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中华实验和临床感染病杂志(电子版) ›› 2021, Vol. 15 ›› Issue (03) : 182 -188. doi: 10.3877/cma.j.issn.1674-1358.2021.03.007

论著

新型冠状病毒肺炎患者早期外周血淋巴细胞亚群及细胞因子特征
廖宝林1, 施海燕1, 刘艳霞1, 赵涵1, 焦倩1, 李丽雅1, 王信1, 曹意1, 何浩岚1,()   
  1. 1. 510060 广州市,广州医科大学附属市八医院感染病中心
  • 收稿日期:2020-07-09 出版日期:2021-06-15
  • 通信作者: 何浩岚
  • 基金资助:
    广州市科技创新委员会健康医疗协同创新重大计划课题项目(No. 201803040002); 广州市高水平临床重点专科传染病专科(No. 2019-2021); "十三五"国家科技重大专项(No. 2017ZX10202102-004-001)

Features of lymphocytes subsets and cytokines in peripheral blood of patients with coronavirus disease 2019 during early phase

Baolin Liao1, Haiyan Shi1, Yanxia Liu1, Han Zhao1, Qian Jiao1, Liya Li1, Xin Wang1, Yi Cao1, Haolan He1,()   

  1. 1. Department of Infectious Diseases, Guangzhou Eighth People’s Hospital, Guangzhou Medical University, Guangzhou 510060, China
  • Received:2020-07-09 Published:2021-06-15
  • Corresponding author: Haolan He
引用本文:

廖宝林, 施海燕, 刘艳霞, 赵涵, 焦倩, 李丽雅, 王信, 曹意, 何浩岚. 新型冠状病毒肺炎患者早期外周血淋巴细胞亚群及细胞因子特征[J/OL]. 中华实验和临床感染病杂志(电子版), 2021, 15(03): 182-188.

Baolin Liao, Haiyan Shi, Yanxia Liu, Han Zhao, Qian Jiao, Liya Li, Xin Wang, Yi Cao, Haolan He. Features of lymphocytes subsets and cytokines in peripheral blood of patients with coronavirus disease 2019 during early phase[J/OL]. Chinese Journal of Experimental and Clinical Infectious Diseases(Electronic Edition), 2021, 15(03): 182-188.

目的

分析新型冠状病毒肺炎(COVID-19)早期(起病14天内)外周血免疫学特征及其与疾病严重程度的关系。

方法

收集2020年1月至3月广州医科大学附属市八医院收治的98例确诊COVID-19患者住院资料,分析COVID-19早期外周血淋巴细胞亚群[CD4+ T、CD8+ T细胞、B细胞、自然杀伤(NK)细胞、NKT细胞与调节性T细胞]和细胞因子[白细胞介素(IL)-2、IL-4、IL-6、IL-10、肿瘤坏死因子-α和干扰素-γ]变化特征,以及上述指标与患者病情严重程度的关系。年龄、起病时间、白细胞、中性粒细胞、淋巴细胞、淋巴细胞亚群和细胞因子水平为非正态分布的计量资料,3组患者间比较采用Kruskal-Wallis检验;性别、基础疾病、吸烟、武汉流行病学史、家庭成员被感染、症状、实验室指标异常比例为计数资料,采用Pearson卡方检验或Fisher确切概率检验;以Logistic回归分析影响疾病严重性的高危因素。

结果

根据病情严重程度将入组患者分为轻型组(17例)、普通型组(73例)和重型组(8例)。早期55.1%(54/98)患者CD4+ T细胞、40.8%(40/98)患者CD8+ T细胞和11.8%(10/85)患者B细胞计数下降。3组患者间CD4+ T细胞(P = 0.010)、CD8+ T细胞(P = 0.003)和B细胞(P = 0.022)计数存在显著差异,以重型组水平最低。3组患者IL-6表达差异有统计学意义(P = 0.001),以重型组水平最高,而其他细胞亚群及细胞因子水平则无显著差异(P均> 0.05)。Logistic回归分析显示,IL-6升高(OR = 12.277、P = 0.024)和年龄> 50岁(OR = 21.823、P = 0.030)分别为发生肺炎和重型化的独立预测因素。

结论

COVID-19早期即明显破坏患者机体细胞和体液免疫应答,与疾病进展密切相关。

Objective

To investigate the immunological features from peripheral blood of patients with coronavirus disease 2019 (COVID-19) in early phase (within 14 days after symptom onset), and its association with the severity of disease.

Methods

Total of 98 patients with COVID-19 in Guangzhou Eighth People’s Hospital, Guangzhou Medical University from January to March 2020 were included. The features of patients of lymphocytes [CD4+ T, CD8+ T, B, natural killer (NK), NKT and Treg cells] and cytokines [interleukin (IL)-2, IL-4, IL-6, IL-10, tumor necrosis factor-α and interferon-γ] were analyzed in early phase, and also their associations with the severity of disease were evaluated. The nonparametric data such as age, days after symptom onset, levels of white blood cells, neutrophil, lymphocytes, lymphocytes subsets and cytokines were analyzed by Kruskal-Wallis test. The categorical variables such as sex, complications, smoke, exposure to Wuhan, family members infected, symptoms, abnormal frequency of laboratory test were analyzed by Pearson’s chi-squared test or Fisher’s exact test. The independent risk factors associated with the severity of disease were analyzed by Logistical regression analysis.

Results

Patients were divided into three groups according to the relevant diagnostic criteria: the mild group (17 cases), the normal group (73 cases) and the severe group (8 cases). Decreased CD4+ T cells, CD8+ T cells and B cells were found in 55.1% (54/98), 40.8% (40/98) and 11.8% (10/85) patients, respectively. The levels of CD4+ T cells (P = 0.010), CD8+ T cells (P = 0.003) and B cells (P = 0.022) were significantly different among the three groups, with the lowest levels in the severe group. The levels of IL-6 (P = 0.001) were different among three groups, with the highest levels in the severe group. Levels of other lymphocytes and cytokines were similar, without significant differences (all P > 0.05). Increased level of IL-6 (OR = 12.277, P = 0.024) was independently associated with pneumonia, and age over 50 years old (OR = 21.823, P = 0.030) was independently associated with the severe case by Logistic regression analysis.

Conclusion

The cellular and humoral immune response was significantly destroyed of patients with COVID-19 in early phase, which was significantly associated with the progression of disease.

表1 COVID-19患者一般特征和感染性指标
指标 轻型组(17例) 普通型组(73例) 重型组(8例) 统计值 P
男性[例(%)] 11(64.7) 43(58.9) 8(100.0) χ2= 5.258 0.072*
年龄[M(P25,P75),岁] 25(21,33) 38(23,50) 58(52,73) Z = 22.225 < 0.001
起病时间[M(P25,P75),d] 3(2,4) 5(2,7) 6.5 (1,11) Z = 3.298 0.192
基础疾病[例(%)] 1(5.9) 17(23.3) 4(50.0) 0.052#
吸烟[例(%)] 1(5.9) 9(12.3) 2(25.0) 0.353#
武汉流行病学史[例(%)] 2(11.8) 6(8.2) 1(12.5) 0.573#
家庭成员被感染[例(%)] 2(11.8) 14(19.2) 2(25.0) 0.799#
发热[例(%)] 6(35.3) 33(45.2) 8(100.0) 0.005#
干咳[例(%)] 6(35.3) 29(39.7) 3(37.5) 0.938#
咳痰[例(%)] 5(29.4) 25(34.2) 6(75.0) χ2= 5.627 0.060*
咽痛[例(%)] 3(17.6) 14(19.2) 1(12.5) 1.000#
乏力[例(%)] 3(17.6) 22(30.1) 4(50.0) 0.258#
腹泻[例(%)] 1(5.9) 5(6.8) 1(12.5) 0.616#
气促[例(%)] 1(5.9) 6(8.2) 4(50.0) 0.011#
胸闷[例(%)] 1(5.9) 8(11.0) 1(12.5) 1.000#
头痛[例(%)] 4(23.5) 9(12.3) 1(12.5) 0.478#
肌肉痛[例(%)] 3(17.6) 11(15.1) 0(0) 0.624#
胸痛[例(%)] 0(0) 2(2.7) 0(0) 1.000#
纳差[例(%)] 2(11.8) 16(21.9) 3(37.5) 0.368#
恶心[例(%)] 1(5.9) 4(5.5) 2(25.0) 0.140#
呕吐[例(%)] 0(0) 2(2.7) 1(12.5) 0.297#
白细胞计数[M(P25,P75),× 109/L] 5.5(4.3,7.2) 5.2(4.4,6.7) 5.2(4.5,6.2) Z = 0.217 0.897
中性粒细胞计数[M(P25,P75),× 109/L] 3.7(2.4,4.9) 3.1(2.5,4.3) 3.9(3.2,5.3) Z = 2.191 0.334
淋巴细胞计数[M(P25,P75),× 109/L] 1.6(1.3,2.0) 1.4(1.0,1.8) 1.0(0.5,1.1) Z = 12.327 0.002
C-反应蛋白升高[例(%)] 0(0.0) 10(13.7) 6(75.0) < 0.001#
D二聚体升高[例(%)] 3(17.6) 27(42.2)a 6(85.7)c 0.006#
降钙素原升高[例(%)] 5(29.4) 27(41.5)b 7(100)c 0.003#
表2 COVID-19患者淋巴细胞亚群特征
指标 参考值 轻型组(17例) 普通型组(73例) 重型组(8例) 统计值 P
CD4+T细胞            
  CD4+T细胞计数(个/μl) 550~1 440 493(386,752) 562 (395,682) 320(252,446) Z = 9.245 0.010
  CD4+T细胞计数下降[例(%)]   9(52.9) 37(50.7) 8(100.0) 0.021
  CD4+T细胞比例(%) 26~61 35.3(31.5,43.1) 37.8(33.7,43.7) 35.6(31.2,41.1) Z = 1.805 0.406
  CD4+T细胞比例下降[例(%)] 1(5.9) 1(1.4) 1(12.5) 0.159
CD8+T细胞            
  CD8+T细胞计数(个/μl) 320~1 250 430(321,534) 353(271,512) 209(135,281) Z = 11.632 0.003
  CD8+T细胞计数下降[例(%)] 4(23.5) 29(39.7) 7(87.5) 0.008
  CD8+T细胞比例(%) 15~44 27.9(24.9,33.0) 26.1(21.1,31.5) 22.3(16.4,29.3) Z = 2.904 0.234
  CD8+T细胞比例下降[例(%)]   0(0) 1(1.4) 1(12.5) 0.186
CD4+/CD8+T            
  CD4+/CD8+T比值 1.4~2.0 1.3(1,1.6) 1.5(1.2,1.9) 1.8(1.2,2.4) Z = 2.246 0.325
  CD4+/CD8+T比值下降[例(%)]   10(58.8) 30(41.4) 3(37.5) 0.394
B细胞            
  B细胞计数(个/μl) 90~560 205(144,271)a 145(110。217)b 107(62。206)d Z = 7.652 0.022
  B细胞计数下降[例(%)] 0(0.0)a 7(10.8)b 3(42.9)d 0.030
  B细胞比例(%) 5~18 12.0(11.2,16.8)a 12.1(9.4,13.8)b 9.7(5.7,16.7)d Z = 1.825 0.402
  B细胞比例下降[例(%)]   0(0.0)a 2(3.1)b 1(14.3)d 0.284
NK细胞            
  NK细胞计数(个/μl) 150~1 100 133(102,276)a 191(133,316)b 271(127,330)d Z = 2.762 0.251
  NK细胞计数下降[例(%)]   8(61.5)a 22(33.8)b 2(28.6)d 0.174
  NK细胞比例(%) 7~40 11.8(5.6,19.3)a 13.6(9.9,20.4)b 25.1(15.9,34.1)d Z = 9.848 0.007
  NK细胞比例下降[例(%)]   4(30.8)a 6(9.2)b 0(0)d 0.090
NKT细胞            
  NKT细胞计数(个/μl) 40~300 27(24,46)a 26(15,45)b 13(11,30)d Z = 4.73 0.094
  NKT细胞比例下降[例(%)]   8(61.5)a 44(67.7)b 7(100.0)d 0.188
  NKT细胞比例(%) 2~13 1.9(1.5,2.9)a 1.9(1.2,2.9)b 2(1.4,2.5)d Z = 0.360 0.835
  NKT细胞比例下降[例(%)]   7(53.8)a 35(53.8)b 3(42.9)d 0.929
Treg细胞            
  Treg细胞比例(%) 3~8 10.5(8.8,11.5)a 9.3(7.3,11.2)c 11.4(9.5,12.6)d Z = 3.517 0.172
  Treg细胞比例升高[例(%)]   12(92.3)a 39(63.9)c 6(85.7)d 0.079
表3 COVID-19患者的细胞因子特征
表4 单因素Logistic回归分析影响COVID-19早期患者疾病严重性的高危因素
表5 多因素Logistic回归分析影响COVID-19早期患者疾病严重性的高危因素
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