Identification of Human Tissues Based on The Tissue-Specific Myosin Isozymes.

本研究制备了人脑和肝脏肌球蛋白重链亚型的单抗，为建立人体组织的鉴定方法奠定了基础。用从人脑和肝脏中提纯的肌球蛋白免疫小鼠，获得了3株具有组织特异性的抗人脑肌球蛋白单抗(HBM1、4和6)和2株组织特异性抗人肝肌球蛋白单抗(HLM1和HL4)。用这些抗体对人非肌肉组织(大脑、小脑、肝、脾、肾和血小板)的肌球蛋白重链亚型进行了免疫印迹，并以主动脉和心肌为对照，在这些组织中发现了2-4种肌球蛋白重链亚型。大脑肌球蛋白重链有四种亚型，分子量分别为200、198、196和194 kDa，肝脏有198、196和194 kDa亚型。由于200 kDa和198 kDa的异构体主要存在于脑中，因此认为它们是脑型的，而196 kDa和194 kDa的异构体则普遍存在于非肌肉组织中，因此它们似乎是非肌型的。另一方面，肝脏中的肌球蛋白重链亚型主要由常见的非肌型组成。单抗HBM 1特异性识别肌球蛋白重链的脑型异构体。因此，HBM-1有可能作为一种免疫学方法用于区分脑组织和其他组织。事实上，使用HBM 1的夹心ELISA法可以在脑挫伤患者的血清中检测到脑型肌球蛋白重链的异构体。

In this study, monoclonal antibodies to the heavy-chain isoforms of myosins in human brain and liver were produced for the development of identification methods of human tissues. Three monoclonal anti-human cerebrum myosin antibodies (HBM 1,4 and 6) and two monoclonal anti-human liver myosin antibodies (HLM 1 and 4) each having distinctive tissue-specificity were obtained by immunizing mice with the myosin purified from the human cerebrum and liver, respectively. And immunoblotting of the heavy-chain isoforms of myosins in human nonmuscle tissues (the cerebrum, cerebellum, liver, spleen, kidney and platelet), and the aorta and cardiac muscle as controls were carried out using these antibodies.Consequently, 2-4 heavy-chain isoforms of myosins were found in these tissues. Myosin heavy-chain of the cerebrum contained four isoforms which showed the molecular masses of about 200,198,196 and 194 kDa, and that of the liver had 198,196 and 194 kDa isoforms. Since 200 and 198 kDa isoforms were mainly found in the brain, they were thought to be brain-type, while 196 and 194 kDa were commonly found in the nonmuscle tissues, therefore, they were seemed to be nonmuscle-type. On the other hand, myosin heavy-chain isoforms in the liver mainly consisted of common nonmuscle-type. Monoclonal antibody HBM 1 recognized specifically the brain-type isoforms of myosin heavy-chain. Therefore, HBM 1 seemed to be useful to discriminate the brain tissue from others by the immunological procedures. Indeed, brain-type isoforms of myosin heavy chain-could be detected in blood sera from bodies with cerebral contusion by sandwich ELISA using HBM 1.