Faculty Profiles - KAWACHI Hiroshi

EFFECT OF TRADITIONAL CHINESE MEDICINE (SAIREI-TO) ON MONOCLONAL ANTIBODY-INDUCED PROTEINURIA IN RATS Reviewed

H KAWACHI, N TAKASHIMA, M ORIKASA, T OITE, F SHIMIZU

PATHOLOGY INTERNATIONAL 44 ( 5 ) 339 - 344 1994.5

Language：English Publishing type：Research paper (scientific journal) Publisher：BLACKWELL SCIENCE

The effects of traditional Chinese medicine (Sairei-to) on monoclonal antibody (mAb) inducing proteinuria were examined. Four hundred, 200 and 100 mg/kg body weight (BW) of Sairei-to and phosphate-buffered saline (PBS) as a control were injected intraperitoneally into four groups of female Wistar rats every day from 5 days before intravenous injection of mAb to the end of the experimental period. The amount of urinary protein excretion was significantly suppressed in roughly a dose-dependent manner. For example, 116.6 +/- 89.7 mg/day of proteinuria was observed in control groups compared to 4.2 +/- 15.2 mg/day in the 400 mg/kg BW of Sairei-to treated group 2 days after mAb injection (P < 0.01).
Statistically significant (P < 0.01) differences were again observed in a repeat experiment (122.1 +/- 53.7 vs 10.2 +/- 10.1 mg/day on the 2nd day) without affecting the glomerular filtration rate.
No significant difference was recognized between the total amount of mAb bound to glomeruli 1 h after mAb injection in Sairei-to-treated and non-treated rats, indicating that Sairei-to pretreatment has no effects on the number or quality of antigenic molecules.
The effect of Sairei-to on a non-immunological model of proteinuria was also examined. No significant reduction of proteinuria by similar Sairei-to treatment was observed in aminonucleoside of puromycin nephropathy.
The authors conclude that mAb-induced proteinuria in rats is significantly suppressed by the traditional Chinese medicine, Sairei-to.

Web of Science

Effect of chlorpromazine on kinetics of injected monoclonal antibody in MoAb-induced glomerular injury Reviewed

N. Takashima, H. Kawachi, T. Oite, S. Nishi, M. Arakawa, F. Shimizu

Clinical and Experimental Immunology 91 ( 1 ) 135 - 140 1993

Language：English Publishing type：Research paper (scientific journal)

The effect of chlorpromazine, one of several calmodulin antagonists that inhibit cytoskeletal movement, on the local kinetics of injected proteinuria-inducing MoAb 5-1-6 was examined to test the hypothesis that proteinuria is inhibited if the antigen recognized by MoAb 5-1-6 or injected MoAb remains on the surface of epithelial foot processes. MoAb 5-1-6 was injected into both chlorpromazine-treated (5 mg/100 g body weight) and untreated rats. As a positive control for the chlorpromazine treatment, anti-Fx1A serum was also injected into other chlorpromazine-treated and untreated rats. Chlorpromazine inhibited neither the change in localization of injected MoAb 5-1-6 nor proteinuria, although it showed an inhibitory effect on redistribution of immune complex and the fixation of complement in passive Heymann glomerulonephritis induced by injection of anti-Fx1A serum. We conclude that the kinetics of bound MoAb 5-1-6 are regulated by a system different from that operating in passive Heymann glomerulonephritis.

Quantitative study of mesangial injury with proteinuria induced by monoclonal antibody 1-22-3 Reviewed

H. Kawachi, T. Oite, F. Shimizu

Clinical and Experimental Immunology 92 ( 2 ) 342 - 346 1993

Language：English Publishing type：Research paper (scientific journal)

Murine MoAb 1-22-3 has already been reported to bind to the mesangial cell surface and to cause transient proteinuria and mesangial morphological changes characterized by mesangiolysis, subsequent mesangial cell proliferation and mesangial matrix increase by a single i.v. injection. In this study, MoAb-induced glomerulopathy was quantitatively analysed. No correlation between the severity of mesangial morphological changes and the degree of proteinuria was detected (r = 0.190). The minimum dose injected to induce abnormal proteinuria was 25 μg. This dose corresponded to 1.79 μg/2 kidneys 30 min after MoAb injection. The highest average level of proteinuria was observed in rats injected with 500 μg of MoAb, and less proteinuria was observed in rats injected with 10.0, 5.0 and 2.0 mg. Although the amounts of kidney-fixing MoAb and the subsequent deposition of rat C3 in the high-dose-injected group were larger than in the 500 μg injected group, the numbers of infiltrating inflammatory cells were the same in both groups. No correlations between the degrees of such mediators and proteinuria were observed.

Nephrotoxic serum nephritis in nude rats: The roles of host immune reactions in the accelerated type Reviewed

T. Sato, H. Kawachi, T. Morioka, N. Takashima, T. Saeki, T. Oite, N. L. Sato, M. Takeuchi, F. Shimizu

Clinical and Experimental Immunology 91 ( 1 ) 131 - 134 1993

Language：English Publishing type：Research paper (scientific journal)

By immunization with rabbit immunoglobulins and the injection of a subnephritogenic dose of rabbit nephrotoxic serum (NTS), accelerated-type nephrotoxic serum nephritis (NTN) was induced in heterozygous (rnu/+) rats but not in athymic nude (rnu/rnu) rats. By transferring rat antibody against rabbit immunoglobulins, marked proteinuria was induced also in nude rats (202.0 ± 98.4 mg/day on day 3) as in rnu/+ rats (122.6 ± 35.3 mg/day on day 3). No marked differences in histological findings could be found between both groups. The most marked increase in the number of intraglomerular infiltrating cells was observed in heterozygous rats indicating that the presence of thymus-derived cells leads to the accumulation of more cells in glomeruli. We conclude that humoral immunity alone is enough to accelerate the pathogenic mechanism which induces glomerular injury with heavy proteinuria in this model.

Divalency of the monoclonal antibody 5-1-6 is required for induction of proteinuria in rats Reviewed

M. Narisawa, H. Kawachi, T. Oite, F. Shimizu

Clinical and Experimental Immunology 92 ( 3 ) 522 - 526 1993

Language：English Publishing type：Research paper (scientific journal)

A single i.v. injection of 3 mg of the F(ab')2 fragment of MoAb 5-1-6 into rats induced immediate proteinuria (128.1 ± 80.7 mg/24 h on day 1) which lasted 1-2 days. In contrast, rats administered 10 mg of the corresponding Fab fragment did not develop abnormal proteinuria even though an equivalent dose of the intact MoAb 5-1-6 far exceeded the nephritogenic dose. The total kidney binding of 125I-Fab fragment was 209.5 ± 34.3 μg/2 kidneys. This exceeded that obtained by injection of 3 mg MoAb 5-1-6 IgG1 (58.9 ± 12.5 μg/2 kidneys at 1 h) and was similar to that obtained following injection of 3 mg F(ab')2 fragment (235.3 ± 16.9 μg/2 kidneys). Immunofluorescence (IF) showed a linear pattern along the glomerular capillary wall at 1 h after the administration of MoAb 5-1-6 IgG1, F(ab')2 or Fab fragment. On day 5, fine to coarse granules were observed scattered in F(ab')2-injected rat glomeruli, whereas granules were densely localized in Fab-injected rat glomeruli. Complement-depleted rats injected with 3 mg of MoAb 5-1-6 IgG1 developed proteinuria with the same time course as non-depleted rats. This observation, together with the ability of F(ab')2 to induce proteinuria, indicates that proteinuria induced by MoAb 5-1-6 is complement-independent. This study suggests that MoAb 5-1-6-induced proteinuria is initiated by cross-linking of the epitopes by divalent MoAb 5-1-6 and is independent of complement activity.

EPITOPE-SPECIFIC INDUCTION OF MESANGIAL LESIONS WITH PROTEINURIA BY A MOAB AGAINST MESANGIAL CELL-SURFACE ANTIGEN Reviewed

H KAWACHI, M ORIKASA, K MATSUI, T IWANAGA, S TOYABE, T OITE, F SHIMIZU

CLINICAL AND EXPERIMENTAL IMMUNOLOGY 88 ( 3 ) 399 - 404 1992.6

Language：English Publishing type：Research paper (scientific journal) Publisher：BLACKWELL SCIENCE LTD

A MoAb 1-22-3 (IgG3) was produced in mice immunized with rat glomeruli. A blotting study indicated the antigen molecule recognized by this MoAb has a molecular weight of about 25 kD, which is the same as that of the Thy 1.1 molecule. This MoAb is capable of inducing the morphological changes similar to those induced by anti-thymocyte serum or the anti-Thy 1.1 MoAb, ER4 and massive proteinuria in rats by a single i.v. injection. Proteinuria started immediately after MoAb 1-22-3 injection and peaked on day 5. Reaction products were detected by immunoelectron microscopy in vitro on the limited mesangial cell surface facing endothelial cells and in vivo in partially lysed mesangial cells 30 min after injection. Unlike the proteinuria-inducing MoAb ER4, reactivity of MoAb 1-22-3 was detected neither on endothelial cells, epithelial cells, nor along the glomerular basement membrane in vivo and in vitro. There is a difference in reactivity toward guinea-pig and rabbit materials between MoAb 1-22-3 and the commercial anti-Thy 1.1 MoAb (OX7). The antigenic determinant of MoAb 1-22-3 is concluded to be a new epitope and only the binding of MoAb 1-22-3 to this epitope proved to lead to an abnormal increase of glomerular capillary wall permeability.

Web of Science

Mesangial sclerotic change with persistent proteinuria in rats after two consecutive injections of monoclonal antibody 1-22-3 Reviewed

H. Kawachi, T. Iwanaga, S. Toyabe, T. Oite, F. Shimizu

Clinical and Experimental Immunology 90 ( 1 ) 129 - 134 1992

Language：English Publishing type：Research paper (scientific journal)

Irreversible mesangial changes with persistent proteinuria were induced in rats given two consecutive injections 2 weeks apart of a MoAb 1-22-3 to rat mesangial cell. The characteristics of the resulting lesions were investigated and compared with those of the reversible change induced by a single injection. At 24 h after the second injection, mesangiolytic changes similar to those after a single injection were evident. The accumulation of macrophage-like cells in glomeruli observed at 1 week after the first injection was not evident during the experimental period after the second injection. Hypercellularity with the characteristics of intrinsic mesangial cell and increased mesangial matrix were already present 1 week after the second injection. And mesangial sclerotic change progressed up to 6 months. Deposition of collagen type I and type III and accumulation of collagen fibril at the ultrastructural level were evident in rats 6 months after the second injection. Proteinuria started immediately and continued for more than 6 months after the second injection. The mesangial sclerotic change with persistent proteinuria described here is considered to be a better model for investigating the mechanism of chronic progression of human mesangial proliferative glomerulonephritis.

A monoclonal antibody (1G10) recognizes a novel human mesangial antigen Reviewed

S. Kagami, K. Okada, M. Funai, K. Matui, T. Oite, H. Kawachi, F. Shimizu, Y. Kuroda

Kidney International 42 ( 3 ) 700 - 709 1992

Language：English Publishing type：Research paper (scientific journal)

We have identified a unique mesangial matrix protein of the human glomerulus by using a monoclonal antibody, 1G10, generated against cultured human glomerular cells. By immunofluorescence, the antigen recognized by 1G10 (1G10 antigen) is present in mesangium and smooth muscle tissue and cannot be detected in any other tissue examined. Immunoelectron microscopy of glomeruli indicated that 1G10 antigen is present exclusively in the mesangial matrix at the endothelial-mesangial interface. The 1G10 antigen is also expressed by cultured mesangial cells, but not by cultured glomerular epithelial cells, umbilical endothelial cells or fibroblasts. 1G10 did not react with the mesangial matrix proteins [fibronectin (FN), laminin (LAM), collagen types I, III, IV, V, and VI (Col I, III, IV, V, VI), heparin sulfate proteoglycan (HSPG), or thrombospondin (TS)] present under normal and diseased states or smooth muscle antigens (myosin, actin), but did react with a 4 M urea extract of renal cortex and a 0.3% deoxycholate extract of isolated glomeruli. Two dimensional immunoblot analysis using the urea extract demonstrated the binding of 1G10 to an ~200 KDa polypeptide with pI 6.0. On one dimensional immunoblot this band did not show cross react with polyclonal antisera to FN, LAM, Col IV, V, VI, HSPG or TS. This mesangial matrix component is trypsin and periodate sensitive, suggesting that it has the character of glycoprotein. In renal biopsy specimens from patients with mesangial proliferative glomerulonephritis (GN) and membranoproliferative GN, the expression of the 1G10 antigen increased along with mesangial hypercellularity or increased accumulation of mesangial matrix, but decreased in completely sclerosed glomeruli. No significant changes in 1G10 antigen expression was observed in membranous GN or minimal change nephrosis compared to normal glomeruli. This study suggests that the 1G10 antigen may not only be a useful marker for the clinical assessment of GN, but may also serve as a potential tool for the study of the pathogenesis of glomerular diseases characterized by cellular proliferation and mesangial matrix expansion.

DOI： 10.1038/ki.1992.337

Quantitative studies of monoclonal antibody 5‐1‐6‐induced proteinuric state in rats Reviewed

H. KAWACHI, K. MATSUI, M. ORIKASA, T. MORIOKA, T. OITE, F. SHIMIZU

Clinical & Experimental Immunology 87 ( 2 ) 215 - 219 1992

Language：English Publishing type：Research paper (scientific journal)

Murine monoclonal antibody (MoAb) 5‐1‐6 was already reported to bind to epithelial cell fool processes and to cause proteinuria in rats. In vivo kinetics of the injected MoAb 5‐1‐6, relationship between the quantity of kidney‐binding antibody and proteinuria. and changes in the amount of antigenic molecule recognized by this MoAb in the proteinuric state were studied. The amount of total kidney‐binding antibody (TK Ab) as determined I h after a 2 mg administration was 50.8 ± 10.4 μg/2 kidneys, and TKAb declined to 1.9 ± 0.4 at day 15. The minimum dose of MoAb required to induce proteinuria was 125 μg as the injected dose. This dose corresponded to 12‐8 μg of TKAb at I h and 0.34 μg of TKAb at day 5. The amount of MoAb 5‐1 ‐6 binding to isolated normal glomeruli was also shown to exceed 147‐7 μg/76000 glomeruli, indicating proteinuria to be induced provided more than 8.7% (= 12.8/147.7) of the critical epitopes is specifically occupied by MoAb. The total amount of MoAb 5‐1‐6 bound to glomeruli in vivo and in vitro was assayed with [125I]‐labelled anti‐mouse IgG. The amount of [125I] anti‐mouse IgG bound to glomeruli was 6.93 ± 0.45 μg/10 000 glomeruli from rat 5 days after this MoAb injection and 26.58 ± 0.66 μg/10000 control glomeruli, indicating the decrease in the number of MoAb 5‐1‐6‐rccognizcd antigen molecules in glomeruli isolated from the rat in proteinuric state induced by this MoAb. Thus, the MoAb 5‐1‐6‐recognized molecule itself may principally function to regulate the permeability of the glomerular capillary wall and the decrease of the molecule may lead to proteinuria. Copyright © 1992, Wiley Blackwell. All rights reserved

DOI： 10.1111/j.1365-2249.1992.tb02977.x

Altered localization of antigen recognized by proteinuriainducing monoclonal antibody in experimental nephrosis Reviewed

Teruo Okasora, Mitsumasa Nagase, Hiroshi Kawachi, Katsuyuki Matsui, Michiaki Orikasa, Tetsuo Morioka, Ichiroh Yamazaki, Takashi Oite, Fujio Shimizu

Virchows Archiv B Cell Pathology Including Molecular Pathology 60 ( 1 ) 41 - 46 1991.12

Language：English Publishing type：Research paper (scientific journal) Publisher：Springer-Verlag

Change in the localization of the antigen recognized by the proteinuria-inducing monoclonal antibody (MA) 5-1-6 in experimental nephrosis was studied by indirect and biotin-avidin immunofluorescence, and immunoperoxidase at light and electron microscopical levels. The proteinuric state was induced by the administration of the aminonucleoside of puromycin (PAN) or adriamycin. The antigen decreased in quantity and/or its distribution changed with an increase in the amount of protein excreted in both experimental models. Recovery from the alterations observed during the development and proteinuria appeared to occur when PAN-induced proteinuria subsided. This antigenic molecule may thus be essential for maintaining the normal permselectivity of glomerular capillary walls. © 1991 Springer-Verlag.

DOI： 10.1007/BF02899526

Nephritogenicity of anti-Engelbreth-Holm-Swarm sarcoma antibody Reviewed

H. Kawachi, T. Nakamura, T. Kazama, E. Yaoita, T. Morioka, K. Matsui, T. Oite, F. Shimizu

Nephron 47 ( 2 ) 150 - 154 1987

Language：English Publishing type：Research paper (scientific journal) Publisher：S. Karger AG

We have examined the antibody activity and nephritogenicity of anti-Engelbreth-Holm-Swarm (EHS) sarcoma antiserum in order to analyze the pathogenesis of the EHS nephropathy which has already been reported by us. An increased amount of urine protein was not recognized in rats injected with a large quantity of anti-EHS sarcoma antiserum. In addition, rats immunized with rabbit IgG before anti-EHS sarcoma antiserum injection developed no abnormal proteinuria, despite positive fluorescent staining for rat IgG as well as rabbit IgG in the glomeruli. From these results we concluded that nephritogenic antibodies could not be produced in rabbits by immunization with EHS sarcoma, which could induce the EHS nephropathy in an active model.

DOI： 10.1159/000184479

Studies on the 'linear pattern' in renal glomeruli demonstrated with immunofluorescence Reviewed

H. Kawachi, T. Oite, F. Shimizu

Nephron 39 ( 1 ) 36 - 39 1985

Language：English Publishing type：Research paper (scientific journal)

We have experimentally induced the 'linear pattern' in immunofluorescence: the linear deposition of endogenous immunoglobulin (Ig) along the glomerular basement membrane (GBM) in rats injected with both protamine and nephrotoxic serum. This Ig was demonstrated to have no specific antibody activity against GBM or rabbit serum. Our findings could be of value in the analysis of the mechanism and pathological meaning of the 'linear pattern' of endogenous Ig in immunofluorescence.