斑秃皮损内朗格汉斯细胞及CD8+T细胞的数量及分布分析

摘要

Objective To analyze the quantity and distribution of Langerhans cells in lesions of alopecia areata (AA),so as to investigate their role in the pathological process of AA as well as their relationship with T lymphocytes.Methods Tissue specimens were obtained from the scalp lesions of AA in 29 patients,including 16 in active phase and 13 in non-active phase.All the specimens were subjected to immunohistochemical staining for CD1a,and 17 specimens to immunohistochemical staining for CD4 and CD8.Fluorescence-based semi-quantitative reverse transcription (RT)-PCR was performed to determine the mRNA expressions of CD1a and granulocytemacrophage colony stimulating factor (GM-CSF) in the full-thickness skin specimens.Statistical analysis was done using Mann Whitney U test and Pearson correlation analysis.Results There was a significant increase in the quantity of CD1a+ Langerhans cells in the epidermis,perivascular and perifollicular areas in the upper and deep dermis of the AA patients compared with the healthy controls (Z =4.354,2.884,4.640,3.217,3.496,all P ＜ 0.01),and in the epidermis,perivascular and perifollicular areas in the deep dermis of AA patients in active phase compared with those in non-active phase (Z =2.457,2.130,1.954,P ＜ 0.05 or =0.05).The relative mRNA expression levels of CD1a and GM-CSF in the patients were similar to the healthy controls in the upper dermis,but significandy higher than the healthy controls in the deep dermis (Z =2.702,2.941,both P ＜ 0.01).A positive correlation was observed between the quantity of perivascular Langerhans cells in the upper dermis and that of perifollicular CD8+ T lymphocytes in the deep dermis in patients with AA (r =0.618,P ＜ 0.05) and in patients with active AA (r =0.795,P =0.01),but not in patients with inactive AA (r =0.304,P ＞ 0.05).Conclusions The number of Langerhans cells is elevated in patients with AA,especially in patients with active AA.The quantity of perivascular Langerhans cells in the upper dermis is positively correlated with that of perifollicular CD8 + T lymphocytes in the deep dermis of active AA lesions,hinting that Langerhans cells play an important role in the progression of AA.%目的 探讨斑秃患者脱发皮损中朗格汉斯细胞在斑秃病理进程中的分布以及与T细胞的关系.方法 对29例斑秃患者(活动期16例,非活动期13例)头皮脱发皮损进行CD1a免疫组化染色,对其中17例斑秃患者行CD4、CD8免疫组化染色.荧光半定量PCR测定局部皮损浅层和深层CD1a和粒细胞巨噬细胞刺激因子(GM-CSF)的mRNA表达水平.结果 斑秃患者表皮和真皮各处包括真皮浅层血管周围、毛囊周围,真皮深层血管周围、毛囊周围CD1a阳性LC数量均较健康对照显著增加(Z=4.354,2.884,4.640,3.217,3.496,均P＜0.01),活动期皮损表皮层、深层血管、深层毛囊CD1a阳性LC数量较非活动期皮损高(Z=2.457,2.130,1.954,P≤0.05).斑秃患者CD1a、GM-CSF mRNA相对表达量在皮损真皮浅层虽与健康对照无差异,但在深层均高于健康对照(Z=2.702,2.941,均P＜0.01).斑秃患者浅层血管周围LC与深层毛囊周围CD8+T细胞数量呈正相关(r=0.618,P＜ 0.05),活动组浅层血管周围LC与深层毛囊周围CD8+T细胞数量分布呈正相关关系(r=0.795,P=0.01),非活动组浅层血管周LC与深层毛囊周围CD8+T细胞数量分布则无相关关系.结论 斑秃患者皮损LC数量增加,且在活动期皮损升高更明显.活动期斑秃皮损中浅层血管周围LC与深层毛囊周围CD8+T细胞数量呈正相关,推测LC在斑秃疾病进展中发挥作用.