Laman J D, Visser L, Maassen C B, de Groot C J, de Jong L A, 't Hart B A, van Meurs M, Schellekens M M
Department of Immunology, Erasmus University Rotterdam and University Hospital Dijkzigt, P.O. Box 1738, 3000 DR, Rotterdam, The Netherlands.
J Neuroimmunol. 2001 Sep 3;119(1):124-30. doi: 10.1016/s0165-5728(01)00356-3.
Experimental autoimmune encephalomyelitis (EAE) induced by immunization of mice with epitopes of the proteolipid protein (PLP), a major myelin constituent, forms a useful model for the study of multiple sclerosis (MS). In addition, MS patients display PLP-specific T- and B-cell responses, suggesting that PLP reactivity is relevant to pathogenesis.Here, the generation and characterization of a panel of mouse monoclonal antibodies (Mab) against PLP139-151, the prominent encephalitogenic sequence in SJL/J mice is described. Five Mab were generated by conventional immunization of an SJL/J mouse and hybridoma generation. These Mab reacted well with the PLP139-151 peptide in ELISA and belonged to the IgG2a and IgG2b subclasses, consistent with CD4+ T helper 1-cell-supported antibody formation. The Mab also efficiently detected PLP peptide-BSA conjugates in Western blot, confirming their multi-assay applicability. The Mab were subsequently used to determine the occurrence of demyelination in brains of MS patients and marmoset monkeys with EAE. Immunohistochemistry on both paraffin and frozen sections demonstrated a homogeneous expression of PLP139-151 in normal myelin, and a complete absence in lesions containing demyelinated areas, confirming that the Mab can be used as a general myelin marker. In active demyelinating MS lesions, the Mab visualized the peptide in the cytoplasm of macrophages containing phagocytosed myelin. In conclusion, this panel of Mab against the encephalitogenic PLP139-151 epitope forms a useful tool for further study of autoantigen expression, demyelination/remyelination and the staging of lesional activity in MS patients, as well as in EAE models in distinct animal species.
用主要髓磷脂成分蛋白脂蛋白（PLP）的表位免疫小鼠诱导的实验性自身免疫性脑脊髓炎（EAE），形成了一种用于研究多发性硬化症（MS）的有用模型。此外，MS患者表现出PLP特异性的T细胞和B细胞反应，这表明PLP反应性与发病机制相关。在此，描述了一组针对PLP139 - 151（SJL/J小鼠中突出的致脑炎性序列）的小鼠单克隆抗体（Mab）的产生和特性。通过对SJL/J小鼠进行常规免疫和杂交瘤生成产生了五种Mab。这些Mab在ELISA中与PLP139 - 151肽反应良好，属于IgG2a和IgG2b亚类，这与CD4 + T辅助1细胞支持的抗体形成一致。这些Mab在蛋白质印迹中也能有效检测PLP肽 - BSA缀合物，证实了它们在多种检测中的适用性。随后，这些Mab被用于确定MS患者和患有EAE的狨猴大脑中脱髓鞘的发生情况。对石蜡切片和冰冻切片进行的免疫组织化学显示，PLP139 - 151在正常髓磷脂中均匀表达，而在含有脱髓鞘区域的病变中完全不存在，证实这些Mab可作为一般的髓磷脂标记物。在活动性脱髓鞘的MS病变中，这些Mab在含有吞噬髓磷脂的巨噬细胞的细胞质中使该肽显色。总之，这组针对致脑炎性PLP139 - 151表位的Mab形成了一种有用的工具，可用于进一步研究MS患者以及不同动物物种的EAE模型中的自身抗原表达、脱髓鞘/再髓鞘化以及病变活动分期。
