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一种新型纤连蛋白受体在小鼠淋巴瘤细胞系RL-male1肝脏浸润中的应用。

Use of a novel fibronectin receptor for liver infiltration by a mouse lymphoma cell line RL-male1.

作者信息

Gazi M H, Ito M

机构信息

Department of Pathology, Yamagata University School of Medicine, Japan.

出版信息

Cancer Res. 1999 Mar 1;59(5):1115-9.

Abstract

The mechanism whereby some lymphomas invade liver extensively has not been fully investigated. There is no basement membrane under the sinusoidal endothelium of the liver, and hepatocytes produce fibronectin (FN); therefore, adhesion to this FN may be particularly important for liver infiltration by lymphoma cells. A mouse lymphoma cell line, RL-male1, adhered to FN. However, this cell line did not express classical FN receptors such as very late antigen (VLA)-4 and VLA-5, as estimated by immunofluorescent staining. We have generated monoclonal antibodies (mAbs) that inhibit adhesion of RL-male1 cells to FN. Western blot and immunoprecipitation analyses showed that the new mAbs recognize a protein with an approximate molecular weight of 55,000 (p55). This antigenic protein was highly purified by immunoprecipitation and processed for microsequencing. From NH2-terminal sequence results, the p55 antigen was not identical to known FN receptors. Radioisotope-labeled RL-male1 cells, when injected i.v. into mice, rapidly infiltrated the liver (30-35% of injected cells), as measured by a gamma counter. Intravenous injection of the new mAbs partially (20%) blocked the infiltration of i.v.-injected lymphoma cells into the liver, whereas control rat IgG and an anti-CD11a mAb did not. These results demonstrate that the mouse lymphoma cell line RL-male1 nses a novel FN receptor for liver infiltration.

摘要

某些淋巴瘤广泛侵袭肝脏的机制尚未得到充分研究。肝脏窦状内皮细胞下方没有基底膜,且肝细胞可产生纤连蛋白(FN);因此,与这种FN的黏附对于淋巴瘤细胞浸润肝脏可能尤为重要。一种小鼠淋巴瘤细胞系RL-male1可黏附于FN。然而,通过免疫荧光染色估计,该细胞系不表达经典的FN受体,如极迟抗原(VLA)-4和VLA-5。我们制备了可抑制RL-male1细胞与FN黏附的单克隆抗体(mAb)。蛋白质印迹和免疫沉淀分析表明,新的mAb识别一种分子量约为55,000的蛋白质(p55)。通过免疫沉淀对这种抗原性蛋白质进行了高度纯化,并进行微量测序处理。根据氨基末端序列结果,p55抗原与已知的FN受体不同。静脉注射放射性同位素标记的RL-male1细胞后,通过γ计数器测量发现,这些细胞迅速浸润肝脏(占注射细胞的30 - 35%)。静脉注射新的mAb可部分(20%)阻断静脉注射的淋巴瘤细胞浸润肝脏,而对照大鼠IgG和抗CD11a mAb则无此作用。这些结果表明,小鼠淋巴瘤细胞系RL-male1利用一种新的FN受体浸润肝脏。

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