通过流式细胞术和免疫细胞化学对人类根尖周炎病变中抗原呈递细胞的表征。

Characterization of antigen-presenting cells in human apical periodontitis lesions by flow cytometry and immunocytochemistry.

作者信息

Lukić A, Vasilijić S, Majstorović I, Vucević D, Mojsilović S, Gazivoda D, Danilović V, Petrović R, Colić M

机构信息

Department of Endodontics, Faculty of Stomatology, University of Belgrade, Belgrade, Serbia and Montenegro.

出版信息

Int Endod J. 2006 Aug;39(8):626-36. doi: 10.1111/j.1365-2591.2006.01125.x.

DOI:10.1111/j.1365-2591.2006.01125.x

PMID:16872457

Abstract

AIM

To analyse phenotypic characteristics of antigen-presenting cells (APC), isolated from human periapical lesions by flow cytometry and immunocytochemistry.

METHODOLOGY

Sixteen periapical lesions were digested for 15 min with 0.05% collagenase. Mononuclear cells, separated from other inflammatory cells by density centrifugation, were processed for flow cytometry and/or immunocytochemistry. Single and double immunostainings were performed using monoclonal antibodies specific for human CD45, CD3, CD19, CD14, HLA-DR, CD1a, CD83 and CD123.

RESULTS

Antigen-presenting cells (HLA-DR(+) cells) represented 32.9 +/- 17.8% of total mononuclear cells. Amongst them, B cells (HLA-DR(+) CD19(+)) were the predominant APC population, followed by activated macrophages (HLA-DR(+) CD14(+)), dendritic cells (DC) (HLA-DR(+) CD14(-) CD19(-) CD3(-)) and activated T cells (HLA-DR(+) CD3(+)). Based on the predominance of T cells (CD3(+)) or B cells and plasma cells (CD19(+) and CD19(lo), respectively) amongst mononuclear cell infiltrates, lesions were divided into T- and B-types. The percentage of DC in T-type lesions (27.1 +/- 6.8% of total HLA-DR(+) cells) was higher, compared with B-type lesions (10.3 +/- 5.2%) (P < 0.01). Within the DC population, the percentages of CD1a (Langerhans cell type) and CD123 (probably plasmacytoid DC type) did not differ significantly between the groups (P > 0.05). However, the percentage of mature DC (CD83(+)) was significantly higher in T-type periapical lesions (P < 0.05).

CONCLUSIONS

Flow cytometry and immunocytochemistry are suitable methods for phenotypic analysis of APC after their isolation from human periapical lesions. APC, that were phenotypically heterogeneous, constituted a significant component of infiltrating cells. Lesions with the predominance of T cells were characterized by a higher proportion of mature DC (HLA-DR(+)CD83(+) cells) than lesions with predominance of B cells/plasma cells.

摘要

目的

通过流式细胞术和免疫细胞化学分析从人根尖周病变中分离出的抗原呈递细胞（APC）的表型特征。

方法

16个根尖周病变用0.05%胶原酶消化15分钟。通过密度离心从其他炎性细胞中分离出单核细胞，进行流式细胞术和/或免疫细胞化学检测。使用针对人CD45、CD3、CD19、CD14、HLA-DR、CD1a、CD83和CD123的单克隆抗体进行单重和双重免疫染色。

结果

抗原呈递细胞（HLA-DR(+)细胞）占单核细胞总数的32.9±17.8%。其中，B细胞（HLA-DR(+) CD19(+)）是主要的APC群体，其次是活化巨噬细胞（HLA-DR(+) CD14(+)）、树突状细胞（DC）（HLA-DR(+) CD14(-) CD19(-) CD3(-)）和活化T细胞（HLA-DR(+) CD3(+)）。根据单核细胞浸润中T细胞（CD3(+)）或B细胞及浆细胞（分别为CD19(+)和CD19(lo)）的优势，病变分为T型和B型。与B型病变（10.3±5.2%）相比，T型病变中DC的百分比（占总HLA-DR(+)细胞的27.1±6.8%）更高（P<0.01）。在DC群体中，两组间CD1a（朗格汉斯细胞型）和CD123（可能是浆细胞样DC型）的百分比无显著差异（P>0.05）。然而，T型根尖周病变中成熟DC（CD83(+)）的百分比显著更高（P<0.05）。