Yamamoto T, Tsuruta J, Kambara T
Department of Allergy, Kumamoto University Medical School, Japan.
Biochim Biophys Acta. 1987 Dec 18;916(3):332-42. doi: 10.1016/0167-4838(87)90178-6.
A rabbit antibody against the light-chain of guinea-pig high-molecular-weight (HMW) kininogen, which was specific to HWM kininogen and did not recognize low-molecular-weight kininogen, was prepared. This antibody demonstrated the presence of HMW kininogen antigen at the interstitial-tissue space in the guinea-pig skin by means of immunohistochemistry. The interstitial-tissue HMW kininogen antigen was extracted from the skin. This antigen molecule in the skin extract behaved identically as HWM kininogen of plasma in slab-polyacrylamide gel electrophoresis under the presence of sodium dodecyl sulfate followed by immunoblotting. Therefore, it was concluded that HMW kininogen was present in the interstitial-tissue fluid in the skin. The amount of HMW kininogen in the skin extract was quantified by a sandwich enzyme-linked immunosorbent assay with the anti-light-chain antibody and a goat anti-guinea-pig HMW kininogen antibody. On the assumption that the interstitial-tissue volume is 50 ml/100 g wet skin tissue, the average concentration of HMW kininogen in the interstitial-tissue fluid of the skin was calculated to be 23% of the plasma concentration. On the other hand, the proportion of intravascular HMW kininogen (derived from blood remaining in the vessels of the harvested skin) in relation to the total HMW kininogen in the skin extract was quantified by measuring the radio-labelled HMW kininogen which had been injected intravenously as a tracer of the intravascular HMW kininogen. About 5% of the total HMW kininogen in the skin extract was calculated to be derived from the intravascular blood volume of the skin, indicating that the majority of the HMW kininogen in the skin extract was derived from the extravascular-tissue space.
制备了一种针对豚鼠高分子量（HMW）激肽原轻链的兔抗体，该抗体对HWM激肽原具有特异性，不识别低分子量激肽原。通过免疫组织化学方法，该抗体证实了豚鼠皮肤间质组织空间中存在HMW激肽原抗原。从皮肤中提取了间质组织HMW激肽原抗原。在十二烷基硫酸钠存在下进行平板聚丙烯酰胺凝胶电泳，随后进行免疫印迹，皮肤提取物中的这种抗原分子表现与血浆中的HWM激肽原相同。因此，得出结论，皮肤间质组织液中存在HMW激肽原。用抗轻链抗体和山羊抗豚鼠HMW激肽原抗体通过夹心酶联免疫吸附测定法定量皮肤提取物中HMW激肽原的含量。假设间质组织体积为50 ml/100 g湿皮肤组织，计算得出皮肤间质组织液中HMW激肽原的平均浓度为血浆浓度的23%。另一方面，通过测量静脉注射作为血管内HMW激肽原示踪剂的放射性标记HMW激肽原，定量皮肤提取物中血管内HMW激肽原（源自收获皮肤血管中残留的血液）相对于总HMW激肽原的比例。计算得出皮肤提取物中总HMW激肽原的约5%源自皮肤的血管内血容量，这表明皮肤提取物中的大多数HMW激肽原源自血管外组织空间。
