Fujioka T, Purev E, Kremlev S G, Ventura E S, Rostami A
Department of Neurology, University of Pennsylvania School of Medicine, 3400 Spruce Street, Philadelphia, PA 19104-4283, USA.
J Neuroimmunol. 2000 Aug 1;108(1-2):181-91. doi: 10.1016/s0165-5728(00)00270-8.
Experimental autoimmune neuritis (EAN) is an animal model that shares clinical, pathological and electrophysiological features with the human disease Guillain-Barré syndrome (GBS). In this study, we isolated and characterized by fluorescent activated cell sorter (FACS) phenotype of the inflammatory cells infiltrating cauda equina (CE) of Lewis rats at the active stage of the disease. We found that at this stage of EAN macrophages (Mphi) and alphabeta T cells were two major populations isolated from CE. We also found that among total cell population isolated from CE, gammadelta T and NK cells composed two small but distinct populations, while B cells were negligible. We characterized phenotype of alphabeta T cells in CE as CD45RC(+)CD8(+) (activated cytotoxic lymphocytes) and CD45RC(-)CD4(+) (memory Th cells). The phenotype of gammadelta T cells was found to be consisted of only CD45RC(+)CD8(+) cells. Both alphabeta and gammadelta T cells in CE expressed a higher level of CD25, CD44 and CD54 activation markers compared to the other tissues. Immunohistochemistry demonstrated that gammadelta T cells existed apart from the intense cellular infiltrate. This is the first report on the isolation and FACS analysis of CE-infiltrating cells, contributing a new and alternative approach to study the inflammatory lesions in EAN. We conclude that both alphabeta and gammadelta T cells have a unique activation/inflammatory phenotype required to traffic through and be retained in the peripheral nerves during EAN.
实验性自身免疫性神经炎(EAN)是一种动物模型,它与人类疾病格林-巴利综合征(GBS)具有临床、病理和电生理特征。在本研究中,我们在疾病活动期通过荧光激活细胞分选仪(FACS)对Lewis大鼠马尾(CE)中浸润的炎性细胞进行分离并鉴定其表型。我们发现在EAN的这个阶段,巨噬细胞(Mphi)和αβ T细胞是从CE分离出的两个主要细胞群。我们还发现,在从CE分离出的总细胞群中,γδ T细胞和NK细胞构成两个小而独特的细胞群,而B细胞数量可忽略不计。我们将CE中αβ T细胞的表型鉴定为CD45RC(+)CD8(+)(活化的细胞毒性淋巴细胞)和CD45RC(-)CD4(+)(记忆性Th细胞)。发现γδ T细胞的表型仅由CD45RC(+)CD8(+)细胞组成。与其他组织相比,CE中的αβ T细胞和γδ T细胞均表达更高水平的CD25、CD44和CD54活化标志物。免疫组织化学显示γδ T细胞存在于密集的细胞浸润区域之外。这是关于CE浸润细胞分离和FACS分析的首次报道,为研究EAN中的炎性病变提供了一种新的替代方法。我们得出结论,在EAN期间,αβ T细胞和γδ T细胞均具有穿越并保留在外周神经中所需的独特活化/炎性表型。