Binding of Griffonia simplicifolia I lectin to rat pulmonary alveolar macrophages and its use in purifying type II alveolar epithelial cells.
Binding of Griffonia simplicifolia I lectin to rat pulmonary alveolar macrophages and its use in purifying type II alveolar epithelial cells.
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Griffonia simplicifolia I 凝集素与大鼠肺泡巨噬细胞的结合及其在纯化 II 型肺泡上皮细胞中的应用。
DOI：
10.1016/0167-4889(86)90035-2
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发表时间：
1986
期刊：
影响因子：
--
通讯作者：
Goldstein,IJ
中科院分区：
文献类型：
--
作者：
Simon,RH;McCoyJr,JP;Chu,AE;Dehart,PD;Goldstein,IJ
本文报道了从G. simplicifoliaseeds与肺组织冷冻切片或支气管肺泡灌洗液中存在的大鼠肺泡巨噬细胞结合。simplicifoliaI-B4不与肺泡上皮细胞结合。我们建立了G. simplicifoliaI-B4通过与这些细胞上存在的末端α-D-吡喃半乳糖残基相互作用而与巨噬细胞结合。通过证明半抗原糖α-d-吡喃半乳糖苷或用咖啡豆α-半乳糖苷酶处理细胞抑制结合，证实了这一点。由于已知鼠层粘连蛋白末端含有α-d-吡喃半乳糖基，并且我们发现抗层粘连蛋白抗血清与大鼠肺泡巨噬细胞结合，我们怀疑G. simplicifoliaI-B4可能与巨噬细胞上的层粘连蛋白结合。为了从大鼠肺中分离肺泡II型上皮细胞，我们开发了一种利用凝集素G的方法。单叶类当蛋白酶衍生的肺细胞悬液与G.当巨噬细胞在单叶状体中聚集时，巨噬细胞凝集并且可以通过尼龙网过滤去除。在组织培养中孵育所得细胞悬浮液后，贴壁细胞为94 ± 2%（S.D.）II型细胞当比较分离的细胞通过反复差分粘附，凝集素制备的II型细胞具有相似的形态和染色特征，形成圆顶在单层和将类似量的棕榈酸酯到二饱和磷脂酰胆碱。我们相信，在本报告中概述的程序提供了一个简单而有效的方法来分离II型肺泡上皮细胞从大鼠肺。
We report that the isolectinGriffonia simplicifoliaI-B4isolated fromG. simplicifoliaseeds binds to rat alveolar macrophages present in frozen sections of lung tissue or bronchoalveolar lavage fluid.G. simplicifoliaI-B4does not bind to alveolar epithelial cells. We established thatG. simplicifoliaI-B4binds to the macrophages via interaction with terminal α-d-galactopyranosyl residues present on these cells. This was substantiated by demonstrating that binding is inhibited either by the haptenic sugar α-d-galactopyranoside or by treating the cells with coffee bean α-galactosidase. Because murine laminin is known to contain terminal α-d-galactopyranosyl end-groups, and because we found that an anti-laminin antiserum binds to rat alveolar macrophages, we suspect thatG. simplicifoliaI-B4may be binding to laminin present on the macrophages. To isolate alveolar type II epithelial cells from rat lungs, we developed a method that utilizes the lectinG. simplicifoliaI. When proteinase-derived suspensions of pulmonary cells are incubated withG. simplicifoliaI, the macrophages agglutinate and can be removed by filtration through nylon mesh. After incubating the resulting cellular suspension in tissue culture, the adherent cells are 94 ± 2% (S.D.) type II cells. When compared to cells isolated by repeated differential adherence, the lectin-prepared type II cells have similar morphology and staining characteristics, form domes in monolayers and incorporate similar amounts of palmitate into disaturated phosphatidylcholine. We believe that the procedure outlined in this report provides a simple and effective method to isolate type II alveolar epithelial cells from rat lungs.