TISSUE DISTRIBUTION AND KININOGEN GENE-EXPRESSION AFTER ACUTE-PHASE INFLAMMATION
TISSUE DISTRIBUTION AND KININOGEN GENE-EXPRESSION AFTER ACUTE-PHASE INFLAMMATION
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DOI：
10.1016/0304-4165(88)90033-5
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发表时间：
1988-03-17
期刊：
影响因子：
--
通讯作者：
CHAO, L
中科院分区：
文献类型：
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作者：
CHAO, J;SWAIN, C;CHAO, L
用缓激肽偶联血蓝蛋白免疫小鼠脾细胞，与小鼠骨髓瘤细胞融合，制备了抗激肽原的单克隆抗体。通过酶联免疫吸附测定（ELISA）和放射免疫测定（RIA）筛选杂交细胞分泌缓激肽抗体。腹水产生并通过缓激肽-琼脂糖亲和柱纯化。在ELISA中，单克隆抗体（IgG 1）与缓激肽、赖氨酸-缓激肽、蛋氨酸-赖氨酸-缓激肽和激肽原结合。此外，该靶向单克隆抗体在Western印迹中识别纯化的低和高分子量牛、人或大鼠激肽原和T-激肽原。松节油诱导的急性炎症后，大鼠激肽原水平显着增加，在肝脏和血清中，以及在灌注的垂体，心脏，肺，肾脏，胸腺和其他组织中，所确定的激肽导向的激肽原抗体在Western印迹分析。结果通过用激肽RIA测量激肽原的激肽当量来证实。在诱导的炎症反应过程中，大鼠激肽原定位与激肽定向的单克隆抗体在肝实质细胞，腺泡细胞和一些颗粒曲管的下颌下腺，并在收集管的肾脏。北方和细胞质斑点印迹分析，使用激肽寡核苷酸探针显示，激肽原mRNA水平在肝脏中，而不是在其他组织中增加后，松节油诱导的炎症。结果表明，大鼠激肽原分布于除肝脏外的多种组织中，急性炎症仅诱导肝脏产生激肽原。靶向激肽原单克隆抗体是研究激肽原或任何含有激肽部分的蛋白质分子的结构、定位和功能的有用试剂。
A kinin-directed monoclonal antibody to kininogens has been developed by the fusion of murine myeloma cells with mouse splenocytes immunized with bradykinin-conjugated hemocyanin. The hybrid cells were screened by an enzyme-linked immunosorbent assay (ELISA) and a radioimmunoassay (RIA) for the secretion of antibodies to bradykinin. Ascitic fluids were produced and purified by a bradykinin-agarose affinity column. The monoclonal antibody (IgG1) bound to bradykinin, Lys-bradykinin, Met-Lys-bradykinin, and kininogens in ELISA. Further, this target-directed monoclonal antibody recognized purified low and high molecular weight bovine, human, or rat kininogens and T-kininogen in Western blotting. After turpentine-induced acute inflammation, rat kininogen levels increased dramatically in liver and serum as well as in the perfused pituitary, heart, lung, kidney, thymus, and other tissues, as identified by the kinin-directed kininogen antibody in Western blot analyses. The results were confirmed by measuring kinin equivalents of kininogens with a kinin RIA. During an induced inflammatory response, rat kininogens were localized immunohistochemically with the kinin-directed monoclonal antibody in parenchymal cells of liver, in acinar cells and some granular convoluted tubules of submandibular gland, and in the collecting tubules of kidney. Northern and cytoplasmic dot blot analyses using a kinin oligonucleotide probe showed that kininogen mRNA levels in liver but not in other tissues increase after turpentine-induced inflammation. The results indicated that rat kininogens are distributed in various tissues in addition to liver and only liver kininogen is induced by acute inflammation. The target-directed kininogen monoclonal antibody is a useful reagent for studying the structure, localization, and function of kininogens or any protein molecule containing the kinin moiety.