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幽门螺杆菌感染儿童胃黏膜中的CD4+和CD8+ T细胞

[CD4+ and CD8+ T cells in gastric mucosa in children infected with Helicobacter pylori].

作者信息

Li Zhong-yue, Chen Fei-bo, Chen Jie

机构信息

Department of Gastroenterology, Children's Hospital Affiliated to Medical College, Zhejiang University, Hangzhou 310003, China.

出版信息

Zhonghua Er Ke Za Zhi. 2005 Jun;43(6):453-6.

Abstract

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 degrees C 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-alpha-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 lymphocyte response between Hp(+)CSG and Hp(-)CSG in children indicated that the local cellular immune reaction may play a critical role in the pathogenesis of Hp infection.

摘要

目的

研究幽门螺杆菌(Hp)感染儿童胃黏膜CD4(+)和CD8(+) T细胞的变化。

方法

对79例有消化道症状的患者进行内镜检查、快速尿素酶试验和组织学检查。44例患者为Hp阳性慢性浅表性胃炎(Hp(+)CSG),35例患者为Hp阴性慢性浅表性胃炎(Hp(-)CSG)。从每位患者获取胃活检标本。从33例患者(12例Hp(+)CSG,21例Hp(-)CSG)获取外周血样本。通过快速尿素酶试验和组织学鉴定Hp感染。当患者这两项检查均为阳性时,确诊为Hp感染。将4块胃窦黏膜标本置于含1 mmol/L二硫苏糖醇(DTT)和1 mmol/L乙二胺四乙酸(EDTA)的汉克平衡盐溶液中。标本在37℃搅拌下用Ⅰ型胶原酶(120 U/ml)处理3小时。通过去除未消化物质收集单核细胞,并用RPMI 1640洗涤3次。分离的胃单核细胞用CD3-异硫氰酸荧光素(FITC)、CD4-藻红蛋白(PE)、CD8-多甲藻叶绿素蛋白(PerCP)染色,并用流式细胞术检测。对黏膜T淋巴细胞进行CD3表达门控。通过直接免疫荧光分析外周血淋巴细胞亚群。

结果

在Hp(-)CSG组中,CD3门控内分离的胃单核细胞百分比为3.26±1.98,在Hp(+)CSG组中为4.37±1.97。Hp(-)CSG组CD3(+)细胞的相对CD4(+)(%)、CD8(+)(%)和CD4(+)/CD8(+)分别为23.74±10.37、47.04±12.00、0.52±0.23,Hp(+)CSG组分别为40.28±11.35、27.91±8.84、1.55±0.52。Hp(+)CSG组的CD4(+)(%)、CD4(+)/CD8(+)显著高于Hp(-)CSG组,CD8(+)(%)低于Hp(-)CSG组(P<0.01)。两组外周血T淋巴细胞亚群无显著差异。

结论

儿童Hp(+)CSG和Hp(-)CSG之间胃T淋巴细胞反应的差异表明,局部细胞免疫反应可能在Hp感染的发病机制中起关键作用。

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