Mehta P D, Kane A, Thormar H
J Immunol. 1976 Dec;117(6):2053-60.
Cellulose acetate and agarose gel electrophoresis of pH 7.4 brain extracts from three patients with subacute sclerosing panencephalitis (SSPE) revealed two to three homogeneous bands in the gamma-globulin region. The IgG concentration in the brain extracts ranged from 6 to 8.5% of the total protein. The IgG was isolated by passing each brain extract twice through gel filtration on Sephadex G-200 column. The Sephadex-isolated IgG preparations, at a concentration of 1 mg/ml, had hemagglutination inhibition (HI) titers ranging from 512 to 800, complement fixation (CF) titers of 64 to 100, and neutralizing titers of 400 to 1024 when tested against envelope, nucleocapsids, and while measles virus, respectively. These IgG preparations were further separated into several fractions by isoelectric focusing (IEF) technique and the reactivities of these eluted fractions were examined against differnt specific measles virus components and against whole virus. The results showed that both slow and fast moving homogeneous IgG had high HI and neutralizing titers. However, CF antibodies were present only in specific IEF-eluted fractions and their profiles differed somewhat from one brain to another. The IgG fractions reacted with both kappa-and gamma-chain antisera, but the reactivity varied from one brain to another. When the polyacrylamide gel electrophoretic (PAGE) patterns of heavy (H) and light (L) chains of SSPE IgG were compared to those of IgG from controls, the H chain patterns of both groups were identical whereas the SSPE brain IgG demonstrated two distinct bands in L chain as compared to a single diffuse band observed in controls. These latter results, in addition to homogeneous bands and selective CF antibody activity in IEF eluted IgG fractions from SSPE brain, fulfill some of the criteria of monoclonality.
对三名亚急性硬化性全脑炎(SSPE)患者pH 7.4的脑提取物进行醋酸纤维素和琼脂糖凝胶电泳,结果显示在γ球蛋白区域有两到三条均匀条带。脑提取物中的IgG浓度占总蛋白的6%至8.5%。通过将每种脑提取物两次通过Sephadex G - 200柱进行凝胶过滤来分离IgG。浓度为1mg/ml的经Sephadex分离的IgG制剂,在分别针对包膜、核衣壳和麻疹病毒进行检测时,血凝抑制(HI)效价范围为512至800,补体结合(CF)效价为64至100,中和效价为400至1024。这些IgG制剂通过等电聚焦(IEF)技术进一步分离成几个组分,并检测这些洗脱组分针对不同特异性麻疹病毒成分和全病毒的反应性。结果表明,慢速和快速移动的均匀IgG均具有高HI和中和效价。然而,CF抗体仅存在于特定的IEF洗脱组分中,并且它们的图谱在不同脑之间略有不同。IgG组分与κ链和γ链抗血清均发生反应,但反应性在不同脑之间有所不同。当将SSPE IgG的重链(H)和轻链(L)的聚丙烯酰胺凝胶电泳(PAGE)图谱与对照IgG的图谱进行比较时,两组的H链图谱相同,而与对照中观察到的单一弥散条带相比,SSPE脑IgG在L链中显示出两条明显的条带。除了SSPE脑IEF洗脱的IgG组分中的均匀条带和选择性CF抗体活性外,这些结果满足了单克隆性的一些标准。