UP (2) Table 1 Clinicopathologic findings of 30 patients with membranous glomerulonephritisa : urinary protein, IF : immunofluorescence findings Fib.:

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1 Search fcr Renal Tubular Epithelial Antigen in Membranous Glomerulonephritis. YOSHIAKI OHTA, AKIO KOYAMA, HIROMI INAGE MITSUHARU NARITA and SHIZUO TOJO. Institute of Clinical Medicine, The University of Tsukuba, Ibaraki, Japan. Key words: membranous glomerulonephritis, immune complex (IC), renal tubular epithelial (RTE) antigen, glomerular immune deposit, circa ating anti- RTE antibody In order to confirm a pathogenic role of renal tubular epithelial (RTE) antigen i n membranous glomerulonephritis, we searched for RTE antigens and anti-rte antibodies in membranous glomerulonephritis patients. RTE antigens were prepared from normal kidneys by a techniq ue adapted from that of Edgington et al. and Naruse et al. Biopsies from 30 patients with membranous glomerulonephritis (22 patients with primary membranous glomerulonephritis and 8 patients with secondary membranous glomerulonephritis) were examined for the presence of RTE antigens in glomerular immune deposits. In no instance was RTE antigen in glomerular immune deposits. Collected sere from 18 patients with membranous glomeru lonephritis (11 patients with primary membranous glomerulonephriti s and 7 patients with secondary membranous glomerulonephritis) were assayed for a nti-rte antibodies by indirect im:inunofluorescence and by radioimmunodiffusion. However, they had no demonstrable circulating anti-rte antibody. T hese studies suggest that if RTE antigen posseses a nephritogenic p otential for human membranous glomerulonephritis, it is probably only rarely expressed.

2 UP (2) Table 1 Clinicopathologic findings of 30 patients with membranous glomerulonephritisa : urinary protein, IF : immunofluorescence findings Fib.: fibrinogen, (gold) : gold therapy RVT: renal vein thrombosis, DM : diabetes mellitus RA : rheumatoid arthritis, SLE : systemic lupus erythematosus

3 Table 2 Clinical and laboratortory data for 18 patients with membranous glomerulonmephritis.

4 Fig. 1 Characterization of RTE antigen. An arrow shows RTE antigen (a zone)

5 日腎誌 第XXⅣ 巻 を 密 着Lautoradiographyを 1393 大田義章他 4名 第11号1984 施 行 して 検 索 した く,抗RTE抗 体(家兔IgG)と 抗 家兔IgGI血清 間 て 形 成 さ れ る 沈 降 線 に一 致し てI標 結 1) RTE抗原の plateに 果 に よ る 感 光(Fig.3で 性状 の 検 討:RTE抗 よ る 電 気 泳 動 上,α 原 はagar gel 領 域 に あ り(Fig.1),radioi- mmunoelectrophoresisに よ る 検 討 で は ヒ ト血 清 成 分は る 険 討:ヒ 抗RTE抗 3)腎切片GBM上 borderに 的 に 螢 光 染 色 さ れ た が,糸 位 尿細 管 さ ら に,こ は 染 色 は み ら れ な か っ た (2)酸 体 に よ る 螢 光 染 色 で は,IgGに の抗 原 光染 に よ る 検 討:Fig.3 の ごと 対 す る 染 色 は 減 弱し,(1) はRTE抗 原 は 認 め ら れ な か っ た 4)患 者 血 清 中 の 抗RTE抗 Section RTE of normal antigen. proximal tubular capillary walls. human Specific 体 法 に よ る抗RTE抗 kidney stained Iluorescence epithelial cells. 識 抗RTE抗 に は 染 色 は み ら れ な か っ た じ 以 上 よ り腎 切片GBMに 色 は 消 失 した 2 処理 に よ と 同 様 に 全 例 で 尿 細 管 上 皮 の 染 色 は み ら れ た が,GBM 体 を 吸 収 した と こ ろ,螢 (2)R adioimmunodiffusion法 記 対像 患者全 の 尿細 管 上 皮 は 染 色 され り抗 原 抗 体 を 解 離 溶 出 した 後 のFITC標 球 体 お よ び 腎 の 他 の 部 分 には 識 抗RTE抗 原 の 検 索:(1)FITC 体 に よ る 螢 光 染 色 で は,上 た が,GBMに 相 当 す る 部 分 のみ が特 異 螢 光 染 色 は み られ な か っ た(Fig.2) Fig. のRTE抗 例 で 正 常 腎 と 同 様 に 腎切片上 ト正 常 腎 ク リオ ス タ ッ ト切 片 のFITC標識 上皮 管 腔 側 のbrush でFITC標 光抗体 法 に よ 体 の 直 接 法 に よ る 螢 光 染 色 で は,近 認 め られ た が, 常 家 兎 血 清 では 沈 降 線上 に 感光は 標 識 抗RTE抗 体 の 性 状 の 検 討:(1)螢 原 認 め ら れ な かった 検 出 さ れ な か っ た 2)抗RTE抗 識RTE抗 矢 印 が 示 す)が negative eontrolの正 との with is observed No (5) fluorescence 体 の 検 索:(1)間 体 の 検 索:上 FITC-conjugated only in is found the on 記 対 象 患者 全 例 で. antibody luminal the 接螢 光抗 layer glomerular to of

6 1394 膜性糸球体腎炎における腎尿細管上皮抗原の関与につ いての検討 Fig. Fig. 3 4 Radioimmunodiffusion of RTE antigen and -RTE antigens on precipitation lines Radioimmunologic antigens search on precipitation for circulating lines. (6) anti-rte anti-rte antibody, antibody. Arrows Arrows show I 125I-RTE

7

8 2) Whitworth, J. A., Leibowitz, S., Kennedy, M. C., Cameron, J. S., Evans, D. J., Glasslock, R. J. and Schoenf eld, L. S.: Absence of glomerular renal tubular epithelial antigen in membranous glomerulonephritis. Gun. Nephrol., 5: , ) Zager, R. A., Couser, W, G., Andrews, B. S., Bolton, W. K. and Pohl, M. A.: Membranous nephropathy: A radioimmunologic search for antirenal tubular epithelial antibodies and circulating immune complexes. Nephron, 24: 10-16, ) Thorpe, L. W. and Cavallo, T.: Renal tubule brush border antigens: Failure to confirm a pathogenic role in human membranous glomerulonephritis. J. Gun. Lab. Immunol., 3: , ) Collins, A. B., Giuseppe, A. A. and McCluskey, R. T.: Lack of evidence for a role of renal tubular antigen in human membranous glomerulonephritis. Nephron, 27: , ) Edgington, T. S., Glassock, R. J., Watson, J. I. and Dixon, F. J.: Characterization and isolation of specific renal tubular epithelial antigens. J.

9 Immunol., 99: , ) Clark, H. F. and Shepard, C. C.: A dialysis technique for preparing fluorescent antibody. Viorology, 20: , ) Narita, M., Wakashin, M. and Wakashin, Y.: A glomerulonephritis with deposition of renal tubular epithelial antigen in glomerular basement membrane after renal homotransplantationn in Cadaveric Renal Transplantation, edited by Iwasaki, Y., Igaku Shoin LTD, Tokyo, , ) Strauss, J., Pardo, V., Koss, M. N., Griswold, W. and McIntosh, R. M.: Nephropathy associated with sickle cell anemia: An autologous immune complex nephritis. I. Studies on nature of glomerular-bound antibody and antigen. Identification in a patient with sickle cell disease and immune deposition glomerulonephritis. Amer. J. Med., 58: , ) Kerpen, H. O., Bhat, J. G., Feiner, H. D. and Baldwin, D. S.: Membranous nephropathy associated with renal cell carcinoma. Evidence against a role of renal tubular or tumor antibodies in pathogenesis. Am. J. Med., 64: , ) Ozawa, T., Boedecker, E. A., Schorr, W., Guggenheim, S. and McIntosh, R. M.: Immune-complex disease with unilateral renal vein thrombosis. Arch. Pafhol. Leb. Med., 100: , ) Schwayder, M., Ozawa, T., Eoedecker, E., Gug genheim, S. and Mclntosh, R. M.: Nephrotic syndrome associated with Fanconi syndrome. Im munopathogemc studies of tubulointerstitial ne phritis with autologous immune-complex glomeru lonephritis. Ann. Intern. Med., 84: , ) Gilboa, N., Durante, D. and McIntosh, R. M.: Glomerular deposition of renal tubular epithelial antigen in patients with systemic lupus erthematosus: Its possible role in lupus nephritis. J. Rheumutol., 4: , ) Droz, D., Noel, L. H., Fouchard, M., Perrone, E. and Parbanel, C.: Presence of renal tubular epithelium antigen in glomeruli of membranous glomerulonephritis: An indicator of renal vein thrombosis? Abstr. 8th. Int. Congr. Nephrol., 253' ) Douglas, M. F. S., Rabideau, D. P., Schwartz, M. M. and Lewis, E. J.: Circulating anti-human proximal tubular brush border antibody in a patient with membranous glomerulonephritis. Abstr. 8th. Int. Congr. Nephrol., 90, 1981.

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