经鼻重组腺相关病毒T-bet基因转染对呼吸道合胞病毒感染小鼠呼吸道炎症的影响  

Effect of Recombinant Adeno Associated Virus Carrying T-bet Gene Transfection on Airway Inflammation of Mice Infected with Respiratory Syncytial Virus

作　　者：张云[1] 韩继敏[1] 王锁英[1] 潘小晶[1] 

机构地区：[1]江苏大学附属医院儿科,江苏镇江212001

出　　处：《实用儿科临床杂志》2011年第4期262-264,共3页Journal of Applied Clinical Pediatrics

摘　　要：目的观察携带T-bet基因的重组腺相关病毒(rAAV-T-bet)转染对呼吸道合胞病毒(RSV)感染模型小鼠呼吸道炎症的影响。方法健康6～8周龄Balb/c小鼠40只。按随机数字表法分为4组:正常对照组(A组)、RSV感染模型组(B组)、模型/对照病毒rAAV载体介导的增强型绿色荧光蛋白(rAAV-eGFP)干预组(C组)及模型/rAAV-T-bet干预组(D组),每组10只。将B组、C组和D组小鼠用滴度为1.26×1010PFU.L-1的RSV液滴鼻,每只100μL;A组用等量人喉癌上皮细胞(Hep-2细胞)培养上清液代替RSV液。于RSV接种第5天,再次麻醉小鼠,A组和B组经鼻滴入293细胞培养上清液;C组经鼻滴入293细胞包装的rAAV-eGFP对照病毒液,D组经鼻滴入293细胞包装的rAAV-T-bet。7 d后处死小鼠,检测各组小鼠支气管肺泡灌洗液(BALF)中细胞成分及IL-4和γ干扰素(IFN-γ)水平,并观察其肺组织病理学变化。结果 1.B组小鼠BALF中细胞总数和淋巴细胞计数百分比分别为(43.91±10.41)×109L-1、(65.15±1.88)%,A组分别为(14.52±3.57)×109L-1、(25.79±1.07)%,A组与B组比较差异均有统计学意义(Pa<0.05);D组小鼠BALF中细胞总数和淋巴细胞计数百分比分别为(24.84±4.35)×109L-1、(29.48±11.97)%,与B组、C组比较差异均有统计学意义(Pa<0.05)。2.B组小鼠BALF中IL-4和IFN-γ水平分别为(74.32±10.06)ng.L-1、(89.36±20.38)ng.L-1,A组分别为(6.01±1.02)ng.L-1、(177.32±18.16)ng.L-1,A组与B组比较差异均有统计学意义(Pa<0.05);D组分别为(49.14±7.59)ng.L-1、(145.28±43.52)ng.L-1,与B组、C组比较差异均有统计学意义(Pa<0.05)。3.A组小鼠呼吸道无明显炎症改变;B组和C组小鼠小支气管、血管黏膜下和周围肺组织有明显炎性细胞浸润,血管壁明显水肿;D组小鼠呼吸道炎症明显减轻。结论 T-bet基因转染能改善RSV感染模型小鼠呼吸道内细胞因子IL-4和IFN-γ异常表达,同时对淋巴细胞在肺内聚集、肺组织炎症有一定抑制作用。Objective To study the effect of recombinant adeno-associated virus with T-bet gene（rAAV-T-bet） transfection on airway inflammation in mice infected with respiratory syncytial virus（RSV）.Methods Forty Balb/c mice（aged 6-8 weeks） were randomly divided into 4 groups：control group（group A）,RSV model group（group B）,model/control recombinant adeno-associated virus carrying enhanced green fluorescent protein（rAAV-eGFP） intervention group（group C）and model/rAAV-T-bet virus intervention group（group D）,10 mice in each group.Under intraperitoneal anesthesia,except group A,other groups were infected with nasal drip of 100 μL RSV which was harvested from Hep-2 cell culturer,rats in group A received Hep-2 cell culturer supernatant.Five days later,anesthetized mice intra-peritoneally once again,rats in group A and group B received nasal drip of 100 μL 293 cell culturer supernatant,rats in group C intranasal instillation of rAAV-eGFP virus and rats in group D intranasal instillation of rAAV-T-bet virus with equal volume as group A.The mice were killed 7 days later,bronchoalveolar lavage fluid（BALF）cell count and differential were studied.IL-4 and interferon-γ（IFN-γ） in BALF were determined.Pathologic changes in lung tissues were observed.Results 1.There were significant differences of the numbers of inflammation cells and the persentage of lymphocytes in BALF in group B[（43.91±10.41）×10^9L^-1,（65.15±1.88）%,respectively]compared with those in group A[（14.52±3.57）×10^9L^-1,（25.79±1.07）%,respectively].The numbers of inflammation cells and lymphocytes in BALF in group D[（24.84±4.35）×10^9L^-1,（29.48±11.97）%,respectively]were also significantly different compared with those in group B and group C（Pa0.05）.2.The IL-4 and IFN-γ levels in BALF in group B[（74.32±10.06） ng·L^-1,（89.36±20.38） ng·L^-1,respectively]were significantly different compared with those in group A[（6.01±1.02） ng·L^-1,（177.32±18.16） ng·L^-1,respectively]（Pa0.

关 键 词：T-BET基因 呼吸道合胞病毒 基因转染 小鼠 

分 类 号：R725.6[医药卫生—儿科]