摘要
目的 探讨抗CD122抗体对小鼠银屑病样皮损的影响及其作用机制.方法 研究时间为2020年7月1日至11月23日.选用56只BALB/c雌性小鼠随机分配为4组:凡士林对照组(空白对照组)、咪喹莫特模型组(模型组)、同型对照抗体实验组(实验组A)、抗CD122抗体实验组(实验组B).采用银屑病面积与严重程度指数(PASI)评分观察银屑病样皮损动态变化;显微镜下观察皮损组织病理改变,并测量表皮层厚度;采用免疫荧光双标法检测皮损中TRM细胞的表型及变化;免疫组织化学法检测皮损中白细胞介素-15(IL-15)、白细胞介素-17A(IL-17A)的表达及变化.计量资料采用独立样本t检验.结果 免疫荧光双标显示,模型组初次(3 d、6 d)TRM细胞数量CD3+CD103+[(16.40±1.43)细胞个数/视野、(29.80±2.28)细胞个数/视野]、CD4+CD103+[(10.80±1.77)细胞个数/视野、(16.20±1.28)细胞个数/视野]、CD8+CD103+[(15.10±1.65)细胞个数/视野、(26.05±1.43)细胞个数/视野]、CD122+CD103+[(16.52±1.43)细胞个数/视野、(26.80±1.40)细胞个数/视野]均高于对照组(均P＜0.01);实验组B初次(3d、6 d)及再次(38 d、41 d)刺激皮损中TRM细胞数量均低于同一时间模型组、实验组A(均P＜0.05),以CD8±TRM细胞降低为主(P＜0.01).免疫组织化学结果显示,模型组再次(38 d、41 d)皮损中IL-15、IL-17A的表达[(115.66±3.69)IOD/area、(121.65±3.36)IOD/area及(104.30± 3.47)IOD/area、(59.87±3.20)IOD/area]均高于对照组[(28.15±2.25)IOD/area、(28.05±2.15)IOD/area及(25.63±2.10)IOD/area、(25.01±2.71)IOD/area],差异均有统计学意义(均P＜0.05);实验组B初次(3 d、6 d)及再次(38 d、41 d)刺激皮损中IL-15、IL-17A的表达均低于同一时间模型组和实验组A(均P＜0.05).结论 TRM细胞、IL-15、IL-17A可能参与小鼠银屑病样皮损的发生和发展,抗CD122抗体可以改善小鼠的银屑病样皮损,该作用可能与其抑制TRM细胞的形成和功能(以CD8+TRM细胞为主),以及降低IL-15、IL-17A的表达有关.
Abstract
Objective To explore the effect and mechanism of anti-CD122 antibody on psoriasis-like skin lesions in mice.Methods The study was conducted from July 1 to November 23,2020.A total of 56 BALB/c female mice were randomly assigned to four groups:a vaseline control group(blank control group),an imiquimod model group(model group),an isotype control antibody experimental group(experimental group A),and an anti-CD122 antibody experimental group(experimental group B).The dynamic changes of psoriasis-like skin lesions were observed using the Psoriasis Area and Severity Index(PASI)score;the pathological changes of skin lesion tissues were observed under the microscope and the thickness of the epidermal layer was measured;the phenotypes and changes of TRM cells in skin lesions were detected using the immunofluorescence double-labeling method;the expressions and changes of interleukin(IL)-15 and IL-17A in skin lesions were detected using the immunohistochemical method.Independent sample t test was used for the measurement data.Results The immunofluorescence double-labeling showed that the number of TRM cells in the initial lesions(3 and 6 days)of the model group[CD3+CD103+:(16.40± 1.43)cell count/field of view,(29.80±2.28)cell count/field of view;CD4+CD103+:(10.80± 1.77)cell count/field of view,(16.20±1.28)cell count/field of view;CD8+CD103+:(15.10± 1.65)cell count/field of view,(26.05±1.43)cell count/field of view;CD122+CD103+:(16.52± 1.43)cell count/field of view,(26.80±1.40)cell count/field of view]were higher than those in control group(all P＜0.01).The number of TRM cells in the initial(3 and 6 days)and re-stimulated(38 and 41 days)lesions of the experimental group B were lower than those in the model group and the experimental group A at the same time(all P＜0.05),with CD8+TRM cells predominantly reduced(P＜0.01).The immunohistochemical results showed that the expressions of IL-15 and IL-17A in the initial lesions(38 and 41 days)of the model group[(115.66±3.69)IOD/area,(121.65±3.36)IOD/area,(104.30±3.47)IOD/area,and(59.87±3.20)IOD/area]were higher than those of the control group[(28.15±2.25)IOD/area,(28.05±2.15)IOD/area,(2 5.63±2.10)IO D/area,and(25.01±2.71)IO D/area],with statistically significant differences(all P＜0.05).The expressions of IL-15 and IL-17A in the initial(3 and 6 days)and re-stimulated(38 and 41 days)lesions of the experimental group B were lower than those of the model group and experimental group A at the same time(all P＜0.05).Conclusions TRM cells,IL-15,and IL-17A might be involved in the occurrence and development of psoriasis-like skin lesions in mice.Anti-CD122 antibody can improve psoriasis-like lesions in mice,and this effect might be related to its inhibition of formation and function of TRM cells,mainly CD8+TRM cells,as well as the reduction of IL-15 and IL-17A expressions.
基金项目
2021年广东省第二人民医院3D打印科研项目(3D-A2021001)
维普
万方数据