CAJ | 학술논문

目的探讨甲型H1N1流行性感冒(流感)患者Th17淋巴细胞表型、比例及其与病毒清除之间的关系.方法将甲型H1N1流感患者70例、季节性流感患者30例、健康对照者68例分别纳入3组.通过细胞内染色,流式细胞技术测定3组人群外周血Th1、Th2、Th17、调节性T细胞(Treg)淋巴细胞比例;ELISA法测定血浆及外周血单个核细胞(PBMC)培养上清液中的IFN-γ、转化生长因子-β(TGF-β)、IL-6水平;RT-PCR检测鼻咽拭子甲型H1N1流感病毒载量.统计学方法采用单因素方差分析和线性相关分析.结果甲型H1N1流感患者Th17淋巴细胞比例为(2.740±0.210)%,较健康对照者的(3.443±0.154)%及季节性流感患者的(3.443±0.277)%显著下降(F=4.242,P＜0.05),而3组间Th1、Th2、Treg淋巴细胞比例无明显差异;甲型H1N1流感患者血浆TGF-β水平为(10±8)ng/mL,较健康者的(43±32)ng/mL及季节性流感患者的(18±10)ng/mL显著下降(F=17.72,P＜0.01);甲型H1N1流感患者PBMC中TGF-β水平为(782±736)pg/mL,较健康者的(1462±315)pg/mL及季节性流感患者的(1481±348)pg/mL显著下降(F=5.730,P＜0.01);Th17淋巴细胞比例与病毒清除时间呈负相关(r=-0.38,P=0.02).结论甲型H1N1流感患者Th17淋巴细胞比例显著降低,且与TGF-β水平密切相关,其降低可能导致机体延长排毒时间.
목적 탐토갑형H1N1류행성감모(류감)환자Th17림파세포표형、비례급기여병독청제지간적관계.방법 장갑형H1N1류감환자70례、계절성류감환자30례、건강대조자68례분별납입3조.통과세포내염색,류식세포기술측정3조인군외주혈Th1、Th2、Th17、조절성T세포(Treg)림파세포비례;ELISA법측정혈장급외주혈단개핵세포(PBMC)배양상청액중적IFN-γ、전화생장인자-β(TGF-β)、IL-6수평;RT-PCR검측비인식자갑형H1N1류감병독재량.통계학방법채용단인소방차분석화선성상관분석.결과 갑형H1N1류감환자Th17림파세포비례위(2.740±0.210)%,교건강대조자적(3.443±0.154)%급계절성류감환자적(3.443±0.277)%현저하강(F=4.242,P＜0.05),이3조간Th1、Th2、Treg림파세포비례무명현차이;갑형H1N1류감환자혈장TGF-β수평위(10±8)ng/mL,교건강자적(43±32)ng/mL급계절성류감환자적(18±10)ng/mL현저하강(F=17.72,P＜0.01);갑형H1N1류감환자PBMC중TGF-β수평위(782±736)pg/mL,교건강자적(1462±315)pg/mL급계절성류감환자적(1481±348)pg/mL현저하강(F=5.730,P＜0.01);Th17림파세포비례여병독청제시간정부상관(r=-0.38,P=0.02).결론 갑형H1N1류감환자Th17림파세포비례현저강저,차여TGF-β수평밀절상관,기강저가능도치궤체연장배독시간.
Objective To investigate the phenotype, frequency of Th17 cells and the association between Th17 cells and viral clearance in patients with H1N1 influenza A. Methods Three groups including 70 confirmed patients with H1N1 influenza A, 30 patients with seasonal influenza as well as 68 healthy subjects as controls were enrolled in this study. The percentages of Th1, Th2, Treg and Th17 lymphocytes in the peripheral blood were determined by intracellular staining and flow cytometry. The levels of interferon-γ (IFN-γ), transforming growth factor-beta (TGF-β),interleukin-6 (IL-6) in plasma and supernatant of the peripheral blood mononuclear cell (PBMC)culture were quantified by enzyme-linked immunosorbent assay (ELISA). Viral load in nasopharyngeal swabs was detected by real time quantitative reverse transcription-polymerase chain reaction (RTPCR). Data were analyzed by one way ANOVA and liner correlation analysis. Results The percentage of Th17 cells in H1N1 influenza A patients was (2. 740±0. 210)%, which the percentage of was significantly decreased compared to healthy subjects (3. 443 ±0. 154)% and seasonal influenza patients (3. 443±0. 277) % (F=4. 242, P＜0. 05); while the percentage of Thl, Th2 and Treg cells were not significantly different among these groups. Moreover, the TGF-β level in plasma of H1N1 influenza A patients was (10±8) ng/mL, which was significantly lower than healthy subjects (43 ±32 ) ng/mL and seasonal influenza patient ( 18 ± 10) ng/mL ( F= 17.72, P＜0.01 ). The TGF-β level in the supernatant of PBMC culture of H1N1 influenza A patients was (782 ± 736) pg/mL, which was significantly lower than healthy subjects (1462±315) pg/mL and seasonal influenza patients (1481 ±348) pg/mL (F=5. 730, P＜0.01). Additionally, the viral clearance period was inversely correlated with the percentage of Th17 cells (r=-0.38, P=0.02). Conclusions The proportion of Th17 cells in patients with H1N1 influenza A is significantly decreased, which is closely correlated with the level of TGF-β. This decrease may results in the delayed viral clearance.