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中华实验和临床感染病杂志(电子版) ›› 2023, Vol. 17 ›› Issue (02) : 84 -91. doi: 10.3877/cma.j.issn.1674-1358.2023.02.003

论著

淋巴细胞精细分型检测在人类免疫缺陷病毒感染者/获得性免疫缺陷综合征患者中的应用
袁瑞, 胡文佳, 桂希恩, 严亚军, 冯玲, 柯亨宁, 熊勇, 杨蓉蓉()   
  1. 430071 武汉市,武汉大学中南医院感染科
  • 收稿日期:2022-10-07 出版日期:2023-04-15
  • 通信作者: 杨蓉蓉
  • 基金资助:
    国家自然科学基金项目(No. 82003511)

Application of refined lymphocyte subsets detection on patients with human immunodeficiency virus infection/acquired immunodeficiency syndrome

Rui Yuan, Wenjia Hu, Xien Gui, Yajun Yan, Ling Feng, Hengning Ke, Yong Xiong, Rongrong Yang()   

  1. Department of Infectious Diseases, Zhongnan Hospital of Wuhan University, Wuhan 430071, China
  • Received:2022-10-07 Published:2023-04-15
  • Corresponding author: Rongrong Yang
引用本文:

袁瑞, 胡文佳, 桂希恩, 严亚军, 冯玲, 柯亨宁, 熊勇, 杨蓉蓉. 淋巴细胞精细分型检测在人类免疫缺陷病毒感染者/获得性免疫缺陷综合征患者中的应用[J]. 中华实验和临床感染病杂志(电子版), 2023, 17(02): 84-91.

Rui Yuan, Wenjia Hu, Xien Gui, Yajun Yan, Ling Feng, Hengning Ke, Yong Xiong, Rongrong Yang. Application of refined lymphocyte subsets detection on patients with human immunodeficiency virus infection/acquired immunodeficiency syndrome[J]. Chinese Journal of Experimental and Clinical Infectious Diseases(Electronic Edition), 2023, 17(02): 84-91.

目的

探讨人类免疫缺陷病毒(HIV)感染后免疫细胞的变化特征,以及其在不同疾病分期的动态变化规律。

方法

采用流式细胞术,对2021年8月17日至2022年9月14日于武汉大学中南医院感染科住院的173例HIV感染者/获得性免疫缺陷综合征(AIDS)患者(研究组)进行淋巴细胞亚群分类计数检测,分别与1 086例健康人群(对照组)进行对比分析。根据CD4+ T淋巴细胞计数进行分期,比较HIV/AIDS患者在不同临床分期淋巴细胞亚群分类计数。计量资料中各类淋巴细胞绝对计数均呈正态分布,以±s表示,HIV/AIDS患者和对照组各类淋巴细胞均值的比较采用两独立样本t检验。

结果

研究组患者T淋巴细胞、B淋巴细胞和NK细胞绝对计数分别为(907 ± 105)个/μl、(128 ± 25)个/μl和(176 ± 16)个/μl,显著低于对照组(t = 8.508、P < 0.001,t = 8.265、P < 0.001,t = 18.552,P < 0.001)。研究组患者CD4+CD28+ T细胞和CD8+CD28+ T细胞绝对计数分别为(123 ± 25)个/μl和(170 ± 48)个/μl (t = 28.522、P < 0.001,t = 5.820、P < 0.001),百分比分别为84.86%和35.41%,均显著低于对照组(t = 2.583、P = 0.011,t = 12.806、P < 0.001)。研究组患者CD3+CD8+HLA-DR+细胞计数和百分比分别为(476 ± 129)个/μl和73.68%,显著高于对照组[(135 ± 98)个/μl和22.38%],差异均有统计意义(t = 8.482、P < 0.001,t = 34.651、P < 0.001)。研究组患者初始和记忆CD4+ T淋巴细胞绝对计数分别为(76 ± 9)个/μl和(152 ± 12)个/μl,均显著低于对照组,差异有统计学意义(t = 19.823、P < 0.001,t = 20.815,P < 0.001)。随着疾病进展,研究组患者CD4+CD28+ T细胞和CD8+CD28+ T细胞绝对计数分别由(540.77 ± 165.54)个/μl和(452.57 ± 135.65)个/μl降至(13.56 ± 33.63)个/μl和(102.96 ± 30.47)个/μl,且CD3+CD8+HLA-DR+细胞百分比由(36.00 ± 17.79)%升至(58.29 ± 13.27)%。

结论

HIV/AIDS患者可出现淋巴细胞数量和功能异常,以及免疫系统的异常激活,且AIDS进展后期患者淋巴细胞功能受损和异常免疫激活更显著。

Objective

To investigate the changing characteristics of the immunocytes after human immunodeficiency virus (HIV) infection and the dynamic changing patterns of different disease stages.

Methods

Lymphocyte subsets of 173 patients with HIV/acquired immunodeficiency syndrome (AIDS) hospitalized in Department of Infectious Diseases, Zhongnan Hospital of Wuhan University from August 17th, 2021 to September 14th, 2022 (research group) were analyzed by flow cytometry, and were compared with 1 086 healthy individuals (control group). AIDS was staged according to the CD4+ T lymphocyte count; The lymphocyte subset classification counts of patients with HIV/AIDS at different AIDS stages were compared. The absolute counts of different types of lymphocytes in the measurement data were normally distributed, expressed by ±s, and the mean values of all types of lymphocytes between patients with HIV/AIDS and control group were compared by two independent samples t-tests.

Results

The absolute counts of T lymphocytes, B lymphocytes and NK cells of patient in research group were (907 ± 105) cells/μl, (128 ± 25) cells/μl and (176 ± 16) cells/μl, respectively, which were significantly lower than those in control group (t = 8.508, P < 0.001; t = 8.265, P < 0.001; t = 18.552, P < 0.001). CD4+CD28+ cells and CD8+CD28+ cells of patient in research group were (123 ± 25) cells/μl and (245 ± 98) cells/μl, which were both significantly lower than those of control group (t = 28.522, P < 0.001; t = 5.820, P < 0.001). CD3+CD8+HLA-DR+ cell counts and percentages of patient of research group were (476 ± 129) cells/μl and 73.68%, respectively, which were both higher than those of control group (t = 8.482, P < 0.001; t = 34.651, P < 0.001). The absolute counts of naïve and memory CD4+ T cells of patient of research group were (76 ± 9) cells/μl and (152 ± 12) cells/μl, respectively, which were significantly lower than those of control group (t = 19.823, P < 0.001; t = 20.815, P < 0.001). As the disease progressed, the absolute counts of CD4+CD28+ cells and CD8+CD28+cells gradually decreased from (540.77 ± 165.54) cells/μl and (452.57 ± 135.65) cells/μl to (13.56 ± 33.63) cells/μl and (102.96 ± 30.47) cells/μl, and the percentages of CD3+CD8+ HLA-DR+ cells increased from (36.00 ± 17.79)% to (58.29 ± 13.27)% of patients in research group.

Conclusions

Abnormalities in lymphocyte count and function, as well as abnormal activation of the immune system are common in patients with HIV/AIDS. In late stage of AIDS, impaired lymphocyte function and abnormal immune activation are more significant.

表1 173例HIV/AIDS患者的基本资料
表2 研究组患者和健康对照组淋巴细胞亚群绝对计数和百分比(±s
表3 研究组患者和健康对照组T淋巴细胞第二信号受体(±s
表4 研究组患者和健康对照组辅助/诱导性T淋巴细胞亚群(±s
表5 不同疾病分期HIV/AIDS患者的淋巴细胞亚群分类计数
指标 AIDS分期   F P
轻度(CD4+ T >350个/μl) 中度(CD4+ T:201~350个/μl) 重度(CD4+ T:51~200个/μl) 终末期(CD4+ T≤50个/μl)
CD3+CD4+/CD3+百分比(%) 30.18 ± 8.95 20.53 ± 8.48 13.63 ± 8.42 2.57 ± 0.82 134.355 < 0.001
CD3+CD4+ T淋巴细胞绝对计数(个/μl) 578.59 ± 201.99 259.61 ± 43.88 110.17 ± 39.48 14.52 ± 4.53 172.896 < 0.001
CD3+CD8+/CD3+百分比(%) 39.01 ± 13.27 49.42 ± 13.21 57.54 ± 15.32 64.64 ± 12.17 33.495 < 0.001
CD3+CD8+ T淋巴细胞绝对计数(个/μl) 865.34 ± 139.39 818.55 ± 228.29 603.66 ± 141.02 376.60 ± 121.53 42.391 < 0.001
CD4+/CD8+ T淋巴细胞比值 0.88 ± 0.21 0.38 ± 0.11 0.30 ± 0.09 0.14 ± 0.03 132.356 < 0.001
CD3+ T淋巴细胞百分比(%) 73.22 ± 9.18 73.40 ± 9.75 74.95 ± 13.28 72.37 ± 12.66 0.456 0.714
CD3+ T淋巴细胞绝对计数(个/μl) 1 282 ± 541.02 969.48 ± 207.62 656.06 ± 152.85 363.02 ± 120.03 103.326 < 0.001
CD16+CD56+ T淋巴细胞百分比(%) 12.73 ± 2.06 11.71 ± 2.01 11.37 ± 1.85 14.44 ± 2.11 3.010 0.390
CD16+CD56+ T淋巴细胞绝对计数(个/μl) 274.52 ± 85.02 163.78 ± 52.76 115.82 ± 37.54 61.02 ± 19.87 54.041 < 0.001
CD19+ T淋巴细胞百分比(%) 12.09 ± 3.13 11.93 ± 2.08 9.20 ± 3.11 8.89 ± 2.16 2.194 0.090
CD19+ T淋巴细胞绝对计数(个/μl) 200.52 ± 62.80 121.32 ± 56.77 69.88 ± 32.01 36.27 ± 10.68 69.415 < 0.001
总淋巴细胞数(个/μl) 1 903.22 ± 457.22 1 349.93 ± 305.64 904.58 ± 251.07 493.31 ± 101.23 110.410 < 0.001
CD4+CD28+/CD4+ T百分比(%) 92.88 ± 7.54 92.78 ± 6.81 89.00 ± 17.70 80.62 ± 15.26 13.621 0.003
CD8+CD28+/CD8+ T百分比(%) 53.84 ± 8.97 43.12 ± 12.98 31.00 ± 9.76 27.84 ± 8.67 23.587 < 0.001
CD4+CD28+ T细胞绝对计数(个/μl) 540.77 ± 165.54 241.11 ± 45.40 99.27 ± 29.72 13.56 ± 33.63 232.433 < 0.001
CD8+CD28+ T细胞绝对计数(个/μl) 452.57 ± 135.65 307.81 ± 99.87 173.11 ± 50.05 102.96 ± 30.47 18.423 < 0.001
CD3+HLA-DR+/CD3+ T(%) 36.00 ± 17.79 47.66 ± 15.72 59.18 ± 16.92 58.29 ± 13.27 22.905 < 0.001
CD3+CD8+HLA-DR+/CD8+ T(%) 57.38 ± 20.71 71.19 ± 18.68 80.31 ± 18.76 81.17 ± 11.78 19.862 < 0.001
CD3+HLA-DR+ T细胞绝对计数(个/μl) 604.09 ± 197.56 596.64 ± 168.92 473.91 ± 146.76 262.53 ± 82.88 6.225 < 0.001
CD3+CD8+HLADR+ T细胞绝对计数(个/μl) 554.24 ± 165.33 636.38 ± 209.26 490.17 ± 139.00 311.09 ± 98.16 3.522 0.016
CD3+CD4+CD45RA+/CD4+ T(%) 35.93 ± 14.70 26.61 ± 16.72 17.64 ± 4.15 18.83 ± 5.84 14.201 < 0.001
CD3+CD4+CD45RO/CD4+ T(%) 64.06 ± 14.70 73.38 ± 16.72 82.35 ± 13.15 81.17 ± 17.84 14.218 < 0.001
CD3+CD4+CD45RA细胞绝对计数(个/μl) 220.95 ± 68.74 68.71 ± 19.20 19.71 ± 5.55 4.25 ± 1.30 77.91 < 0.001
CD3+CD4+CD45RO细胞绝对计数(个/μl) 357.61 ± 107.12 190.89 ± 49.21 90.46 ± 23.96 14.26 ± 3.22 272.409 < 0.001
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