中华皮肤科杂志 ›› 2022, Vol. 55 ›› Issue (9): 778-783.doi: 10.35541/cjd.20210776

• 论著 • 上一篇    下一篇

白细胞介素18对斑秃患者自然杀伤细胞活性的调控

赵鹏    秦小卫    秦俊霞    梁丽丽    张馨中    高杰   

  1. 山西省人民医院皮肤科，太原  030012
  • 收稿日期:2021-10-22 修回日期:2022-05-25 发布日期:2022-09-02
  • 通讯作者: 秦小卫 E-mail:sxsrmyypfk@126.com
  • 基金资助:
    山西省科技厅重点研发计划项目（201803D31158）

Regulatory effect of interleukin-18 on natural killer cell activity in patients with alopecia areata

Zhao Peng, Qin Xiaowei, Qin Junxia, Liang Lili, Zhang Xinzhong, Gao Jie   

  1. Department of Dermatology, Shanxi Provincial People′s Hospital, Taiyuan 030012, China
  • Received:2021-10-22 Revised:2022-05-25 Published:2022-09-02
  • Contact: Qin Xiaowei E-mail:sxsrmyypfk@126.com
  • Supported by:
    Key Research and Development Program from Shanxi Provincial Science and Technology Agency（201803D31158）

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摘要： 【摘要】 目的 观察斑秃患者外周血自然杀伤（NK）细胞亚群和白细胞介素18（IL-18）变化，评估IL-18对NK细胞活性的调控。方法 收集2019年12月至2021年1月在山西省人民医院就诊的67例斑秃患者和25例健康志愿者，分离外周血单个核细胞（PBMC）和血浆，采用流式细胞仪检测PBMC中NK细胞亚群比例，酶联免疫吸附试验检测血浆IL-18水平。使用重组人IL-18刺激PBMC，建立PBMC与721.221细胞、K562细胞、P815-Ab细胞共培养体系，检测NK细胞中CD107a表达细胞比例和CD16表达细胞荧光强度，评估NK细胞功能。组间比较采用t检验或配对t检验。结果 斑秃组CD56+CD16-NK细胞比例（8.12% ± 3.14%）低于对照组（10.78% ± 4.08%，t = 3.33，P = 0.001），CD56+CD16+NK细胞比例（46.08% ± 15.21%）高于对照组（32.14% ± 10.45%，t = 4.22，P < 0.001），CD56-CD16+NK细胞比例（28.81% ± 8.65%）与对照组（27.09% ± 7.62%）比较差异无统计学意义（t = 0.88，P = 0.383）。斑秃组血浆IL-18水平（112.0 ± 23.72 pg/ml）高于对照组（99.34 ± 15.15 pg/ml，t = 2.48，P = 0.015）。斑秃组NK细胞与721.221细胞、K562细胞和P815-Ab细胞共培养后表达CD107a细胞比例（9.53% ± 1.70%、5.15% ± 1.35%、6.50% ± 1.64%）均高于对照组（5.00% ± 1.17%、4.40% ± 1.09%、5.13% ± 1.36%，均P < 0.05）。P815-Ab细胞刺激后，斑秃组CD16阳性NK细胞荧光强度（151.10% ± 59.30%）低于对照组（221.90% ± 93.56%，t = 4.31，P < 0.001）。IL-18刺激后，与721.221细胞共培养的NK细胞CD107a表达比例高于未刺激组（14.47% ± 2.67%、9.93% ± 1.94%，t = 6.00，P < 0.001）；与K562细胞和P815-Ab细胞共培养的NK细胞CD107a表达比例与未刺激组比较差异无统计学意义（P > 0.05）。 结论 斑秃患者IL-18通过增加自然杀伤毒性受体介导的细胞毒性，增强NK细胞活性。

关键词: 斑秃, 白细胞介素18, 自然杀伤T细胞, 抗体依赖细胞细胞毒性, 溶酶体相关膜蛋白质1

Abstract: 【Abstract】 Objective To investigate changes of natural killer （NK） cell subsets and interleukin-18 （IL-18） level in peripheral blood of patients with alopecia areata, and to assess the regulatory effect of IL-18 on NK cell activity. Methods A total of 67 patients with alopecia areata （alopecia areata group） and 25 healthy volunteers （control group） were collected from Shanxi Provincial People′s Hospital between December 2019 and January 2021. Peripheral blood mononuclear cells （PBMCs） and plasma were isolated. The percentage of NK cell subsets was investigated by flow cytometry, and plasma IL-18 level was measured by enzyme-linked immunosorbent assay. PBMCs were stimulated with recombinant human IL-18, and co-culture systems of PBMCs with 721.221 cells, K562 cells and P815-Ab cells were established separately. NK cell function was assessed by determining the percentage of CD107a-expressing NK cells and fluorescence intensity of CD16+ NK cells. Comparisons between groups were performed using t test or paired t test. Results Compared with the control group, the alopecia areata group showed significantly decreased percentage of CD56+CD16－ NK cells（8.12% ± 3.14% vs. 10.78% ± 4.08%, t = 3.33, P = 0.001）, but significantly increased percentage of CD56+CD16+ NK cells （46.08% ± 15.21% vs. 32.14% ± 10.45%, t = 4.22, P < 0.001）, and there was no significant difference in the percentage of CD56-CD16+ NK cells between the alopecia areata group and control group （28.81% ± 8.65% vs. 27.09% ± 7.62%, t = 0.88, P = 0.383）. The plasma IL-18 level was significantly higher in the alopecia areata group than in the control group （112.0 ± 23.72 pg/ml vs. 99.34 ± 15.15 pg/ml, t = 2.48, P = 0.015）. After co-culture with 721.221 cells, K562 cells and P815-Ab cells, the percentage of CD107a-expressing NK cells was significantly higher in NK cells from the alopecia areata group （9.53% ± 1.70%, 5.15% ± 1.35%, 6.50% ± 1.64%, respectively） than in those from the control group （5.00% ± 1.17%, 4.40% ± 1.09%, 5.13% ± 1.36%, respectively, all P < 0.05）. After the stimulation with P815-Ab cells, the alopecia areata group showed significantly decreased fluorescence intensity of CD16+ NK cells （151.10% ± 59.30%） compared with the control group （221.90% ± 93.56%, t = 4.31, P < 0.001）. After IL-18 stimulation, the percentage of CD107a-expressing NK cells significantly increased in the co-culture system of NK cells with 721.221 cells compared with the unstimulated co-culture system （14.47% ± 2.67% vs. 9.93% ± 1.94%, t = 6.00, P < 0.001）, while there was no significant difference between the IL-8-stimulated co-culture system of NK cells with K562 cells or P815-Ab cells and the unstimulated co-culture systems （both P > 0.05）. Conclusion IL-18 could enhance NK cell activity in patients with alopecia areata, likely by promoting natural cytotoxicity receptor-mediated cytotoxicity.

Key words: Alopecia areata, Interleukin-18, Natural killer T-cells, Antibody-dependent cell cytotoxicity, Lysosomal-associated membrane protein 1