Matsumoto Y, Hara N, Tanaka R, Fujiwara M
J Immunol. 1986 May 15;136(10):3668-76.
The rat central nervous system (CNS) during experimental allergic encephalomyelitis (EAE) was analyzed immunohistochemically from the preclinical to recovery stage by using monoclonal antibodies specific for rat T lymphocyte subsets and Ia antigen. Through combination of the avidin-biotin technique and carefully selected fixative, cells with dendritic morphology (DC) and infiltrating mononuclear cells were clearly and intensely demonstrated in the CNS parenchyma during EAE. In normal and complete Freund's adjuvant (CFA)-injected controls, there were no inflammatory foci. Ia (OX3)-positive parenchymal cells were not detected, whereas W3/25 stained DC that were located mainly in the white matter and W3/13 stained axons. At the preclinical stage, 11 days after CNS/CFA sensitization, a few clusters of Ia+ DC were detected in some sections of the spinal cord. The number of Ia+ DC increased as clinical signs developed (P less than 0.001). In rats with a clinical score of 1 or 2, Ia+ DC were mainly located in the perivascular region and closely associated with infiltrating T lymphocytes. However, at moribund state (score 3), Ia+ DC were evenly distributed in gray and white matter on almost all sections of the spinal cord. In recovered rats, the numbers of inflammatory foci and Ia+ DC were less than those in clinical EAE rats (P less than 0.001). Rats without clinical signs throughout the course also contained a few clusters of Ia+ DC. Double immunofluorescent staining with OX3 and anti-glial fibrillary acidic protein (GFAP) antiserum demonstrated that Ia+ DC were negative for GFAP. Their morphology and distribution were similar to those of nucleoside diphosphatase-positive cells, suggesting that Ia+ DC are microglia. In contrast to DC, no astrocytes or endothelial cells express detectable levels of Ia antigen in control and clinical EAE rats. These findings suggest that brain cells other than Ia+ DC may not be involved in the local immune interaction. Ia+ DC may play a significant role in antigen presentation in the CNS with EAE.
采用针对大鼠T淋巴细胞亚群和Ia抗原的单克隆抗体，通过免疫组织化学方法对实验性变态反应性脑脊髓炎（EAE）大鼠从临床前期到恢复期的中枢神经系统（CNS）进行分析。通过抗生物素蛋白-生物素技术与精心挑选的固定剂相结合，在EAE期间，CNS实质内具有树突状形态的细胞（DC）和浸润的单核细胞清晰且强烈地显示出来。在正常和注射完全弗氏佐剂（CFA）的对照中，没有炎症病灶。未检测到Ia（OX3）阳性实质细胞，而W3/25染色的DC主要位于白质中，W3/13染色的是轴突。在临床前期，即CNS/CFA致敏后11天，在脊髓的一些切片中检测到少数Ia + DC簇。随着临床症状的出现，Ia + DC的数量增加（P小于0.001）。临床评分为1或2的大鼠中，Ia + DC主要位于血管周围区域，并与浸润的T淋巴细胞密切相关。然而，在濒死状态（评分3）时，Ia + DC在脊髓几乎所有切片的灰质和白质中均匀分布。在恢复的大鼠中，炎症病灶和Ia + DC的数量少于临床EAE大鼠（P小于0.001）。整个过程无临床症状的大鼠也含有少数Ia + DC簇。用OX3和抗胶质纤维酸性蛋白（GFAP）抗血清进行双重免疫荧光染色表明，Ia + DC对GFAP呈阴性。它们的形态和分布与核苷二磷酸酶阳性细胞相似，表明Ia + DC是小胶质细胞。与DC相反，在对照和临床EAE大鼠中，没有星形胶质细胞或内皮细胞表达可检测水平的Ia抗原。这些发现表明，除Ia + DC外的脑细胞可能不参与局部免疫相互作用。Ia + DC可能在EAE的CNS抗原呈递中起重要作用。
