IgE-FcεRI交联促进过敏性哮喘小鼠动脉粥样硬化的发生发展

目的:观察过敏性哮喘加速小鼠动脉粥样硬化(AS)的发生和发展是否与Th2细胞及白细胞介素4(IL-4)有关,以及免疫球蛋白E(IgE)-Fc ε受体I(FcεRI)交联激活巨噬细胞途径在其中发挥的作用。方法:取6周龄的ApoE~(-/-)小鼠,以卵清蛋白致敏和激发建立过敏性哮喘模型,并分为对照组、哮喘安慰剂组和哮喘IL-4单克隆抗体干预组,分别干预8周,干预结束后处死小鼠,油红O染色检测主动脉根部斑块面积,流式细胞术检测脾脏Th2细胞比例,real-time PCR检测IL-4、IL-6、单核细胞趋化蛋白1(MCP1)和巨噬细胞炎症蛋白1α(MIP1α)的mRNA表达水平,ELISA法检测血清IL-4和IgE的含量。结果:与对照组相比,过敏性哮喘ApoE~(-/-)小鼠主动脉根部AS病变显著加重,并伴有体内Th2细胞和IL-4水平的增高,同时斑块处IgE和FcεRIα的表达显著增高,MCP-1、MIP-1α和IL-6的mRNA表达也显著增加;IL-4单克隆抗体干预8周后,主动脉根部AS病变缓解的同时,增高的IgE和FcεRIα表达被显著抑制,巨噬细胞相关炎性因子的表达水平也显著降低。结论:过敏性哮喘显著加速ApoE~(-/-)小鼠AS病变进展,此作用与体内Th2细胞和IL-4水平增加,以及IgE-FcεRI交联激活巨噬细胞途径有关。     

Fcγ和Fcε受体在小鼠三叉神经节的表达

目的 检测正常小鼠三叉神经节(TG)是否表达免疫球蛋白G(IgG)和免疫球蛋白E(IgE) Fc段Fcγ受体和Fcε受体,及其在过敏小鼠TG中的变化.方法 通过腹腔注射OVA和铝剂,建立小鼠过敏性结膜炎(ACJ)模型.EHSA检测血清总IgE.用Westernblot和免疫荧光检测Fcγ受体和Fcε受体的表达.结果 小鼠TG表达Fcγ受体和Fcε受体.其中IgG激活型高亲和力受体FcγRI和抑制型低亲和力受体FcγRⅡ只表达在小鼠TG神经元上,而IgG激活型低亲和力受体FcγRⅢ表达在小鼠TG中的卫星胶质细胞上.IgE激活型高亲和力受体FcεR Ⅰ表达在小鼠TG神经元上,而IgE低亲和力受体FcεRⅡ同时表达在小鼠TG神经元和卫星胶质细胞上.与正常小鼠比较,ACJ小鼠的血清总IgE水平升高,TG的FcεR Ⅰ和FcγRⅡ表达增加(P＜0.05).而ACJ小鼠的FcεRⅡ和FcγRⅠ表达下降(P＜0.05).FcγRⅢ在正常和ACJ小鼠TG中的表达无显著差别.结论 小鼠TG表达的Fcγ和Fcε受体可能参与ACJ及其他过敏性疾病的发生和发展.     

支气管哮喘与过敏性鼻炎患者对蟑螂变应原特异性IgE反应的对比研究

为比较支气管哮喘与过敏性鼻炎患者对不同种属蟑螂的过敏反应程度,探讨不同种属蟑螂间可能存在的交叉抗原性及其程度,用ELISA法检测支气管哮喘与过敏性鼻炎患者血清中对三种蟑螂(美洲大蠊、黑胸大蠊和德国小蠊)变应原的特异性IgE抗体(sIgE).结果显示支气管哮喘患者对美洲大蠊、黑胸大蠊和德国小蠊变应原的sIgE阳性率分别为23.5%、16.0%和14.0%,美洲大蠊sIgE阳性率比对德国小蠊为高,有显著性差异(P=0.015),虽然美洲大蠊sIgE阳性率比对黑胸大蠊高、黑胸大蠊sIgE阳性率比对德国小蠊高,但差异均无统计学意义(P=0.060;P=0.575);哮喘患者中美洲大蠊与黑胸大蠊sIgE反应符合率74.0%,美洲大蠊与德国小蠊sIgE反应符合率73.5%,黑胸大蠊与德国小蠊sIgE反应符合率85.0%.过敏性鼻炎患者对美洲大蠊、黑胸大蠊和德国小蠊sIgE阳性率分别为24.8%、17.6%和15.8%,美洲大蠊sIgE阳性率比对德国小蠊高,有显著性差异(P=0.040),虽然美洲大蠊sIgE阳性率比对黑胸大蠊高、黑胸大蠊sIgE阳性率比对德国小蠊高,但差异均无统计学意义(P=0.106,P=0.658);过敏性鼻炎患者对中美洲大蠊与黑胸大蠊sIgE反应符合率为73.9%,对美洲大蠊与德国小蠊sIgE反应符合率为75.2%,对黑胸大蠊与德国小蠊sIgE反应符合率为86.1%.过敏性鼻炎患者对美洲大蠊、黑胸大蠊和德国小蠊sIgE阳性率均比支气管哮喘患者高,但差异均无统计学意义(P=0.764;P=0.688;P=0.638).提示支气管哮喘与过敏性鼻炎患者对三种蟑螂的过敏反应程度基本相当,三种蟑螂间可能存在一定程度的交叉抗原性成分.     

巨噬细胞极化在过敏性哮喘中的作用

过敏性哮喘(allergic asthma,AA)是常见的慢性呼吸道疾病,严重危害人们的身体健康.巨噬细胞是存在于肺组织中最丰富的免疫细胞群,参与了过敏性哮喘的发病过程.在受到不同变应原刺激后,巨噬细胞表现出不同的活化状态,并发挥不同的免疫功能,这一过程称之为巨噬细胞极化(mac-rophge polarization).这里主要讨论巨噬细胞及巨噬细胞极化在过敏性哮喘发病过程中的作用,以及巨噬细胞极化过程中表观遗传学变化的新概念,包括miRNAs、DNA甲基化和组蛋白修饰等,它们可能通过调节细胞信号和标志基因的表达来调控巨噬细胞极化,从而影响了过敏性哮喘发病过程,为过敏性哮喘的免疫治疗策略提供了新的见解.     

沉默Fcgr3降低SiO2诱导的小鼠肺泡巨噬细胞系MH-S炎性因子表达

目的 探讨沉默免疫球蛋白G Fc段受体Ⅲ(FcγRⅢ)对SiO₂诱导的小鼠肺泡巨噬细胞系MH-S炎性因子表达的影响。方法 构建Fcgr3 shRNA的慢病毒质粒;人胚肾细胞系293T培养包装产生慢病毒;慢病毒感染沉默MH-S中的FcγRⅢ,并筛选稳定感染的细胞。定量多聚酶链反应(qPCR)和蛋白免疫印迹法(Western blot)检测MH-S巨噬细胞系中FcγRⅢ的mRNA和蛋白表达。将MH-S细胞分为MH-S+sh-NC、MH-S+SiO₂+sh-NC、MH-S+sh-Fcgr3和MH-S+SiO₂+sh-Fcgr3,其中SiO₂处理条件为100 mg/L连续刺激12 h。通过qPCR检测炎性因子转录水平,酶联免疫吸附测定(ELISA)检测细胞培养上清中的炎性因子的含量,包括肿瘤坏死因子(TNF-α)、白介素1β(IL-1β)和白介素6(IL-6)。结果 包装慢病毒并感染细胞后建立稳定沉默Fcgr3的MH-S细胞系,稳定转染细胞中FcγRⅢ的转录与蛋白表达水平较对照组降低(P<0.05)。...     

小儿过敏性哮喘与过敏性鼻炎的相关性

小儿过敏性哮喘与过敏性鼻炎在临床上较为常见,为了临床上更好的诊断,预防,治疗这两种疾病,本文从流行病学,临床症状,解剖学特点,发病特点,炎性介质以及临床治疗等方面简要论述两种疾病的相关性,认为除临床症状外,两种疾病在多方面存在着较大的相似性。     

过敏性哮喘的免疫治疗研究进展

过敏性哮喘是一种顽固性疾患,多数在婴幼儿时期发病,若得不到有效治疗将会伴随终身。大部分哮喘患者都存在过敏现象或者有过敏性鼻炎,有过敏性鼻炎的哮喘患者发病前兆会有打喷嚏、流鼻涕、鼻痒、眼痒、流泪等症状。由于症状与呼吸道感染或炎症相似,大人缺乏相关知识,往往在早期忽视治疗,也极有可能被误诊[1]。     

TAP1基因和FcεRIβ基因多态性与哮喘及其表型的相关性研究

目的：为研究TAP1基因和FcεRIβ基因多态性与哮喘及其表型的关系。方法：选择TAP1基因中微卫星DNA标志-TAP1及FcεRIβ基因中RFLP位点-RsaI,采用Amp-FLP和RFLP方法在散哮喘病人中进行分析。并检测其血清IgE水平,过敏原发皮试及气道高反应性,应用相关分析观察TAP1及RsaI等位基因与哮喘及其表型的关系。结果：RsaI位点等位基因片段与哮喘及气道高反应性相关,未见TAP1与哮喘或其表型相关。结论：FcεRIβ基因可能是哮喘发病的一个候选基因,TAP1基因可能与哮喘的发病无关。     

巨噬细胞载脂蛋白E的分泌与动脉粥样硬化

载脂蛋白Ｅ在体内参与血脂的转运和代谢,可由多种器官和组织分泌,包括许多组织中的巨噬细胞。动脉粥样硬化斑块内巨噬细胞载脂蛋白Ｅ的分泌受众多因素调节,并通过几种不同的局部作用机制对动脉粥样硬化的发生、发展产生重要影响。     

Wnt/β-catenin通路在动脉粥样硬化发生发展中的作用

Atherosclerosis is a multifactorial process associated with endothelial cell injury and dysfunction, inflammation, oxidative stress, cell proliferation, angiogenesis and so on, all of which play a crucial role in atherosclerosis. Wnt/β-catenin signaling pathway is highly conservative in the development of body, abnormal activation of which is related to types of diseases including cancer. Accumulating studies have shown that Wnt/β-catenin signaling pathway is involved in inflammation, oxidative stress and so on. This article would make a review about the role of Wnt/β-catenin signaling pathway in atherosclerosis based on the pathogenic mechanisms underlying atherosclerosis as mentioned above.     

A VLP‐based vaccine against interleukin‐1α protects mice from atherosclerosis

Interleukin (IL)‐1α is a potent proinflammatory cytokine that has been implicated in the development of atherosclerosis. We investigated whether a vaccine inducing IL‐1α neutralizing antibodies could interfere with disease progression in a murine model of atherosclerosis. We immunized Apolipoprothin E (ApoE)‐deficient mice with a vaccine (IL‐1α‐C‐Qβ) consisting of full‐length, native IL‐1α chemically conjugated to virus‐like particles derived from the bacteriophage Qβ. ApoE^?/? mice were administered six injections of IL‐1α‐C‐Qβ or nonconjugated Qβ over a period of 160 days while being maintained on a western diet. Atherosclerosis was measured in the descending aorta and in cross‐sections at the aortic root. Macrophage infiltration in the aorta was measured using CD68. Expression levels of VCAM‐1, ICAM‐1, and MCP‐1 were quantified by RT‐PCR. Immunization against IL‐1α reduced plaque progression in the descending aorta by 50% and at the aortic root by 37%. Macrophage infiltration in the aorta was reduced by 22%. Inflammation was also reduced in the adventitia, with a decrease of 54% in peri‐aortic infiltrate score and reduced expression levels of VCAM‐1 and ICAM‐1. Active immunization targeting IL‐1α reduced both the inflammatory reaction in the plaque as well as plaque progression. In summary, vaccination against IL‐1α protected ApoE^?/? mice against disease, suggesting that this may be a potential treatment option for atherosclerosis.     

Targeted IgA Fc receptor I (FcαRI) therapy in the early intervention and treatment of pristane‐induced lupus nephritis in mice

The Fc receptor I for IgA (FcαRI) down‐regulates humoral immune responses and modulates the risk of autoimmunity. This study aimed to investigate whether FcαRI targeting can affect progression of pristine‐induced lupus nephritis. In the first experiment (early intervention), four groups of animals were evaluated: untreated FcαRI/FcRγ transgenic (Tg) mice and Tg mice administered control antibody (Ctr Fab), saline and anti‐FcαRI Fab [macrophage inflammatory protein (MIP)‐8a], respectively, three times a week for 29 weeks, after being injected once intraperitoneally with 0·5 ml pristane. In the second experiment, antibody injection started after the onset of nephritis and was carried out for 2 months, with similar groups as described above. MIP‐8a improved proteinuria, decreased the amounts of glomerular injury markers, serum interleukin (IL)‐6, IL‐1 and monocyte chemoattractant protein (MCP)‐1, and F4/80 macrophages in the interstitium and glomeruli, in both experiments. When MIP‐8a was used as early intervention, a decrease in mouse serum anti‐nuclear antibody (ANA) titres and reduced deposition of immunoglobulins in glomeruli were observed. This effect was associated with reduced serum titres of immunoglobulin (Ig)G2a but not IgG1, IgG2b and IgG3. Furthermore, pathological analysis showed lower glomerular activity index and less fibronectin in MIP‐8a treated mice. This study suggests that FcαRI targeting could halt disease progression and lupus activation by selective inhibition of cytokine production, leucocyte recruitment and renal inflammation. Our findings provide a basis for the use of FcαRI as a molecular target for the treatment of lupus.     

Pristimerin attenuates ovalbumin-induced allergic airway inflammation in mice

Pristimerin has been shown to possess antiinflammatory activity. However, its potential use for asthma induced by airway inflammation has not yet been studied. First, we established a ovalbumin (OVA)-induced allergic asthma mice model. BALB/c mice were immunized and challenged by OVA. Treatment with pristimerin caused a marked reduction in the levels of OVA-specific IgE, immune cells, and IL-4, IL-5, IL-13 secretion. Histological studies using H&E staining were used to study the alterations in lung tissue. These results were similar to those obtained with dexamethasone treatment. We then investigated which signal transduction mechanisms could be implicated in pristimerin activity by Western blot. The data showed that pristimerin could inhibit MAPKs and NF-κB inflammatory pathways.     

薯蓣皂苷减轻卵清蛋白诱导的过敏性哮喘小鼠的气道炎症

目的探讨薯蓣皂苷对卵清蛋白(ovalbumin,OVA)诱导的过敏性哮喘小鼠中过敏性支气管炎的影响及机制。方法24只小鼠随机分为对照组、OVA组、OVA+30 mg/kg薯蓣皂苷组和OVA+60 mg/kg薯蓣皂苷组,每组纳入6只小鼠。全自动生化仪检测各组支气管肺泡灌洗液(bronchoalveolar lavage fluid,BALF)中嗜中性粒细胞、嗜酸性粒细胞和单核细胞的数量;ELISA法检测BALF中促炎因子IL-1β、IL-4、IL-5和TNF-α的含量;PAS染色法观察肺组织黏液分泌情况并对黏液分泌程度进行评分;免疫组化法观察肺组织p-NF-κB p65的表达和分布情况;Western blotting法检测肺组织中p-IκB和NF-κB p65的蛋白表达水平。结果薯蓣皂苷可降低过敏性哮喘小鼠BALF中炎性细胞数量,同时降低BALF中促炎因子IL-1β、IL-4、IL-5和TNF-α的水平以及肺部黏液的分泌和NF-κB的活化水平。结论薯蓣皂苷能减轻过敏性哮喘小鼠的气道炎症,且其抗炎作用与抑制NF-κB活化有关。     

Development of atherosclerosis in Balb/c apolipoprotein E-deficient mice

BACKGROUND: Since its creation in 1992 by gene inactivation via gene targeting, the apolipoprotein E "knockout" mouse has become the most widely used rodent model for the study of atherosclerosis. Commercially available apolipoprotein E(-/-) mice are bred on a C57BL/6J background. The goal of the present study was to investigate the development of atherosclerosis in apolipoprotein E-deficient mice generated on a Balb/c background. METHODS: We compared serum cholesterol concentrations and the development of atherosclerotic lesions in heterozygous Balb/c [apolipoprotein E(+/-)] mice fed regular rodent chow, Balb/c apolipoprotein E-deficient mice fed regular chow, and Balb/c apolipoprotein E-deficient mice fed a high-fat diet for up to 30 weeks. Expression of the chemokine JE (murine homologue of MCP-1), as well as the adhesion molecules E-selectin, intercellular adhesion molecule-1, and vascular cell adhesion molecule-1, in the aortas of knockout mice fed a high-fat diet was measured by enzyme-linked immunosorbent assay. RESULTS: Balb/c apolipoprotein E-deficient mice develop atherosclerotic lesions in a reproducible temporal and morphological pattern. Total serum cholesterol concentrations in Balb/c apolipoprotein E-deficient mice fed regular chow or a high-fat diet, respectively, closely parallel those reported for C57BL/6J apolipoprotein E-deficient mice. The expression of all three adhesion molecules in the aorta follows a similar temporal pattern, peaking in the first 15 weeks, whereas JE concentrations peak around 23 weeks. CONCLUSION: The availability of Balb/c apolipoprotein E-deficient mice will facilitate the study of atherosclerosis in a mouse strain that can concomitantly develop other pathological states that are not readily inducible in mice with the C57BL/6J background.     

TLR4 Asp299Gly、Thr399Ile基因多态性对变应性哮喘的发病及IgE水平的影响

目的　探讨TLR4 (toll likereceptor 4 )Asp2 99Gly、Thr399Ile基因多态性对变应性哮喘的发病和血浆IgE水平的影响。方法　利用聚合酶链反应 限制性片段长度多态性分析技术 (PCR RFLP) ,对 1 97例变应性哮喘患者和 1 5 6例健康人进行TLR4的Asp2 99Gly、Thr399Ile两位点的基因型检测。同时利用免疫发光法检测血浆IgE的水平。结果　 1 97例变应性哮喘患者TLR4基因Asp2 99Gly位点Asp Asp、Asp Gly和Gly Gly的基因型频率为 0 .81 7、0 .1 4 7和 0 .0 36 ,与正常对照组相比差异无统计学意义 (χ2 =0 .0 32 ,P =0 .984 ) ;但变应性哮喘患者Gly Gly、Asp Gly基因型血浆总IgE对数值 ( x±s:2 .6 1 5± 0 .6 0 0 1 ,n =36 )与Asp Asp基因型血浆总IgE对数值 ( x±s:2 .2 4 0± 0 .6 894 ,n =1 6 1 )相比较高 ,差异有统计学意义 (P =0 .0 0 2 )。TLR4基因Thr399Ile位点Thr Thr、Thr Ile和Ile Ile的基因型频率为 0 .970、0 .0 2 0和 0 .0 1 0 ,与正常对照组相比差异无统计学意义 (χ2 =0 .6 2 0 ,P =0 .733) ;变应性哮喘患者Ile Ile、Thr Ile基因型血浆总IgE对数值 ( x±s:2 .4 1 7± 0 .4 4 2 3,n =6 )与Thr Thr基因型血浆总IgE对数值 ( x±s:2 .30 5± 0 .6 94 9,n =1 91 )相比差异无统计学意义 (P =0 .5 71 )。     

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     

Oxidized high‐density lipoprotein accelerates atherosclerosis progression by inducing the imbalance between treg and teff in LDLR knockout mice

High density lipoprotein (HDL) dysfunction has been widely reported in clinic, and oxidation of HDL (ox‐HDL) was shown to be one of the most common modifications in vivo and participate in the progression of atherosclerosis. But the behind mechanisms are still elusive. In this study, we firstly analyzed and found strong relationship between serum ox‐HDL levels and risk factors of coronary artery diseases in clinic, then the effects of ox‐HDL in initiation and progression of atherosclerosis in LDLR knockout mice were investigated by infusion of ox‐HDL dissolved in chitosan hydrogel before the formation of lesions in vivo. Several new evidence were shown: (i) the serum levels of ox‐HDL peaked early before the formation of lesions in LDLR mice fed with high fat diet similar to oxidative low density lipoprotein, (ii) the formation of atherosclerotic lesions could be accelerated by infusion of ox‐HDL, (iii) the pro‐atherosclerotic effects of ox‐HDL were accompanied by imbalanced levels of effector and regulatory T cells and relative gene expressions, which implied that imbalance of teff and treg might contribute to the pro‐atherosclerosis effects of ox‐HDL.