地塞米松对呼吸道合胞病毒感染哮喘加重小鼠气道TSLP分泌及炎症的影响

目的:探讨地塞米松(DEX)对呼吸道合胞病毒(RSV)感染哮喘加重小鼠胸腺基质淋巴细胞生成素(TSLP)分泌及气道炎症的影响。方法:雌性BALB/c小鼠32只,随机分成4组,分别为磷酸盐缓冲液(PBS)对照组、鸡卵白蛋白(OVA)组、OVA/RSV组、OVA/RSV/DEX组;应用OVA腹腔注射致敏、OVA气道雾化结合RSV滴鼻激发哮喘,地塞米松1mg/kg肌肉注射;无创肺功能检测各组小鼠气道反应性;ELISA法检测小鼠血清IL-4、IL-5、IL-13、IFNγ-和气管灌洗液(BALF)TSLP含量;小鼠肺组织病理观察炎症反应,免疫组化观察小鼠气道上皮细胞TSLP表达水平。结果:无创肺功能检测显示地塞米松抑制RSV感染哮喘加重小鼠气道反应性的增高,OVA/RSV/DEX组小鼠Penh值明显低于OVA/RSV组(P<0.01);OVA/RSV/DEX组小鼠血清IL-4、IL-5、IL-13、IFNγ-浓度[分别为(86.78±27.04)、(227.66±40.87)、(194.65±73.27)和(17.33±3.06)pg/ml]和BALF中TSLP浓度[(1 873±10)pg/ml],均明显低于OVA/RSV组[分别为(274.2±103.7)、(293.3±46.1)、(330±93.5)、(30.1±5.7)、(2 127±46)pg/ml](P<0.01);病理观察显示地塞米松显著减轻RSV感染哮喘小鼠气道炎症细胞浸润;免疫组化染色证实地塞米松抑制RSV感染哮喘小鼠气道上皮细胞TSLP表达。结论:地塞米松可以抑制RSV感染哮喘加重小鼠气道上皮细胞表达TSLP,减轻RSV感染哮喘加重小鼠气道炎症反应。     

胸腺基质淋巴生成素受体及其抗体在哮喘小鼠气道炎症反应中的作用

目的 探讨胸腺基质淋巴生成素受体(TSLPR)及其抗体在实验性哮喘小鼠气道炎症反应中的作用以及对气道树突状细胞(DCs)成熟和活化的影响.方法 BALB/c小鼠随机分成A、B、C三组.B和C组小鼠腹腔注射卵清白蛋白(OVA)致敏,A组腹腔注射PBS作为正常对照.B和C组小鼠在OVA激发哮喘发作前分别吸入非特异性IgG和TSLPR IgG.采集各组小鼠支气管肺泡灌洗液(BALF)进行细胞分类计数,采用ELISA定量检测BALF中IL-4、IL-5、IFN-γ和IL-10浓度.采集各组小鼠肺组织标本进行病理学检查,采用流式细胞术分别检测各组小鼠淋巴结和肺组织中DCs数量和表型.结果 与A组小鼠比较,8和C组小鼠BALF中各种细胞因子水平均明显升高(P<0.01).C组小鼠BALF中IL-4和IL-5水平低于8组(P<0.05,P<0.01),IFN-γ和IL-10水平高于8组(P<0.05,P<0.01).C组小鼠BALF中细胞总数、嗜酸性粒细胞和淋巴细胞数也明显低于B组(P<0.01).B组小鼠支气管周围有大量炎性细胞浸润以及杯状细胞增生,黏液分泌增强,C组小鼠仅见微弱的炎性细胞浸润和杯状细胞增生.B组小鼠膈淋巴结中DCs数量以及肺组织中DCs的I-Ad、CD40、CD80和CD86表达水平均高于C组小鼠(P<0.05).结论 TSLP/TSLPR具有促哮喘效应并与其调节气道DCs活性的作用密切相关,TSLPR抗体干预可明显减弱TSLP/TSLPR上述作用,故有作为抗哮喘药物的前景.     

抗CD86单克隆抗体对哮喘小鼠TH1/TH2细胞因子的影响

目的观察抗CD86单克隆抗体对卵蛋白干粉(OVA)致敏并刺激小鼠TH1和TH2细胞因子比例的变化,为防治哮喘提供实验依据.方法经OVA致敏的雌性Balb/c于激发前腹腔注射抗CD86单克隆抗体及同型对照IgG2α 50 μg,末次激发48 h后收集支气管肺泡灌洗液(BALF),以酶联免疫吸附法(ELISA)测定BALF中白细胞介素-4(IL-4)、白细胞介素-5(IL-5)和干扰素-γ(INF-γ)水平.结果以OVA致敏并激发的哮喘对照组小鼠BALF中IL-4、IL-5水平升高[分别为(64.23±3.91)pg/mL、(379.84±73.02)pg/mL],与PBS对照组[分别为(22±2)pg/mL、(245.75±50.01)pg/mL]比较,差异有显著性(P＜0.05);OVA致敏并激发的哮喘对照组小鼠BALF中INF-γ水平降低[(98.73±16.41)pg/mL],与PBS对照组[(218.35±48.63)pg/mL]比较,差异有显著性(P＜0.05).抗CD86单克隆抗体组小鼠BALF中IL-4、IL-5水平降低[分别为(35.78±4.88)pg/mL、(222.98±58.68)pg/mL],而INF-γ水平升高[(206.92±47.8)pg/mL],与哮喘对照组及同型对照IgG2α组(98.73±16.41)pg/mL、(102.32±15.49)pg/mL比较差异有显著性(P＜0.05).结论抗CD86单克隆抗体通过阻断共刺激信号,纠正TH1/TH2失衡,从而达到抗气道炎症的作用.     

玉屏风散对小鼠哮喘合并人偏肺病毒感染的干预机制探讨

目的:建立人偏肺病毒(Human metapneumovirus,hMPV)感染的哮喘小鼠模型,验证玉屏风散对hMPV诱导的哮喘小鼠气道高反应性和炎症的疗效并初步探讨其机理。方法:随机将48只雌性BALB/c小鼠分为对照组(A组)、OVA致敏激发哮喘组(B组)、偏肺病毒哮喘组(C组)、小剂量玉屏风组(D组)、中剂量玉屏风组(E组)、大剂量玉屏风组(F组),采用卵蛋白致敏和激发,hMPV滴鼻方法建立哮喘人偏肺病毒感染模型并给予玉屏风治疗,采用动物体描箱法测定气道反应性;采用支气管肺泡灌洗液(BALF)进行细胞分类计数;肺组织病理切片HE染色观察炎性细胞浸润;采用流式细胞术分别测定肺部细胞因子IFNγ-和IL-4、IL-17。结果:(1)随吸入乙酰甲胆碱浓度增加各组气道反应性明显增加,C组比B组气道反应性显著升高(P<0.05);F组气道反应性明显降低,低于C组(P<0.05);(2)各组BALF中白细胞总数及炎症细胞数都比对照组显著升高,C组BALF中白细胞总数、嗜酸性粒细胞与B组、F组相比显著升高(P<0.05);(3)各哮喘组的炎症评分都显著高于对照组。在细支气管周围炎、肺泡炎方面B组和F组比C组明显减轻(P<0.05)。(4)A、B组IFNγ-无差异(P>0.05),各剂量组IFNγ-,IFNγ-/IL-4均显著高于B、C组(P<0.05);哮喘组、治疗组IL-4显著高于正常组(P<0.01),但各组间无明显差异;与B组相比,C组IL-17水平明显升高(P<0.05);与C组相比,E、F组IL-17水平明显降低。结论:大剂量玉屏风散可以抑制感染hMPV哮喘小鼠的气道高反应性及气道炎症,且改善Th1/Th2失衡状态。     

HDM通过肺泡巨噬细胞TLR4诱导哮喘小鼠气道炎症的机制研究

目的:研究HDM通过肺泡巨噬细胞(AM)Toll样受体4(TLR4)的高表达及诱导AM的活化,探讨其对哮喘小鼠气道炎症的影响.方法:BALB/c小鼠随机分为哮喘模型组(OVA)A,HDM处理组(HDM+OVA)B,对照组(生理盐水)C.用卵白蛋白(OVA)致敏与激发建立小鼠哮喘模型;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和IFN-γ的含量,实时定量PCR法测定AM的TLR4的表达,流式细胞技术(FCM)检测AM的CD80、CD86的表达.结果:与A组相比,B组BALF上清中IL-4、IL-5和IL-13的水平显著增高(P<0.05),而IFN-γ差异无统计学意义(P>0.05),与A组相比,AM的TLR4mRNA表达明显增高(P<0.05),CD80的表达差异无统计学意义,CD86的表达水平显著增高,差异有统计学意义(P<0.05).结论:HDM通过AM的TLR4的高表达诱导AM的活化,加重哮喘的气道炎症.     

A20重组蛋白通过NF-κB/TGF-β1/CTGF信号通路抑制哮喘小鼠气道重构的初步实验研究

目的探讨A20重组蛋白对支气管哮喘小鼠气道重构及NF-κB信号通路的影响。方法 40只雄性清洁级Balb/c小鼠,随机数字表法分为4组,每组10只,分别为:生理盐水对照组;卵蛋白(OVA)哮喘组;A20重组蛋白治疗3 d组;A20重组蛋白治疗7 d组。在末次激发24 h后所有小鼠取左肺组织行苏木精-伊红(HE)染色及PAS染色。取右肺组织分别用酶联免疫吸附法(ELISA),RT-PCR和Western blote检测支气管肺泡灌洗液(BALF)中IL-4、IL-5、TNF-α及IFN-γ含量以及肺组织中结缔组织生长因子(Connective tissue growth factor,CTGF)、转化生长因子-β1(trailsforming growth factor-β1,TGF-β1)的mRNA表达和核转录因子(nuclear factor kappa B,NF-κB)的表达。结果哮喘模型组小鼠与对照组相比较BALF中炎症细胞计数、IL-4、IL-5、TNF-α水平增高,而IFN-γ水平降低;肺组织CTGF、TGF-β1的转录和表达水平以及NF-κB表达水平均显著高于对照组(P<0.01)。A20重组蛋白3 d和7 d干预组小鼠与哮喘模型组相比较BALF中炎症细胞计数、IL-4、IL-5、TNF-α水平,CTGF、TGF-β1的转录和表达水平,NF-κB表达水平均显著降低,而BALF中IFN-γ水平明显上升,具有显著差异(P<0.05),但2个不同治疗组之间上述各指标间差异无统计学意义(P>0.05)。结论 A20重组蛋白可抑制哮喘小鼠气道重构的发生,其机制有可能是通过抑制NF-κB/TGF-β1/CTGF信号通路而实现的。     

雾化吸入干扰素γ阻断GATA-3表达防治哮喘的实验研究

目的 研究雾化吸入IFN-γ对支气管哮喘（哮喘）防治作用及其机制。方法 以Balb/c小鼠采用卵蛋白（OVA）、氢氧化铝建立哮喘模型（C组）,于23d至30d雾化吸入IFN-γ 12μg 1次/d,31d时取肺泡灌洗液（BALF）测定细胞成分,白细胞介素4（IL-4）、白细胞介素5（IL-5）浓度,取血测定血浆IgE水平,观察肺组织病理学变化及GATA-3的表达。设正常对照组（A组）和哮喘模型PBS处理组（B组）。每组小鼠12只。结果 ①C组BALF中的嗜酸性粒细胞（EOS）数量明显低于B组（P〈0．01）;②C组BALF中的IL-4和IL-5的水平明显低于B组（P〈0．01）;③C组血浆中总IgE和OVA特异性IgE水平显著低于B组（P〈0．01）;④B组支气管平滑肌肥厚,黏膜充血、水肿,黏膜层增厚,并有EOS为主的炎性细胞浸润,管腔内可见黏液栓,支气管壁周围有EOS为主的炎症细胞浸润,而C组小鼠的上述炎症性改变则明显减轻;⑤免疫组化显示B组肺组织中GATA-3的表达明显增加,而C组GATA-3的表达明显减少。结论 雾化吸入IFN-γ可抑制哮喘鼠IL4、IL5的合成,抑制气道炎症,抑制EOS在气道内的炎性浸润及降低血浆总LgE和OVA特异性LgE水平,其机制可能与IFN-γ阻断GATA-3的表达,从而继发抑制Th2型反应有关。     

干扰素-γ质粒基因转染对哮喘小鼠支气管肺泡灌洗液炎症细胞的影响

目的　观察气道内干扰素 -γ(IFN -γ)基因转染对哮喘小鼠支气管肺泡灌洗液 (BALF)嗜酸性粒细胞、中性粒细胞和淋巴细胞的影响 .方法　C5 7BL/ 6小鼠 4 0只 ,随机分为 4组 ,每组 10只 ,分别为正常对照组、哮喘组、模型空质粒干预组 (空质粒组 )和模型干扰素质粒干预组 (干扰素组 ) .卵白蛋白 (OVA)抗原溶液腹腔注射致敏 ,滴鼻激发造模 .对照组用生理盐水代替OVA ;空质粒组和干扰素组分别经鼻滴入空质粒或重组干扰素质粒 .观察各组实验小鼠的哮喘症状以及BALF中IFN -γ水平和各类炎症细胞的变化 .结果　哮喘组BALF中IFN -γ水平比正常对照组显著降低 ;哮喘小鼠干扰素质粒气道内转导后哮喘症状明显减轻 ,BALF中IFN -γ水平显著升高 ,同时嗜酸性粒细胞、中性粒细胞和淋巴细胞显著减少 .结论　气道内转染干扰素质粒能有效改善哮喘小鼠的症状和气道炎症 .     

IL-27对哮喘小鼠气道炎症的影响

目的研究IL-27对卵白蛋白(OVA)激发哮喘小鼠气道炎症的影响。方法 24只雌性BALB/c小鼠随机分为生理盐水组、哮喘组及IL-27组,每组8只。应用OVA建立哮喘模型,IL-27组小鼠应用1μgIL-27(溶于50μlPBS中)滴鼻给药,观察3组小鼠肺组织病理改变,计数支气管肺泡灌洗液(BALF)中嗜酸性粒细胞;ELISA法测定小鼠BALF中IL-4和IFN-γ浓度,RT-PCR测定肺组织T-bet mRNA的表达量。结果 IL-27组小鼠肺组织炎症反应明显轻于哮喘组小鼠;IL-27组小鼠BALF中嗜酸性粒细胞计数为(2.21±0.33)×107/L明显低于哮喘组的(12.82±2.17)×107/L(P0.01);IL-27组小鼠BALF中IL-4浓度为(20.4±3.2)μg/L,明显低于哮喘组的(61.3±13.1)μg/L(P0.05);IL-27组小鼠BALF中IFN-γ浓度为(50.3±6.3)μg/L,明显高于哮喘组的(11.1±3.3)μg/L(P0.05);IL-27组小鼠肺组织T-bet mRNA表达量(吸光度积分比值)为(0.268±0.048),明显高于哮喘组的(0.130±0.012)(P0.05)。结论 IL-27可能通过增强T-bet mRNA的表达增强Th1反应,减少BALF中嗜酸性粒细胞数量,进而减轻了哮喘小鼠肺组织炎症反应。     

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.     

The role of L-carnitine in treatment of a murine model of asthma

Leukotrienes, one of the mediators of inflammation in asthma, have a strong bronchoconstrictive effect. L-carnitine has been reported to influence respiratory functions. It has also been reported that L-carnitine inhibits leukotriene synthesis. To evaluate the effects of L-carnitine on oxygen saturation, urine leukotriene E4 levels and lung histopathology in a murine model of asthma, high IgE responder BALB/c mice (n = 24) were systemically sensitized to ovalbumin and chronically challenged with low particle mass concentrations of aerosolized ovalbumin, and then they were divided into 3 groups (study groups A, B, and C) each including eight mice. After methacholine-induced bronchoconstriction, the mice in groups A and B were given intraperitoneal L-carnitine (250 and 125 mg/kg, respectively), while the mice in group C were given placebo. Oxygen saturation of the mice was measured by pulse oxymeter before and after methacholine and after L-carnitine/ placebo application. In addition, urine leukotriene E4 levels were measured before asthma development, and 24-h after L-carnitine injection in asthmatic mice. Inflammation in the lung tissues of the sacrificed animals was scored histopathologically to determine the effect of L-carnitine on tissue level. A control group of non-sensitized mice (n = 8) treated with placebo only was used for comparison of urine leukotriene E4 levels and of histopathological parameters. Oxygen saturation of the mice in the study groups tended to decrease after methacholine and to improve after L-carnitine injection, although these changes were not significant at all time points. Urine leukotriene E4 levels of all 3 study groups increased significantly after asthma development. The rate of increment was smallest in the group given the highest L-carnitine dose (group A). Inflammation at the tissue level was also mildest in group A, and severest in the group that was not given carnitine (group C). All of the study groups and the control group differed significantly with respect to inflammation scores. In conclusion, L-carnitine improved oxygen saturation, and decreased urine leukotriene E4 levels and inflammation in lung tissues in the present murine model of asthma.     

Induction of Experimental Antiphospholipid Antibody Syndrome in PL/J Mice Following Immunization With β2GPI

PROBLEM: Immunization with β₂-glycoprotein I (β₂GPI) induces antiphospholipid antibodies (aPL) in normal mice and rabbits. Recently we reported early onset of autoimmunity in MRL/++ mice following immunization with β₂GPI. There is a close association between aPL with thrombosis, recurrent fetal loss, and intrauterine growth retardation. In this study we evaluated the effect of β₂GPI-induced aPL on pregnancy outcomes in an inbred strain of mice (PL/J). METHOD: Three groups of seven-week-old female PL/J mice (12 per group) were studied. Group A was immunized with β₂GPI and group B with ovalbumin; group C was not immunized. After two booster injections, the mice were tested for aPL, anti-DNA by ELISA, and for ANA by indirect immunofluorescence. Platelet count and pregnancy outcomes were studied at the age of 14 weeks. RESULTS: The aPL and anti-DNA levels were higher at 12 and 14 weeks in group A; the optical densities (OD) were 1.72±0.6 and 0.699±0.25 for group A, 0.091 ±0.040 and 0.230±0.47 for group B, and 0.0435±0.003 and 0.119±0.026 for group C (comparing group A with groups B and C combined, P<0.001). ANA titers rose in groups A and B by age, but they were significantly higher at 14 weeks in group A. The mean titers were 1/286, 1/90, and 1/16 for A, B, and C, respectively (P<0.001). The platelet counts were not significantly different among the three groups. The litter size was significantly smaller in group A, as evidenced by the numbers of viable fetuses among the mice that became pregnant in each group: 0.75, 2.45, and 5.5 in groups A, B, and C, respectively. Seven pregnant mice in group A had complete resorption, seven pregnant mice in group B showed focal (partial) resorption areas, and only one mouse in group C had complete resorption of the embryos, as shown by histopathological studies, although the fecundity rate was similar in the three groups. CONCLUSION: Our data suggest a pathogenic role for β₂GPI-induced aPL in the development of experimental models of APS in PL/J mice.     

雾化吸入灭活草分枝杆菌降低哮喘小鼠核因子κB及细胞间黏附分子1的表达

 目的:研究雾化吸入灭活草分枝杆菌对支气管哮喘小鼠气道炎症,以及哮喘肺组织中核因子κB(NF-κB)、细胞间黏附分子1(ICAM-1)和血管细胞黏附分子1(VCAM-1)的影响,探讨雾化吸入灭活草分枝杆菌防治哮喘的机制。方法:将24只雄性BALB/c小鼠按随机数字表法分为3组,每组8只：正常对照组（A）、哮喘模型组（B）和治疗组（C）。以鸡卵清蛋白致敏制造小鼠支气管哮喘模型。C组在激发后给予雾化吸入灭活草分枝杆菌治疗5 d,每天1次。各组动物处死后提取肺组织和支气管肺泡灌洗液（BALF）。进行病理HE染色及AB-PAS染色观察气道炎症浸润及黏液分泌情况,并行病理半定量分析。对BALF中炎症细胞进行分类计数。实时荧光定量PCR检测肺组织NF-κB、ICAM-1和VCAM-1的mRNA表达水平。结果:治疗组嗜酸性粒细胞比例低于模型组(P<0.05),气道炎症病变及黏液分泌情况较模型组减轻(P<0.05, P<0.01)。哮喘模型组的肺组织中NF-κB mRNA含量与正常组相比显著升高(P<0.01),而治疗组肺组织中的NF-κB mRNA 含量明显低于模型组(P<0.05);模型组的ICAM-1 mRNA 水平比正常组高(P<0.05),但治疗后明显降低(P<0.01);VCAM-1的 mRNA水平在各组间无显著差异。相关性检验发现小鼠肺组织中VCAM-1的mRNA与ICAM-1的mRNA呈明显正相关(r=0.84,P<0.01),但NF-κB的mRNA与ICAM-1的mRNA、VCAM-1的mRNA无明显相关性(均P>0.05)。结论:雾化吸入草分枝杆菌对支气管哮喘小鼠气道炎症及黏液分泌有抑制作用;NF-κB参与哮喘发病过程,雾化吸入灭活草分枝杆菌降低哮喘小鼠的NF-κB水平。同时雾化吸入灭活草分枝杆菌可降低黏附分子尤其是ICAM-1的表达,是其控制炎症的另一个重要机制。     

地龙对哮喘模型小鼠肺组织α-SMA及纤维蛋白的抑制作用

目的： 探讨地龙对哮喘小鼠肺组织平滑肌肌动蛋白-α（α-SMA）及纤维连接蛋白（FN）的影响。方法： BALB/c小鼠80只,随机分为对照组（A组）、哮喘组（B组）、地龙大剂量治疗组（C组）及地龙小剂量治疗组（D组）各20只。以腹腔注射0.02%鸡卵清蛋白和1%鸡卵清蛋白雾化吸入建立慢性哮喘模型。治疗组在每次雾化激发前给予地龙干预。肺泡灌洗计数各炎症细胞,ELISA检测血清总IgE水平,分离右肺固定,石蜡切片,免疫组化,用Leica QWIN V3分析系统,计算表达阳性结果;取左肺置液氮中保存,留作RT-PCR,用AlphaImager 2200半定量分析系统分析。结果： 与对照组相比,哮喘组气道平滑肌肌动蛋白-α（α-SMA）及纤维连接蛋白（FN）阳性表达显著升高,哮喘组α-SMA mRNA及FN mRNA的表达上调;地龙大剂量治疗组的α-SMA及FN的阳性表达、α-SMA mRNA及FN mRNA的表达均显著低于哮喘组;地龙小剂量治疗组的α-SMA及FN的阳性表达、α-SMA mRNA及FN mRNA的表达亦低于哮喘组,但无显著差异。结论： 地龙可抑制慢性哮喘模型小鼠肺组织中α-SMA及FN表达,提示抑制α-SMA及FN的表达可能是地龙抑制哮喘气道重构的重要机制之一。     

T-bet基因转染对哮喘小鼠气道炎症的影响

目的： 观察气道内T-bet基因转染对哮喘小鼠气道炎症的影响。方法: C57BL/6小鼠40只,随机分为4组,每组10只,分别为正常对照组（A组）、哮喘模型组（B组）、空质粒干预组(C组) 和T-bet质粒干预组(D组)。卵白蛋白(OVA) 抗原溶液腹腔注射致敏,滴鼻造模。正常对照组用生理盐水代替OVA,空质粒干预组和T-bet质粒干预组OVA激发48 h前,分别经鼻滴入50 μg空质粒和重组T-bet质粒。观察各组实验小鼠的肺组织炎症以及BALF中各类炎症细胞以及IL-4、IFN-γ水平的变化。 结果: Western blotting检测发现,小鼠气道转染pcDNA3-T-bet质粒48 h后肺组织T-bet蛋白表达显著增加。pcDNA3-T-bet质粒转染能较好抑制给药后48 h OVA激发的哮喘小鼠气道炎症（包括炎症细胞浸润,上皮细胞损伤、黏液分泌、血管壁水肿及管腔缩窄）;下调小鼠BALF中Th2因子IL-4并上调Th1因子IFN-γ水平。 结论: 气道内转染T-bet质粒能有效改善哮喘小鼠的气道炎症。     

白介素17单克隆抗体对变应性鼻炎小鼠气道炎症的作用

目的 探讨白介素17单克隆抗体（IL-17mAb）的不同给予剂量及方式在变应性鼻炎小鼠气道炎症中的作用。方法 将48只小鼠采用随机数字表法分为A、 B、 C、 D、 E、 F组,每组8只。分别于第0、 7、 14 d将20 μg卵清蛋白（OVA）加2 mg铝佐剂腹腔注射处理A、C、D、E及F组小鼠,间隔7 d,第22天开始进行鼻腔激发,每天每侧鼻孔各给予OVA 10 μl（共500 μg）滴鼻,连续7 d。A、C、D、E组小鼠于每次OVA鼻腔激发前1 h分别给予生理盐水、100 ng IL-17mAb、500 ng IL-17mAb、5 μg IL-17mAb滴鼻,F组小鼠于每次OVA鼻腔激发前4 h给予5 μg IL-17mAb腹腔注射,B组小鼠于相同时间点给予等量生理盐水腹腔注射及滴鼻。所有小鼠于最后1次激发后评估鼻部症状学变化,Diff-Quik染色观察鼻腔灌洗液（NLF）中嗜酸性粒细胞浸润情况,ELISA方法检测血清及NLF中IL-6、IL-10水平,鼻黏膜组织行甲苯胺蓝染色观察肥大细胞。结果 ４周末A组所有小鼠症状学评分均＞5分,提示造模成功。F组小鼠的挠鼻及喷嚏次数均少于A组（P＜0.05）;F组小鼠NLF中嗜酸性粒细胞数、血清IL-6水平低于A组,血清及NLF中IL-10水平均高于A组（P＜0.05）;E组小鼠血清中IL-10水平高于A组（P＜0.05）;A组小鼠鼻黏膜组织中肥大细胞数多于B组,统计学意义显著（P＜0.01）;F组小鼠鼻黏膜组织中肥大细胞数少于A组,但差异无统计学意义（P＞0.05）;F组小鼠鼻黏膜组织中肥大细胞数与B组比较,差异无统计学意义（P＞0.05）。结论 高剂量的（5 μg）IL-17mAb腹腔注射处于激发阶段的变应性鼻炎小鼠促使小鼠变应性鼻炎症状明显减轻,鼻腔灌洗液嗜酸性粒细胞减少。促使变应性鼻炎小鼠血清中IL-6表达降低,血清中及鼻腔灌洗液中IL-10表达升高,因此推测这些细胞因子的变化可能抑制Th17/促进Treg的分化,进而对变态反应产生抑制作用。     

Effects of adjuvants on the function of antigen presenting cells in the production of IgE in mice

In an examination of the effects of adjuvants on the production of IgE, amounts of mRNAs for cytokines in antigen presenting cells(APCs) were assayed by RT-PCR and expressions of surface molecules on the cells were analyzed by flowcytometry when primed with antigen plus adjuvant. When mice were primed with ovalbumin plus alum, the levels of total and specific IgE were higher than those of mice primed with ovalbumin plus CFA. The APCs from mice primed with ovalbumin plus alum expressed higher levels of IL-1 mRNA than those in APCs from mice primed with ovalbumin plus CFA. B7 molecules were more expressed on the surface of the APCs from mice primed with ovalbumin plus CFA. The results suggested that these modulations of the functions of APCs affected the induction of helper T cell subsets in mice primed with different adjuvant.     

Special consideration is required for the component-resolved diagnosis of egg allergy in infants

BackgroundThere are few reports regarding differences in reactivity to the major egg allergens according to children's age, although component-resolved diagnosis is gradually being used.ObjectiveTo investigate differences in reactivity to major egg allergens among various age groups of children with egg allergy.MethodsTwenty-seven patients diagnosed with egg allergy were included. Egg allergy was defined as a convincing history of reproducible symptoms within 2 hours of egg consumption and an egg white–specific IgE level of at least 0.35 kU_A/L. Patients were divided into 3 age groups: younger than 12 months (group A, 7 subjects), 12 to 23 months (group B, 8 subjects), and at least 24 months (group C, 12 subjects). Immunoblotting and enzyme-linked immunosorbent assay investigated IgE reactivity toward ovalbumin, ovomucoid, and ovotransferrin in eggs.ResultsImmunoblotting analysis showed that all patients in group A reacted to ovalbumin, whereas reactions to other proteins were not detected. All patients in group B displayed a reaction to ovalbumin and ovomucoid. IgE binding to ovotransferrin was shown in 3 patients in group B. All patients in group C displayed reactivity to ovalbumin, 5 patients showed a reaction to ovomucoid, and 8 patients displayed a reaction to ovotransferrin. As a patient's age increased, specific IgE binding to ovalbumin and ovotransferrin increased (P = .011 and .004).ConclusionIgE reactivity to egg allergens differs according to children's ages.     

B Cells Carrying Surrogate Receptors in their Membranes Process and Present Antigen to Specific Murine T Cells

A palmitate-conjugate derivative of ovalbumin which can be inserted into the membrane of B cells has been prepared. The ability of these cells to act as antigen-presenting cells for specific T lymphocytes obtained from immunized mice was tested. It was found that the conjugates were more efficiently processed and presented than the naive form of the antigen. Palmitate-conjugated antibodies specific to ovalbumin were also inserted into the cell membrane of normal B lymphocytes. These cells were pulsed with the antigen and tested as antigen-presenting cells for T cells obtained from immunized mice. The antibody-decorated B cells presented ovalbumin more efficiently than non-decorated controls. Whether antibody-decorated, antigen-pulsed B cells could prime T cells in viro was investigated. Some priming activity was found.     

Lung responsiveness to antigen in sensitised mice of different strains

A new model of the isolated perfused lung from different strains of mice was developed. Lungs from Swiss, Balb/C and CBA mice actively sensitised to ovalbumin were challenged intratracheally (i.t.) by antigen on day 14. In Swiss mice instillation of ovalbumin led to the release of leukotriene (LT) C₄ significantly above basal values. Conversely, lungs from Balb/C and CBA mice were unresponsive to ovalbumin in terms of production of LTC₄. All strains failed to release histamine when challenged with antigen. Intratracheal instillation of platelet-activating factor (PAF), to lungs from non-sensitised animals, induced the release of comparable amounts of LTC₄, irrespective of the strain. In contrast, i.t. administration of fMLP to lungs from Swiss mice elicited release of significantly higher amounts of LTC₄ as compared to Balb/C and CBA mice. In separate experiments, ovalbumin was injected into the paws and anaphylactic oedema was evaluated. Balb/C and CBA required 1 μg to show an oedema formation, whereas the dose of ovalbumin for Swiss mice to develop a similar response was at least 30-fold lower. In conclusion, antigen provocation induced release of LTC₄ from lungs from Swiss mice but not from Balb/C or CBA. This difference may be accounted for by strain-dependent factors, such as antibody production and requires further investigation.