IL—3,IL—5和GM—CSF受体研究的进展

ＩＬ－３，ＩＬ－５和ＧＭ－ＣＳＦ是体内十分重要的造血细胞生长因子，近年来在其受体的分子生物学方面国外进行了大量的研究。     

雷公藤对哮喘豚鼠IL-5、GM-CSF基因转录的影响及作用机制

探讨雷公藤哮喘ＩＬ－５、ＧＭ－ＣＳＦ基因转录的影响及其机制,方法：采用卵蛋白（ＯＡ）致敏复制哮喘豚鼠模型,用原位杂交就雷公藤甲素对其肺组织和外周血单个核细胞ＩＬ－５、ＧＭ－ＣＳＦｍＲＮＡ表达的影响进行检测。并通过凝胶电泳迁移试验（ＥＭＳＡ）检测ＴＰ对伴刀豆蛋白或佛波脂（ＰＭＡ）刺激的人Ｔ细胞活化蛋白－１（ＡＰ－１）的ＤＮＡ结合活性的影响。结果：①哮喘豚鼠肺组织和ＰＢＭＣ－ＩＬ－５、ＧＭ－ＣＳＦｍＲ     

重组人GM-CSF／IL-3融合蛋白(G3)的分离纯化

Ｇ３是由基因工程技术构建、在大肠杆菌中高效表达获得的由ＧＭ－ＣＳＦ和ＩＬ－３组成的融合蛋白,能刺激造血细胞的增殖、分化和调节免疫应答。Ｇ３的生物学活性与ＧＭ－ＣＳＦ和ＩＬ－３相比有明显提高［１］。本文用抗Ｇ３单抗（ＭＧ３１）分别与ＣＮ－Ｂｒ－Ｓｅｐｈａｒｏｓｅ４Ｂ和ｒＰｒｏｔｅｉｎＡ－ＳｅｐｈａｒｏｓｅＦＦ偶联制备免疫亲和色谱（ＩＡＣ）柱,对Ｇ３进行纯化,取得了满意的结果。１　材料和方法１．１　ＭＧ３１单抗［２］的纯化　饱和硫酸铵溶液沉淀法。１．２　ＭＧ３１－ＣＮＢｒ－Ｓｅｐｈａｒｏｓｅ４Ｂ柱（…     

新疆伊犁地区维吾尔、汉族不同人群变应性鼻炎患者IL-4基因甲基化比较研究

目的探讨IL-4基因启动子区甲基化在维吾尔、汉族两个民族变应性鼻炎(AR)患者中的差异。方法采用甲基化特异性聚合酶链反应(MSP),病例组维吾尔族、汉族AR患者各31例,健康对照组维吾尔族、汉族各31例,检测其外周血IL-4基因甲基化状态。结果维吾尔族、汉族AR患者IL-4甲基化率均高于健康对照组,有统计学意义(P<0.05);维吾尔族、汉族AR患者IL-4甲基化率相比较,无统计学差异(P>0.05)。维吾尔族、汉族健康对照组未甲基化率相比较,有统计学差异(P<0.01)。结论 IL-4基因高甲基化是导致AR患病的一个重要原因.汉族人群比维吾尔族易患AR。易感人群     

用GM-CSF和 IL-4在体外诱导高纯度CD14+树突状细胞

本研究改进传统的树突状细胞（DC）诱导方法，用GM－CSF（150ng/ml）和IL－4（80U／ml），在体外经7d从健康人外周血中诱导出了大量高纯度DC，其高表达HLA－I、HLA－II类分子，共刺激分子和黏附分子，同时还高表达其前体单核细胞的特异性标志CD14分子，显示出成熟 DC的特征。这些CD14^ DC能强烈诱导同种异体淋巴细胞的增殖，其内吞能力在第3天达最高，之后明显下降。此结果丰富了DC的类型，并为CD14^ DC的深入研究和应用奠定了基础。     

新疆维、汉人群变应性鼻炎患者血清IL-4和IFN-γ水平及其与鼻粘膜嗜酸性粒细胞浸润的相关性

目的:探讨新疆维、汉人群变应性鼻炎(Allergic rhinitis,AR)患者血清白细胞介素4(Interleukin-4,IL-4)和干扰素γ(Interferon gamma,IFN-γ)水平与鼻粘膜嗜酸性粒细胞(Eosionphils,EOS)浸润的相关性。方法:采用酶联免疫吸附法(Enzyme-linked immunosorbnent assay,ELISA)测定59例维吾尔族AR患者、50例汉族AR患者外周血清IL-4和IFN-γ水平,高倍显微镜下观察病理涂片并计数AR患者鼻粘膜EOS计数。结果:维、汉人群AR患者血清IL-4和IFN-γ水平均无显著差异(t值分别为1.304和1.408,P均>0.05);维、汉人群AR患者IL-4水平与鼻粘膜嗜酸性粒细胞计数均呈正相关(r分别为0.828和0.691,P均<0.01),IFN-γ水平与鼻粘膜EOS计数均呈负相关(r分别为-0.712和-0.764,P均<0.01)。结论:维、汉人群AR患者的血清IL-4和IFN-γ水平均无显著差异,AR患者鼻粘膜EOS增多与血清IL-4分泌过多而IFN-γ分泌受抑制有关。     

IL-4和IL-4受体基因多态性与成人变应性哮喘的关系

目的　研究白细胞介素 4 (IL 4 )、IL 4受体α链的 2个基因多态性位点与中国成人变应性哮喘的关系。方法　采用病例对照方法 ,用聚合酶链反应 限制性片段长度多态性方法 (PCR RFLP)对IL 4启动子区C - 5 89T和IL 4Rα链Q5 76R进行基因分型。结果　IL 4C - 5 89T与中国成人变应性哮喘无关 ,然而 ,变应性哮喘组IL 4Rα链 5 76R R频率显著性高于对照组 (χ2 =9.36 9,P <0 .0 1;OR =3.797) ,且与血浆高IgE相关。结论　IL 4Rα链 5 76R R基因型是中国成人变应性哮喘的基因危险因子     

甲亢患者IL-4、IL-10水平检测的临床意义

目的 :探讨IL - 4、IL - 10水平变化在甲亢的临床意义。方法 :选择甲亢未治疗组 6 4例、甲亢经1 31 I治疗组 39例和正常对照组 35例 ,用化学发光法测定FT3、FT4 、TSH的含量 ,用放射免疫分析测定IL - 4、IL - 10含量。结果 :甲亢未治疗组患者血清IL - 4、IL - 10水平均高于正常对照组 (p <0 0 1、p <0 0 5 )。1 31 I治疗组患者血清IL - 4、IL - 10水平均低于未治疗组 (p <0 0 1、p <0 0 5 )。甲亢1 31 I治疗组患者血清IL - 4、IL - 10与正常对照组差异无显著性 (p>0 0 5 )。结论 :IL - 4、IL - 10在甲亢的发病机理中起重要作用 ,判断疗效有一定意义。     

变应性鼻炎和哮喘患者血清IL-5、IL-15及IL-18的水平研究

目的探讨IL-5、IL-15和IL-18在变应性鼻炎、支气管哮喘、变应性鼻炎合并哮喘疾病中的作用。方法采用双抗体夹心ELISA测定法对33例支气管哮喘患者、35例变应性鼻炎患者、35例变应性鼻炎合并哮喘的患者及35例正常健康查体者血清中IL-5、IL-15和IL-18的水平进行检测。结果支气管哮喘、变应性鼻炎、变应性鼻炎合并哮喘的患者血清中IL-5、IL-15和IL-18水平较正常对照组升高（P〈0．01）,IL-5、IL-15,IL-18水平在变应性鼻炎合并哮喘组均高于鼻炎组与和哮喘组;鼻炎组IL-5水平高于哮喘组（P=0．003）,哮喘组IL-18水平高于鼻炎组（P=0．001）。结论IL-5、IL-15和IL-18参与了过敏性鼻炎和哮喘的发病过程;变应性鼻炎合并哮喘的炎症程度较高;哮喘和鼻炎因发病部位不同炎症反应也有不同。     

神经生长因子和白血病抑制因子上调哮喘大鼠脾淋巴细胞IL-4、IL-5的表达

目的:探讨神经生长因子(NGF)、白血病抑制因子(LIF)对哮喘大鼠脾淋巴细胞IL-4、IL-5表达的影响。方法:16只大鼠随机分为对照组(A组)8只,哮喘组(B组)8只,采用卵蛋白和氢氧化铝腹腔内注射致敏,2周后给予1%卵蛋白雾化吸入激发哮喘。(1%卵蛋白/生理盐水)雾化吸入2周后,24h内体外分离培养对照组大鼠和哮喘组大鼠脾淋巴细胞;通过RT-PCR半定量法和ELISA法测定2组大鼠脾淋巴细胞表达Th2细胞因子IL-4、IL-5mRNA转录水平和分泌蛋白的基础水平,观察外源性NGF和LIF体外干预后淋巴细胞IL-4和IL-5表达随干预浓度的变化。结果:(1)哮喘组IL-4、IL-5mRNA表达及蛋白水平明显高于对照组(P0.05);(2)在外源性不同浓度的NGF、LIF干预下,IL-4、IL-5mRNA和蛋白表达量分别比前一梯度低浓度干预组有显著升高(P0.05),NGF、LIF上调IL-4、IL-5mRNA和蛋白表达呈浓度依赖性。结论:在哮喘大鼠中,NGF、LIF可能通过上调Th2类细胞因子IL-4、IL-5mRNA表达和蛋白分泌,参与促进和放大Th2类细胞因子免疫失衡效应。     

Immunohistochemical detection of GM-CSF, IL-4 and IL-5 in a murine model of allergic bronchopulmonary aspergillosis

Background Granulocyte macrophage-colony stimulating factor (GM-CSF), interleukin-4 (IL-4) and IL-5 are important in tissue eosinophil accumulation and high IgE production in allergic inflammatory reaction. Objective We examine lung GM-CSF, IL-4 and IL-5 expression in a murine model of allergic bronchopulmonary aspergillosis (ABPA) characterized by eosinophil and lymphocyte lung infiltration and elevated serum IgE level. Methods C57BL/6 mice were intranasally treated three times a week for 1, 2 or 3 week(s) with Aspergillus fumigatus (Af) antigen or saline and were sacrificed on days 7, 14 and 21. Immunohistochemical analyses for GM-CSF, IL-4 and IL-5 were performed on lung sections. Results Af treatment induced a remarkable pulmonary eosinophil influx. Increased numbers of lung T lymphocytes and GM-CSF positive cells were observed on days 14 and 21. IL-4 and IL-5 positive cells were increased significantly only on day 14. Immunostained serial sections showed that most (≥98%) cytokine positive cells were CD3 positive. Few eosinophils (<2% of cytokine positive cells) were immunoreactive for GM-CSF and IL-5. Significant correlations were found between the number of GM-CSF and IL-5 positive cells, and the number of eosinophils in Af-treated lung (r = 0.62, P < 0.05 and r = 0.52, P < 0.05, respectively), and between the number of IL-4 positive cells and the serum total IgE level (r = 0.64, P<0.01). Conclusions Our data suggest a role for T lymphocyte GM-CSF, IL-4 and IL-5 in Af-induced mouse pulmonary eosinophilia and increased serum IgE production and further support the importance of T helper (TH) cells in the pathogenesis of ABPA.     

IL-3 in dendritic cell development and function: a comparison with GM-CSF and IL-4

Dendritic cells (DC) develop in vivo from hematopoietic precursor cells. This process can be mimicked in vitro by growth factor stimulation. Among those factors granulocyte-macrophage colony-stimulating factor (GM-CSF) is the best known and most widely used for generation of rodent and human DC of the myeloid lineage. GM-CSF is often combined with interleukin-4 (IL-4) to suppress macrophage (Mph) outgrowth in cultures of human cells, but this does not apply to the mouse, and detailed analyses on the role of IL-4 are rare. Despite evidence for the importance of GM-CSF for DC development derived from in vitro data, GM-CSF-deficient mice are largely normal with respect to their DC populations. This raised the interest in other growth factors for DC. IL-3 can also support DC growth in vitro, but has been neglected for some years. Now it has been revived by a series of publications. In this review, some new features of myeloid DC regarding their early developmental stages, the GM-CSF/IL-4-interplay, and the role of IL-3 are summarized.     

Identification of a fourth ancient member of the IL-3/IL-5/GM-CSF cytokine family,KK34, in many mammals

The related cytokine genes IL-3, IL-5 and GM-CSF map to the (extended) T_H2 cytokine locus of the mammalian genome. For chicken an additional related cytokine gene, KK34, was reported downstream of the IL-3 plus GM-CSF cluster, but hitherto it was believed that mammalian genomes lack this gene. However, the present study identifies an intact orthologue of chicken KK34 gene in many mammals like cattle and pig, while remnants of KK34 can be found in human and mouse. Bovine KK34 was found to be transcribed, and its recombinant protein could induce STAT5 phosphorylation and proliferation of lymphocytes upon incubation with bovine PBMCs. This concludes that KK34 is a fourth functional cytokine of the IL-3/IL-5/GM-CSF/KK34-family (alias IL-5 family) in mammals.While analyzing KK34, the present study also made new identifications of cytokine genes in the extended T_H2 cytokine loci for reptiles, birds and marsupials. This includes a hitherto unknown cytokine gene in birds and reptiles which we designated “IL-5famE”. Other newly identified genes are KK34, GM-CSF(-like), IL-5, and IL-13 in reptiles, and IL-3 in marsupials.     

IL-5, IL-8 and GM-CSF immunostaining of sputum cells in bronchial asthma and chronic bronchitis

Background: Granulocyte-macrophage colony stimulating factor (GM-CSF) and inlerleukin (IL)-5 or IL-8 have been .suggested to play an important role in the pathogenesis of eosinophilic airway inflammation in bronchial asthma or neutrophilic airway inflammation in chronic bronchitis, respectively, However, GM-CSF and IL-8 have biological activities to either eosinophils or neutrophils. Objective: To investigate the contribution of these cytokines to airway inflammation, we compared the cellular differential and immunolocalization of GM-CSF, IL-5 and IL-8 in sputum cells from patients with bronchial asthma and chronic bronchitis. Methods: Cytospins of sputum cells from 12 patients with bronchial asthma and 12 with chronic bronchitis were subjected to cellular differential counting and immunocytochemistry with antihuman GM-CSF, IL-5 and IL-8 antibody. Results: The predominant cells in bronchial asthma were eosinophils and lymphocytes, while those in chronic bronchitis were neutrophils. All cytokines examined were detected in either bronchial asthma or chronic bronchitis, although the percentage of GM-CSF and IL-5 positive cells in bronchial asthma (53.4 ± 6.0 [mean±sfm ]% and 9.7 ± 2.8%, respectively) was significantly higher than that in chronic bronchitis (11.4±2.5%; P < 0.001 and 1.7plusmn;0.3%; P < 0.007. respectively). In contrast, the percentage of IL-8 positive cells in chronic bronchitis (23.8 ± 7.0%) was significantly higher than that in bronchial asthma (7.7 ± 1.9%; P < 0.04). The cells positive for IL-5 were lymphocytes in bronchial asthma and chronic bronchitis. The cells positive for GM-CSF in bronchial asthma were predominantly eosinophils. while those in chronic bronchitis were monocytes/macrophages and neutrophils. In contrast, neutrophils are mainly positive for IL-8 in chronic bronchitis, while monocytes/macrophages and bronchial epithelial cells are positive for IL-8 in bronchial asthma. Conclusion: The immunochcmical comparison of GM-CSF and IL-8 localization in sputum cells between bronchial asthma chronic bronchitis suggests the differential regulation and roles of these cytokines in eosinophilic vs neutrophilic airway inflammation, resulting in the development of different types of airway inflammation.     

Effects of IL-5, granulocyte/macrophage colony-stimulating factor (GM-CSF) and IL-3 on the survival of human blood eosinophils in vitro.

The mechanisms that could affect the lifespan of eosinophils after they have left the bone marrow, and their capacity to respond to activation factors were studied by examining the effects of IL-5, GM-CSF and IL-3 on purified human blood eosinophils in culture. All three cytokines prolonged the lifespan of the majority of blood eosinophils. This effect was dose-dependent: IL-5 greater than GM-CSF greater than IL-3. Light density eosinophils from most patients had a longer lifespan in culture than did normal density eosinophils, with or without the three cytokines. Eosinophil death in the absence of these cytokines occurred by apoptosis. Eosinophils from two patients did not survive when cultured with IL-5, although they survived in the presence of IL-3 or GM-CSF. IL-5, GM-CSF and IL-3 induced the expression of the activation epitope on the eosinophil ribonucleases recognized by monoclonal antibody EG2. We conclude that small amounts of IL-5, GM-CSF and IL-3 prevented programmed cell death in human blood eosinophils and induced the expression of the activation forms of eosinophil ribonucleases. We suggest that differences in the capacity of normal and light density eosinophils to survive in culture, and in the ability of eosinophils from some patients to respond to IL-5 could account for variations in the severity of disease seen in patients with persistent eosinophilia.     

支气管哮喘患者血清IL-10、IL-5和ECP的变化及其意义

目的 通过对支气管哮喘患者和正常对照组的血清IL-10, IL-5和 ECP水平的测定以及相互关系的研究,探讨它们在支气管发病中的作用.方法 应用Pharmacia UniCAP系统和ELISA方法分别测定支气管哮喘患者和正常对照组的血清E CP和IL-10、IL-5水平.结果 正常对照组和发作期的支气管哮喘患者的 ECP水平分别为(3.97±2.13) μg/L和(21.76±12.08) μg/L.正常对照组,支气管哮喘ECP 升高组和支气管哮喘ECP正常组的血清IL-5水平分别为(10.90±4.41) pg/mL,(20.62± 15.7 4) pg/mL和(9.24±7.16) pg/mL.正常对照组和支气管哮喘ECP升高组之间IL-5水平有显著差异(P＜0.01),正常对照组和支气管哮喘ECP正常组之间IL-5水平无差异(P＞0.05) ,支气管哮喘ECP正常组和ECP升高组之间IL-5水平有显著差异(P＜0.01),ECP与IL-5 明显相关 ,两者之间的相关系数r=0.465(P＜0.01).正常对照组,支气管哮喘ECP升高组和支气管哮喘ECP正常组的血清IL-10水平分别为(38.28±15.17) pg/mL,(22.76±15.25) pg/mL 和(42.32±14.61) pg/mL,支气管哮喘ECP正常组和ECP升高组之间IL-10水平有显著差异( P＜0. 01).结论 支气管哮喘的发生与嗜酸粒细胞有密切的关系,ECP和IL-5可反映嗜酸粒细胞的活化程度,在支气管哮喘发作时嗜酸粒细胞处于激活状态,易于释放蛋白颗粒;IL-5对嗜酸粒细胞有调控作用.支气管哮喘组的血清IL-10水平较正常对照组低,提示支气管哮喘患者IL-10分泌减少,不能有效抑制炎症或促炎症细胞因子的合成及释放,亦即不能有效抑制炎症反应,可能是导致或加重气道炎症的原因之一.     

Serum levels of soluble IL-2 receptor, IL-4 and IgE-binding factors in childhood allergic diseases.

The serum levels of soluble IL-2 receptor (sIL-2R), IL-4 and IgE-binding factors were examined in children with allergic diseases, and compared with those in non-allergic controls of the same age and sex. The results showed age-related decreases in the serum levels of sIL-2R and IgE-binding factors, but not in that of IL-4 in both allergic and non-allergic individuals. Significant elevation of sIL-2R was observed in sera from children with atopic eczema or history of an anaphylactic reaction to food, as compared with that in non-allergic controls. The serum concentration of IL-4 was elevated in all allergic groups, including cases of atopic eczema, bronchial asthma and anaphylaxis to food, compared with non-allergic controls, and was correlated significantly with the serum level of IgE (r = 0.59). The IgE-binding factor levels in sera from patients aged 6-10 years with bronchial asthma, or patients aged 1-5 years with a history of food anaphylaxis were elevated as compared with those in non-allergic controls of same age. There was no significant correlation between the serum levels of IgE-binding factors and IgE. Since sIL-2R is released by activated T cells, the present study is in favour of T cell activation causing allergic skin disorders. The serum levels of IL-4 as well as IgE did not differ among allergic patients of different clinical categories. The role of IgE in atopic eczema and other allergic diseases is not clearly established; however, it seems likely that IL-4 is deeply involved in the increased production of IgE seen in allergic individuals. The possible involvement of IgE-binding factors in the age-related changes of clinical manifestations in childhood allergic diseases was also discussed.