外周血淋巴细胞亚群检测在系统性硬化症治疗中的应用

doi:10.19723/j.issn.1671-167X.2021.04.017

摘要/Abstract

摘要：

目的: 检测免疫抑制剂治疗中系统性硬化症 (systemic sclerosis, SSc) 患者外周血T淋巴细胞、B淋巴细胞及自然杀伤 (natural killer,NK) 细胞的表达水平,分析其与临床实验室指标之间的相关性,进而探讨外周血淋巴细胞亚群检测在SSc治疗中的意义。方法: 采用流式细胞术检测使用免疫抑制剂的32例SSc患者(SSc组)和 30例健康对照(healthy control,HC)组外周血T、CD4⁺T、CD8⁺T、B、NK细胞数量及比例,比较SSc组与HC组外周血淋巴细胞亚群的差异,分析外周血淋巴细胞亚群与SSc其他实验室及临床指标之间的相关性。结果: 与HC组相比,SSc组中T、CD4⁺T、CD8⁺T、B、NK细胞数量均明显减少 (P<0.05), 同时,NK细胞占淋巴细胞的百分比也明显降低(P=0.004); 此外,使用免疫抑制剂的SSc患者中65%以上外周血存在各淋巴细胞亚群细胞数量减少。CD4⁺T淋巴细胞数量降低组与正常组相比,其出现雷诺现象的比例明显升高(P=0.024), 红细胞沉降率和C-反应蛋白也明显升高(P<0.001,P=0.018); CD8⁺T淋巴细胞数量降低组与正常组相比,红细胞沉降率明显升高(P=0.022);B淋巴细胞数量降低组与正常组相比,发生指尖溃疡的风险明显增高(P=0.019); NK细胞数量降低组与正常组相比,发生指尖溃疡的风险明显增高(P=0.033),而体内免疫球蛋白(immunoglobulin,Ig)M水平明显降低(P=0.049)。相关性分析可见,红细胞沉降率与总T淋巴细胞(r=-0.455,P=0.009)、CD4⁺T淋巴细胞(r=-0.416,P=0.018)、CD8⁺T淋巴细胞(r=-0.430,P=0.014)、B细胞(r=-0.366,P=0.039)数量呈负相关。结论: ⁺T、CD8⁺T、B及NK细胞数量明显减少,某些淋巴细胞亚群的变化可能与雷诺现象、指尖溃疡的发生有关,与红细胞沉降率、C-反应蛋白呈明显负相关,使用免疫抑制剂治疗SSc中应定期检测外周血淋巴细胞亚群的细胞数量。

关键词: 系统性硬化症, T淋巴细胞, B淋巴细胞, 自然杀伤细胞

Abstract:

Objective: To explore the significance of lymphocytes in systemic sclerosis (SSc), by detecting the levels of T lymphocytes, B lymphocytes and natural killer (NK) cells, and analyzing the correlation between the lymphocytes and clinical laboratory indexes. Methods: The numbers and proportion of T, CD4⁺T, CD8⁺T, B, and NK cells were detected by flow cytometry in peripheral blood of 32 SSc patients who had taken immunosuppressive drugs and 30 healthy controls (HC). The comparison of the lymphocyte subsets in SSc with them in the HC groups, and the correlation between the lymphocytes and other clinical and laboratory indicators were analyzed by the relevant statistical analysis. Results: Compared with the HC group, the numbers of T, CD4⁺T, CD8⁺T, and NK cells in peripheral blood of SSc group,who had taken immunosuppressive drugs, were significantly decreased (P<0.05). More-over, the proportion of NK cells in peripheral blood of the SSc group was also significantly lower than that in the HC group (P=0.004). In addition, all the lymphocyte subsets were decreased in peripheral blood of more than 65% of the SSc patients who had taken immunosuppressive drugs. Compared with CD4⁺T normal group, the positivity of Raynaud’s phenomenon, erythrocyte sedimentation rate (ESR) and C-reactive protein (CRP) was significantly increased in CD4⁺T reduction group, respectively (P=0.024, P<0.001, P=0.018). ESR was higher in CD8⁺T reduction group than CD8⁺T normal group (P=0.022). The prevalence of fingertip ulcer was significantly increased in B cell decrease group (P=0.019). Compared with NK cell normal group, the prevalence of fingertip ulcer was significantly increased in NK cell lower group (P=0.033), IgM was remarkablely decreased yet (P=0.049). The correlation analysis showed that ESR was negatively correlated with the counts of T lymphocytes (r=-0.455, P=0.009), CD4⁺T lymphocytes (r=-0.416, P=0.018), CD8⁺T lymphocytes (r=-0.430, P=0.014), B cells (r=-0.366, P=0.039). Conclusion: The number of CD4⁺T, CD8⁺T, B, and NK cells significantly decreased in peripheral blood of SSc patients who had used immunosuppressive drugs, some lymphocyte subsets might be related with Raynaud’s phenomenon and fingertip ulcer, and reflected the disease activity by negatively correlated with ESR and CRP; the numbers of lymphocyte subsets in peripheral blood should be detected regularly in SSc patients who had taken immunosuppressive drugs.

Key words: Systemic sclerosis, T lymphocytes, B lymphocytes, Natural killer cells

中图分类号:

R593.25

马向波,张学武,贾汝琳,高颖,刘洪江,刘玉芳,李英妮. 外周血淋巴细胞亚群检测在系统性硬化症治疗中的应用[J]. 北京大学学报（医学版）, 2021, 53(4): 721-727.

MA Xiang-bo,ZHANG Xue-wu,JIA Ru-lin,GAO Ying,LIU Hong-jiang,LIU Yu-fang,LI Ying-ni. Application of lymphocytes test in peripheral blood of patients with systemic sclerosis during the treatment[J]. Journal of Peking University (Health Sciences), 2021, 53(4): 721-727.

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Group	Number of lymphocytes/(×10⁶/L)
Group	T cells	CD4⁺T cells	CD8⁺T cells	B cells	NK cells
HC	1 539(1 296-1704)	835(678-926)	566(477-721)	228(169-293)	262(216-336)
SSc	1 010(554-1368)	639(355-803)	347(205-529)	141(73-262)	143(59-179)
P	<0.001	0.017	0.001	0.022	<0.001

Group	Percentage of cells/%
Group	T cells	CD4⁺T cells	CD8⁺T cells	B cells	NK cells
HC	73.61±6.36	41.41±7.28	29.65±6.28	11.29±3.83	14.27±4.79
SSc	76.62±10.14	48.05±11.35	28.80±9.60	12.16±6.88	10.02±5.43
P	0.099	0.017	0.260	0.573	0.004

Items	Reduction group (n=14)	Normal group (n=18)	P
Age/years, $\bar{x}±s$	51.64±14.56	49.72±13.17	0.699
Women, n(%)	13 (92.86)	18 (100)	1.000
Disease duration/years, M(P₂₅, P₇₅)	8.50 (1, 28)	3.75 (0.5, 20)	0.085
Raynaud’s phenomenon, n(%)	14 (100)	12 (66.67)	0.024
Diffuse cutaneous changes, n(%)	13 (92.86)	12 (66.67)	0.104
Fingertip ulcer, n(%)	3 (21.43)	4 (22.22)	1.000
Pulmonary interstitial fibrosis, n(%)	12 (83.71)	11 (61.11)	0.235
ANA, n(%)	13 (92.86)	16 (88.89)	1.000
Scl-70, n(%)	5 (35.71)	6 (33.33)	1.000
ESR/(mm/h), M(P₂₅, P₇₅)	25 (18.00, 41.50)	9.00 (7.50, 14.50)	<0.001
CRP/(mg/L), M(P₂₅, P₇₅)	7.93 (1.76, 14.72)	2.21 (0.47, 3.14)	0.018
IgG/(g/L), M(P₂₅, P₇₅)	14.25 (10.20, 19.70)	11.05 (9.65, 15.40)	0.180
IgM/(g/L), M(P₂₅, P₇₅)	0.81 (0.64, 1.05)	0.85 (0.66, 1.04)	0.955
IgA/(g/L), $\bar{x}±s$	2.11±1.28	2.24±1.49	0.798
C3/(g/L), $\bar{x}±s$	0.85±0.24	0.89±0.17	0.583
C4/(g/L), $\bar{x}±s$	0.19±0.08	0.19±0.07	0.979

	Reduction group (n=15)	Normal group (n=17)	P
Age/years, $\bar{x}±s$	47.76±13.79	53.73±13.10	0.221
Women, n(%)	15 (100)	16 (94.12)	0.531
Disease duration/years, M(P₂₅, P₇₅)	4 (0.5, 20)	6.5 (1, 28)	0.189
Raynaud’s phenomenon, n(%)	12 (70.59)	14 (93.33)	0.178
Diffuse cutaneous changes, n(%)	13 (76.47)	12 (80)	0.355
fingertip ulcer, n(%)	3 (17.65)	4 (26.66)	0.678
Pulmonary interstitial fibrosis, n(%)	12 (70.59)	11 (73.33)	1.000
ANA, n(%)	14 (93.33)	15 (88.24)	1.000
Scl-70, n(%)	5 (33.33)	6 (35.29)	1.000
ESR/(mm/h), M(P₂₅, P₇₅)	23.00 (11.50, 27.50)	9.00 (7.50, 17.00)	0.022
CRP/(mg/L), M(P₂₅, P₇₅)	7.15 (0.71, 12.83)	2.43 (0.55, 3.14)	0.142
IgG/(g/L), M(P₂₅, P₇₅)	13.50 (9.30, 16.40)	11.20 (9.50, 15.70)	0.551
IgM/(g/L), M(P₂₅, P₇₅)	0.77 (0.52, 0.94)	0.91 (0.72, 1.18)	0.370
IgA/(g/L), $\bar{x}±s$	1.96±1.27	2.39±1.48	0.387
C3/(g/L), $\bar{x}±s$	0.87±0.20	0.88±0.21	0.940
C4/(g/L), $\bar{x}±s$	0.20±0.07	0.18±0.08	0.520

	Reduction group (n=10)	Normal group (n=22)	P
Age/years, $\bar{x}±s$	56.00±12.61	48.09±13.57	0.129
Women, n(%)	10 (100)	21 (95.45)	1.000
Disease duration/years, M(P₂₅, P₇₅)	10.5 (1, 28)	4 (0.5, 20)	0.052
Raynaud’s phenomenon, n(%)	10 (100)	16 (72.73)	0.142
Diffuse cutaneous changes, n(%)	10 (100)	15 (68.18)	0.069
fingertip ulcer, n(%)	5 (50)	2 (9.09)	0.019
Pulmonary interstitial fibrosis, n(%)	8 (80)	15 (68.18)	0.681
ANA, n(%)	8 (80.00)	21 (95.45)	0.224
Scl-70, n(%)	4 (40.00)	7 (31.82)	0.703
ESR/(mm/h), M(P₂₅, P₇₅)	21.00 (10.00, 26.00)	10.50 (7.50, 20.00)	0.092
CRP/(mg/L), M(P₂₅, P₇₅)	6.78 (0.97, 9.9)	2.50 (0.55, 5.19)	0.325
IgG/(g/L), M(P₂₅, P₇₅)	12.25 (10.15, 14.4)	11.70 (9.50, 15.70)	0.889
IgM/(g/L), M(P₂₅, P₇₅)	0.81 (0.52, 0.94)	0.87 (0.72, 1.18)	0.562
IgA/(g/L), $\bar{x}±s$	2.79±1.74	1.91±1.12	0.092
C3/(g/L), $\bar{x}±s$	0.90±0.21	0.86±0.20	0.639
C4/(g/L), $\bar{x}±s$	0.21±0.07	0.19±0.08	0.491