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度普利尤单抗治疗特应性皮炎患者的白细胞、B和T淋巴细胞评估以及B淋巴细胞上CD200和CD23的表达——一项试点研究

Evaluation of Leukocytes, B and T Lymphocytes, and expression of CD200 and CD23 on B lymphocytes in Patients with Atopic Dermatitis on Dupilumab Therapy-Pilot Study.

作者信息

Čelakovská Jarmila, Čermáková Eva, Boudková Petra, Andrýs Ctirad, Krejsek Jan

机构信息

Department of Dermatology and Venereology, Faculty Hospital and Medical Faculty of Charles University, 50002, Hradec Králové, Czech Republic.

Department of Medical Biophysics, Medical Faculty of Charles University, 50002, Hradec Králové, Czech Republic.

出版信息

Dermatol Ther (Heidelb). 2023 May;13(5):1171-1192. doi: 10.1007/s13555-023-00918-y. Epub 2023 Apr 25.

Abstract

BACKGROUND

There are a lot of studies that describe the change in quantity of T cells in patients with atopic dermatitis (AD) compared with healthy subjects. Other components of lymphocytes such as B cells are not examined as well as T cells.

OBJECTIVE

We focus on immunophenotyping of B cells with their subsets (memory, naïve, switched, non-switched) and the expression of CD23 and CD200 markers in patients with AD with and without dupilumab therapy. We also evaluate the count of leukocytes and their subsets, T lymphocytes (CD4, CD8), natural killer (NK) cells, and T regulatory cells.

METHODS

A total of 45 patients suffering from AD were examined: 32 patients without dupilumab treatment (10 men, 22 women, average age 35 years), 13 patients with dupilumab treatment (7 men, 6 women, average age 43.4 years), and 30 subjects as a control group (10 men, 20 women, average age 44.7 years). Immunophenotype was examined by flow cytometry in which monoclonal antibodies with fluorescent molecules were used. We compared the absolute and relative count of leukocytes and their subsets, T lymphocytes (CD4 , CD8), NK cells, T regulatory cells, absolute and relative count of B lymphocytes (memory, naïve, non-switched, switched, transient), and expression of CD23 and CD200 activation markers on B cells and on their subsets in patients with AD and control group. For statistical analysis we used nonparametric Kruskal-Wallis one-factor analysis of variance with post hoc by Dunn's test with Bonferroni modification of significance level.

RESULTS

In patients with AD with and without dupilumab therapy we confirmed the significantly higher count of neutrophils, monocytes, and eosinophils; there was no difference in absolute count of B cells, NK cells and transitional B cells compared with control subjects. We confirmed higher expression of activation marker CD23 on total, memory, naïve, non-switched, and switched B lymphocytes and higher expression of CD200 on total B lymphocytes in both groups of patients with AD compared with controls. In patients without dupilumab therapy we confirmed significantly higher count of relative monocytes, relative eosinophils, and higher expression of CD200 on memory, naïve, and non-switched B lymphocytes compared with controls. In patients with dupilumab therapy we confirmed significantly higher expression of CD200 on switched B lymphocytes, higher count of relative CD4 T lymphocytes, and lower count of absolute CD8 T lymphocytes compared with controls.

CONCLUSION

This pilot study shows higher expression of CD23 on B lymphocytes and on their subsets in patients with AD with and without dupilumab therapy. The higher expression of CD200 on switched B lymphocytes is confirmed only in patients with AD with dupilumab therapy.

摘要

背景

有许多研究描述了特应性皮炎(AD)患者与健康受试者相比T细胞数量的变化。淋巴细胞的其他成分如B细胞并未像T细胞那样得到充分研究。

目的

我们聚焦于AD患者在接受和未接受度普利尤单抗治疗时B细胞及其亚群(记忆性、幼稚型、转换型、未转换型)的免疫表型分析以及CD23和CD200标志物的表达情况。我们还评估白细胞及其亚群、T淋巴细胞(CD4、CD8)、自然杀伤(NK)细胞和调节性T细胞的计数。

方法

共检查了45例AD患者:32例未接受度普利尤单抗治疗(10例男性,22例女性,平均年龄35岁),13例接受度普利尤单抗治疗(7例男性,6例女性,平均年龄43.4岁),以及30名作为对照组的受试者(10例男性,20例女性,平均年龄44.7岁)。通过流式细胞术检查免疫表型,其中使用了带有荧光分子的单克隆抗体。我们比较了AD患者和对照组中白细胞及其亚群、T淋巴细胞(CD4、CD8)、NK细胞、调节性T细胞的绝对计数和相对计数,B淋巴细胞(记忆性、幼稚型、未转换型、转换型、短暂型)的绝对计数和相对计数,以及B细胞及其亚群上CD23和CD200激活标志物的表达。对于统计分析,我们使用非参数Kruskal-Wallis单因素方差分析,并通过Dunn检验进行事后检验,对显著性水平进行Bonferroni校正。

结果

在接受和未接受度普利尤单抗治疗的AD患者中,我们证实中性粒细胞、单核细胞和嗜酸性粒细胞的计数显著更高;与对照组相比,B细胞、NK细胞和过渡性B细胞的绝对计数没有差异。与对照组相比,两组AD患者中总B淋巴细胞、记忆性B淋巴细胞、幼稚型B淋巴细胞、未转换型B淋巴细胞和转换型B淋巴细胞上激活标志物CD23的表达更高,总B淋巴细胞上CD200的表达更高。在未接受度普利尤单抗治疗的患者中,与对照组相比,我们证实相对单核细胞、相对嗜酸性粒细胞的计数显著更高,记忆性、幼稚型和未转换型B淋巴细胞上CD200的表达更高。在接受度普利尤单抗治疗的患者中,与对照组相比,我们证实转换型B淋巴细胞上CD200的表达显著更高,相对CD4 T淋巴细胞的计数更高,绝对CD8 T淋巴细胞的计数更低。

结论

这项初步研究表明,在接受和未接受度普利尤单抗治疗的AD患者中,B淋巴细胞及其亚群上CD23的表达更高。仅在接受度普利尤单抗治疗的AD患者中证实转换型B淋巴细胞上CD200的表达更高。

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