Development of an animal molel for Periarteritis nodosa-like vasculitis and identification of its

在过去的一年里，我们报道了自身免疫性MRL/lpr小鼠，核结合蛋白（Nuc）基因缺陷，已知可以增加抗dna抗体的产生，发生结节性动脉周围炎（PN）样anca阳性坏死性血管炎伴月牙性肾小球肾炎（CrGN）的频率很高。其特点是nuc缺失的MRL/lpr小鼠（-/-）在血清中产生的免疫复合物（IC）是野生MRL/lpr小鼠（+/+）的3倍。因此，我们用SDS-PAGE分析了天然条件下-/-小鼠肾脏中IC的成分。电泳转移到PVDF膜上的45 kd区域的主要条带被处理用于氨基酸测序。经二维电泳证实为精氨酸琥珀酸合成酶（ASS）。如上次会议所示，IC也在一定程度上包含actin。从染色强度判断，ASS的含量是actin的10倍。因此，我们制备了用于ELISA的重组小鼠ASS。正如预期的那样，-/-小鼠的抗ass抗体水平显著高于+/+小鼠。值得注意的是，该水平与早期病变的存在（相当于人类经典PN的I至II期）密切相关，但与血管炎（III期和IV期）和/或GN的严重程度无关。本实验在动物模型中揭示了一种新的抗原抗体系统，该系统将用于早期预测人类多血管炎（MPA）等Pie致命疾病。

In the previous year, we have reported that autoimmune MRL/lpr mice, rendered genetically deficient for nucleobindin (Nuc), which is known to augment anti-DNA antibody production, developed periarteritis nodosa (PN)-like ANCA-positive necrotizing vasculitis accompanying crescentic glomerulonephritis (CrGN) at high frequency. The characteristic was that the Nuc-deficient MRL/lpr mice(-/-) produced immune complexes (IC) in the serum 3 times as much as those in wild MRL/lpr mice (+/+). Thus, we analyzed the components of IC in kidneys of the -/- mice by SDS-PAGE under native conditions. A major band at 45 kd region on the gel, which was electrophoretically transferred onto a PVDF membrane, was processed for aminoacid sequencing. It turned out to be arginino-succinate synthase (ASS), and was further confirmed by 2-dimensional electrophoresis. IC also contained actin to some extent as shown in the previous meeting. The content of ASS was 10 times as much as that of actin, as far as judged from staining intensity. Accordingly, we produced recombinant mouse ASS for ELISA.As was expected, the level of anti-ASS antibodies in the -/- mice was significantly higher than those of the +/+ mice. Of note was that the level was associated well with the presence of early lesions comparable to the stage from I to II of human classical PN but not with the severity of vasculitis (stage III and IV) and/or GN.A novel antigen-antibody system revealed in this experiment using animal model for PN or microscopic polyangiitis (MPA) will be of use for an early prediction of Pie fatal disease such as MPA in humans.