吸入白细胞介素-5对支气管哮喘患者外周血嗜酸性粒细胞数及其活化状态的影响

随机选择8例哮喘患者雾化吸入人重组IL-5,分别吸入前、吸入后2 h、24 h和48 h进行外周血有核细胞计数和分类,以放射免疫法测定血清中嗜酸性阳离子蛋白(ECP)及总IgE水平.结果,吸入IL-5后嗜酸性粒细胞(EOS)占细胞总数的百分比、EOS绝对细胞数以及ECP水平均随时间而明显升高,至24 h达最高值,48 h后仍维持在较高水平;IL-5在整个实验过程中对血清总IgE水平无明显影响.表明,吸入IL-5不仅可以促使循环中EOS数明显增多,且还能招致其活化从而参与哮喘的发病过程.     

白介素-16与支气管哮喘关系的研究

白介素—16(IL—16)作为一种淋巴细胞趋化因子，参与了哮喘发病的早期调节，通过与细胞表面的CD4分子结合引起CD4^ 细胞、嗜酸性粒细胞和单核细胞的趋化性活动并在活化中发挥重要作用，参与哮喘气道炎症形成．     

胸腺基质淋巴生成素对增塑剂DEHP诱导性哮喘的介导作用（综述）

胸腺基质淋巴生成素（Thymic stromal lymphopoietin,TSLP）是一种新型的细胞因子,也是过敏反应起始的重要因子,能通过诱导树突细胞表达协同刺激分子CD40、CD80,促进树突细胞招募Th2细胞的趋化因子,进而诱导Th2炎症细胞的大量生成,在哮喘发生过程中发挥重要的作用.增塑剂邻苯二甲酸二乙基己酯（Di-（2-ehtylhexyl）phthalate,DEHP）在过敏性哮喘的发生过程中具有＂佐剂作用＂,并与气道炎症的关系密不可分.研究表明,在成功构建的小鼠过敏性哮喘模型中,TSLP在DEHP致哮喘作用过程中也发挥着重要的介导作用.     

观察雷公藤多甙联合舒利迭治疗哮喘的临床疗效及探讨其治疗机制

观察雷公藤多甙(TP)联合舒利迭(SERETIDE)与单纯吸入舒利迭对哮喘患者的治疗效果,并比较两种治疗后患者外周血中CD+4CD+25调节性T(Treg)细胞的变化.选取西京医院呼吸内科门诊已确诊为哮喘的患者42例,随机分成甲、乙两组.甲组给予舒利迭(沙美特罗替卡松)吸入治疗,乙组给予舒利迭吸入治疗联合服用雷公藤多甙片治疗.采用流式细胞技术检测患者治疗前后外周血中CD+4CD+25Treg细胞,同时测定哮喘患者肺功能并填写ACQ评分表.结果甲乙两组哮喘患者治疗前外周血中CD+4CD+25 Treg细胞占CD+4T细胞的百分比分别为1.45±0.56,1.44±0.58(P＞0.05)无显著性差异.甲乙两种方法治疗4周后哮喘患者外周血中CD+4CD+25Treg细胞占CD+4T细胞的百分比分别为5.35±0.54,6.97±0.87 (P<0.05);FEV1 占预计值百分比分别为98.8±13.8,103.6±16.8(P<0.05),ACQ评分表平均得分0.81±0.5,0.28±0.3(P<0.05).发作期患者外周血CD+4CD+25 Treg细胞百分率与FEV1呈显著正相关(r=0.598 2,P<0.01).雷公藤多甙片联合舒利迭可有效治疗哮喘的发作,疗效明显优于单纯吸入舒利迭.     

木犀草素抑制哮喘大鼠气道炎症的机制研究

目的探讨木犀草素抑制哮喘大鼠气道炎症的可能机制.方法取健康清洁级Wistar大鼠60只,随机分为木犀草素组、哮喘组、对照组,每组各20只.木犀草素组、哮喘组建立大鼠哮喘模型,建模过程中每次激发后1h给予哮喘组和对照组腹腔注射生理盐水,木犀草素组腹腔注射1 mg/kg的木犀草素.光镜下观察大鼠肺组织病理变化,测定支气管肺泡灌洗液中的细胞数及IL-4水平;免疫组化法检测p38MAPK,PPARγ蛋白表达情况;RTPCR检测p38MAPK,PPARγ的相对表达量.结果 3组大鼠支气管基底膜周径之间差异无统计学意义(P0.05);哮喘组大鼠平滑肌厚度、管壁厚度均明显高于对照组及木犀草素组大鼠(P0.05).哮喘组大鼠细胞总数、嗜酸粒细胞数、中性粒细胞数均明显高于木犀草素组和对照组(P0.05);木犀草素组细胞总数、嗜酸粒细胞数、中性粒细胞数均明显高于对照组(P0.05);哮喘组IL-4含量明显高于木犀草素组及对照组(P0.05);哮喘组大鼠p38MAPK蛋白表达明显高于对照组,PPARγ蛋白表达明显低于对照组(P0.05);木犀草素组大鼠p38MAPK蛋白表达明显低于哮喘组,PPARγ蛋白表达明显高于哮喘组(P0.05).哮喘组大鼠p38MAPK mRNA相对表达量明显高于对照组,木犀草素组表达量明显低于哮喘组(P0.05);哮喘组大鼠PPARγmRNA相对表达量明显低于对照组,木犀草素组表达量明显高于哮喘组(P0.05).结论木犀草素可能通过影响PPARγ表达和p38MAPK信号通路而发挥抑制哮喘大鼠气道炎症的作用.     

人Lrg在真核细胞内亚细胞定位的检测

在真核细胞内,进行人Lrg蛋白亚细胞定位的检测。采用绿色荧光蛋白载体plRES^2-EGFP。构建plRES2-EGFP-Lrg重组载体,稳定转染到真核细胞内,荧光显微镜进行人Lrg蛋白亚细胞定位的检测;同时利用间接免疫方法,检测人Lrg蛋白在几种细胞内的亚细胞定位。发现与EGFP融合的人Lrg蛋白表达于胞浆;间接免疫荧光实验同样表明Lrg是一种胞浆蛋白。说明人Lrg蛋白可能定位于胞浆。上述结果为阐明人Lrg的功能奠定了初步基础。     

正常人和AITD患者对碘致甲状腺细胞凋亡的不同敏感性

目的:探讨碘对体外培养的Grave s病(GD)、桥本氏甲状腺炎(HT)、正常人甲状腺滤泡上皮细胞(TEC)凋亡及相关蛋白表达的影响并明确正常人和自身免疫性甲状腺疾病(AITD)患者甲状腺细胞对碘的敏感性.方法:分离培养正常、GD和HT甲状腺细胞,给予不同浓度碘化钠(NaI)干预,流式细胞仪检测细胞凋亡率,免疫组化检测各组Fas/Fasl表达的水平;比较各组细胞对碘致凋亡和凋亡蛋白表达的敏感性.结果:NaI可以使正常、GD、HT患者甲状腺细胞凋亡率增加(P<0.05),并使相关的凋亡蛋白及基因Fas/Fasl表达增加,但HT和GD甲状腺细胞凋亡和Fas/Fasl表达率较正常甲状腺细胞增加更明显,以HT患者甲状腺细胞加高碘培养后凋亡更多(P<0.05).结论:与正常人相比,AITD患者对高碘导致的甲状腺细胞凋亡更敏感.     

诱导痰中IL-8和中性粒细胞在支气管哮喘发病中作用的研究

目的探讨哮喘发病过程中嗜中性粒细胞和IL-8的作用及相互关系.方法以确诊哮喘的病人为研究对象,急性发作期10例、缓解期10例,同时选10例正常人作对照,进行诱导痰的细胞分类检查,IL-8,MAD检测.结果急性发作期哮喘病人诱导痰的嗜中性粒细胞数目、IL-8水平、MAD水平明显高于缓解期(P<0.05)和正常人(P<0.01).结论哮喘病人诱导痰IL-8明显增高,对嗜中性粒细胞有强大的趋化作用,促使其进入气道并参与嗜中性粒细胞脱颗粒、产生活性氧自由基及花生四烯酸代谢产物等物质损伤气道,引起气道炎症和气道高反应性,在哮喘发作或重度哮喘时能起到很重要的作用.     

嗜酸性粒细胞的凋亡及对哮喘的意义

嗜酸性粒细胞（EOS）在支气管哮喘中起到重要作用，是参与哮喘气道炎症关键的效应细胞，其募集入肺并释放损伤性介质是气道炎症的中心环节。而在支气管哮喘炎症消退过程中，细胞凋亡是炎性细胞消除的主要方式。     

竞争性RT-PCR检测哮喘患者IL-8表达水平

利用反向ＰＣＲ法,缺失人ＩＬ－８ｃＤＮＡ内部ｂｐ６邓列,构建得到ＩＬ－８突变内标分子。用已知最的该内标分子,通过竞争性ＲＴ－ＰＣＲ法,检测支气管哮喘患者外周血ＩＬ－８基因的表达情况,发现患者Ｌ－８ｍＲＮＡ的表达水平明显高于正常人对照组,这一结果初步表明,利用竞争性ＲＴ－ＰＣＲ技术对支气管哮喘患者进行早期基因诊断,具有一定的可行性。     

精神分裂症患者T淋巴细胞上CD226分子的表达研究

目的 通过检测T细胞上CD226分子的表达情况来阐明SCH患者T细胞的活化状态及功能．方法 将精神分裂症患者和正常对照群的外周血单个核细胞各分为植物血凝素(PHA)刺激组与非刺激组,用双色免疫荧光染色和流式细胞术检测T细胞上CD226分子的表达百分率,观察PHA刺激对T细胞表达CD226的影响．结果 1)无论是否给予PHA刺激,SCH患者外周血T淋巴细胞上CD226阳性百分率均显著高于正常对照组．2)给予PHA刺激后,SCH患者和正常对照者外周血T淋巴细胞上CD226分子的表达均显著增高;在升高幅度上,SCH患者CD226表达增长值显著低于正常对照组．3)在患者与对照组二者之间,未发现CD4,CD8 2个T细胞亚群之间CD226表达百分率存在显著性差异．4)精神分裂症患者CD226的表达与性别无关．结论 SCH患者外周血T淋巴细胞上CD226分子的表达百分率显著升高,细胞敏感性增强．在SCH患者体内。大量T淋巴细胞已处于活化状态．SCH患者体内存在某种非正常刺激因素。     

支气管哮喘患者吸入白细胞介素-5所致细胞间粘附因子增高的研究

采用随机、双盲及设置对照组的办法进行研究。在吸入ＩＬ５之前，对８例不吸烟的哮喘患者及６例非哮喘过敏性疾病患者测定其痰及血清的ｓＩＣＡＭ１；吸入ＩＬ５后，于２ｈ、２４ｈ、４８ｈ及７２ｈ取上述标本分别测定其ｓＩＣＡＭ１的水平。结果示对照组患者的ｓＩＣＡＭ１在整个研究过程中都没有明显的改变。但吸入ＩＬ５后支气管哮喘患者于痰中ｓＩＣＡＭ１出现明显的增高，于４８ｈ达最高峰，但持续不超过７２ｈ。同样的结果亦出现在血清中。研究表明ＩＬ５可能是导致支气管哮喘患者痰及血的ｓＩＣＡＭ１增高的原因，对非哮喘过敏疾病患者则无此作用。     

Soluble Jagged 1/Fc chimera protein induces the differentiation and maturation of bone marrow-derived dendritic cells

A soluble Jagged 1/Fc chimera protein （Jagged 1/Fc） was directly used to induce differentiation and maturation of bone marrow-derived dendritic cells （DCs） in mice in vitro. A model of inducing and amplifying DCs in vitrowas established. The effect of Jagged 1/Fc on morphology of DCs induced by both rmGM-CSF and rmlL-4 was observed under a confocal microscope. A fluorescein-labeled monoclonal antibody staining combined with flow cytometry was applied to detect the effect of Jagged 1/Fc on the expression of CD11c, MHC-Ⅱ, CD86, CD80 and CD40 molecules on the surface of DCs. The results showed that Jagged 1/Fc did not affect the morphological properties of DC differentiation induced by both rmGM-CSF and rmlL-4. But it could promote the differentiation and maturation of DCs induced by both. The effect of it was strikingly different in the expression profile of co-stimulating molecules and the morphologic properties of DCs from lipopolysaccharide （LPS）. The levels of MHC-Ⅱ and CD40 molecule expression on the surface of DCs stimulated by Jagged 1/Fc were significantly lower than those stimulated by LPS, and the level of CD80 expression on the surface of DCs induced by Jagged 1/Fc was near to that induced by LPS. Jagged 1/Fc had no influence on the expression of CD86 molecule on the surface of DCs. Jagged 1/Fc when used alone could not maintain the growth, differentiation and maturation of DCs. All the findings indicate that Jagged 1/Fc influences the differentiation and maturation of DCs, which is not markedly similar to LPS, providing important evidence for its development and application as a novel immunosuppressant.     

氢气对实验性小鼠过敏性鼻炎减轻作用的研究

目的 观察氢气能否减轻小鼠过敏性鼻炎.方法 BALB/c雌鼠24只,随机分成正常组、模型组、氮氧组、氢氧组.除正常组外,其余三组小鼠,前14 d内,每2天一次,腹腔注射溶有50μg卵白蛋白(ovalbumin,OVA)+2 mg氢氧化铝[Al(OH)3]的生理盐水0.3 mL;第15～21天给予两鼻腔内滴10μL内含2...     

吸入白细胞介素-5对支气管哮喘患者外周血嗜酸性粒细胞数及其活化状态的影响

随机选择８例哮喘患者雾化吸入人重组ＩＬ－５,分别吸入前,吸入后２ｈ,２４ｈ和４８ｈ进行外周血有核细胞计烽和分类,以放射免疫法测定血清中嗜酸性阳离子蛋白（ＥＣＰ）及总ＩｇＥ水平,结果,吸入ＩＬ－５后嗜酸性粒细胞（ＥＯＳ）占细胞总数的百分比,ＥＯＳ绝对细胞数以及ＥＣＰ水平均随时间而明显升高,至２４ｈ达最高值,４８ｈ后仍维持在较高水平,ＩＬ－５在整个实验过程中对血清总ＩｇＥ水平无明显影响,表明,吸入ＩＬ     

Activated human eosinophils generate SRS-A leukotrienes following IgG-dependent stimulation

Eosinophils, a class of granular leukocytes, are prominent in many inflammatory processes, particularly in asthma, certain allergic diseases and during infections with helminthic parasites. Following incubation with the Ca ionophore A23187 (refs 1-4) (a non-physiological agent which circumvents membrane calcium-gating mechanisms), eosinophils generate large amounts of sulphidopeptide leukotrienes, potent inducers of smooth muscle constriction and mucus production. These are now known to represent the activity previously termed 'slow-reacting substance of anaphylaxis' (SRS-A) but attempts to identify a physiological stimulus for SRS-A production by eosinophils have so far been unsuccessful. The cells contain recognized receptors for IgG (Fc) and it is known that they adhere to, and can be activated by, contact with the surface of large organisms such as helminthic larvae. We show here that eosinophils, particularly when activated, produce sulphidopeptide leukotrienes after contact with large particles coated with IgG.     

Enhancement of T cell immune response to lymphoma cells by CD28/CD80 alternative pathway

The aim of this study is to determine whether myeloma and lymphoma tumor cells can function as efficient antigen presenting cells (APC) to enhance the co-stimulation of T cells. The expression and function of T cell activation-related molecules, especially CD80, CD28, CD40 and CD40 ligand (CD40L), were studied on nine human myeloma cell lines (HMCL) and two B lymphoma cell lines. In the case of myeloma cell lines, the cells generally lacked CD80 antigen and expressed a heterogeneous CD40, and the expressions of CD40 and CD80 molecules could not be induced by either CD28 stimulation or CD40 ligation. Conversely, in the two B lymphoma cell lines, tumor cells expressed both CD80 and CD40 to some extent. CD28 stimulation could obviously increase the expression of CD80, CD40 and some adhesion molecules, and therefore generate a more efficient anti- tumor cell immunity. In conclusion, CD28 stimulation combined with CD40 antibody or soluble CD40 ligand may be a promising immunotherapeutic approach to B lymphoma.     

特异性脱敏法治疗柳树花粉引起的 过敏性鼻炎的实验研究

目的 建立柳树花粉引起的过敏性鼻炎小鼠模型,并对其进行特异性脱敏治疗。 方法 提取柳树花粉蛋白, 对小鼠腹腔注射并滴鼻激发,建立过敏性鼻炎小鼠模型;通过递增变应原剂量对小鼠皮下注射,观察小鼠行为学症 状、鼻黏膜组织形态学变化和检测血清 sIgE 的含量,探索特异性脱敏疗法的效果。 结果 治疗前小鼠出现搔鼻、打 喷嚏、流鼻涕等过敏症状,鼻黏膜中嗜酸性粒细胞明显多于对照组,血清 sIgE 含量高于激发前,成功建立了过敏性 鼻炎小鼠模型;治疗后小鼠过敏症状 逐 渐 减 轻 至 消 失,鼻 黏 膜 中 嗜 酸 性 粒 细 胞 逐 渐 减 少,血 清 sIgE 含 量 减 少。 结论 柳树花粉能够建立过敏性鼻炎小鼠模型,且特异性脱敏法可将其治疗。     

Engagement of the CD4 molecule influences cell surface expression of the T-cell receptor on thymocytes

The intrathymic differentiation process by which precursor cells derived from the bone marrow develop into immuno-competent T lymphocytes is poorly understood. Most thymocytes express both CD4 and CD8 accessory molecules, yet little is known about either the function of these molecules or the responsiveness of the CD4+8+ double positive thymocytes that bear them. Here, we address the possibility that CD4 engagement influences T-cell receptor (TCR) expression on developing thymocytes. We engaged CD4 molecules on murine thymocytes by in vivo injection of an anti-CD4 monoclonal antibody, which reduced the surface expression of CD4 on CD4+ thymocytes. More importantly, CD4 engagement also affected TCR expression on CD4+ thymocytes, but the effect on CD4+8+ double positive and CD4+8- single positive thymocytes was very different. CD4+8+ thymocytes responded to CD4 engagement by dramatically increasing surface expression of TCR, whereas CD4+8- thymocytes decreased surface expression of TCR. These results demonstrate that the effect of CD4 engagement on TCR expression is dependent upon the developmental state of the responding thymocyte, and, most interestingly, results in increased TCR expression by double positive thymocytes.