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目的 研究抗肾小球基底膜 (GBM)抗体的抗原决定簇及其与临床表现的关系。方法 以抗Ⅳ型胶原α3 链非胶原区 1[α3(Ⅳ )NC1]单抗 (Mab3)和抗α5(Ⅳ )NC1单抗 (Mab5) ,及辣根过氧化物酶标记的亲和层析纯化的抗GBM自身抗体 (APab HRP) ,作为识别GBM上不同抗原决定簇的探针 ,应用竞争性ELISA法测定 33例抗GBM抗体的抗原决定簇 ,同时初步探讨其与临床表现的关系。结果33例患者血清均可抑制Mab3,30 / 33例可抑制APab HRP ,11/ 33例可抑制Mab5。但不同血清对同一探针 ,同一血清对不同探针的抑制率均不平行。单纯抗GBM抗体组 (2 7例 )对APab HRP的平均抑制率为 5 5 %± 18% ,而抗GBM抗体伴抗中性粒细胞胞浆抗体阳性组 (6例 )仅为 32 %± 2 2 % ,两组相比差异有显著意义 (P =0 .0 1)。在有或无肺出血患者之间 ,对各种探针抑制率的差异均无显著意义(P >0 .0 5 )。结论 抗GBM抗体血清均识别位于α3(Ⅳ )NC1上的主要抗原决定簇。但不同患者所识别的抗原决定簇并不完全一致 ,少数血清可识别α5(Ⅳ )NC1。是否合并ANCA ,所识别的抗原决定簇可能存在差异。
Objective To study the antigenic determinants of anti-glomerular basement membrane (GBM) antibodies and their relationship with clinical manifestations. Methods The anti-collagen type 1 α3 chain non-collagen 1 (α3 (Ⅳ) NC1] monoclonal antibody (Mab3) and anti-α5 (Ⅳ) NC1 monoclonal antibody (Mab5), and horseradish peroxidase-labeled affinity chromatography Purified anti-GBM autoantibody (APab HRP) was used as a probe to identify different antigenic determinants in GBM. The anti-GBM antibody epitopes of 33 anti-GBM antibodies were assayed by competitive ELISA and the relationship with clinical manifestations was also discussed. Results All the 33 patients could inhibit Mab3 in serum and 30/33 of them could inhibit APab HRP, while 11 of 33 patients could inhibit Mab5. However, the same probe with different serum, the same serum for different probe inhibition rate are not parallel. The anti-GBM antibody group (27 cases) had an average inhibition rate of APA HRP of 5 5% ± 18%, while the anti-GBM antibody anti-neutrophil cytoplasmic antibody positive group (6 cases) was only 32% ± 2 2%, there was significant difference between the two groups (P = 0.01). There was no significant difference in the inhibitory rates of various probes between patients with or without pulmonary hemorrhage (P> 0.05). Conclusion Anti-GBM antibody sera all recognize major epitopes on α3 (Ⅳ) NC1. However, the antigenic determinants identified by different patients are not exactly the same. A few serums can recognize α5 (Ⅳ) NC1. Whether or not ANCA is combined, the identified epitopes may differ.