摘要目的 探讨H7N9禽流感患者外周血T淋巴细胞亚群及相关细胞因子的变化及临床意义.方法 2013年4月至2015年4月在苏州大学附属第一医院救治的H7N9禽流感患者23例作为H7N9感染组,选取同期在该院门诊体检健康者20名作为健康对照组.采用流式细胞术检测外周血T淋巴细胞亚群CD3+CD4+T淋巴细胞、CD3+CD8+T淋巴细胞、Treg、Th17、CD3+CD8-γ干扰素+(Th1)、CD3+CD8-IL-4+(Th2)百分率的变化,采用Bio-PlexProhumancytokineGroupⅠ27-plexpanel和GroupⅡ21-plexpanel测定血清中相关细胞因子[IL-8、IL-18、干扰素诱导蛋白(IP)-10、巨噬细胞炎性蛋白(MIP)-1b]水平的变化.采用Kolmogorov-Smirnov检验行正态性检验,两组间比较应用两独立样本t检验.结果 治疗前,H7N9感染组外周血CD3+CD4+T淋巴细胞百分率[(27.90±10.19)%比(38.75±6.78)%,t=-2.726,P=0.012]、CD3+CD8+T淋巴细胞百分率[(14.82±7.72)%比(22.79±6.12)%,t=-2.556,P=0.018]明显低于健康对照组,Th17百分率[(2.64±1.40)%比(0.29±0.21)%,t=4.668,P<0.001]、Th2百分率[(2.24±2.00)%比(0.35±0.25)%,t=2.626,P=0.014]较健康对照组明显升高.治疗后好转15例(好转组),治疗后外周血CD3+CD4+T淋巴细胞[(36.54±9.20)%比(25.86±7.22)%,t=-3.339,P=0.007]、CD3+CD8+T淋巴细胞百分率[(22.70±5.68)%比(15.08±8.80)%,t=-2.811,P=0.017]、Treg百分率[(6.08±1.70)%比(3.26±1.64)%,t=-6.670,P<0.001]较治疗前明显升高,Th17百分率[(2.12±1.28)%比(2.76±1.33)%,t=2.220,P=0.048]、Th1细胞百分率[(4.00±2.13)%比(6.97±1.81)%,t=4.407,P=0.001]较治疗前明显下降;死亡8例(死亡组),治疗前后上述免疫学指标的差异均无统计学意义(均P>0.05).治疗前,H7N9感染组IL-8[(23.19±14.35)比(12.78±6.76)ng/L,t=2.277,P=0.035]、IL-18[(230.55±230.18)比(72.80±27.91)ng/L,t=2.348,P=0.036]、IP-10[(28870.55±41815.22)比(1356.13±1093.10)ng/L,t=2.371,P=0.035]、MIP-1b[(197.74±119.87)比(118.51±41.86)ng/L,t=2.198,P=0.043]水平较健康对照组明显增高.结论 H7N9禽流感患者存在严重的T淋巴细胞失衡,监测H7N9禽流感患者外周血T淋巴细胞变化,对评估临床预后具有重要的参考价值,在H7N9禽流感发展过程中存在着细胞因子风暴,可能是引起多脏器功能衰竭的主要原因.

abstractsObjective To investigate the changes and clinical significance of peripheral blood T lymphocyte subsets and cytokines in influenza A (H7N9) virus infection patients.Methods Twenty-three patients with H7N9 virus infection who received treatment at the First Affiliated Hospital of Soochow University from April 2013 to April 2015 were enrolled as case group.Twenty healthy subjects in the outpatient clinic during the same period were selected as control group.The percentages of T lymphocyte subsets in the peripheral blood including CD3+CD4+ T cells, CD3+CD8+ T cells, regulatory T lymphocytes (Treg), Th17, CD3+CD8-interferon (IFN)-γ+ (Th1) cells and CD3+CD8-IL-4+ (Th2) cells were detected by flow cytometry.The changes of plasma cytokines including interleukin (IL)-8, IL-18, interferon-inducible protein (IP)-10, macrophage inflammatory protein (MIP)-1b were quantified by Bio-Plex Pro human cytokine Group Ⅰ 27-plex panel and Group Ⅱ 21-plex panel.The normality test was performed by Kolmogorov-Smirnov test.Two independent samples t test were used to compare the two groups.Results The percentages of CD3+CD4+ and CD3+CD8+ T cells before treatment in case group were significantly lower than control group ([27.90±10.19]% vs [38.75±6.78]%, t=-2.726, P=0.012;[14.82±7.72]% vs [22.79±6.12]%, t=-2.556, P=0.018), while the percentages of Th17 and Th2 before treatment in case group were significantly higher than control group ([2.64±1.40]% vs [0.29±0.21]%, t=4.668, P<0.001;[2.24±2.00]% vs [0.35±0.25]%, t=2.626, P=0.014).After treatment, 15 cases achieved improvement, among which the percentages of CD3+CD4+ T cells, CD3+CD8+ T cells and Treg after treatment were significantly higher than those before treatment ([36.54±9.20]% vs [25.86±7.22]%, t=-3.339, P=0.007;[22.70±5.68]% vs [15.08±8.80]%, t=-2.811, P=0.017;[6.08±1.70]% vs [3.26±1.64]%, t=-6.670, P<0.001), while the percentages of Th17 and Th1 cells after treatment were significantly lower than those before treatment ([2.12±1.28]% vs [2.76±1.33]%, t=2.220, P=0.048;[4.00±2.13]% vs [6.97±1.81]%, t=4.407, P=0.001).A total of 8 patients died with no significance differences of all the above mentioned immunological parameters after treatment compared with before treatment (all P>0.05).Before treatment, levels of IL-8, IL-18, IP-10 and MIP-1b of case group were significantly higher than those in control group (IL-8: [23.19±14.35] vs [12.78±6.76] ng/L, t=2.277, P=0.035;IL-18:[230.55±230.18] vs [72.80±27.91] ng/L, t=2.348, P=0.036;IP-10:[28 870.55±41 815.22] vs [1 356.13±1 093.10] ng/L, t=2.371, P=0.035;MIP-1b: [197.74±119.87] vs [118.51±41.86] ng/L, t=2.198, P=0.043).Conclusions Patients with H7N9 virus infection exhibit an imbalance of T lymphocyte subsets.It is very important to monitor the changes of T lymphocyte subsets in those patients for clinical prognosis.A storm of cytokines could exist during H7N9 virus infection, which may be the main reason for multiple organ failure.