Recognition of murine integrin beta 1 by a rat anti-stromal cell monoclonal antibody.
Recognition of murine integrin beta 1 by a rat anti-stromal cell monoclonal antibody.
复制标题
大鼠抗基质细胞单克隆抗体对鼠整合素β1的识别。
DOI：
10.1089/hyb.1994.13.409
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发表时间：
1994
期刊：
影响因子：
--
通讯作者：
Gimble,JM
中科院分区：
文献类型：
--
作者：
Wu,X;Miyake,K;Medina,KL;Kincade,PW;Gimble,JM
先前对大鼠单克隆抗体KMI6的研究已将其抗原在体内定位于骨髓基质细胞的一个离散亚群。通过氨基酸序列分析和抗血清与禽类整合素β1的交叉反应，KMI6抗原已被鉴定为小鼠整合素β1的同源物。Western blot检测小鼠整合素β1的相对组织丰度。虽然固定小鼠造血组织的免疫过氧化物酶染色显示细胞内β1丰富，但通过免疫荧光和流式细胞术评估，很少有淋巴造血原代来源的细胞表面阳性。胎儿红细胞是唯一的例外。相反，该抗原在几种培养细胞的Unes表面很容易检测到，并与多种α链相关。表面标记的小鼠整合素β1的生化特性与其对应的人相似，在减少条件下或在n -聚糖酶处理后表现出改变的电泳迁移。抗体对蛋白质的识别对糖基化状态、二价阳离子、洗涤剂或转移到硝化纤维素膜不敏感。然而，在Western blot中，由于蛋白的减少，表位丢失，这表明它是构象依赖的。这些数据表明，尽管KMI6表位分布广泛，但其表面表达可能在淋巴造血组织中受到限制。
Previous studies with the rat monoclonal antibody KMI6 had localized its antigenin vivoto a discrete subpopulation of marrow stromal cells. The KMI6 antigen has now been identified as the murine homolog of integrin β1by amino acid sequence analysis and by cross-reactivity with antiserum to the avian integrin β1. The relative tissue abundance of murine integrin β1was determined by Western blot. Although immunoperoxidase staining of fixed murine hematopoietic tissues demonstrated an abundance of intracellular β1, few primary-derived cells of lymphohematopoietic origin were surface positive as assessed by immunofluorescence and flow cytometry. Fetal erythroblasts provided the only exception. In contrast, the antigen was readily detected on the surface of several cultured cell Unes in association with a variety of α chains. The biochemical properties of the surface labeled murine integrin β1were similar to those of its human counterpart, exhibiting an altered electrophoretic migration under reduced conditions or followingN-glycanase treatment. The antibody recognition of the protein was insensitive to glycosylation state, presence of divalent cations, detergents, or transfer to a nitrocellulose membrane. However, on Western blot, the epitope was lost on reduction of the protein, suggesting that it is conformation dependent. These data indicate that although KMI6 epitope is widely distributed, its surface expressionin vivomay be restricted within lymphohemopoietic tissues.