Mills B G, Singer F R
Clin Orthop Relat Res. 1987 Apr(217):16-25.
This study evaluates previous viral antigen data obtained from fixed tissue sections and cells grown in culture from bone affected by Paget's disease. Finding antigens to both respiratory syncytial virus (RSV) and measles virus (MV) in the same osteoclasts of ten patients could not be explained on the basis of any previously known cross-reactivity. Therefore, possible causes for these observations were sought. Monoclonal antibodies to viral proteins of RSV and MV were used to label proteins. Polyclonal antibodies that were monospecific and were produced exclusively in nonhuman species were used to rule out nonspecific reaction with human proteins. Antivimentin antibody was used to test the possibility of cross-reactivity with a cytoskeletal protein, as a second antibody F(ab')2 conjugated to fluorescein was used to rule out nonspecific reactivity with Fc receptors. Electron microscopy was used to evaluate bone cell cultures derived from Paget's bone in comparison with Paget's osteoclasts. Results showed that the pattern of monoclonal viral antibody labeling followed different patterns in different patients. Nonspecific reactivity was ruled out by significant negative and positive controls. Cross-reactivity with vimentin could not account for the positive immunofluorescent results because of an entirely different pattern of fluorescence in the same samples of live and fixed cells after colchemid treatment. It was concluded that specific viral antigens are present in osteoclasts and in cells grown from Paget's bone and that the present data are compatible with the possibility that Paget's disease of bone is a slow virus infection.
本研究评估了从佩吉特病受累骨骼的固定组织切片以及培养的细胞中获取的既往病毒抗原数据。在10例患者的同一破骨细胞中发现呼吸道合胞病毒(RSV)和麻疹病毒(MV)的抗原,这无法基于任何先前已知的交叉反应来解释。因此,探寻了这些观察结果的可能原因。使用针对RSV和MV病毒蛋白的单克隆抗体标记蛋白质。使用单特异性且仅在非人类物种中产生的多克隆抗体来排除与人类蛋白质的非特异性反应。使用抗波形蛋白抗体检测与细胞骨架蛋白交叉反应的可能性,使用与荧光素偶联的第二抗体F(ab')2来排除与Fc受体的非特异性反应。与佩吉特破骨细胞相比,使用电子显微镜评估源自佩吉特骨的骨细胞培养物。结果显示,单克隆病毒抗体标记模式在不同患者中呈现不同模式。通过显著的阴性和阳性对照排除了非特异性反应。由于秋水仙酰胺处理后活细胞和固定细胞的相同样本中荧光模式完全不同,与波形蛋白的交叉反应无法解释阳性免疫荧光结果。得出的结论是,破骨细胞和源自佩吉特骨的细胞中存在特异性病毒抗原,并且目前的数据与骨佩吉特病是一种慢病毒感染的可能性相符。