幽门螺杆菌感染儿童胃窦黏膜CD4^+、CD8^+的研究  

CD4^+ and CD8^+ T cells in gastric mucosa in children infected with Helicobacter pylori

作　　者：李中跃[1] 陈飞波[1] 陈洁[1] 

机构地区：[1]浙江大学医学院附属儿童医院消化科,杭州310003

出　　处：《中华儿科杂志》2005年第6期453-456,共4页Chinese Journal of Pediatrics

摘　　要：目的研究幽门螺杆菌(Hp)感染儿童胃窦黏膜T淋巴细胞CD4+、CD8+及CD4+/CD8+的变化。方法流式细胞仪(FCM)直接免疫荧光法测定79例慢性浅表性胃炎(CSG)患儿(Hp+44例,Hp-35例)胃窦黏膜及其中33例(Hp+12例,Hp-21例)患儿外周血的T淋巴细胞CD4+、CD8+变化。所有患儿均行内镜检查并取胃窦黏膜作快速尿素酶试验、组织病理学检查及胃窦黏膜单个核细胞提取。33例儿童同时抽取外周静脉血2ml。提取的胃窦黏膜及外周血单个核细胞经异硫氰酸荧光素标记CD3(CD3-FITC)、藻红蛋白标记CD4(CD4-PE)、多甲藻(黄)素叶绿素蛋白标记CD8(CD8-PerCP)单克隆抗体染色后行流式细胞术测定,其中胃窦黏膜T淋巴细胞的检测以CD3设门。结果(1)胃窦黏膜CD3+细胞的检出率分别为:Hp-CSG组(3·26±1·98)%,Hp+CSG组(4·37±1·97)%。(2)胃窦黏膜CD3+细胞中CD4+,CD8+的相对百分比及CD4+/CD8+值分别为:Hp-CSG组为(23·74±10·37)%,(47·04±12·00)%,0·52±0·23,Hp+CSG组为(40·28±11·35)%,(27·91±8·84)%,1·55±0·52。Hp+CSG组胃窦黏膜CD4+细胞相对百分比、CD4+/CD8+比值明显高于Hp-CSG组,CD8+细胞相对百分比则低于Hp-CSG组(P均<0·01)。(3)两组外周血T淋巴细胞CD3+(%)、CD4+(%)、CD8+(%)、CD4+/CD8+的比较差异无统计学意义。结论儿童Hp+CSG的T淋巴细胞变化为局部胃窦黏膜的细胞免疫应答所致。Objective To study the changes of gastric mucosal CD4 + and CD8 + T cells in Helicobacter pylori (Hp) infected children.Methods Seventy nine patients with digestive tract symptoms were assessed by endoscopy, rapid urease test and histology. Forty four patients had Hp positive chronic superficial gastritis (Hp +CSG) and 35 patients had Hp negative chronic superficial gastritis (Hp -CSG). Gastric biopsy specimens were obtained from each patient. Peripheral blood samples were obtained from 33 patients (12 with Hp +CSG, 21 with Hp -CSG). Hp infection was identified by rapid urease test and histology. Hp infection was confirmed when a patient was positive for both of these tests. Four pieces of gastric antrum mucosal specimens were placed in Hank′s balanced salt solution containing 1 mmol/L dithiothreitol (DTT) and 1 mmol/L ethylenediamine tetraacetic acid(EDTA). The specimens were treated with collagenase type Ⅰ (120 U/ml) for three hours at 37℃ with agitation. The mononuclear cells were collected by removing undigested material and washed three times with RPMI 1640. Isolated gastic mononuclear cells were stained with CD3-FITC (fluorescein isothiocyanate), CD4-PE (R-phycoerthrin), CD8-PerCP (Peridinin-chlorophyll-α-protein) and measured by flow cytometry. Mucosal T lymphocytes were gated for the expression of CD3. Peripheral blood lymphocyte subsets were analysed by direct immunofluorescence.Results The percentage of isolated gastric mononuclear cells within the CD3 gate were 3.26±1.98 in Hp -CSG, 4.37±1.97 in Hp +CSG. Relative CD4 + (%), CD8 + (%) and CD4 +/CD8 + of the CD3 + cells respectively were 23.74±10.37, 47.04±12.00, 0.52±0.23 in Hp -CSG group, 40.28± 11.35, 27.91±8.84, 1.55±0.52 in Hp +CSG group. CD4 +(%), CD4 +/CD8 + in Hp +CSG group were significantly higher than those of Hp -CSG group and CD8 +(%) was lower than those of Hp -CSG group ( P <0.01). There were no significant differences in peripheral blood T lymphocyte subsets between the two groups. Conclusion The difference of gastric T lym

关 键 词：胃窦黏膜 幽门螺杆菌感染 CD4^+/CD8^+比值 儿童 CD4^+/CD8^+值 CD3^+细胞 幽门螺杆菌(Hp) 异硫氰酸荧光素标记 外周血单个核细胞 外周血T淋巴细胞 慢性浅表性胃炎 快速尿素酶试验 组织病理学检查 CD4^+细胞 CD8^+细胞 

分 类 号：R725.7[医药卫生—儿科]