夏洪波+陳珺汝+郭錦錦

[摘要]目的 檢測(cè)兒童新甲型（H1N1）流感疫苗接種后其遠(yuǎn)期疫苗活化Th1/Th2細(xì)胞因子分泌水平的變化情況。方法 選取本院2009年12月～2010年1月收治的自愿接受新甲型H1N1流感疫苗接種的31名健康兒童，取3 ml外周血分離淋巴細(xì)胞，分別設(shè)新甲型H1N1流感疫苗刺激組（實(shí)驗(yàn)組）和正常細(xì)胞組（對(duì)照組），進(jìn)行細(xì)胞培養(yǎng)后，采用流式CBA法及ELISA法檢測(cè)Th細(xì)胞因子分泌水平。結(jié)果 流式CBA技術(shù)檢查顯示，試驗(yàn)組的TNF-α、IL-10水平明顯低于對(duì)照組，差異有統(tǒng)計(jì)學(xué)意義（P<0.05）。ELISA方法檢測(cè)顯示，兩組的TNF-α、IL-4水平比較，差異無統(tǒng)計(jì)學(xué)意義（P>0.05）；實(shí)驗(yàn)組的IL-10水平顯著低于對(duì)照組，差異有統(tǒng)計(jì)學(xué)意義（P<0.05）。對(duì)照組和實(shí)驗(yàn)組流式CBA方法檢測(cè)的IL-10 水平明顯高于ELISA方法檢測(cè)結(jié)果，差異有統(tǒng)計(jì)學(xué)意義（P<0.05），且兩種方法檢測(cè)結(jié)果的相關(guān)性具有統(tǒng)計(jì)學(xué)意義，分別得到回歸方程Y=206.59+0.082X和Y=136.70+0.168X。結(jié)論 新甲流疫苗再次刺激未能很好地活化Th1及Th2細(xì)胞，即未產(chǎn)生較好的Th1型細(xì)胞免疫反應(yīng)和較好的體液免疫，提示新甲流疫苗特異性免疫記憶功能欠佳；流式CBA方法與傳統(tǒng)ELISA方法檢測(cè)細(xì)胞因子均具有較高的靈敏度且具有相同的結(jié)果規(guī)律。

[關(guān)鍵詞]新甲型H1N1流感疫苗；Th1/ Th2細(xì)胞因子；兒童；免疫記憶

[中圖分類號(hào)] R511.7 [文獻(xiàn)標(biāo)識(shí)碼] A [文章編號(hào)] 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.