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人扁桃体树突状细胞的分离与鉴定

Isolation and characterization of human tonsil dendritic cells.

作者信息

Hart D N, McKenzie J L

机构信息

Department of Haematology, Christchurch Hospital, New Zealand.

出版信息

J Exp Med. 1988 Jul 1;168(1):157-70. doi: 10.1084/jem.168.1.157.

Abstract

Human dendritic cells were isolated from tonsils by density gradient separation followed by FACS IV sorting with mAbs to remove contaminating cell populations. The resulting dendritic cell population consisted of large cells with plentiful basophilic cytoplasm, lacking in granules but containing a prominent Golgi apparatus and numerous mitochondria. The cell membrane was irregular, and marked cell protrusions were obvious when stained with anti-HLA class II reagents. Their nuclei were irregular and often indented with a visible nucleolus. These cells were not phagocytic and stimulated autologous and allogeneic lymphocytes more effectively than other tonsil cell types in MLR. Phenotypic analysis of these cells confirmed that they expressed the leucocyte common antigen and stained strongly for HLA-class II antigens. Tonsil dendritic cells also coexpressed the LFA-1 alpha and LFA-1 beta chains but did not stain with a wide variety of anti-monocyte or anti-macrophage antibodies. The cells also lacked Fc and complement receptors and failed to stain with CD1 antibodies. Extensive testing with mAbs revealed only a few positive reactions, and these were consistent with reports of these antibodies staining interdigitating cells in tissue sections. This established that tonsil dendritic cells belong to the unique haemopoietic cell lineage of dendritic cells. No cytoplasmic staining of IL-1 alpha or IL-1 beta was demonstrated, although these lymphokines were readily detected in activated monocytes.

摘要

通过密度梯度分离从扁桃体中分离出人类树突状细胞,随后用单克隆抗体进行荧光激活细胞分选(FACS IV)以去除污染的细胞群体。所得的树突状细胞群体由大细胞组成,细胞质丰富且呈嗜碱性,无颗粒,但含有明显的高尔基体和大量线粒体。细胞膜不规则,用抗II类 HLA 试剂染色时可见明显的细胞突起。它们的细胞核不规则,常有凹陷且可见核仁。这些细胞不具有吞噬作用,在混合淋巴细胞反应(MLR)中比其他扁桃体细胞类型更有效地刺激自体和异体淋巴细胞。对这些细胞的表型分析证实它们表达白细胞共同抗原,且对II类 HLA 抗原染色强烈。扁桃体树突状细胞还共表达淋巴细胞功能相关抗原-1(LFA-1)的α链和β链,但不被多种抗单核细胞或抗巨噬细胞抗体染色。这些细胞也缺乏Fc和补体受体,且不能被CD1抗体染色。用单克隆抗体进行的广泛检测仅显示少数阳性反应,且这些反应与这些抗体在组织切片中对交错突细胞染色的报道一致。这证实扁桃体树突状细胞属于独特的树突状细胞造血细胞谱系。虽然在活化的单核细胞中很容易检测到白细胞介素-1α或白细胞介素-1β,但未观察到这些细胞因子的细胞质染色。

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