Complement C3 fragments in urine: detection in systemic lupus erythematosus patients by western blotting.

R H Kelly, A B Carpenter, K S Sudol, S P Jagarlapudi, S Manzi
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Abstract

We characterized urinary excretion of C3 fragments among patients with systemic lupus erythematosus (SLE) as a possible indicator of renal involvement. 28 patients, representing a broad range of disease activity were admitted to our study. Urinary proteins were separated on 4-20% gradient SDS-PAGE gels, under reducing conditions, and transblotted to nitrocellulose. Western blots were developed with a polyvalent goat-anti-human C3d antiserum, and an alkaline phosphatase-conjugated rabbit anti-goat IgG. Three patterns were obtained: 1) no bands detected; 2) bands suggesting the presence of intact C3; and 3) samples with additional low molecular (< 4 x 10(4)) bands. The 12 patients with no C3 bands had minimal disease activity (e.g. fatigue, arthralgia, arthritis, rash, oral ulcers). The seven patients with intact C3 patterns also had minimally active disease. Their primary clinical findings included fatigue, pleurisy, renal disease which had been treated, hemolytic anemia, and arthritis. Patients with low molecular weight C3 fragments in their urine formed two sub-sets, based upon their presenting features. The first group had severe disease and contained all patients with active lupus nephritis (n = 4), while the second consisted of non-renal patients with primary clinical findings of moderate disease activity (e.g. thrombocytopenia, pneumonitis, arthritis). Our results suggest urinary excretion of low molecular weight C3 fragments correlates with active renal disease, but is a variable finding among SLE patients with non-renal manifestations of disease activity.

免疫印迹法检测系统性红斑狼疮患者尿液补体C3片段。
我们将系统性红斑狼疮(SLE)患者尿液中C3片段的排泄作为肾脏受累的可能指标。28例患者被纳入我们的研究,他们代表了广泛的疾病活动。尿蛋白在4-20%梯度SDS-PAGE凝胶上分离,在还原条件下,转印迹到硝化纤维素上。用多价山羊抗人C3d抗血清和碱性磷酸酶偶联兔抗山羊IgG进行Western blot检测。得到三种模式:1)未检测到条带;2)提示C3完整存在的条带;3)样品具有额外的低分子(< 4 × 10(4))波段。没有C3带的12例患者的疾病活动度最低(如疲劳、关节痛、关节炎、皮疹、口腔溃疡)。7例C3模式完整的患者也有最小活动性疾病。他们的主要临床表现包括疲劳、胸膜炎、治疗后的肾脏疾病、溶血性贫血和关节炎。尿中含有低分子量C3片段的患者根据其表现特征分为两个亚组。第一组病情严重,包括所有活动性狼疮性肾炎患者(n = 4),而第二组由非肾脏患者组成,主要临床表现为中度疾病活动性(如血小板减少症、肺炎、关节炎)。我们的研究结果表明尿中低分子量C3片段的排泄与活动性肾脏疾病相关,但在有疾病活动性非肾脏表现的SLE患者中是一个可变的发现。
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