骨髓与支气管哮喘嗜酸性粒体细胞增多症的关系研究进展

产生于骨髓的嗜酸性粒细胞（Eos)在哮喘气道炎症中发挥重要作用。骨髓的造血机构成了哮喘气道炎症形成过程的活性组分，如骨髓嗜酸性粒细胞及其祖细胞数量的增加；骨髓细胞表达致炎因子如IL－5mRNA、蛋白质和IL－5受体等。治疗哮喘的药物如糖皮质激素能抑制骨髓嗜酸性粒细胞形成。骨髓和肺组织一样，可能成为抗哮喘治疗的另一种重要靶器官.     

059 IL-13与支气管哮喘

支气管哮喘是由多种细胞特别是肥大细胞、嗜酸性粒细胞和T淋巴细胞参与的慢性气道炎症.哮喘的发生过程中有多种细胞因子参与,白介素-13(IL-13)是近年来新克隆的淋巴因子.IL-13作为Th2型细胞因子成员,具有多种生物学功能.通过激活嗜酸性粒细胞,减少其凋亡,促进IgE分泌等机制,参与哮喘炎症的维持,诱导气道高反应性及小气道结构重建.     

椒目油A2通过抑制EOS浸润和骨髓粒系动员减轻哮喘小鼠气管炎症反应

目的： 观察椒目油A2对哮喘支气管嗜酸性粒细胞（EOS）浸润与CD34+细胞及骨髓粒系动员的影响。方法: 腹腔注射 20% 氢氧化铝［Al(OH)3］+10% 卵蛋白（OVA） 混合液致敏与雾化吸入1% OVA诱发BALB/c 小鼠哮喘模型。在激发10 d并同时给药治疗后,小鼠被处死,瑞氏染色检测肺灌洗液（BALF）炎性细胞及骨髓造血细胞系变化,原位杂交（ISH） 检测肺组织和骨髓白细胞介素-5（IL-5）mRNA和嗜酸性粒细胞趋化因子（EON）mRNA 表达,免疫组化检测肺组织和骨髓中IL-5、EON的蛋白表达,HE和荧光染色观察肺组织炎症细胞浸润和CD34+细胞。结果: 椒目油A2和泼尼松显著地减轻肺组织支气管炎性反应的病变程度如组织肿胀、重构、增生、上皮细胞脱落等,抑制变应源致敏刺激机体所致EOS 增多和浸润, 明显地减少哮喘小鼠气管周围及骨髓中EOS密度,抑制骨髓中幼粒细胞向嗜酸性粒细胞分化,同时支气管区及骨髓中CD34+IL-5、 CD34+IL-5R、 CD34+CCR-3细胞数显著减少,这与肺组织内IL-5、IL-4、GM-CSF等细胞因子水平降低有关。结论: 椒目油A2减轻实验性哮喘小鼠气道炎症反应,其机制可能与其抑制支气管EOS增多和骨髓粒系动员相关。     

气道内应用IL-12抑制抗原诱导的过敏性气道反应

目的探讨气道内应用白细胞介素 12 (IL- 12 )对抗原诱导的过敏性反应的影响。方法 C5 7BL/ 6小鼠经 OVA免疫建立哮喘模型 ,在主动免疫及抗原激发阶段气道内应用 IL- 12 ,观察肺泡灌洗液细胞成份、肺部淋巴细胞产生细胞因子、外周血 Ig E水平变化。结果 1致敏阶段应用 IL- 12可明显抑制嗜酸性粒细胞浸润、肺淋巴细胞对 IL- 5的分泌以及血浆总 Ig E和抗原特异性 Ig E的水平 ;2激发阶段应用 IL- 12可明显抑制嗜酸性粒细胞的浸润 ,抑制肺淋巴细胞产生 IL- 4、IL- 5 ,增加其产生 IFN- γ,但对抗原特异性 Ig E无明显影响 ;3如致敏和激发阶段均应用 IL- 12 ,则明显抑制肺淋巴细胞产生 IL- 4、IL- 5 ,增强 IFN- γ产生 ,抑制气道内嗜酸性粒细胞的浸润及血浆 Ig E的升高。结论气道应用 IL- 12对抗原诱导的气道过敏性炎症有明显调节作用 ,且与应用时机有关 ,为 IL- 12治疗哮喘提供依据     

气道内应用IL—2抑制抗原诱导的过敏性气道反应

目的 探讨气道内应用白细胞介素12（IL-12）对抗原诱导的过敏性反应的影响。方法 C57BL/6小鼠经OVA免疫建立哮喘模型,在主动免疫及抗原激发阶段气道内应用IL-12,观察肺泡灌洗液细胞成份、肺部淋巴细胞产生细胞因子、外周血IgE水平变化。结果 ①致敏阶段应用IL-12可明显抑制嗜酸性粒细胞浸润、肺淋巴细胞对IL-5的分泌以及血浆总IgE和抗原特异性IgE的水平;②激发阶段应用IL-12可明显抑制嗜酸性粒细胞的浸润,抑制肺淋巴细胞产生IL-4、IL-5,增加其产生IFN-γ,但对抗原特性IgE无明显影响;③如致敏和激发阶段均应用IL-12,则明显抑制肺淋巴细胞产生IL-4、IL-5,增强IFN-γ产生,抑制气道内嗜酸性粒细胞的浸润及血浆IgE的升高。结论 气道应用IL-12对抗原诱导的气道过敏性炎症有明显调节作用,且与应用时机有关,为IL-12治疗哮喘提供依据。     

Thl7淋巴细胞在哮喘小鼠气道炎症中的初步研究

目的:初步探索Th17细胞及其分泌的炎症介质在哮喘小鼠气道炎症中的作用机制.方法:20只小鼠随机均分为哮喘组和正常对照组.哮喘组用卵白蛋白(OVA)致敏与激发建立小鼠哮喘模型.正常对照组致敏与激发均以生理盐水代替.HE染色观察小鼠气道及肺组织病理变化;光学显微镜下观察小鼠支气管肺泡灌洗液(BALF)中细胞分类及计数;酶联免疫吸附试验(ELISA)检测小鼠BALF上清中IL-4、IL-5、IL-13、IFN-γ及IL-17的含量,流式细胞技术(FCM)检测小鼠外周血Th1、Th2及Th17淋巴细胞占CD4+T淋巴细胞百分率情况.结果:哮喘组小鼠BALF中细胞总数和中性粒细胞、嗜酸性粒细胞、淋巴细胞百分率均显著高于对照组(P<0.05),BALF上清中IL-4、IL-5、IL-13及IL-17的水平显著增高(P<0.05),而IFN-γ差异无统计学意义(P>0.05),外周血Th2、Th17细胞明显增高(P<0.05),而Th1细胞无明显变化.结论:Th17细胞及其分泌的炎症介质可促进中性粒细胞及嗜酸性粒细胞在气道内聚集,加重哮喘气道炎症,可能与哮喘气道重塑密切相关.     

抗白介素5单克隆抗体抑制哮喘小鼠嗜酸性粒细胞迁移

目的 观察抗白介素5(IL-5)单克隆抗体对哮喘小鼠嗜酸性粒细胞(Eos)从骨髓到气道迁移的影响.方法 15只雄性C57BL/6小鼠.常规法复制哮喘,并在每次卵白蛋白(Ova)激发前2 h鼻腔内滴入抗IL-5单克隆抗体,同时设立阴性对照组,即以生理盐水代替Ova 和抗IL-5抗体.在末次抗原激发后12 h测定支气管肺泡灌洗液(BALF)、外周血(PB)和骨髓(BM)中炎症细胞数以及肺组织内Eos数.结果 Ova抗原激发后12 h小鼠的BALF、PB和BM及细支气管和肺组织中的Eos数显著增加(P＜0.01, P＜0.05).抗IL-5单克隆抗体则使上述变化显著减轻(P＜0.05,P＜0.01).结论 抗IL-5单克隆抗体能显著抑制哮喘小鼠嗜酸性粒细胞的迁移.     

嗜酸性粒细胞募集在支气管哮喘发病中的作用

支气管哮喘是一种气道慢性炎症性疾病,伴有嗜酸性粒细胞增多、杯状细胞肥大、黏液分泌增多、可逆性的气道阻塞及对吸入变应原和非特异性刺激的高反应性等特点,其中嗜酸性粒细胞募集和随后的激活在支气管哮喘发病中起着十分重要的作用。本文就嗜酸性粒细胞募集在支气管哮喘发病机制中的研究进展作一综述,为支气管哮喘的治疗提供一线新的曙光。     

microRNAs:支气管哮喘的潜在新靶点

正支气管哮喘(简称哮喘)是以持续的气道炎症、气道高反应性和气道重塑为特征的慢性呼吸道疾病,有多种细胞及细胞组分的参与,包括嗜酸性粒细胞、中性粒细胞、T淋巴细胞及肥大细胞等炎症细胞,气道上皮细胞、平滑肌细胞等气道结构细胞和IL-4、IL-9、IL-13、FOXP3转录因子、TNF-α等细胞组分~([1])。哮喘临床出现反复发作的呼吸道症状,如反复的喘息、气急、胸闷、咳嗽等~([2])。据统计,全球     

支气管哮喘患者血清骨桥蛋白和嗜酸细胞趋化因子表达及临床意义

支气管哮喘是慢性气道炎症性疾病,嗜酸性粒细胞在气道炎症反应中起重要作用[1]。现认为细胞因子网络失衡是发病的基础,而Th2细胞因子、嗜酸细胞趋化因子(eotaxin)共同诱导的嗜酸性粒细胞募     

The role of the bone marrow in allergy and asthma

The above studies have begun to address the fundamental question of the mechanisms of bone marrow involvement and response to allergen challenge in allergic asthmatics. Further studies in this area should complement our investigations in human asthma — which suggest that a particular bias toward differentiation of Eo-Baso progenitors characterizes the atopic state — as well as our findings in the dog model of allergen-induced airway hyperresponsiveness, which indicate that the bone marrow responds to inhalation of allergen or corticosteroids. Taken in the context of previous indications that IgE and bronchial responsiveness may both be transferrable through bone marrow transplantation (log), these findings indicate a physiologic role for the bone marrow in allergic inflammation. Likewise, these concepts provide a basis for making certain predictions regarding management and novel therapeutic interventions in atopy and asthma.     

Is rhinosinusitis a cause of asthma?

There is a great deal of evidence of an association between rhinosinusitis and asthma. However, it is less clear whether rhinosinusitis is a direct trigger for asthma or the two conditions are simply manifestations of a common underlying process. Evidence for a role for rhinosinusitis as a trigger for asthma includes many examples of improvement in asthma once concomitant rhinosinusitis is treated medically or surgically. Possible mechanisms for this relationship include naso-pharyngo-bronchial reflexes, postnasal drip, abnormal breathing, and the local production of inflammatory mediators that trigger pulmonary inflammation via the bone marrow. On the other hand, evidence exists that rhinosinusitis and asthma are manifestations of a common process. For example, there are similarities between the histopathological changes in the epithelium in chronicrhinosinusitis and asthma. The bone marrow may provide the link between the upper and lower airways in creating a common disease. A second possible mechanism for a common disease is response to staphylococcal enterotoxins. Although evidence exists to suggest that rhinosinusitis either triggers asthma or represents a local manifestation of a shared disorder, the key to reconciling this apparent controversy is to consider that rhinosinusitis is not just a single, uniform disease. Current evidence suggests that rhinosinusitis with neither polyps nor eosinophilic inflammation acts as a direct trigger for asthma, whereas rhinosinusitis with both polyps and eosinophilic inflammation shares underlying mechanisms with asthma. Clearly, however, there is considerable overlap between the different, complex mechanisms that link rhinosinusitis to asthma.     

骨髓间充质干细胞培养上清液治疗支气管哮喘及对肺部炎症的影响

背景：医学界普遍认为支气管哮喘是一种与Th2相关的免疫反应疾病,目前尚缺乏理想的治疗方法。骨髓间充质干细胞作为一种成体干细胞,不仅具有增殖和多向分化能力,还具有低免疫原性和免疫调节能力。 目的：探讨骨髓间充质干细胞培养上清液对支气管哮喘小鼠肺部炎症的影响。 方法：将20只实验小鼠随机分为对照组和实验组,每组10只,在第0天及第14天小鼠腹腔注射卵清蛋白溶液致敏,并在第24-26天雾化吸入卵清蛋白溶液激发。实验组从第24天开始,在每次激发前2 h,腹腔注射制备好的骨髓间充质干细胞培养上清液2 mL,对照组腹腔注射生理盐水。末次激发后麻醉致小鼠安乐死亡,采取血清、肺泡灌洗液及肺脏组织进行研究。 结果与结论：①对照组小鼠肺组织结构发生异常,黏膜下层和肌层内能够看见大量嗜酸性粒细胞及单核细胞浸润,经骨髓间充质干细胞培养液治疗后,实验组小鼠肺部炎症较轻。②实验组白细胞总数、嗜酸性粒细胞数、嗜酸性粒细胞百分比显著低于对照组(P < 0.01)。③实验组肺泡灌洗液以及血清中白细胞介素17水平显著低于对照组(P < 0.05);肺泡灌洗液以及血清中白细胞介素4水平与对照组比较差异无显著性意义(P > 0.05)。上述结果提示支气管哮喘小鼠腹腔注入骨髓间充质干细胞培养上清液能够减轻肺部炎性病理反应严重程度,降低肺泡灌洗液以及血清中相关的炎症指标水平。  中国组织工程研究杂志出版内容重点：干细胞;骨髓干细胞;造血干细胞;脂肪干细胞;肿瘤干细胞;胚胎干细胞;脐带脐血干细胞;干细胞诱导;干细胞分化;组织工程      

Monotypic immunoglobulin E plasma cells in an allogeneic bone marrow transplant recipient

A patient with acute lymphoblastic leukaemia in first remission received a bone marrow transplant from his HLA-identical brother. The patient had a remote history of asthma and the bone marrow donor had allergic asthma. The patient developed acute graft-versus-host disease and died 2 months after transplantation. At autopsy there were high numbers of plasma cells in lymphoid tissues. The majority of this cell population was of polytypic IgG, IgM or IgA origin, but there was a significant contribution by monotypic IgE-lambda-containing cells, varying from 10% in the appendix to 35% in lymph node. The serum IgE level in the patient was less than 0.5 IU/ml before transplantation, and 8.5 IU/ml 1 month thereafter. In the donor the value was about 400 IU/ml. In the donor only, specific IgE antibodies to various allergens were detectable. The bone marrow of the donor contained 0.4% plasma cells, of which 36% were IgE positive (chi/lambda ratio 1/11). These findings are compatible with literature data on elevations in serum IgE level following bone marrow transplantation. We suggest that the IgE-lambda plasma cell population is of donor origin.     

Anti-IL-5 (mepolizumab) therapy induces bone marrow eosinophil maturational arrest and decreases eosinophil progenitors in the bronchial mucosa of atopic asthmatics

BACKGROUND: Eosinophils develop from CD34(+) progenitors under the influence of IL-5. Atopic asthmatic individuals have increased numbers of mature eosinophils and eosinophil pro-genitors within their bone marrow and bronchial mucosa. We have previously reported that anti-IL-5 monoclonal antibody treatment decreases total bone marrow and bronchial mucosal eosinophil numbers in asthma. OBJECTIVE: Using an anti-IL-5 monoclonal antibody, we examined the role of IL-5 in eosinophil development within the bone marrow and bronchial mucosa in asthma. METHODS: Blood, bone marrow, and airway mucosal biopsy specimens were examined before and after anti-IL-5 (mepolizumab) treatment of asthmatic individuals in a double-blind, placebo-controlled trial. Numbers of mature and immature eosinophils were measured by histologic stain (bone marrow myelocytes, metamyelocytes, and mature eosinophils), flow cytometry (bone marrow and blood CD34(+)/IL-5Ralpha(+) cells), enumeration of bone marrow-derived eosinophil/basophil colony-forming units in methylcellulose culture, and sequential immunohistochemistry and in situ hybridization (bronchial mucosal CD34(+)/IL-5Ralpha mRNA(+) cells). RESULTS: Mepolizumab decreased mature eosinophil numbers in the bone marrow by 70% (P =.017) in comparison with placebo and decreased numbers of eosinophil myelocytes and metamyelocytes by 37% (P =.006) and 44% (P =.003), respectively. However, mepolizumab had no effect on numbers of blood or bone marrow CD34(+), CD34(+)/IL-5Ralpha(+) cells, or eosinophil/basophil colony-forming units. There was a significant decrease in bronchial mucosal CD34(+)/IL-5Ralpha mRNA(+) cell numbers in the anti-IL-5 treated group (P =.04). CONCLUSION: These data suggest that anti-IL-5 therapy might induce partial maturational arrest of the eosinophil lineage in the bone marrow. The reduction in airway CD34(+)/IL-5 mRNA(+) cell numbers suggests that IL-5 might also be required for local tissue eosinophilopoiesis.     

Functional Role of FcγRIIB in the Regulation of Mesenchymal Stem Cell Function

Mesenchymal stem cells (MSCs) derived from bone marrow are plural-potent stem cells with immune regulatory functions. We aimed to evaluate role of FcγRIIB in the regulation of bone marrow-derived MSC function. MSCs were prepared from mouse bone marrow derived from wild-type (WT) or FcγRIIB-deficient (FcγRIIB-/-) mice. MSCs were co-cultured with bone marrow-derived dendritic cells (BMDCs), and BMDC maturation and function were evaluated by flow cytometric analysis and carboxyfluorescein succinimidyl ester-labeled OT-II T-cell addition. An acute asthma model was established by aeresol ovalbumin challenge in mice. Mice received WT or FcγRIIB-/- MSC therapy. Lung function was evaluated by histological examination and cytokine production measurement. mRNA and protein expression levels of target genes were examined by real-time quantitative polymerase chain reactionor western blotting. We found that MSCs derived from bone marrow exhibit a high level of FcγRIIB expression. FcγRIIB deficiency impaired the suppressive function of MSCs, as FcγRIIB deficiency efficiently reversed the inhibitory effect of MSCs on BMDC maturation and function. Additionally, FcγRIIB-/-MSCs were less potent at suppressing asthma in model mice, possibly through reduced expression of Smad2, Smad3, Cox-2, and prostaglandin E2 in FcγRIIB-/-MSCs. FcγRIIB might play an essential role in regulating the inhibitory effects of MSCs derived from bone marrow.     

哮喘豚鼠模型肺和骨髓中CCR3及CD34的表达及意义

目的:观察哮喘豚鼠肺和骨髓组织CCR3及EOS不同时相表达的变化, 探讨哮喘发病的可能机制.方法:用卵白蛋白(OVA)和生理盐水致敏法制备豚鼠哮喘和对照模型, 分为正常对照组(N)及哮喘组(A、B、C、D、E), 每组8只, 于激发后30 min, 6 h, 12 h, 24 h, 48 h处死, 瑞氏染色计数骨髓涂片和外周血涂片中EOS比例, 免疫组织化学技术检测骨髓中CCR3及CD34蛋白的表达;制备豚鼠肺组织病理标本, 苏木精-伊红染色, 免疫组化检测肺组织中CCR3和CD34的表达.结果:(1)哮喘组骨髓和外周CCR3与CD34及EOS表达明显增加;(2)OVA激发后, 骨髓和外周CCR3、CD34及EOS表达:30 min变化不明显(肺内CD34表达增加), 6 h(肺内CD34表达下降)、12 h达到峰值, 24 h开始下降, 48 h恢复正常水平;(3)骨髓的变化早于外周, CCR3的表达高峰早于CD34表达与EOS的增多高峰, 且CCR3、CD34和EOS互呈线性相关(肺组织内除外).结论:哮喘豚鼠肺组织和骨髓之间存在骨髓CD34 祖细胞、EOS细胞到循环再到肺组织的通路, 骨髓和肺组织CCR3表达增强, 为CD34 细胞、EOS从骨髓快速募集到肺组织提供了可能.     

Transfer of allergen-specific IgE-mediated hypersensitivity with allogeneic bone marrow transplantation

We investigated whether allergen-specific IgE-mediated hypersensitivity is transferred by bone marrow transplantation. Twelve patients, 14 to 47 years of age, undergoing allogeneic bone marrow transplantation for the treatment of hematologic cancer were selected, along with their donors, by a screening questionnaire for a history of atopy in the donor. We evaluated these donor-recipient pairs before transplantation and at several points afterward for immediate skin-test reactivity to 17 allergens. For allergens for which pretransplantation skin tests had been positive in the donors and negative in the recipients, 20 of 46 post-transplantation skin tests were positive in 8 of the 11 recipients who survived for more than one year after transplantation. For allergens for which both donors and recipients had had negative skin tests before transplantation, only 6 of 256 tests (2.3 percent) were positive in the recipients after transplantation. Long-term transfer of donor-derived mite-specific IgE was demonstrated by radioallergosorbent testing in two recipients. Seven recipients either acquired or had an exacerbation of allergic rhinitis, and two recipients without a history of asthma had asthma one year after transplantation. We conclude that allergen-specific IgE-mediated hypersensitivity is adoptively transferred by bone marrow transplantation from donor to recipient by B cells with allergen-specific memory.     

Interleukin-17 orchestrates the granulocyte influx into airways after allergen inhalation in a mouse model of allergic asthma

Interleukin (IL)-17 is produced by activated memory CD4(+) cells and induces cytokines and chemokines that stimulate neutrophil generation and recruitment. Here, we investigated the involvement of IL-17 in the bronchial influx of neutrophils in experimental allergic asthma. Inhalation of nebulized ovalbumin (OVA) by sensitized mice with bronchial eosinophilic inflammation resulting from chronic OVA exposure induced early IL-17 mRNA expression in inflamed lung tissue, concomitant with a prominent bronchial neutrophilic influx. Anti-IL-17 monoclonal antibodies (mAb) injected before allergen inhalation strongly reduced bronchial neutrophilic influx, in a manner equally as potent as the anti-inflammatory dexamethasone. Remarkably, anti-IL-17 mAb significantly enhanced IL-5 levels in both BAL fluid and serum, and aggravated allergen-induced bronchial eosinophilia. In another series of experiments, anti-IL-17 mAb were given repeatedly during the inhalatory challenge phase with OVA of sensitized mice. This treatment regimen abated bronchial neutrophilia in parallel with reduction of bone marrow and blood neutrophilia. In addition, anti-IL-17 mAb treatment elevated eosinophil counts in the bone marrow and bronchial IL-5 production, without alteration of allergen-induced bronchial hyperresponsiveness. In summary, our results demonstrate that IL-17 expression in airways is upregulated upon allergen inhalation, and constitutes the link between allergen-induced T cell activation and neutrophilic influx. Because neutrophils may be important in airway remodeling in chronic severe asthma, targeting IL-17 may hold therapeutic potential in human asthma.