Dubois-Dalcq M, Worthington K, Gutenson O, Barbosa L H
Lab Invest. 1975 Apr;32(4):518-26.
A hamster neuroadapted strain of subacute sclerosing panencephalitis (SSPE) virus induces in the newborn animal an acute encephalitis with ultrastructural features similar to those observed earlier in measles encephalitis and characteristic of a productive viral infection. The direct immunoperoxidase technique, using labeled SSPE gamma-globulin, was applied to formaldehyde-fixed thick sections. These were either chopped after perfusion of the animal or cryocut after quick freezing of the tissue and then fixed. The latter procedure showed good correlation with light microscopic immunoperoxidase or fluorescent antibody staining on paired thin cryostat sections, whereas the former procedure resulted in easier handling of the sections and better ultrastructural preservation, especially when the lysine-periodate-formaldehyde fixative was used. Under electron microscopy the plasma membrane of the nerve cells was often labeled on its inner side in areas where nucleocapsids accumulate to form buds. The membrane antigen appeared to be more focal than in productive SSPE infection in vitro. Within the cytoplasm, most of the tubular inclusions were stained with the label located on the external granular coat of the nucleocapsid. Nonstructural antigen was also detected in the rough endoplasmic reticulum membrane (often on the side of a tubular inclusion), in the cytoplasmic matrix, and sometimes in the nucleus and on the nuclear membrane of giant cells. It thus seems that SSPE gamma-globulin reacts with viral proteins close to the site of synthesis. Because of the high specificity and reproducibility of this method for the detection of intracellular viral antigens, its use is recommended for further studies of conventional and nonconventional viral central nervous system diseases in which an immune response is present.
一种仓鼠神经适应性亚急性硬化性全脑炎(SSPE)病毒株可在新生动物中引发急性脑炎,其超微结构特征与先前在麻疹脑炎中观察到的相似,具有生产性病毒感染的特征。使用标记的SSPEγ球蛋白的直接免疫过氧化物酶技术应用于甲醛固定的厚切片。这些切片要么在动物灌注后切碎,要么在组织快速冷冻后冷冻切片,然后固定。后一种方法与配对薄冷冻切片上的光镜免疫过氧化物酶或荧光抗体染色显示出良好的相关性,而前一种方法使切片处理更容易,超微结构保存更好,尤其是在使用赖氨酸-高碘酸盐-甲醛固定剂时。在电子显微镜下,神经细胞的质膜在内侧经常在核衣壳聚集形成芽的区域被标记。膜抗原似乎比体外生产性SSPE感染中的更具局灶性。在细胞质内,大多数管状内含物被位于核衣壳外部颗粒包膜上的标记物染色。在粗面内质网膜(通常在管状内含物的一侧)、细胞质基质中,有时在巨细胞的细胞核和核膜中也检测到非结构抗原。因此,SSPEγ球蛋白似乎在病毒合成位点附近与病毒蛋白发生反应。由于该方法检测细胞内病毒抗原具有高特异性和可重复性,建议将其用于对存在免疫反应的传统和非传统病毒性中枢神经系统疾病的进一步研究。