Knobler R L, Dubois-Dalcq M, Haspel M V, Claysmith A P, Lampert P W, Oldstone M B
J Neuroimmunol. 1981 Mar;1(1):81-92. doi: 10.1016/0165-5728(81)90010-2.
Demyelination may be induced by several different pathogenetic mechanisms. We have been utilizing mouse hepatitis virus (MHV) to study virus-induced demyelination in the central nervous system (CNS). To learn whether the different disease phenotypes in 4-week-old mice, caused by wild type (a model for fatal encephalomyelitis) or mutant ts8 (a model for primary demyelination), is due to an altered cellular tropism, we have developed an immunolabeling technique to evaluate critically the localization of MHV antigens in the unique cells of the CNS. Using mouse derived L-cells and primary neuronal cells in vitro, we determined an appropriate fixative (4% paraformaldehyde and 0.5% glutaraldehyde) that both preserved MHV antigenicity and cell structure. These studies in vitro showed the presence of MHV antigens on the surface of cells. Utilizing immunoperoxidase labeling as developed, we studied the localization of MHV antigens in vivo. MHV antigens associated with wild type (wt) virus were localized in neuronal cells as well as oligodendrocytes, which might account for the encephalomyelitis and primary demyelination, respectively. In contrast, MHV antigens associated with ts8 were localized rarely in neurons but commonly in oligodendrocytes. This might account for the uncommon occurrence of fatal encephalomyelitis, but the frequent presence of primary demyelination. Of interest was the finding of viral antigens during MHV infection in the cytoplasmic processes of oligodendrocytes surrounding intact myelin sheaths. We conclude that the different disease phenotypes caused by wt and mutant ts8 reflect differences in the cellular tropism of the two viruses for cells in the CNS.
脱髓鞘可能由几种不同的致病机制引起。我们一直在利用小鼠肝炎病毒（MHV）研究中枢神经系统（CNS）中病毒诱导的脱髓鞘。为了了解由野生型（一种致命性脑脊髓炎模型）或突变体ts8（一种原发性脱髓鞘模型）引起的4周龄小鼠的不同疾病表型是否归因于细胞嗜性的改变，我们开发了一种免疫标记技术来严格评估MHV抗原在中枢神经系统独特细胞中的定位。利用体外培养的小鼠来源的L细胞和原代神经元细胞，我们确定了一种合适的固定剂（4%多聚甲醛和0.5%戊二醛），它既能保留MHV抗原性又能保持细胞结构。这些体外研究表明细胞表面存在MHV抗原。利用所开发的免疫过氧化物酶标记，我们研究了体内MHV抗原的定位。与野生型（wt）病毒相关的MHV抗原定位于神经元细胞以及少突胶质细胞，这可能分别解释了脑脊髓炎和原发性脱髓鞘。相比之下，与ts8相关的MHV抗原很少定位于神经元，但通常定位于少突胶质细胞。这可能解释了致命性脑脊髓炎不常见，但原发性脱髓鞘频繁出现的原因。有趣的是，在MHV感染期间，在完整髓鞘周围的少突胶质细胞的细胞质突起中发现了病毒抗原。我们得出结论，wt和突变体ts8引起的不同疾病表型反映了这两种病毒对中枢神经系统细胞的细胞嗜性差异。
