儿童新甲型（H1N1）流感疫苗接种后远期疫苗活化Th1/Th2细胞因子分泌水平分析

夏洪波+陈珺汝+郭锦锦

[摘要]目的 檢测儿童新甲型（H1N1）流感疫苗接种后其远期疫苗活化Th1/Th2细胞因子分泌水平的变化情况。方法 选取本院2009年12月～2010年1月收治的自愿接受新甲型H1N1流感疫苗接种的31名健康儿童，取3 ml外周血分离淋巴细胞，分别设新甲型H1N1流感疫苗刺激组（实验组）和正常细胞组（对照组），进行细胞培养后，采用流式CBA法及ELISA法检测Th细胞因子分泌水平。结果 流式CBA技术检查显示，试验组的TNF-α、IL-10水平明显低于对照组，差异有统计学意义（P<0.05）。ELISA方法检测显示，两组的TNF-α、IL-4水平比较，差异无统计学意义（P>0.05）；实验组的IL-10水平显著低于对照组，差异有统计学意义（P<0.05）。对照组和实验组流式CBA方法检测的IL-10 水平明显高于ELISA方法检测结果，差异有统计学意义（P<0.05），且两种方法检测结果的相关性具有统计学意义，分别得到回归方程Y=206.59+0.082X和Y=136.70+0.168X。结论 新甲流疫苗再次刺激未能很好地活化Th1及Th2细胞，即未产生较好的Th1型细胞免疫反应和较好的体液免疫，提示新甲流疫苗特异性免疫记忆功能欠佳；流式CBA方法与传统ELISA方法检测细胞因子均具有较高的灵敏度且具有相同的结果规律。

[关键词]新甲型H1N1流感疫苗；Th1/ Th2细胞因子；儿童；免疫记忆

[中图分类号] R511.7 [文献标识码] A [文章编号] 1674-4721（2017）03（a）-0011-05

[Abstract]Objective To test the changes of secretion levels of long-term vaccine activation of Th1/Th2 cytokine after new influenza A （H1N1） vaccination in children.Methods A total of 31 healthy children who were on their own willing for new influenza A （H1N1） vaccination from December 2009 to January 2010 in our hospital were selected，lymphocyte cells were separated from 3 ml peripheral blood for culture，and then were divided into new influenza A （H1N1） stimulation group （the experimental group） and normal cell group （the control group）.The levels of Th cytokine secretion were measured by flow CBA and enzyme-linked immunosorbent assay （ELISA）.Results The technical examination of the flow CBA showed that the levels of TNF-α and IL-10 in the experimental group was significantly lower than those in the control group，with significant difference （P<0.05）.The ELISA method displayed no significant difference in the level of TNF-α and IL-4 between the experimental group and the control group （P>0.05）.The level of IL-10 in the experimental group was significantly lower than that in the control group，which was in statistical differences （P<0.05）.The levels of IL-10 in both the control group and the experimental group tested by flow CBA was significantly higher than those by the ELISA test，with statistical differences （P<0.05）.The correlation of test outcomes by the two methods was displayed a statistical significance in equation of Y=206.59+0.082X and Y=136.70+0.168X respectively.Conclusion Re-stimulation of new influenza A H1N1 vaccine cannot activate Th1 and Th2 cells in a great success，that′s to say，no satisfactory Th1 cellular immune response and humoral immunity suggesting that the specific immune memory function of the new influenza A H1N1 is poor.The higher sensitivity and the same test results are obtained by flow CBA and the traditional ELISA method in detecting cytokines.