Yoshida H, Wakashin Y, Ueda S, Azemoto R, Iesato K, Yamamoto S, Mori T, Ogawa M, Mori Y, Wakashin M
First Department of Internal Medicine, School of Medicine, Chiba University, Japan.
Kidney Int. 1990 May;37(5):1286-94. doi: 10.1038/ki.1990.113.
Immunopathogenicity of trypsin-solubilized or non-solubilized renal tubular basement membrane (TBM) of the Lewis (LEW) rat was investigated. Autoimmune tubulointerstitial nephritis (TIN) was induced in BALB/c mice by immunization with trypsin-solubilized LEW rat TBM, while immunization with non-solubilized TBM did not produce the disease. Based on this preliminary experiment we studied the characterization of immunogenic and nephritogenic TBM antigen of the LEW rat. TIN was characterized by severe mononuclear cell infiltrates with multi-nucleated giant cells in the interstitium, tubular destruction and intensive IgG and C3 deposits along the TBM. Anti-TBM antisera and eluate from the nephritic mouse kidneys reacted with the TBM of normal LEW rat kidney by immunofluorescence. LEW rat TBM was also detected immunofluorescently by using antisera from BALB/c mice immunized with autologous trypsin-solubilized TBM. A competitive inhibition test revealed a higher titer of anti-TBM antibody in the eluate than in the adsorption-treated antisera per microgram IgG. Immunoblotting showed one reactive band with a molecular weight of 45,000 daltons, and the blotting patterns in tryptic TBM of the Brown Norway (BN) and LEW rats appeared similar. Amino acid analysis of nephritogenic LEW rat tryptic TBM showed that it contained no hydroxyproline and hydroxylysine, suggesting that this TBM preparation was not collagenous. These findings suggest that tryptic digestion contributes to the release of nephritogenic antigen from the LEW rat TBM and that this antigen system might participate in the immune system involved in the anti-TBM associated TIN that is well known to be induced by non-digested TBM of TBM antigen positive animals.
研究了Lewis（LEW）大鼠胰蛋白酶溶解或未溶解的肾小管基底膜（TBM）的免疫致病性。用胰蛋白酶溶解的LEW大鼠TBM免疫BALB/c小鼠可诱发自身免疫性肾小管间质性肾炎（TIN），而用未溶解的TBM免疫则不会引发该疾病。基于此初步实验，我们研究了LEW大鼠免疫原性和致肾炎性TBM抗原的特性。TIN的特征为间质中出现严重的单核细胞浸润并伴有多核巨细胞、肾小管破坏以及沿TBM密集的IgG和C3沉积。抗TBM抗血清以及来自肾炎小鼠肾脏的洗脱液通过免疫荧光与正常LEW大鼠肾脏的TBM发生反应。使用用自体胰蛋白酶溶解的TBM免疫的BALB/c小鼠的抗血清，也可通过免疫荧光检测到LEW大鼠TBM。竞争性抑制试验显示，每微克IgG的洗脱液中抗TBM抗体的滴度高于吸附处理后的抗血清。免疫印迹显示一条分子量为45,000道尔顿的反应带，并且Brown Norway（BN）大鼠和LEW大鼠的胰蛋白酶处理的TBM中的印迹模式相似。对致肾炎性LEW大鼠胰蛋白酶处理的TBM进行氨基酸分析表明，其不含羟脯氨酸和羟赖氨酸，这表明该TBM制剂不是胶原性的。这些发现表明，胰蛋白酶消化有助于从LEW大鼠TBM中释放致肾炎性抗原，并且该抗原系统可能参与了与抗TBM相关的TIN的免疫系统，众所周知，这种TIN是由TBM抗原阳性动物的未消化TBM诱导的。
