Perentes E, Rubinstein L J
Acta Neuropathol. 1986;70(3-4):284-8. doi: 10.1007/BF00686085.
The immunoreactivity of purified mouse myeloma IgM immunoglobulins (mouse IgM) to human myelin sheaths and astroglial cells was evaluated with the peroxidase-antiperoxidase method on paraffin-embedded tissues from human gliomas and areas of multiple sclerosis, and from normal human cerebrum, spinal cord and spinal nerve roots. The mouse IgM reacted positively with central and peripheral myelin sheaths and, as shown independently by others, with the cytoplasm of neoplastic and reactive astroglia. Parallel immunostaining of successive sections with an anti-glial fibrillary acidic protein (GFAP) serum and/or the anti-Leu 7 monoclonal antibody was of considerable assistance in identifying the immunoreactive elements and in distinguishing specific from non-specific immunostaining of myelin sheaths and astroglia. Pretreatment with normal human serum inhibited the non-specific binding by mouse IgM without altering GFAP and Leu 7 reactivities. The non-specific binding of mouse IgM to human myelin sheaths and astroglia can therefore be overcome, and the specificity of mouse IgM monoclonal antibodies retained, by the parallel immunostaining of successive sections with mouse IgM. If non-specific binding by mouse IgM is found to occur, it can then be inhibited by preincubation with normal human serum without loss of specific antigenicity.
采用过氧化物酶-抗过氧化物酶法,对来自人类胶质瘤、多发性硬化区域以及正常人类大脑、脊髓和脊神经根的石蜡包埋组织进行检测,评估纯化的小鼠骨髓瘤IgM免疫球蛋白(小鼠IgM)对人髓鞘和星形胶质细胞的免疫反应性。小鼠IgM与中枢和外周髓鞘呈阳性反应,并且,正如其他人独立证明的那样,与肿瘤性和反应性星形胶质细胞的细胞质呈阳性反应。用抗胶质纤维酸性蛋白(GFAP)血清和/或抗Leu 7单克隆抗体对连续切片进行平行免疫染色,对于识别免疫反应性成分以及区分髓鞘和星形胶质细胞的特异性与非特异性免疫染色有很大帮助。用人正常血清预处理可抑制小鼠IgM的非特异性结合,而不改变GFAP和Leu 7的反应性。因此,通过用小鼠IgM对连续切片进行平行免疫染色,可以克服小鼠IgM与人髓鞘和星形胶质细胞的非特异性结合,并保留小鼠IgM单克隆抗体的特异性。如果发现小鼠IgM存在非特异性结合,那么可以通过与人正常血清预孵育来抑制,而不会丧失特异性抗原性。