调节性B淋巴细胞在视神经脊髓炎谱系疾病患者外周血中的表达和意义

doi:10.12114/j.issn.1007-9572.2023.0031

摘要/Abstract

摘要： 背景调节性B淋巴细胞（Breg）是一类具有负向免疫调节作用的细胞，在多种自身免疫性疾病中表达异常，视神经脊髓炎谱系疾病（NMOSD）系自身免疫性疾病，目前关于Breg在NMOSD患者外周血中的表达和意义的研究较少。目的研究Breg在NMOSD患者外周血中的表达和意义。方法选取2019年6月—2021年12月新疆医科大学第一附属医院神经内科收治的NMOSD患者55例作为NMOSD组，选取同期本院体检正常的健康志愿者50例作为对照组。运用流式细胞仪检测两组研究对象外周血中CD19+CD24hiCD38hi Breg、CD19+CD24hiCD27+ Breg的表达。采用酶联免疫吸附测定（ELISA）法检测和比较两组研究对象血清中白介素10（IL-10）、白介素35（IL-35）和转化生长因子β1（TGF-β1）的水平。结果 NMOSD组患者CD19+CD24hiCD38hi Breg、CD19+CD24hiCD27+ Breg细胞表型比例低于对照组（t=6.235、12.131，P<0.05）。NMOSD组患者IL-10、IL-35、TGF-β1水平均低于对照组（t=4.249、7.316、2.057，P<0.05）。结论 Breg及其细胞因子在NMOSD患者中表达下降，提示负性免疫调节在NMOSD疾病的发生和进展中发挥作用。

关键词: 视神经脊髓炎, 视神经脊髓炎谱系疾病, B淋巴细胞，调节性, 细胞因子类, 免疫调节, 白细胞介素类, 淋巴细胞表型, 酶联免疫吸附测定

Abstract:

Background

Regulatory B-lymphocytes (Breg) are a group of cells with negative immunoregulatory effect, which have been detected of abnormal expression in the researches of various autoimmune diseases. Neuromyelitis optica spectrum disorders (NMOSD) belong to the category of autoimmune diseases, there are few studies on the expression and significance of Breg in the peripheral blood of patients with NMOSD.

Objective

To explore the expression and significance of Breg in peripheral blood of patients with NMOSD.

Methods

A total of 55 patients with NMOSD admitted to the Department of Neurology of the First Affiliated Hospital of Xinjiang Medical University from June 2019 to December 2021 were selected as the NMOSD group, and 50 healthy volunteers in the same period were selected as the control group. The expression of CD₁₉⁺CD₂₄^hiCD₃₈^hi Breg and CD₁₉⁺CD₂₄^hiCD₂₇⁺ Breg in peripheral blood of patients in the two groups was detected using flow cytometry, the levels of interleukin 10 (IL-10), interleukin 35 (IL-35) and transforming growth factor β1 (TGF-β1) in the serum of patients in the two groups were compared and measured by enzyme-linked immunosorbent assay (ELISA) .

Results

The proportions of CD₁₉⁺CD₂₄^hiCD₃₈^hi Breg and CD₁₉⁺CD₂₄^hiCD₂₇⁺ Breg cell phenotypes were lower in patients in the NMOSD group than in the control group (t=6.235, 12.131, P<0.05). The levels of IL-10, IL-35 and TGF-β1 were lower in patients in the NMOSD group than in the control group (t=4.249, 7.316, 2.057, P<0.05) .

Conclusion

The reduced expression of Breg and their cytokines in patients with NMOSD suggests a role for negative immune regulation in the development and progression of NMOSD disease.

Key words: Neuromyelitis optica, Neuromyelitis optica spectrum disorders, B-lymphocytes, regulatory, Cytokines, Immunomodulation, Interleukins, Lymphocyte phenotypings, Enzyme-linked immunosorbent assay

何丹,马建华,马娟,等. 调节性B淋巴细胞在视神经脊髓炎谱系疾病患者外周血中的表达和意义[J]. 中国全科医学, 2023, 26(20): 2513-2517. DOI: 10.12114/j.issn.1007-9572.2023.0031.
HE Dan,MA Jianhua,MA Juan, et al. Expression and Significance of Regulatory B-lymphocytes in Peripheral Blood of Patients with Neuromyelitis Optica Spectrum Disorders[J]. Chinese General Practice, 2023, 26(20): 2513-2517. DOI: 10.12114/j.issn.1007-9572.2023.0031.

图/表 3

表1 两组研究对象CD19+CD24hiCD38hi Breg、CD19+CD24hiCD27+ Breg细胞表型比例比较（±s，%）

Table 1 Comparison of the proportions of CD19+CD24hiCD38hi Breg and CD19+CD24hiCD27+ Breg phenotypes of patients between the two groups

组别	例数	CD₁₉⁺CD₂₄^hiCD₃₈^hi Breg	CD₁₉⁺CD₂₄^hiCD₂₇⁺ Breg
对照组	50	9.19±2.95	23.05±4.70
NMOSD组	55	5.82±2.59	13.21±3.45
t值		6.235	12.131
P值		<0.001	<0.001

图1 两组研究对象外周血CD19+CD24hiCD38hi、CD19+CD24hiCD27+表达散点图注：A为设门CD19，B为对照组-CD19+CD24hiCD38hi，C为NMOSD组-CD19+CD24hiCD38hi，D为设门CD19，E为对照组-CD19+CD24hiCD27+，F为NMOSD组-CD19+CD24hiCD27+；SSC=侧向散射光，FSC=前向散射光，APC=别藻蓝蛋白，PE=人藻红蛋白，NMOSD=视神经脊髓炎谱系疾病。

Figure 1 Scatter graph of CD19+CD24hiCD38hi and CD19+CD24hiCD27+ expression in peripheral blood of patients in the two groups

表2 两组研究对象细胞因子IL-10、IL-35、TGF-β1水平比较（±s，ng/L）

Table 2 Comparison of the levels of cytokines including IL-10，IL-35 and TGF- β1 in patients between the two groups

组别	例数	IL-10	IL-35	TGF-β1
对照组	50	51.41±12.35	251.24±56.89	10.81±5.41
NMOSD组	55	42.80±7.63	181.19±38.49	8.84±4.41
t值		4.249	7.316	2.057
P值		<0.001	<0.001	0.042

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