＣＤ８＾＋、ＩｇＭ和ｓＩＬ－６Ｒ显著升高常规治疗指标明显好转,与正常对照组比较,仍有显著差异（Ｐ〈０．０５）;九代组治疗后有所恢复,但与正常对照组比较均无显著性差异（Ｐ〉０．０５）。结论：九代清源口服液具有较强的免疫调节作用,可明显增强ＲＲＩ患儿的细胞和体液免疫功能。     

雷公藤对哮喘豚鼠外周血淋巴细胞IL-5、粒细胞-巨噬细胞集落刺激因子mRNA及CD4+、CD8+表达的影响

目的：探讨哮喘豚鼠白介素－ ５（ＩＬ－ ５） 、粒细胞－ 巨噬细胞集落刺激因子（ ＧＭ － ＣＳＦ） 及ＣＤ４ ＋ 和ＣＤ８ ＋ Ｔ 细胞在哮喘发病中的作用及雷公藤的干预作用。方法：实验分为哮喘组、雷公藤治疗组（ 处理组） 和对照组,每组各１０ 只豚鼠,采用原位杂交方法检测豚鼠外周血淋巴细胞的ＩＬ－ ５ 、ＧＭ－ ＣＳＦ ｍ ＲＮＡ 表达;应用免疫细胞化学方法检测外周血淋巴细胞ＣＤ４ ＋ 和ＣＤ８ ＋ 的表达。结果：哮喘组外周血淋巴细胞ＩＬ－ ５ 、ＧＭ－ ＣＳＦ－ ｍ ＲＮＡ 表达均明显高于处理组和对照组（ Ｐ＜ ０－０１） ,而处理组和对照组无显著差异。哮喘组外周血淋巴细胞ＣＤ４ ＋ 表达明显高于对照组和处理组（ Ｐ＜０－０１） ,ＣＤ８ ＋ 表达低于对照组和处理组（ Ｐ＜ ０－０５） 。结论：哮喘豚鼠外周血淋巴细胞ＩＬ－ ５ 、ＧＭ － ＣＳＦ ｍ ＲＮＡ 表达增高、ＣＤ４ ＋ 表达增高,ＣＤ８ ＋ 表达降低,而雷公藤甲素可降低外周血淋巴细胞ＩＬ－ ５ 、ＧＭ － ＣＳＦ ｍＲＮＡ 表达,并可提高ＣＤ８ ＋ 表达,降低ＣＤ４ ＋ 表达。表明雷公藤可能在抗哮喘气道炎症中发挥一定作用     

外周血T细胞亚群及活化状态改变与过敏性哮喘的关系

利用ＡＰＡＡＰ法了２０例中重症过敏性哮喘发作患者外周血Ｔ细胞来及其活化状态改变，并与２０例缓解期过敏性哮喘患者和２０例健康对照者进行了比较。结果表明：Ｔ细胞亚群的改变在３组间差异无显著性（Ｐ〉０．０５），而反映ＣＤ４Ｔ细胞活化的ＣＤ２５和ＨＬＡ－ＤＲ的表达在中重度发作期嗜同于其他两组（Ｐ均〈０．０１），且分别与血清总ＩｇＥ呈正相关（ｒ＝０．３８５５，０．３５５４，Ｐ均〈０．０５），与第１秒用力呼气     

IL-2等6种细胞因子与儿童支气管哮喘关系的研究

观察急性发作期、激素治疗后缓解期支气管哮喘患儿血浆细胞因子的变化,探讨儿童支气管哮喘与细胞因子的关系。采用免疫分析技术对64例急性发作期支气管哮喘患儿、45例缓解期患儿及20例正常儿童血浆IL-2、sIL-2R、IL-4、IL-5、IL-8、TNF-α、IgE等细胞因子水平进行测定;用逆转录聚合酶键反应技术（RT-PCR）对外周淋巴细胞IL-4、IL-5mRNA表达进行定量分析。结果:（1）发作期哮喘患儿血浆IL-2、sIL-2R、IL-4、IL-5、IL-8、TNF-α和IgE水平均明显高于正常对照组（P值均<0.001）,以IL-4、IL-5和IgE变化最为明显,缓解期均明显下降,但IL-4、IL-5及IgE水平仍高于正常水平（P<0.01）。（2）发作期患儿外周淋巴细胞IL-4、IL-5mRNA表达增强,治疗缓解后减弱,同时血浆IL-4、IL-5分别与IL-4、IL-5mRNA呈明显正相关关系。结论:（1）本研究结果提示细胞因子的释放与哮喘的发作密切相关。（2）外周淋巴细胞IL-4、IL-5强表达以及血浆IL-4、IL-5高水平提示哮喘发作期Th2亚群的激活;同时IL-8和TNF-α的升高表明中性粒细胞和单核巨噬细胞可能参与哮喘的发作。（3）糖皮质激素抑制多种细胞因子的释放,可能是其治疗哮喘的主要机制。     

支气管哮喘患儿内皮素-1及相关炎性细胞因子测定

目的:探讨内皮素及相关炎性细胞因子在小儿哮喘发病机制中的作用。方法:分别用放射免疫分析及酶免法测定了42例哮喘患儿治疗前后内皮素-1、白细胞介素-5、白细胞介素-6及白细胞介素-8水平。结果:发作期组内皮素水平高于缓解组及对照组差异非常显著(P值均<0 01),缓解组较发作组水平下降显著但仍显著高于对照组(P<0 05)。3种细胞因子水平则发作期组均显著高于缓解组及对照组,统计差异极显著(P值均<0 01),缓解组前2种细胞因子仍显著高于对照组(P值均<0 05);而后一种细胞因子已下降至近对照组水平(P>0 05)。ET与IL-5呈显著正相关(发作期r=0 560,P<0 01;缓解期r=0 435,P<0 01)。结论:内皮素与细胞因子以不同方式参与了小儿哮喘的发病机制,其测定对于评价炎症程度及疗效、指导临床治疗有重要意义。     

FcεRⅡ／CD23在支气管哮喘发病中作用的研究

本文以急性发作期病人和缓解期病人及正常人各２０例为研究对象，探讨ＩＬ－４－ＦｃεＲⅡ／ＣＤ２３－ＩｇＥ在支气管哮喘中的调节作用。结果表明，急性发作期病人的ＩＬ－４分泌细胞、ＦｃεＲⅡ／ＣＤ２３阳性淋巴细胞和血清总ＩｇＥ水平均明显高于缓解期病人和正常对照组。在急性发作时，ＩＬ－４和ＣＤ２３之间存在着正相关。进一步表明病人体内ＩＬ－４水平升高可以诱导淋巴细胞表面ＦｃεＲⅡ／ＣＤ２３分子表达的增强，并在ＩｇＥ抗体形成中起着重要作用。     

IL-6、IL-8、TNF-α及IFN-α在儿童哮喘不同病期的变化

检测哮喘发作期（35例）、稳定期（18例）及正常对照组（28例）血清IL-6、IL-8、TNF-α和IFN-α水平。结果表明哮喘发作期IL-6、IL-8、TNF-α水平明显增高,而IFN-α显著降低;至稳定期血清IL-6、IL-8、TNF-α恢复到对照组水平,唯IFN-α仍低于正常儿。血清THN-α与IL-6、IL-8、IFN-α间存在正相关关系。提示哮喘患儿存在多种细胞因子产生和释放异常。细胞因子网络失调可能是哮喘发病的分子生物学基础。     

支气管哮喘患儿治疗前后血清GM-CSF、IL-8和IL-6联检的临床意义

目的：探讨了支气管哮喘患儿血清GM—CSF、IL-8和IL-6水平的变化及意义。方法：应用放射免疫分析对32例支气管哮喘患儿进行了血清GM—CSF、IL-8和IL-6水平测定,并与30例正常健康儿作比较。结果：支气管哮喘患儿血清GM—CSF、IL-8和IL-6水平非常显著地高于正常儿组（P〈0．01）。经治疗3个月后则与正常儿比较无显著性差异（P〉0．05）。结论：血清GM—CSF、IL-8和IL-6水平的异常升高足支气管哮喘患儿发病的病理因素之一．有重要的临床价值。     

急性期川崎病Th17细胞变化初探

目的 探讨Th17细胞在川崎病(Kawasaki disease,KD)免疫发病机制中的作用.方法 急性期KD患儿60例,正常同年龄对照组32例,KD患儿分别于静脉丙种球蛋白(IVIG)治疗前后直接取血备检.采用荧光定量PCR(real-time PCR)检测CD4+T细胞IL-17A/F、转录因子ROR-γt、Foxp3及PBMC IL-6、TGF-β、IL-23p19、IL-27p28、IL-27EBI3、IFN-γ等mRNA表达;酶联免疫吸附试验检测血浆中IL-6、TGF-β、IL-23、IL-27、IFN-γ的蛋白浓度;流式细胞术检测外周血CD4+CD25+调节性T细胞(Tr)的比例.结果 急性期KD患儿CD4+T细胞高表达IL-17A及IL-17F(P<0.01),IVIG治疗后明显降低(P<0.01);急性期KD患儿IL-17A/IL-17F与红细胞沉降率(ESR)、C反应蛋白(CRP)、白细胞数目(WBC)呈正相关(IL-17A:0.70,0.85,0.80,P<0.01;IL-17F:0.63,0.65,0.69,P<0.01);急性期KD患儿Th17细胞转录因子ROR-γt及前炎症细胞因子IL-6转录水平明显高于对照组(P<0.01),IVIG治疗后显著降低(P<0.01);TGF-β、IL-23p19、IL-27p28、IL-27EBI3 mRNA水平及血浆蛋白浓度与对照组比较差异无统计学意义(P>0.05);血浆IFN-γ浓度显著升高,mRNA水平无变化;急性期KD患儿CD4+ CD25+ Tr细胞比例明显低于正常对照组(P<0.01),其转录因子Foxp3表达亦明显降低(P<0.01).结论 急性期KD患儿Th17细胞过度活化可能参与了KD免疫发病机制.     

Purified excretory-secretory component of filarial parasite enhances Fc epsilon RII/CD23 expression on human splenic B and T cells and IgE synthesis while potentiating T-helper type 2-related cytokine generation from T cells.

The CD23-bearing cells are known to be involved in multiple biological activities, including IgE synthesis and IgE-dependent cytotoxicity to parasites. The factors that regulate interleukin-4 (IL-4)-induced IgE synthesis in helminthic infection were analysed by using an excretory-secretory component (ESC) of Dirofilaria immitis (DI). Human splenic B and T cells significantly enhanced the expression of low-affinity Fc receptors for IgE (Fc epsilon RII/CD23) by stimulation with ESC, either acting alone or in synergy with IL-4. On B cells, ESC potentiated the CD23 expression in synergy with IL-4, whereas ESC alone was unable to modulate CD23 expression. In contrast, ESC directly induced CD23 expression on T cells by acting alone and no further enhancement was observed in the presence of IL-4. Furthermore, IL-4-induced IgE synthesis by splenic mononuclear cells (SMNC) was greatly enhanced in the presence of ESC. Of particular interest, T cells primed by ESC significantly produced a set of cytokines including IL-3, IL-4, IL-5 and IL-6. Inasmuch, IL-4-induced IgE synthesis in helminthic infection may be selectively modulated by parasite protein(s) acting on the generation of T-helper type 2 (Th2)-related cytokines.     

Peripheral blood CD4+ and CD8+ T cell type 1 and type 2 cytokine production in atopic asthmatic and normal subjects

BACKGROUND: Increased production of IL-4 and IL-5 and decreased production of IFN-gamma by CD4+ T cells has been implicated in asthma pathogenesis. However, CD8+ T cells also produce type 1 and type 2 cytokines and the relative roles of CD4+ and CD8+ T cell cytokine production in asthma have not been previously studied. OBJECTIVE: To determine the production of the type 1 and type 2 cytokines by CD4+ and CD8+ T cell subsets in asthmatic and normal subjects. METHODS: Intracellular cytokine staining for IL-4, -5, -10, -13 and IFN-gamma was analysed in peripheral blood CD4+ and CD8+ T cells from 24 atopic asthmatic and 20 normal subjects. RESULTS: Both subsets of T cells produced all cytokines studied and there were no significant differences between CD4+ and CD8+ T cells in their capacity to produce either type 1 or type 2 cytokines. There were significantly increased frequencies of IFN-gamma-positive CD4+ (13.1 +/- 2.4%, vs. 7.3 +/- 1.4%) and CD8+ (20.0 +/- 2.9%, vs. 9.6 +/- 2.1%) T cells in asthmatic subjects compared with normal subjects (P < 0.05), but not in frequencies of CD4+ or CD8+ T cells staining positively for IL-4, -5, -10 or -13. CONCLUSION: The frequencies of peripheral blood CD8+ T cells producing type 1 and type 2 cytokines are comparable with the frequencies of CD4+ T cells. There was an increased frequency of IFN-gamma producing CD4+ and CD8+ T cells in asthmatic compared with normal subjects. Further studies investigating T cells derived from the airways and investigating various stages within the disease process are required to further elucidate the importance of type 2 and type 1 T cell cytokine production in the pathogenesis of human allergic disease.     

Contribution of IL-18-induced innate T cell activation to airway inflammation with mucus hypersecretion and airway hyperresponsiveness

Human bronchial asthma is characterized by airway hyperresponsiveness (AHR), eosinophilic airway inflammation, mucus hypersecretion and high serum level of IgE. IL-18 was originally regarded to induce T(h)1-related cytokines from Th1 cells in the presence of IL-12. However, our previous reports clearly demonstrated that IL-18 with IL-2 promotes Th2 cytokines production from T cells and NK cells. Furthermore, IL-18 with IL-3 stimulates basophils and mast cells to produce Th2 cytokines. Thus, we examined the capacity of IL-2 and IL-18 to induce AHR, airway eosinophilic inflammation and goblet cell metaplasia. Intranasal administration of IL-2 and IL-18 induces AHR, mucus hypersecretion and eosinophilic inflammation in the lungs of naive mice. CD4+ T cells are prerequisite for this IL-2 plus IL-18-induced bronchial asthma, because CD4+ T cells-depleted or Rag-2-deficient (Rag-2-/-) mice did not develop bronchial asthma after IL-2 plus IL-18 treatment. Both STAT6-/- mice and IL-13-neutralized wild-type mice failed to develop AHR, goblet cell metaplasia and airway eosinophilic inflammation, while IL-4-/- mice almost normally developed, suggesting that IL-13 is a major causative factor and IL-4 mainly enhances the degree of AHR and eosinophilic inflammation. Both IL-4 and IL-13 equally induce eotaxin in mouse embryonic fibroblasts. However, only IL-13 blockade inhibited asthma symptoms, suggesting that IL-13 but not IL-4 is produced abundantly and plays a critical role in the pathogenesis of bronchial asthma in this model. As airway epithelial cells store robust IL-18, IL-18 might be critically involved in pathogen-induced bronchial asthma, in which pathogens stimulate epithelial cells to produce IL-18 without IL-12 induction.     

Nitric oxide blocks inflammatory cytokine secretion triggered by CD23 in monocytes from allergic, asthmatic patients and healthy controls.

BACKGROUND: In allergic asthma, monocytes/macrophages may be activated to produce inflammatory cytokines through triggering of the low-affinity IgE receptor (CD23). Elevated airway levels of nitric oxide (NO) are associated with asthmatic exacerbations. Our previous work suggested that NO may function in an anti-inflammatory capacity by downregulating endotoxin-stimulated cytokine production by alveolar macrophages and matured monocytes. OBJECTIVE: The purpose of this study was to determine the effect of NO on CD23-triggered cytokine production by monocytes from asthmatic patients and healthy controls. METHODS: Monocytes were obtained from normal volunteers (n = 13) and asthmatic patients with atopy (n = 8). Monocyte cultures were treated with interleukin-4 (IL-4) and granulocyte macrophage colony-stimulating factor (GM-CSF) for 24 hours to upregulate CD23 expression. Cultures were stimulated by anti-CD23 and treated with DETA NONOate [2,2-(hydroxynitrosohydrazonon)-bis-ethanamine] releases NO in culture with t(1/2) of 20 hours at 37 degrees C for 24 hours. Cell free culture supernatants were collected and assayed by enzyme-linked immunoadsorbent assay for macrophage inflammatory protein-1-alpha (MIP-1) and IL-6. RESULTS: NO inhibits MIP-1 secretion triggered by CD23 activation of IL-4- and GM-CSF-matured monocytes (percentage of MIP-1 suppression = 52 +/- 11 of monocytes from asthmatic patients; percentage = 55 +/- 8 healthy controls). The inhibitory effect of NO was not cytokine-specific, as similar results were obtained with IL-6 (50 +/- 9% IL-6 suppression, asthmatic patients; 66 +/- 20%, healthy controls). CONCLUSIONS: The results demonstrate for the first time an inhibitory effect of NO on cytokine production stimulated by CD23 receptor activation. We suggest that NO may be upregulated as a potent anti-inflammatory agent in the asthmatic lung.