哮喘气道炎症中嗜酸性粒细胞凋亡与IL—5、IL—10关系的研究

目的 动态观察哮喘动物模型气道嗜酸性细胞凋亡在炎症中的意义和调控机理。研究IL-5和IL-10对凋亡的调节作用。方法 卵白蛋白（OVA）腹腔注射与雾化吸入诱发BALB/c小鼠哮喘发作，在激发后0、8、24、48、96小时及7、14天行BAL，ELISA法测定IL-5、IL-10浓度。CD15、CD49d双色标记嗜酸细胞（EOS），流式细胞仪上检测EOS凋亡率。结果 OVA激发后电镜观察显示肺内有明显炎性细胞浸润和细胞凋亡。哮喘组EOS凋亡水平与对照大体相似，EOS凋亡率与IL-10/IL-5比值密切相关。结论 IL-5和IL-10分别上行和下行调节哮喘气道炎症。气道局部有EOS凋亡现象，并受到IL-5、IL-10相互作用的调节。     

哮喘气道炎症中嗜酸性粒细胞凋亡与 IL-5、IL-10关系的研究

目的　动态观察哮喘动物模型气道嗜酸性细胞凋亡在炎症中的意义和调控机理，研究IL-5和IL-10对凋亡的调节作用。方法　卵白蛋白（OVA）腹腔注射与雾化吸入诱发BALB/c小鼠哮喘发作。在激发后0、8、24、48、96小时及7、14天行BAL，ELISA法测定IL-5、IL-10浓度。CD15、CD49d双色标记嗜酸细胞(EOS)，流式细胞仪上检测EOS凋亡率。结果　OVA激发后电镜观察显示肺内有明显炎性细胞浸润和细胞凋亡。哮喘组EOS凋亡总体水平与对照大体相似。EOS凋亡率与IL-10/IL-5比值密切相关。结论　IL-5和IL-10分别上行和下行调节哮喘气道炎症。气道局部有EOS凋亡现象，并受到IL-5、IL-10相互作用的调节。     

哮喘动物模型气道中白细胞介素6的动态变化及其意义

目的：研究炎症促进因子白细胞介素6（IL-6）的变化趋势、相互关系和对炎症反应的调节作用。方法卵蛋白腹腔注射与雾化吸入诱发小鼠哮喘发作,动态观察哮喘动物气道炎症的改变。在激发后0,8,24,48,96h及7,14d行支气管肺泡灌洗,测定回收液中细胞总数与分类计数,酶联免疫吸附（ ELLSA）法测定IL-6的浓度。结果卵白蛋白激发后气道出现明显的炎症反应。8 h～7 d支气管肺泡灌洗回收液中IL-6水平的动态变化呈近似正态分布曲线,各个时相与对照组比较差异均有显著性（第7天P＜0．05,其余时相,P＜0．01）。嗜酸细胞计数与IL-6数值呈密切正相关（r＝0．8,P＜0．01）。结论 IL-6上行调节哮喘气道炎症。     

1,25-二羟维生素D3对哮喘大鼠调节性T淋巴细胞和气道炎症的影响

目的探讨1,25-二羟维生素D3[1,25(OH)2D3]对哮喘大鼠调节性T淋巴细胞(即CD4 CD25 Foxp3 T细胞)及气道炎症的影响。方法28只Wistar大鼠用卵白蛋白作为致敏原制备大鼠哮喘模型,然后随机分为四组(n=7)。治疗组1于激发前1 h给予口服1,25(OH)2D3;治疗组2于激发前1 h给予皮下注射地塞米松;治疗组3则于激发前1 h给予以上两药联合应用;组4于激发前1 h不用药(哮喘对照组)。分别测定各组大鼠支气管肺泡灌洗液(BALF)中IL-4和IL-10水平,计数总细胞数和嗜酸性粒细胞数;流式细胞仪检测外周血和脾脏单个核细胞中Treg与CD4 T细胞的比值;检测肺组织中IL-10和Foxp3mRNA表达。以未造模Wistar大鼠作为正常对照组(n=7)。结果哮喘对照组与正常对照组比较各项指标均有显著差异。与哮喘对照组比较,各哮喘治疗组大鼠BALF中细胞总数及嗜酸性粒细胞计数明显减少,IL-4水平降低而IL-10水平增高;外周血和脾脏单个核细胞中Treg与CD4 T细胞比值明显增高;肺组织中IL-10 mRNA和Foxp3 mRNA表达增加。各哮喘治疗组间及其与正常对照组间各项检测指标比较均无显著差异。结论哮喘大鼠给予1,25(OH)2D3可增加外周血和脾脏Treg比例和Foxp3mRNA表达水平。提示1,25(OH)2D3对Treg具有调节作用,对哮喘可能有潜在的治疗作用。     

CD4T细胞活化与白细胞介素5释放在过敏性哮喘发病中的作用

目的了解特异性过敏原刺激下ＣＤ４Ｔ细胞活化以及白细胞介素５（ＩＬ５）释放在过敏性哮喘发病中作用。方法对１２例过敏性哮喘（ＡＡ组）、９例过敏性非哮喘（ＡＮ组）患者以及１０例正常对照者（Ｎ组）用过敏原屋尘螨提取液（ＨＤＭ）进行全肺吸入激发,测定激发前后支气管肺泡灌洗（ＢＡＬ）液和周围血单个核细胞（ＰＢＭＣ）的ＣＤ４ＣＤ２５细胞、嗜酸粒细胞（ＥＯＳ）、ＩＬ５、嗜酸细胞阳离子蛋白（ＥＣＰ）水平。结果ＡＡ组迟发气道反应（ＬＡＲ）发生率明显高于ＡＮ组,差异有非常显著意义（Ｐ＜００１）,此种差异与ＢＡＬ液中的ＥＯＳ计数（ｒ＝０５３９,Ｐ＜００５）、ＣＤ４ＣＤ２５细胞、ＩＬ５及ＥＣＰ水平相关。结论ＣＤ４Ｔ细胞活化与哮喘状态有关,也与过敏状态有关;特异过敏原刺激是过敏性哮喘病人ＣＤ４Ｔ细胞活化的重要原因;ＩＬ５是ＥＯＳ选择性活化因子,它主要在气道局部调控ＥＯＳ的聚集和活化     

IL-17A抑制Th2细胞分化减轻哮喘小鼠气道嗜酸性粒细胞炎症

目的 研究IL-17A对哮喘小鼠Th2细胞分化及其相关炎症的作用.方法 24只C57BL/6J小鼠按随机数字表法分为对照组、哮喘组和IL-17A处理组(n=8).哮喘组和IL-17A处理组予以卵清蛋白(ovalbumin,OVA)致敏及激发.每次雾化激发前1h,IL-17A处理组给予重组小鼠IL-17A气道滴入.各步对照均予以生理盐水.末次激发后24h处死小鼠,收集支气管肺泡灌洗液(bronchoalveolar lavage fluid,BALF)行细胞总数及分类计数.ELISA检测BALF中IL-4、IL-5、IFN-γ、IL-17A的浓度.HE和PAS染色及半定量评分评估小鼠肺部病理变化.流式细胞术检测脾脏和支气管淋巴结Th细胞分化.免疫磁珠分选健康小鼠幼稚CD4+T细胞,用Th2极化培养基体外培养,并给予IL-17A或等量PBS干预,检测Th2细胞的增殖、凋亡和分化.结果 哮喘组较对照组,BALF中细胞总数、嗜酸性粒细胞数及其比例(P＜0.05)、IL-4、IL-5、IL-17A浓度均显著增高(P＜0.05),IFN-γ浓度显著下降(P＜0.05);支气管、血管周围炎症细胞浸润和杯状细胞化生明显加重(P＜0.01);脾脏和淋巴结Th2细胞分化比例显著增高(P＜0.05).IL-17A处理组较哮喘组,BALF中的细胞总数[(26.00±5.43)×104/mLvs(58.40 ±26.93)×104/mL,P＜0.05]、嗜酸性粒细胞数[(8.04±1.98)×104/mL vs(31.95±12.28)×104/mL,P＜0.05]及其比例[(29.93 ±3.03)％ vs(53.47 ±6.62)％,P＜0.01]显著降低,而中性粒细胞数及其比例无明显变化;BALF中Th2相关因子IL-4浓度[(9.86 ±2.77) pg/mL vs(28.13 ±4.62) pg/mL,P＜0.01]、IL-5浓度[(7.30 ±0.50) pg/mL vs(10.50±1.10) pg/mL,P＜0.01]均显著降低;支气管、血管周围炎症细胞浸润减轻,HE染色半定量评分降低[(2.00 ±0.51)vs(3.12 ±0.64),P＜0.05],杯状细胞化生减少[(0.80 ±0.45)vs(2.40 ±0.55),P＜0.01];脾脏[(2.24±0.44)％vs(4.82±1.83)％,P＜0.01]和淋巴结[(7.05±0.58)％vs(10.57±1.35)％,P＜0.05]中Th2细胞分化比例显著减少.极化培养的幼稚CD4+T细胞,予IL-17A干预后,诱导分化的Th2细胞比例显著减少(P＜0.05),而增殖和凋亡无显著变化.结论 IL-17A有抑制Th2细胞分化,减轻哮喘小鼠气道嗜酸性粒细胞炎症的作用.     

Aerosol administration of dexamethasone and methotrexate attenuated Th2 reaction and eosinophil infiltration of the lung in ovalbumin sensitized mice

ＡｅｒｏｓｏｌａｄｍｉｎｉｓｔｒａｔｉｏｎｏｆｄｅｘａｍｅｔｈａｓｏｎｅａｎｄｍｅｔｈｏｔｒｅｘａｔｅａｔｅｎｕａｔｅｄＴｈ２ｒｅａｃｔｉｏｎａｎｄｅｏｓｉｎｏｐｈｉｌｉｎｆｉｌｔｒａｔｉｏｎｏｆｔｈｅｌｕｎｇｉｎｏｖａｌｂｕｍｉｎｓｅｎｓｉｔｉｚｅ...     

系统性红斑狼疮患者血清IL-10与T细胞凋亡的关系研究

目的:研究系统性红斑狼疮(SLE)患者血清IL-10与T细胞凋亡的关系.方法:采用ABC-ELISA法检测SLE患者血清IL-10含量;三色荧光流式细胞术检测患者外周血中CD3 、CD4 、CD8 T细胞亚群及其Fas、FasL的表达和早期凋亡,并分析其与血清IL-10水平的相关性.结果:SLE患者血清IL-10水平明显升高,并与SLE活动指数(SLEDAI)和血清中抗ds-DNA抗体呈正相关;外周血中T细胞CD4 亚群比例下降、CD4 /CD8 的比值降低,CD3 、CD4 、CD8 T细胞Fas、FasL的表达均增高,T细胞尤其是CD4 亚群凋亡增加,以上变化以活动性SLE更为显著;SLE患者血清IL-10水平与CD4 /CD8 细胞比值呈负相关,与T细胞FasL表达异常增加和CD4 T细胞亚群凋亡显著增多呈正相关.结论:SLE异常增高的IL-10促使T细胞高表达FasL,诱导CD4 T细胞凋亡,参与了SLE的病理进程.     

支气管哮喘淋巴细胞亚群变化的观察

应用抗人T细胞单克隆抗体CD系列对支气管哮喘发作期、缓解期及脱敏治疗半年以上者共63例进行了检测及比较分析,发现支气管哮喘发作期外周血CO_3、CD_8细胞减少,CD_4/CD_3比值增高。在缓解期和脱敏治疗后CD_8、CD_4/CD_8比值均恢复,CD_3仍较正常减低。外源性嗜喘发作期T细胞亚群改变较内源性哮喘更明显,并且其血清总IgE与CD_8变化呈负相关,与CD_4/CD_8比值变化呈正相关。本文结果指示T细胞亚群的异常变化在支气管哮喘发病中起着重要作用。     

Inhibition of allergic responsiveness in a murine asthma model via IFN-γ transgene expression

Objective To investigate adenoviral vector mediated exogenous gene expression in mouse lungs and the effect of mIFN-γ transgene expression on allergen-induced pulmonary eosinophil infiltration in a murine asthmatic model. Methods LacZ marker gene was transduced into CD-1 mouse airway epithelial cells by installation of a replication-deficient adenovirus with LacZ gene (AdCMVLacZ) 5×10[9]plaque forming unit (pfu) in the intratrachea or nostril.C57 mice were sensitized intraperitoneally and challenged by aerosol with ovalbumin (OVA) to produce an asthmatic model.AdCMVmIFNγ 5×10[9] pfu was administered via nostril in asthmatic mice 48 h before OVA challenge.Sera, bronchial alveolar lavage (BAL) and lungs were recovered 48 h after OVA challenge.Results After administration with AdCMVLacZ by intratracheal installation or nose-drop, the lungs revealed a high level of widespread LacZ transduction with X-gal staining, mainly along airways.IFN-γ via adenoviral vector transduction could be overexpressed both in vitro and in vivo (1624.7±1321.5 pg/ml in BAL 96 h after AdCMVIFNγ infection).In AdCMVIFNγ treated asthmatic models, histological evaluation revealed marked suppression of eosinophil peribronchial and perivascular infiltration; the recoverable percentage of eosinophils in BAL was an average of 9.00%±4.58%, which was a statistically significant decrease versus that of the positive control group (75.13%±6.85%) (P<0.001).The total cell number in BAL ((145±55.6)×10[3 cells/ml) in AdCMVmIFNγ treated mice also was tremendously reduced compared to the positive control group ((216.6±71.1)×10[3 ]cells/ml).Conclusions Adenoviral vector was able to overexpress exogenous gene in murine lungs.IFN-γ overexpression via adenoviral vector in pulmonary epithelia in vivo can abrogate allergen-induced eosinophilic infiltration in lungs in an asthmatic model, which may suggest a new preventively therapeutic method for cytokine immunogenetic transfer in allergic asthma.     

皮肌炎/多发性肌炎相关间质性肺病患者支气管肺泡灌洗液淋巴细胞分析

目的 探讨皮肌炎/多发性肌炎相关性间质性肺病(DM/PM-ILD)患者支气管肺泡灌洗液(BALF)中淋巴细胞计数及不同淋巴细胞亚群的分布特征,为进一步研究DM/PM-ILD发病机制提供理论依据。 方法 2013年1月-9月安徽省立医院收治的16例DM/PM-ILD患者行经支气管肺泡灌洗,获得BALF,并行总细胞计数及分类细胞计数。流式细胞术检测BALF及外周血中淋巴细胞亚群(CD3+总T细胞、CD3+CD4+ T细胞、CD3+CD8+ T细胞、CD3-CD16+CD56+ NK细胞、CD3-CD19+ B细胞),计算每种淋巴细胞亚群百分比。比较DM/PM-ILD患者与正常对照BALF中细胞总数及各淋巴细胞亚群百分比。比较DM/PM-ILD患者BALF与外周血各淋巴细胞亚群百分比。 结果 DM/PM-ILD患者外周血不同淋巴细胞百分比大多均正常;BALF细胞总数较正常对照显著增多,明显淋巴细胞占优势(每例均>15%)。DM/PM-ILD患者BALF中CD3+总T细胞和CD8+ T细胞百分比较正常对照显著增加,并且较外周血中百分比显著增加;DM/PM-ILD患者BALF中NK细胞百分比较正常对照显著下降,B细胞百分比与正常对照差异无统计学意义,两者均较外周血中显著下降。 结论 DM/PM-ILD患者BALF中存在明显的淋巴细胞计数增多和淋巴细胞亚群比例失调,其中CD8+ T细胞明显增加,可能在DM/PM-ILD形成中发挥重要作用。      

支气管哮喘患者外周血CD34+细胞、白细胞介素-5和嗜酸粒细胞的变化及其意义

目的　探讨外周血CD34 细胞、白细胞介素 - 5 (IL -5 )和嗜酸粒细胞 (EOS)在支气管哮喘发病中的作用。方法　选择中、重度哮喘急性发作期患者 30例 ,常规取血测定CD34 细胞、IL 5和EOS ;用二丙酸倍氯米松 (必可酮 ) 5 0 0 μg/d吸入治疗 2周后 ,再次取血测定外周血EOS计数、IL 5水平及CD34 细胞数目。同时观察其咳嗽频度、气喘、胸闷、鼻塞、饮食及睡眠等改变。结果　哮喘患者外周血EOS、IL 5及CD34 细胞数明显高于正常组 (P <0 . 0 1 ) ;吸入二丙酸倍氯米松治疗 2周后 ,哮喘患者外周血EOS、IL 5及CD34 细胞数明显下降 (P <0 . 0 1 )。哮喘患者血清IL 5水平与EOS数呈显著正相关 (r=0 .92 ,P <0 .0 1 ) ,与CD34 细胞数亦呈显著正相关 (r =0 . 90 ,P <0 . 0 1 )。结论外周血EOS、IL- 5及CD34 细胞在哮喘急性发作时增加 ,提示在肺组织和骨髓造血之间有相关通路。     

儿童支气管哮喘96例血清IL-13的研究

目的:探讨血清IL-13水平与儿童哮喘的关系。方法:用ELISA法检测血清IL-13、总IgE水平,采用荧光酶联免疫法测定嗜酸性粒细胞阳离子蛋白(ECP)水平,同时计数外周血嗜酸性粒细胞(EOS),并对检测结果进行统计学处理。结果:哮喘组患儿血IL-13水平与正常对照组比较差异有显著性(P<0.001),哮喘组血IL-13水平与ECP、总IgE及EOS计数均呈正相关。结论:血IL-13水平与儿童哮喘发病间可能存在着密切的关系。     

Effects of glucocorticoid and cysteinyl leukotriene 1 receptor antagonist on CD34+ hematopoietic cells in bone marrow of asthmatic mice

Background Corticosteroids remain the most effective therapy available for asthma. They have widespread effects on asthmatic airway inflammation. However, little is known about the effects of corticosteroids on the production of bone marrow inflammatory cells in asthma. This study observed the effects of glucocorticoid and cysteinyl leukotriene 1 receptor antagonist on CD34+ hematopoietic cells, so as to explore the possible effectiveness of a bone marrow-targeted anti-inflammatory strategy. Methods Balb/c mice were sensitized and challenged with ovalbumin (OVA) to establish an asthmatic model. For two consecutive weeks, asthmatic mice were challenged with OVA while being given either prednisone, montelukast, prednisone plus montelukast, or sterile saline solution. The mice were killed 24 hours after the last challenge with OVA, and bronchoalveolar lavage fluid (BALF), peripheral blood, and bone marrow were collected. Eosinophils in peripheral blood and BALF, and nucleated cells in BALF, peripheral blood, and bone marrow were counted. The percentages of CD34+ cells, CD4+ T lymphocytes and CD8+ T lymphocytes among nucleated cells in peripheral blood and bone marrow were counted by flow cytometry. Immunocytochemistry and in situ hybridization were employed to detect expression of CD34 and interleukin (IL)-5Rα mRNA (CD34+ IL-5Rα mRNA+ cells) among bone marrow hematopoietic cells. Results Compared with the sterile saline solution group, the number of eosinophils in BALF and peripheral blood, CD34+ cells in peripheral blood and bone marrow, and CD34+ IL-5Rα mRNA+ cells in bone marrow of mice from the prednisone and prednisone plus montelukast groups were significantly lower （P&lt;0.01）. The number of eosinophils in BALF from the montelukast group was also significantly lower （P&lt;0.05）. Conclusions The results suggest that, in this asthmatic mouse model, prednisone probably inhibits proliferation, differentiation, and migration of CD34+ cells in bone marrow, blocks eosinophilopoiesis in bone marrow, and interferes with eosinophil migration into peripheral blood and subsequent recruitment in the airway. In addition, montelukast may suppress eosinophil infiltration into the lungs of asthmatic mice. However, a significant inhibitory effect of montelukast on the proliferation and migration of CD34+ cells and a cooperating effect with prednisone on bone marrow of asthmatic mice were not observed.     

转染IL-10的CD_4~+T细胞治疗哮喘小鼠分子机制的研究

目的:研究携带小鼠IL-10基因的重组腺病毒表达载体修饰的CD4+T细胞对哮喘小鼠气道NF-κB表达的影响,从而探讨IL-10治疗哮喘的可能分子机制。方法:BALB小鼠随机分成6组,正常对照组(A组,),哮喘模型组(B组),IL-10基因修饰CD4+T细胞治疗组(C组),LacZ基因修饰的CD4+T细胞治疗组(D组),未经任何基因修饰的CD4+T细胞治疗组(E组),生理盐水治疗组(F组),应用卵白蛋白激发的哮喘模型。应用流式细胞仪分离小鼠外周血CD4+T细胞,基因转染采用重组腺病毒表达载体。应用肺组织标本冰冻切片免疫组化染色以确定肺气道NF-κB的表达。结果:IL-10基因转染的CD4+T细胞在第7天表达IL-10最高,并可以持续维持高水平表达40d左右。C组与D组和E组NF-κB表达经统计学处理差异均具有显著性(P<0.05)。结论:IL-10基因转染的CD4+T细胞使哮喘小鼠气道NF-κB表达下降,提示IL-10治疗哮喘可能通过抑制NF-κB传导有关。     

吸入脂多糖对哮喘小鼠气道炎症和气道黏液分泌的影响

目的 观察哮喘小鼠吸入脂多糖 （LPS,作为刺激原）其气道炎症和气道黏液分泌的变化 。 方法 30只清洁级BALB/c小鼠随机分为哮喘组 (AST组)、LPS哮喘组 （LAS组）和正常组 (NS组),每组10只。哮喘组用卵清白蛋白 （OVA）致敏和激发制作哮喘模型,LPS哮喘组在哮喘模型的基础上加用LPS (50 μg/mL)雾化吸入30 min,正常组用生理盐水代替OVA。检测各组小鼠支气管肺泡灌洗液 (BALF)细胞总数和细胞分类计数,采用ELISA检测BALF中的白细胞介素4 (IL-4)和肿瘤坏死因子-α （TNF-α）水平,HE染色观察肺部病理学改变,用阿尔辛蓝-过碘酸雪夫 (AB-PAS)染色气道杯状细胞,免疫组织化学法检测肺组织中黏蛋白5ac (Muc5ac)的表达,荧光定量RT-PCR检测Muc5ac mRNA在肺内的表达。并分析Muc5ac蛋白表达与各指标的相关性。 结果 AST及LAS组小鼠较NS组在BALF中的细胞总数、嗜酸性粒细胞、单核细胞、淋巴细胞百分比,IL-4和TNF-α水平、肺组织AB-PAS阳染面积, Muc5ac蛋白和mRNA表达明显升高,其差异均有统计学意义 (P均<0.05)。LAS组较AST组上述气道炎症（除单核细胞数外）和气道黏液高分泌指标明显增高,差异均有统计学意义 (P<0.05)。气道Muc5ac蛋白表达与BALF中细胞总数、嗜酸性粒细胞数、IL-4、TNF-α水平、气道AB-PAS染色阳性着色面积均呈正相关（P均<0.05）。 结论 OVA致敏和激发的哮喘小鼠出现以嗜酸性粒细胞、淋巴细胞浸润为主的气道炎症及杯状细胞增生的气道黏液高分泌,且气道炎症和气道黏液高分泌关系密切。LPS可使气道炎症和气道黏液高分泌加重,可能与LPS激发了体内炎症介质的生成、活化有关。     

川芎嗪对哮喘大鼠Th2型细胞因子作用的研究

目的 探讨川芎嗪对哮喘大鼠白细胞介素-4（IL-4）、白细胞介素-13（IL-13）水平的影响及其防治哮喘的作用机制。方法 60只SD大鼠随机分为正常对照组、哮喘模型组、地塞米松组、小剂量及大剂量川芎嗪组、联合用药组，以卵蛋白致敏和激发制备大鼠哮喘模型，用酶联免疫吸附试验（ELISA）检测支气管肺泡灌洗液（BALF）中IL4、IL—13的水平。结果 哮喘模型组BALF中IL-4、IL—13水平均高于正常对照组（P〈0．001）；地塞米松组和川芎嗪治疗各组均显著低于哮喘模型组（P〈0．001）；川芎嗪大剂量组和联合用药组降低IL-4和IL—13的作用与地塞米松组无显著性差异（P〉0．05）；川芎嗪大剂量组降低IL-4的作用与川芎嗪小剂量组无显著性差异（P〉0．05），川芎嗪大剂量组降低IL—13的作用优于川芎嗪小剂量组（P〈0．05）；BALF中IL-4和TL—13水平呈显著正相关（r=0．714，P〈0．01）。结论 川芎嗪通过抑制IL-4和IL—13的水平而抑制哮喘气道炎症。     

调节性B细胞在卵巢癌病人外周血中表达及其临床意义

目的探究调节性B细胞(Bregs)在卵巢癌病人外周血中的表达频率及其与FIGO分期间的关系,并分析其与调节性T细胞(Tregs)及IL-10之间的相关性。方法将卵巢癌(23例)和卵巢良性肿瘤病人(26例)纳入研究,流式细胞术检测外周血CD19+CD24hiCD38hiBreg和CD4+CD25+Treg细胞表达频率及血清IL-10表达水平。并分析两种细胞与FIGO分期、IL-10浓度的关联及两者间的相关性。结果卵巢癌病人外周血中Bregs、Tregs百分比高于卵巢良性肿瘤病人(P < 0.01),并与FIGO分期密切相关,卵巢癌Ⅲ期+Ⅳ期病人高于Ⅰ期+Ⅱ期(P < 0.01);卵巢癌病人血清IL-10高于卵巢良性肿瘤病人(P < 0.01)。相关性分析显示:卵巢癌病人外周血Breg细胞与Treg细胞比例呈正相关关系(P < 0.05);卵巢癌病人血清IL-10与Bregs、Tregs细胞均呈正相关关系(P < 0.05)。结论Bregs和Tregs在卵巢癌病人外周血中所占比例显著升高,能够通过分泌IL-10参与卵巢癌免疫反应,这可能为卵巢癌的免疫治疗提供新靶点。     

MP感染对哮喘患儿BALF中T细胞功能及细胞因子水平的影响分析

目的:探讨肺炎支原体(MP)感染对哮喘患儿支气管灌洗液(BALF)中T细胞功能、细胞因子水平的影响作用.方法:选取本院儿科2013年4月~2015年9月收治81例哮喘患儿作为研究对象,其中合并MP感染的哮喘患儿39例(MP组)、42例哮喘患儿MP感染阴性(非MP组),30例因上呼吸道感染的非哮喘、非MP感染患儿作为对照组,采用流式细胞仪检测三组BALF中国T细胞亚群、细胞因子水平.结果:MP组和非MP组患儿的FEV1%测定值显著的低于对照组;三组患儿BALF中T细胞、细胞因子检测结果,MP组患儿的CD3+、CD4+、CD4+/CD8+、INF-γ值均显著的低于非MP组和对照组,CD8+、IL-4、IL-5、IL-4/INF-γ显著的高于非MP组和对照组;非MP组患儿的CD3+、CD4+、CD4+/CD8+、INF-γ值均显著的低于对照组,CD8+、IL-4、IL-5、IL-4/INF-γ显著的高于对照组.结论:MP感染会进一步降低哮喘患儿的T细胞功能、炎症因子水平增高,这可能导致哮喘患儿病情加重有关.     

CD8+CD28-T细胞在哮喘小鼠发病机制中的作用及地塞米松对其的影响

目的 探讨CD8+CD28-T细胞在哮喘发病机制中的作用及地塞米松对该细胞的影响.方法 30只BALB/c小鼠随机分为哮喘组、地塞米松组、正常对照组,各10只.哮喘组和地塞米松组给予卵白蛋白致敏后雾化吸入卵白蛋白溶液,地塞米松组每次雾化吸入前腹腔注射地塞米松1 mg/kg,各组分别于末次雾化激发后测定小鼠的气道反应性;对支气管肺泡灌洗液(BALF)行细胞总数、嗜酸性粒细胞(EOS)计数;取肺组织作HE染色病理切片;测BALF中IgE含量;流式细胞仪检测小鼠血、BALF中CD8+CD28-T细胞占淋巴细胞百分比;分析BALF中IgE、EOS计数与血液中CD8+CD28-T细胞百分比的相关性.结果 哮喘组、地塞米松组气道反应性明显高于正常对照组.哮喘组BALF中细胞总数和EOS计数分别为(5.56±4.06)× 102/L和(3.29±2.23)× 102/L,均明显高于地塞米松组[(2.59±1.69)× 102/L,P=0.044和(1.11±0.73)×102/L,P=0.008]及正常对照组[(0.91±0.65)×102/L,P=0.003和(0.43±0.37)× 102/L,P=0.001)];而后两组之间差异均无统计学意义(均P>0.05).哮喘组、地塞米松组、正常对照组BALF中IgE含量分别为(23.85±5.97)g/L、(13.15±2.22)g/L、(6.54±1.03)g/L,三组间差异有统计学意义(F=38.558,P=0.000).哮喘组、地塞米松组、正常对照组CD8+CD28-T细胞百分比在外周血中分别为(18.68±4.12)%、(13.43±2.90)%、(8.43±4.60)%;在BALF中分别为(1.25±0.40)%、(0.66±0.49)%、(0.21±0.19)%,组间差异均有统计学意义(F=11.837,P=0.001;F=12.885,P=0.000).哮喘组BALF中IgE含量和EOS计数与外周血中CD8+CLY28-T细胞百分比均呈正相关(r=0.864,P=0.012和r=0.804,P=0.029).结论 CD8+CD28-T细胞数量与哮喘小鼠气道炎症有明显相关性,地塞米松可有效抑制哮喘气道炎症并可能抑制了CD8+CD28-T细胞的表达和功能.

Abstract：

Objective To explore whether or not CD8+ CD28- T cell play a pathogenic role in asthma and detect the effects of dexamethasone ( DXM ). Methods A total of 30 mice were randomly divided into 3 groups: asthmatic group, DXM group and control group ( n = 10 each). The asthmatic and DXM groups were sensitized twice and inhaled ovalbumin. The DXM Group received an intraperitoneal injection of DXM lmg/kg before inhaling ovalbumin. After successful modeling, 3 mice were selected randomly from each group to measure the airway responsiveness. Also a bronchoalveolar lavage cytological study was performed and lung tissue sections were prepared for histopathologic examination to evaluate the airway inflammation. The content of IgE in bronchoaleolar lavage fluid ( BALF) was detected with a murine IgE ELISA kit. And the fractions of CD8 + CD28- T cell of peripheral blood and BALF were tested by flow cytometry to analyze the correlation between IgE, eosinophils ( EOS) of BALF and CD8 + CD28 - T cell of blood. Results The airway hyperresponsiveness in asthmatic and DXM groups were significantly higher than that in the control group. The number of total cells and EOS of BALF in the asthmatic group [ ( 5. 56 ±4. 06) × 102/L; (3. 29 ±2. 23) × 102/L] were significantly higher than that in control group [ (0. 91 ±0.65)×102/L, P = 0.003; (0.43 ±0.37) × 102/L, P = 0.001] and DXM group [(2.59 ±1.69) ×102/L, P =0.044; (1. 11 ±0.73) ×l02/L, P = 0.008]; while the DXM group was insignificantly higher than the control group (P=0. 234, P=0. 363). There were significant differences in the contents of IgE of BALF for the asthmatic, DXM and control groups [ (23. 85 ±5. 97) g/L, (13. 15 ±2.22) g/L, (6.54±1. 03) g/L, F = 38. 558, P = 0. 000 ] . The percentages of CD8 + CD28- T cell in peripheral blood in asthmatic and DXM groups [ (18. 68 ±4. 12)% and ( 13.43 ± 2. 91) % ] were significantly higher than those in control mice [ (8. 43 ± 4. 60) % , both P < 0. 05 ]. The percentages of CD8 + CD28 - T cell of BALF in asthmatic group and DXM group [(1.25±0. 40)% and (0. 66 ± 0. 49) % ] were also significantly higher than those in control mice [ (0. 21 ± 0. 19) % , both P < 0. 05 ]. The percentages of CD8 + CD28 - T cell of blood and BALF in the DXM mice were significantly lower than those in asthmatic group. The correlations between IgE ( r = 0. 864, P = 0. 012), EOS ( r = 0. 804, P = 0.029) and CD8 + CD28- T cell were significant. Conclusion The fraction of CD8 + CD28- T cell is closely correlated with the inflammation of asthmatic airway. The airway hyperresponsiveness and inflammation in asthmatic mice may be relieved by DXM through its effect of inhibiting the expression of CD8 + CD28- T cell.