摘要
目的探讨支气管哮喘（哮喘）患儿外周血自然杀伤T淋巴细胞（NKT细胞）数量及体外扩增活化后产生细胞因子水平的变化。方法选取2009年2月至2012年3月在上海交通大学医学院附属仁济医院儿科门诊确诊的哮喘患儿52例（哮喘组），其中轻度间歇26例，轻度持续14例，中度持续12例。另选20例体检健康儿童作为健康对照组。收集所有研究对象外周静脉血标本，采用密度梯度离心法分离获得外周血单个核细胞，应用免疫荧光标记和流式细胞术检测哮喘组和健康对照组儿童外周血NKT细胞和CD4＋NKT细胞的比例，观察NKT细胞和CIM＋NKT细胞与过敏指标总IgE、嗜酸性细胞阳离子蛋白（ECP）的相关性。经α-半乳糖神经鞘胺醇扩增培养活化后用ELISA法检测外周血细胞因子IFN-γ、IL4、IL-10水平。比较哮喘组和健康对照组之间的差异。结果哮喘组患儿的NKT细胞和CD4＋NKT细胞比例均较健康对照组显著下降（P均〈0．01）；在不同病情哮喘患儿中，NKT细胞和CD4＋NKT细胞的比例无明显变化（P〉0．05）。NKT细胞、CD4＋NKT细胞的比例与总IgE、ECP均无明显相关性（P均〉0．05）。扩增培养活化后，哮喘组患儿NKT细胞分泌的IL-4水平较健康对照组明显升高（P〈0．01），IL-10水平显著下降（P〈0．01），而IFN-γ水平与健康对照组比较无统计学差异（P〉0．05）。结论哮喘发病可能与NKT细胞和CD4＋NKT细胞数量的低下及功能严重失调相关。
Objective To study the changes of natural killer T cells （ NKT cells） in peripheral blood and the ability of NKT cells to proliferate in response to alpha-galactosyl ceramide （ot-Galcer） in children with asthma. Me- thods Fifty-two asthmatic children who were diagnosed by Pediatric Clinic of Renji Hospital Affiliated to Shanghai Jiaotong University Medical College from Feb. 2009 to Mar. 2012 were selected as asthmatic group （26 cases in inter- mittent group, 14 cases in mild persistent group and 12 eases in moderate persistent group）. Twenty healthy children were selected as healthy control group. Peripheral blood mononuclear cells were collected by way of density gradient eentrifugation method from blood samples. The ratio of peripheral blood NKT and CD4＋NKT cells were assessed by im- munofluorescence and flow cytometry assays. The relationships between the NKT cells number, CD4＋ NKT cells and atopic indexes such as the total IgE,eosinophil cationic protein （ECP） and the severity of asthma were investigated. The levels of IFN-γ, IL-4, IL-10 in peripheral blood were measured by enzyme-linked immunosorbent assay after proli-ferate in response to α-Galcer. The data were analyzed by using statistics software. Results Compared with healthy control group, the ratio of peripheral blood NKT and CD4＋ NKT cells in asthmatic patients were significantly decreased （all P 〈0.01 ） ; There were no significant differences in asthmatic patients among the subgroups in terms of severity （P 〉 0.05 ）. There was no significant correlation between the NKT cells, CD4＋NKT cells and atopic indexes such as the total IgE and ECP（ all P 〉 0.05 ）. The level of IL-4 in serum secreted by NKT cells in asthmatic group was signifi- cantly higher than that of healthy control group （P 〈 0.01 ） ;The level of IL-10 in serum secreted by NKT cells in asth- matic group was decreased significantly （P 〈0.01 ） ;There was no significant difference in IFN-γ secreted by NKT cells between the asthmatic group and healthy control group （ P 〉 0.05 ）. Conclusion The pathogenesis of asthma may be related to the ratio and dysfunction of NKT ceils and CD＋ NKT cells.
出处
《中华实用儿科临床杂志》
CAS
CSCD
北大核心
2013年第10期775-777,共3页
Chinese Journal of Applied Clinical Pediatrics
基金
上海交通大学医学院横向课题（2010HX003）
