【摘要】：目的：建立呼吸道合胞病毒(RSV)感染裸鼠模型,探討T細(xì)胞缺陷對RSV感染所致氣道炎癥和氣道高反應(yīng)性的影響。 方法：SPF級6-8周雌性裸鼠及BALB/c鼠隨機(jī)分為對照組和RSV感染組,在10%水合氯醛3μl/g麻醉下,予以1.5×108PFU/ml RSV或DMEM培養(yǎng)基100μl滴鼻后第3、5、7及14天評估氣道高反應(yīng)性,并檢測支氣管肺泡灌洗液(BALF)中細(xì)胞總數(shù)和白細(xì)胞分類計(jì)數(shù),HE染色檢測肺部炎癥病理,ELSIA法檢測肺泡灌洗液中細(xì)胞因子水平。并在第5天Q-PCR檢測裸鼠膽堿能受體M2(M2R)及Toll樣受體3(TLR3)的mRNA表達(dá)。 結(jié)果：RSV感染后在裸鼠肺部復(fù)制持續(xù)時間更長,病毒滴度更高。裸鼠RSV感染組和BALB/c RSV感染組支氣管肺泡灌洗液中細(xì)胞總數(shù)、肺部炎癥病理評分均明顯高于對照組(P0.05)。支氣管肺泡灌洗液中淋巴細(xì)胞比例增加,巨噬細(xì)胞比例減低；HE染色顯示肺部炎癥以淋巴細(xì)胞浸潤,血管壁、支氣管壁及肺泡間隔增厚為主,其中BALB/cRSV感染后炎癥較裸鼠RSV感染持續(xù)更久。裸鼠及BALB/c鼠肺功能檢測顯示感染后氣道反應(yīng)性均較其對照組增加,以BALB/c鼠RSV感染后第5天氣道反應(yīng)性最高,與同期裸鼠RSV感染組比較有統(tǒng)計(jì)學(xué)差異(P0.05)。細(xì)胞因子IFN-γ在BALB/c RSV感染后明顯增加,而在裸鼠RSV感染后與其對照組比較沒有顯著改變。裸鼠RSV感染后TLR3mRNA均明顯高于對照組(P0.05),而IL-4、IL-10等細(xì)胞因子以及膽堿能受體M2與對照組比較無顯著差異(P0.05)。 結(jié)論：裸鼠感染RSV后病毒復(fù)制持續(xù)時間更長,但肺部炎癥持續(xù)時間較BALB/c鼠短,氣道反應(yīng)性顯著低于BALB/c RSV感染組,BALF中IFN-γ水平明顯低于BALB/c鼠,提示RSV感染后氣道炎癥與氣道高反應(yīng)性不完全平行,T細(xì)胞及其分泌的細(xì)胞因子(如[FN-γ等)可能在RSV感染后氣道病理變化中起重要作用。
[Abstract]:Aim: to establish a nude mouse model of respiratory syncytial virus (RSV) (RSV) infection and to investigate the effect of T cell deficiency on airway inflammation and airway hyperresponsiveness induced by RSV infection. Methods female nude mice and BALB/c mice of 6-8 weeks were randomly divided into control group and RSV infected group. Under 10% chloral hydrate 3 渭 l / g anesthesia, the airway hyperresponsiveness was evaluated by 1.5 脳 108PFU/ml RSV or 100 渭 l DMEM medium on the 3rd day and 14th day after nasal drip. The total number of cells in bronchoalveolar lavage fluid (BALF) and white blood cell count (WBC) in bronchoalveolar lavage fluid (BALF) were detected by HE staining. The level of cytokines in alveolar lavage fluid was detected by ELSIA method. The mRNA expression of cholinergic receptor M2 (M 2R) and Toll like receptor 3 (TLR3) in nude mice was detected by Q-PCR on day 5. Results the replication duration was longer and the virus titer was higher in nude mice after infection. The number of cells in bronchoalveolar lavage fluid (BALF) and the pathological score of pulmonary inflammation in nude mice infected with RSV and BALB/c RSV were significantly higher than those in control group (P0.05). The proportion of lymphocytes in bronchoalveolar lavage fluid increased, and the proportion of macrophages decreased. He staining showed that the inflammation of lung was mainly lymphocytic infiltration, thickening of blood vessel wall, bronchial wall and alveolar septum. Inflammation after BALB/cRSV infection lasted longer than RSV infection in nude mice. Lung function test of nude mice and BALB/c mice showed that the airway reactivity was higher in BALB/c mice than in the control group, and the response of the 5th weather tract was the highest in BALB/c mice after RSV infection, which was significantly different from that in RSV infected nude mice at the same time (P0.05). The cytokine IFN- 緯 increased significantly after BALB/c RSV infection, but had no significant change after RSV infection in nude mice compared with the control group. The TLR3mRNA of nude mice after RSV infection was significantly higher than that of the control group (P0.05), but IL-4,IL-10 and cholinergic receptor M2 had no significant difference compared with the control group (P0.05). Conclusion: the duration of viral replication in nude mice infected with RSV is longer, but the duration of pulmonary inflammation is shorter than that of BALB/c mice, and the airway reactivity is significantly lower than that of BALB/c mice infected with BALB/c RSV. These results suggest that airway inflammation and airway hyperresponsiveness are not completely parallel to T cells and their cytokines (such as [FN- 緯]) after RSV infection may play an important role in the pathological changes of airway after RSV infection.
【學(xué)位授予單位】：重慶醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2012
【分類號】：R725.1

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1 周娟;王莉佳;崔玉霞;劉雪梅;楊錫強(qiáng);;裸鼠呼吸道合胞病毒感染的動物模型[J];Virologica Sinica;2006年03期

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