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A Study of Antigen Presenting Dysfunction of Dendritic Cell from Peripheral Blood in the Patients with Connective Tissue Diseases

结缔组织病患者外周血树突状细胞抗原呈递功能障碍的研究

基本信息

批准号：
60570299
负责人：
SUZUKI Teruhiko
金额：
$ 1.02万
依托单位：
Saitama Medical School
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1985
资助国家：
日本
起止时间：
1985 至 1986
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-60570299/
关键词：
Dendritic Cell Systemic Lupus Erythematosus Mixed Lymphocyte Reaction クラスター

项目摘要

We succeeded in preparation of dendritic cell(DC) from human peripheral blood. In brief, mononuclear cells were obtained by Ficoll-Hypaque gradients from 50ml heparinized peripheral blood, and then cultured in Petri dish. Non-adherent cells were removed and adherent cells were cultured overhight. The DC were detached from the dish. The DC fraction was cultured in anti-human IgG coated plastic dish for the purpose of excluding of macrophage and B cell. Monoclonal antibodies to 3C10, Leul and Leul4 plus complement were added for killing each contaminating cells if needed. DC possessed characteristic prosseses and on its surface HLA-DR and C3bireceptor were present, but Fc-receptor, surface Ig and OKT 3 antigen absent. Functional study proved that DC was "active stimulator" in mixed lymphocyte reaction (MLR). Reduced MLR was well documentated in the patients with systemic lupus erythematosus(SLE). We examined SLE-DC function using MLR assay. New findings were obtained that major cause of reduced MLR in the patients with SLE resulted in impaired function of SLE-DC and not in T cells as before reported by several authors. DC formed characteristic cell aggregates with autologous or allogneic T cells (cluster). T cells formed cluster were mainly OKT4 positive. Cultured cluster supernatant was proved to contain some factors to generate SAC-stimulated B cells into antibody secreting cells(BSF). Supernatant from cluster formed by SLE-DC showed very low grade BSF activities, indicating that SLE-DC had functional abnormalities.

我们成功地从人外周血中制备了树突状细胞（DC）。简而言之，通过Ficoll-Hypaque梯度从50ml肝素化的外周血中获得单核细胞，然后在培养皿中培养。去除非贴壁细胞并将贴壁细胞培养在过高的温度下。 DC 与盘分离。将DC级分培养在抗人IgG包被的塑料皿中，以排除巨噬细胞和B细胞。如果需要，添加针对 3C10、Leul 和 Leul4 加补体的单克隆抗体以杀死每种污染细胞。 DC具有特征性的结构，其表面存在HLA-DR和C3双受体，但不存在Fc受体、表面Ig和OKT 3抗原。功能研究证明DC是混合淋巴细胞反应(MLR)的“主动刺激剂”。系统性红斑狼疮 (SLE) 患者的 MLR 降低已得到充分证实。我们使用 MLR 测定检查了 SLE-DC 功能。新的发现表明，SLE 患者 MLR 降低的主要原因是 SLE-DC 功能受损，而不是像之前几位作者报道的那样导致 T 细胞功能受损。 DC与自体或异体T细胞形成特征性细胞聚集体（簇）。形成簇的T细胞主要为OKT4阳性。培养簇上清液被证明含有一些使SAC刺激的B细胞生成抗体分泌细胞(BSF)的因子。 SLE-DC形成的簇的上清液显示出非常低等级的BSF活性，表明SLE-DC具有功能异常。