Faculty Profiles - KAWACHI Hiroshi

Papers - KAWACHI Hiroshi

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Role of mesangial cell damage in progressive renal disease Reviewed

F Shimizu, H Kawachi, M Orikasa

KIDNEY & BLOOD PRESSURE RESEARCH 22 ( 1-2 ) 5 - 12 1999

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

Mesangial cell proliferation and matrix accumulation are considered to contribute to the development of glomerulosclerosis. To investigate the pathological role played by mesangial cell damage in progressive renal disease appropriate progressive models initiated by a mesangial cell injury should be developed, Monoclonal antibody (mAb) 1-22-3 (IgG3) recognizes a critical epitope of the Thy-1.1 molecule on the mesangial cell surface, binding of which induces more severe mesangial cell injury than in the case of OX-7, a commercially obtainable anti-Thy-1.1 mAb, The mAb 1-22-3 has enabled us to develop irreversible models of renal damage induced by two consecutive injections, by a single injection into unilaterally nephrectomized rats, or by a single simultaneous injection with another mAb, in addition to the corresponding reversible model. Detailed examinations using the combination of both types of models, all of which are initiated by an immune mechanism directed against the identical epitope on the Thy-1.1 molecule, are expected to provide new insights into mechanisms of irreversible renal injury initiated by mesangial cell damage and also to develop novel and rational therapeutic approaches to progressive, primarily mesangial diseases.

DOI： 10.1159/000025903

単クローン抗体により惹起されるモデルを用いた糸球体腎炎の発症, 進展機序の解析.

河内裕

新潟医学会雑誌 112 219 - 225 1998.12

.Effect of Sairei-to on irreversible glomerular sclerotic lesions in rats. Reviewed

Li,P, Kawachi,H, Orikasa,M, Shi,Z.S, Shimizu,F

Nephrology 4 49 - 59 1998.12

腎とアポトーシス Reviewed

河内裕, 清水不二雄

医学のあゆみ 187 486 - 489 1998.12

Altered anionic GBM components in monoclonal antibody against slit diaphragm-injected proteinuric rats Reviewed

Yoshihide Fujigaki, Mitsumasa Nagase, Sumi Hidaka, Katsuyuki Matsui, Mikiko Shirai, Hitonari Nosaka, Hiroshi Kawachi, Fujio Shimizu, Akira Hishida

Kidney International 54 ( 5 ) 1491 - 1500 1998

Language：English Publishing type：Research paper (scientific journal) Publisher：Blackwell Publishing Inc.

Background. We previously reported that monoclonal antibody (mAb) 5-1-6 bound to renal filtration slits induces massive proteinuria without causing ultrastructural changes in the glomerulus. This study evaluated the underlying mechanisms of the increase in glomerular permeability. Methods. The distribution of endogenous albumin and IgG in the glomerular basement membrane (GBM) was studied in in situ drip-fixed glomeruli of Munich-Wistar rats by use of immunogold immunocytochemistry in the presence and absence of mAb 5-1-6. The density of foot process glycocalyx components was estimated by labeling with Limax fluvus lectin- or Helix pomatia lectin-gold complexes. Anionic sites in the GBM were examined by labeling with cationic gold at pH 2.0 or 7.4. Carboxyl groups, which also furnish all anionic charge to the GBM, were examined by specific biotinylation and colloidal gold probe methods. In addition, the infusion-staining of anionic sites was performed by use of ruthenium red in both Munich-Wistar and Wistar rats. Results. The urinary excretion of albumin and IgG was increased markedly in the treated rats, indicating a non-selective barrier defect. In the control rats, albumin and IgG molecules were mainly located along the inner half of the GBM, and to a lesser degree in the lamina rara externa. In the treated rats, the albumin and IgG moieties were more equally distributed throughout the width of the GBM. Newly appearing, small dense peaks at the outer side of the GBM were evident, indicating a barrier function of outer zone of the GBM and/or epithelial cell layer. No intergroup differences in the density of lectin binding sites on foot processes were seen. The reduction in the number of ruthenium red-positive anionic sites and cationic gold (pH 2.0)-labeled anionic sites in the lamina rara externa was significant in the treated rats at day 3, indicating a possible alteration of charged proteoglycan in the lamina rara externa. No such changes were seen with cationic gold (pH 7.4)- labeled anionic sites in the GBM. The density of labeled carboxyl groups was significantly reduced in the treated rats relative to the controls. Conclusions. These results show that the injection of mAb 5-1-6 induced a perturbation of the charge- and probably the size-selective glomerular filtration barrier. The observed reduction in the levels of various negatively charged substances resulted in massive proteinuria, implying that alteration of target antigens can affect the integrity of the GBM constituents maintaining the normal barrier function.

DOI： 10.1046/j.1523-1755.1998.00131.x

Comparative nephritogenicity of two monoclonal antibodies that recognize different epitopes of rat Thy-1.1 molecule Reviewed

H. Nakayama, T. Oite, H. Kawachi, T. Morioka, H. Kobayashi, M. Orikasa, M. Arakawa, F. Shimizu

Nephron 78 ( 4 ) 453 - 463 1998

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

The pathophysiological role of the Thy-1.1 molecule expressed on rat mesangial cells with regard to mesangial cell dysfunction and injury remains unknown. The mechanism of Thy-1.1-associated injury has now been investigated with two monoclonal antibodies, 1-22-3 and OX7, that recognize different epitopes of Thy-1.1. Mesangiolysis and mesangial cell proliferation were more marked in rats injected with 1-22-3 than in those treated with OX7. Immunostaining for rat complement component C3 and also C9 was similar in the kidneys of rats 1 h after injection of either antibody. Alpha smooth muscle actin was first detected 3 days after injection of 1-22-3 and peaked on day 5
type I collagen staining showed a mesangial pattern on days 5 and 10. The staining for alpha smooth muscle actin and type I collagen was less intense in OX7-treated rats than in the 1-22-3-injected rats. The amounts of mRNAs encoding collagen types I and III peaked 5 days after injection of 1-22-3 and 10 days after injection of OX7. Rats injected with 1-22-3 developed proteinuria that was already marked on day 1 and peaked at 150 mg/day on day 3, whereas OX7 induced a low grade of proteinuria with large interindividual variability on day 3. Immunostaining for rat C3 in the normal rat kidneys, incubated in vitro with 1-22-3 or OX7 followed by incubation with normal rat fresh serum as a complement source, as well as the levels of serum complement activity, CH50, 30 min after injection of 1-22-3 or OX7 were similar, suggesting that the difference in the nephritogenicity of these two antibodies is not attributable to a difference in their complement-fixing activities, but rather may result from the difference in epitope specificities. The epitope recognized by 1-22-3 thus appears to be important in the initiation and progression of antibody-induced nephritis.

DOI： 10.1159/000044975

Immunohistochemical identification of type B intercalated cells in the rat kidney by a monoclonal antibody Reviewed

Haruko Yanase, Michiaki Orikasa, Fujio Shimizu, Hiroshi Kawachi, Toshihiko Iwanaga

Archives of Histology and Cytology 61 ( 2 ) 151 - 161 1998

Language：English Publishing type：Research paper (scientific journal) Publisher：Japan Society of Histological Documentation

We recently produced monoclonal antibodies using macrophagic cells derived from cultured rat glomeruli as the antigen. One of the antibodies, named OS-3, was found to detect a cell population scattered in collecting ducts of the rat kidney as well as macrophages in various tissues. The present study deals with the cellular and subcellular localization of immunoreactivities with OS-3 in the kidney and other organs of rats. Double immunostaining using OS-3 and an antiserum against either calbindin or epidermal cytokeratin showed that OS-3-immunoreactive cells exist exclusively in both the connecting segment and cortical collecting duct, and differ from calbindin- or cytokeratinpositive epithelial cells. Ultrastructurally, OS-3- immunoreactive cells appeared spherical in shape with few cytoplasmic microprojections on the narrow apical surface. Their relatively dark cytoplasm contained numerous mitochondria and a developed tubulo-vesicular system. The intense immunoreactivity was selectively localized in the basolateral membrane exhibiting shallow but complicated infoldings. Distribution and ultrastructural properties of the OS-3-immunoreactive cells showed that they were type B intercalated cells, which are engaged in the regulation of the acid-base balance mainly by secreting HCO3 -. Another positive staining with OS-3 was found in the macula densa and some epithelial cells of Bowman's capsule, the former monitoring Cl- concentrations in the urine. Immunoblotting of extracts from the rat kidney demonstrated a protein band immunoreactive to OS-3 at a molecular weight of 43 kDa. Aside from the kidney, a specific and intense immunoreactivity with OS-3 was also found in the epithelial cells of the pancreatic excretory duct and in the secretory cells of the salivary, pyloric and duodenal glands, all of which are HCO3 - secreting cells. These immunohistochemical findings imply that OS-3 is useful for the detection of type B intercalated cells and recognizes a functional molecule involved in the production/secretion of HCO3 - or transport of Cl- .

DOI： 10.1679/aohc.61.151

Quantification of apoptotic cells using isolated glomeruli Reviewed

S Omori, T Morioka, M Wakasugi, H Kawachi, M Arakawa, F Shimizu, T Oite

NEPHRON 77 ( 4 ) 474 - 478 1997.12

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

In order to detect apoptotic cells in the kidney, the terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end-labeling (TUNEL) method is used in tissue sections. But the number of apoptotic cells per glomerulus in several experimental models of nephritides is about 1 per single tissue section even at peak levels. In this study, we have reported that the TUNEL method and immunostaining of cell-specific markers to a whole isolated glomerulus in combination with laser scan microscopy are potentially useful methods for the analysis of cell turnover within glomeruli.

Slit diaphragm-reactive nephritogenic MAb 5-1-6 alters expression of ZO-1 in rat podocytes Reviewed

H Kawachi, H Kurihara, PS Topham, D Brown, MA Shia, M Orikasa, F Shimizu, DJ Salant

AMERICAN JOURNAL OF PHYSIOLOGY-RENAL PHYSIOLOGY 273 ( 6 ) F984 - F993 1997.12

Language：English Publishing type：Research paper (scientific journal) Publisher：AMER PHYSIOLOGICAL SOC

Monoclonal antibody (MAb) 5-1-6 identifies a 51-kDa protein (p51) on rat podocyte foot processes and causes severe complement-and leukocyte-independent proteinuria when injected into rats. In the studies reported here, we used various immunohistological techniques to define the precise location of p51 and its relationship to ZO-1, a known component of the podocyte slit diaphragm in adult rat glomeruli. Our results demonstrate that p51 and ZO-1 lie close to each other on opposite sides of the podocyte plasma membrane at the point of insertion of the slit diaphragm: ZO-1 on the cytoplasmic face and p51 on the slit diaphragm and adjoining outer leaflet of the plasma membrane bordering the filtration slits. In addition to their geographic proximity, there appears to be a relationship between p51 and ZO-1.After MAb 5-1-6 injection, there was a progressive decline in stainable ZO-1 in the podocytes of heavily proteinuric rats. In addition, Western blot analysis of glomerular lysates showed that the decline in staining was due to a loss of immunoreactive ZO-1 rather than redistribution or diffusion of the protein. Simultaneously, the distribution of glomerular-bound MAb 5-1-6 became more clumped, apparently because of partial endocytosis into a lysosomal compartment, while the slit diaphragms remained morphologically intact. These findings suggest that MAb 5-1-6 alters the molecular composition of the slit diaphragm and thereby affects the glomerular permeability barrier.

Creation of an in vivo cytosensor using engineered mesangial cells. Reviewed

Kitamura M, Kawachi H

J Clin Invest 100 1394 - 1399 1997.12

Suppressive effects of Sairei-to on monoclonal antibody 1-22-3-induced glomerulonephritis: Analysis of effective components Reviewed

Ping Li, Hiroshi Kawachi, Tetsuo Morioka, Michiaki Orikasa, Takashi Oite, Zhen Sheng Shi, Fujio Shimizu

Pathology International 47 ( 7 ) 430 - 435 1997

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

The effects of traditional Chinese medicine (Sairei-to) on experimental glomerulonephritis induced in rats by monoclonal antibody (mAb) 1-22-3 injection was examined. The level of proteinuria in the Sairei-to-treated group was significantly lower than that in the PBS treated group. This suppressive effect was caused by the major component of Sairei-to, Syo- saiko-to but not by another component, Gorei-san. The suppressive effect of Syo-saiko-to was identified in its components (Bupleuri radix, Pinelliae tuber and Zingiberis rhizoma), but not in the other combined components (Ginseng radix and Zizyphi fructus). Further study revealed that the suppressive effects of the combined components were mainly derived from Bupleuri radix. It was demonstrated that the actual active ingredient is probably Salkosaponin-d. Light microscopy revealed that Sairei-to and its effective components suppressed the proliferation of mesangial cells end mesangial matrix expansion. Semiquantitative morphological studies of glomerular lesions on the eighth day showed that Syo-saiko-to and its combined components (Bupleuri radix, Zingiberis rhizoma and Pinelliae tuber) suppressed mesangial matrix expansion significantly compared with phosphate- buffered saline control groups (matrix score: 28.0±19.1 vs 102.3±14.1
30.9±30.1 vs 102.3±14.1, P&lt
0.005, respectively). It was concluded that Salkosaponin-d, as well as Bupleuri radix, Syo-saiko-to and Sairei-to can suppress proteinuria and morphological changes in the rat glomeruionephritis model induced by mAb 1-22-3.

DOI： 10.1111/j.1440-1827.1997.tb04520.x

Slit diaphragm-reactive nephritogenic MAb 5-1-6 alters expression of ZO-1 in rat podocytes Reviewed

Hiroshi Kawachi, Hidetake Kurihara, Peter S. Topham, Dennis Brown, Michael A. Shia, Michiaki Orikasa, Fujio Shimizu, David J. Salant

American Journal of Physiology 273 ( 6 ) F984 - F993 1997

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

Monoclonal antibody (MAb) 5-1-6 identifies a 51-kDa protein (p51) on rat podocyte foot processes and causes severe complement- and leukocyte-independent proteinuria when injected into rats. In the studies reported here, we used various immunohistological techniques to define the precise location of p51 and its relationship to ZO-1, a known component of the podocyte slit diaphragm in adult rat glomeruli. Our results demonstrate that p51 and ZO-1 lie close to each other on opposite sides of the podocyte plasma membrane at the point of insertion of the slit diaphragm: ZO-1 on the cytoplasmic face and p51 on the slit diaphragm and adjoining outer leaflet of the plasma membrane bordering the filtration slits. In addition to their geographic proximity, there appears to be a relationship between p51 and ZO-1. After MAb 5-1-6 injection, there was a progressive decline in stainable ZO-1 in the podocytes of heavily proteinuric rats. In addition, Western blot analysis of glomerular lysates showed that the decline in staining was due to a loss of immunoreactive ZO-1 rather than redistribution or diffusion of the protein. Simultaneously, the distribution of glomerular-bound MAb 5-1-6 became more clumped, apparently because of partial endocytosis into a lysosomal compartment, while the slit diaphragms remained morphologically intact. These findings suggest that MAb 5-1-6 alters the molecular composition of the slit diaphragm and thereby affects the glomerular permeability barrier. glomerular epithelial cell
foot process
tight junction
proteinuria
51-kilodalton protein Copyright © 1997 the American Physiological Society.

Quantification of apoptotic cells using isolated glomeruli Reviewed

Saori Omori, T. Morioka, M. Wakasugi, H. Kawachi, M. Arakawa, F. Shimizu, T. Oite

Nephron 77 ( 4 ) 474 - 478 1997

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

In order to detect apoptotic cells in the kidney, the terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end-labeling (TUNEL) method is used in tissue sections. But the number of apoptotic cells per glomerulus in several experimental models of nephritides is about 1 per single tissue section even at peak levels. In this study, we have reported that the TUNEL method and immunostaining of cell-specific markers to a whole isolated glomerulus in combination with laser scan microscopy are potentially useful methods for the analysis of cell turnover within glomeruli. © 1997 S. Karger AG, Basel.

DOI： 10.1159/000190327

Candesartan prevents the progression of mesangioproliferative nephritis in rats Reviewed

T. Nakamura, J. E. Obata, M. Onizuka, H. Kimura, S. Ohno, Y. Yoshida, H. Kawachi, F. Shimizu

Kidney International, Supplement 51 ( 63 ) S226 - S228 1997

Language：English Publishing type：Research paper (international conference proceedings)

We previously reported a new animal model of progressive glomerulonephritis induced by a single intravenous injection of the anti- Thy-1 monoclonal antibody MoAb 1-22-3 into uninephrectomized rats (Clin Exp Immunol 102:181-185, 1995). We examined the effects of angiotensin II (Ang II) receptor antagonist (candesartan) on the clinical features and morphological lesions of this new model. By 10 weeks after induction of nephritis, untreated rats had developed hypertension, massive proteinuria, renal dysfunction, and severe glomerular injury, while uninephrectomized control rats had not. There was a significant increase in levels of glomerular protein and cortical mRNA for transforming growth factor-β (TGF- β) and type I and type III collagens in untreated nephritic rats. Ten week treatments with candesartan and hydralazine significantly reduced blood pressure (BP) to an equal extent. Candesartan, but not hydralazine, prevented proteinuria, normalized renal function, and ameliorated glomerular injury. Candesartan also reduced levels of glomerular protein and cortical mRNA for TGF-β and type I and type III collagens, while hydaralazine did not. These findings suggest that candesartan prevents progression to end-stage renal failure by modulating the effects of Ang II at least in part on the production of TGF-β and type I and type III collagens, and not merely on systemic BP.

Detailed analysis of phenotypes of macrophages infiltrating glomeruli in rat anti-thy1 nephritis Reviewed

H. Kobayashi, M. Orikasa, M. Naito, K. Kawasaki, T. Oite, H. Kawachi, A. Yamashita, M. Takeya, H. Kobayashi, F. Shimizu

Nephron 77 ( 3 ) 333 - 339 1997

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

A phenotypic analysis of infiltrating macrophages in rat anti-Thy1 glomerulonephntis induced by monoclonal antibody (mAb) 1-22-3 was carried out using recently reported macrophage-specific mAbs. This was combined with a more detailed quantitative analysis, counting positive cells in isolated glomeruli, to obtain more information on the roles played by macrophages in glomerulonephritis. In normal glomeruli a small number of ED1- or OX-3(anti-Ia)-positive cells but almost no ED2-, TRPM-3- or Mar-3-positive cells were observed. ED1-positive cells increased from 2 h and peaked between days 3 and 7 after mAb injection. TRPM-3-positive cells increased from day 3 and peaked on day 7, later than ED1. The numbers of OX-3-positive cells changed in parallel with those of ED1-positive cells. Mar-3, which stained blood monocytes and ED2, which is an indicator of tissue-fixed resident macrophages, did not react with glomerular infiltrating macrophages. In a double staining study, about 40% of ED1- or OX-3-positive cells costained with TRPM-3 on day 3 and the percentage increased on day 7, but hardly any cells were positive for TRPM-3 alone. This results in two different phenotypes (ED1+, ED2−, OX-3+, Mar-3−, TRPM-3− and ED1+, ED2−, OX-3+, Mar-3−, TRPM-3+) of infiltrating macrophages. We conclude that in rat anti-Thyl glomerulonephntis, monocytes/ macrophages may infiltrate the mesangium, rapidly changing their phenotype (Mar-3+ to Mar-3−) and resulting in a gradual shift to TRPM-3-positive, activated macrophages. © 1997 S. Karger AG, Basel.

DOI： 10.1159/000190297

Roles of TGF-β and latent TGF-β-binding protein in glomerulosclerosis induced by two consecutive injections of monoclonal antibody 1-22-3 in Rats Reviewed

Masaru Nakayama, Seiya Okuda, Kiyoshi Tamaki, Fujio Shimizu, Hiroshi Kawachi, Takashi Ando, Taihei Yanagida, Masatoshi Fujisliima

Nephron 76 ( 1 ) 82 - 89 1997

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

The present study demonstrated the elevated synthesis and gene expressions of transforming growth factor β (TGF-β) or latent TGF-β binding protein (LTBP) in an irreversible glomerulosclerosis rat model induced by two consecutive injections of monoclonal antibody (MoAb) 1-22-3. The rats were intravenously injected with 500 #x03BC
g of MoAb 1-22-3 either once or twice at an interval of 2 weeks. The rats were sacrificed at 24 h, 1 week, 2 weeks or 16 weeks after the last injection. At 24 h, the mesangiolytic changes in the rats with two injections of MoAb 1-22-3 were similar to those in the rats with one injection. The glomerular matrix score in the rats with two injections was significantly higher than that in the rats with one injection at weeks 1 2 or 16. An increased LTBP localization in the glomeruli of the rats at week 1 after either one or two injections was detected in the segmentally expanded mesangial matrix. Moreover, LTBP in the glomeruli of rats at week 1 after two injections appeared to be more strongly stained in the enlarged mesangial matrix than that in the rats after one injection. A TGF-β bioassay using mink lung epithelial cells revealed that the total TGF-β in the glomerular culture conditioned medium in the rats at week 1 after two injections was significantly larger than that in the rats after one injection. A Northern blotting analysis of the glomeruli showed that both the expressions of TGF-β and LTBP mRNA in the rats after two injections were higher than those in the rats after one injection. These findings suggested that the elevated TGF-β or LTBP may thus be related to the irreversible glomerulosclerosis that was induced by two injections of MoAb 1-22-3 into rats. © 1997 S. Karger AG, Basel.

DOI： 10.1159/000190145

Structural continuity of filtration slit (slit diaphragm) to plasma membrane of podocyte Reviewed

Y Fujigaki, T Morioka, K Matsui, H Kawachi, M Orikasa, T Oite, F Shimizu, Batsford, SR, A Vogt

KIDNEY INTERNATIONAL 50 ( 1 ) 54 - 62 1996.7

Language：English Publishing type：Research paper (scientific journal) Publisher：NATURE PUBLISHING GROUP

Murine monocional antibody 5-1-6 was reported to bind to the slit membrane and closely related structures in rat renal glomeruli; it induced heavy, reversible proteinuria and appeared to redistribute onto the plasma membrane of epithelial cells after binding at the original target sites. This phenomenon of antigenic movement has not been analyzed in detail to date. In addition to normal kidneys we also studied localization of the antigen recognized by monoclonal antibody 5-1-6 in protamine sulfate-perfused rat kidneys, in which slit diaphragms are known to be functionally modified. Isolated glomeruli as well as ultrathin kidney cryosections were labeled by the immunogold technique to clarify the relation between this antigen and the slit diaphragm. Sequential localization of injected monoclonal antibody was visualized using a post-embedding immunogold method in rats 2 hours to 12 daps after injection of antibody. Ultrastructural immunoqold labeling demonstrated that under normal conditions antigenic molecules were expressed mainly in the area beneath the slit diaphragms. Occasionally labeling was found at the base of the foot process, facing the glomerular basement membrane. After protamine sulfate treatment antigenic sites were dislocated due to the lifting and disruption of slit diaphragms, indicating that this antigen is associated with slit diaphragms. Injected antibody was localized at the filtration slits at 2 hours, and by 12 hours it had moved onto the apical plasma cell membrane of foot process. In addition, from 3 days onwards patch or cap-like formation on the plasma cell membrane of podocytes was seen. Possible shedding of antibody from podocyte cell surface membrane was occasionally encountered, but internalization of antibody was a minor event. Elution experiments in isolated glomeruli al day 3 indicated that antigen and antibody were both localized on the podocyte cell surface membrane, suggesting redistribution of immune complexes. In conclusion, filtration slits (slit diaphragms) and the apical membrane of foot process of podocytes demonstrate structural continuity as revealed by the movement of the antigen recognized by monoclonal antibody 5-1-6 as antigen-antibody complexes.

PROGRESSIVE RENAL LESIONS INDUCED BY ADMINISTRATION OF MONOCLONAL-ANTIBODY-1-22-3 TO UNILATERALLY NEPHRECTOMIZED RATS Reviewed

QL CHENG, M ORIKASA, T MORIOKA, H KAWACHI, XM CHEN, T OITE, F SHIMIZU

CLINICAL AND EXPERIMENTAL IMMUNOLOGY 102 ( 1 ) 181 - 185 1995.10

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

A new animal model of progressive glomerulosclerosis was developed by administering a single i.v, injection of MoAb 1-22-3 to unilaterally nephrectomized rats. Renal morphological analysis revealed that glomerular lesions characterized by mesangial cell proliferation and mesangial matrix expansion were induced in about 95% of the glomeruli. Approximately 20% of the glomeruli of the unilaterally nephrectomized rats showed sclerosis or segmental sclerosis by week 6 after MoAb injection and crescent formation was observed in some glomeruli (ca 4%). Cellular infiltration was also noted in some parts of the interstitium. Increased expression of transforming growth factor-beta (TGF-beta) was observed in the unilaterally nephrectomized rats treated with MoAb 1-22-3, but we could not demonstrate pathological involvement of platelet-derived growth factor (PDGF), even though early-stage mesangial cell proliferation was observed. The mechanism of mesangial cell proliferation in this model remains to be elucidated. The relatively short period of time needed to induce the sclerotic changes is considered to be a great advantage of this model for clarifying the mechanisms involved in the chronic progression of mesangial proliferative glomerulonephritis.

Strain variation in susceptibility to the development of monoclonal antibody 5-1-6-induced proteinuria in rats Reviewed

D. Gollner, H. Kawachi, T. Oite, M. Oka, M. Nagase, F. Shimizu

Clinical and Experimental Immunology 101 ( 2 ) 341 - 345 1995

Language：English Publishing type：Research paper (scientific journal) Publisher：Blackwell Publishing Ltd

Susceptibility to the development of MoAb 5-1-6-induced proteinuria was investigated in four different rat strains, i.e. Brown-Norway (BN), Lewis (LEW), Sprague-Dawley (SD) and Wistar. An intravenous injection of 5 mg of MoAb 5-1-6 to female 7-week-old rats of a given strain induced massive proteinuria in BN, LEW and Wistar rats. However, SD rats developed almost no proteinuria. A similar tendency was observed in the second experiment, in which the injected dose of MoAb was adjusted according to the body weight of each rat (3 mg/100 g body weight). Immunofluorescence (IF) and immunoelectron microscopy (IEM) revealed no differences between the binding patterns of the MoAbs to normal rat kidneys derived from each strain. Quantitative study using 125I-labelled MoAb showed that there was no significant difference in the amount of antibody bound to the kidney 1 h and 5 days after injection between two rat strains, LEW and SD. Localization of 5-1-6 in vivo and its kinetics were investigated. In IF a linear-like pattern along capillary walls was observed 2 h after injection in both LEW and SD strains. This linear-like pattern was shifted to a granular pattern in proteinuric LEW rats 6 days after injection, whereas it remained linear-like in non-proteinuric SD rats. IEM confirmed this difference in the localization of injected MoAb 6 days after injection to LEW and SD rats also at the ultrastructural level. We conclude that there is a clear-cut strain difference in the development of proteinuria induced by MoAb 5-1-6. SD rats were less susceptible to MoAb-induced glomerular injury than BN, LEW and Wistar rats. Although the exact reason for strain variation in susceptibility to MoAb-induced proteinuria remains to be clarified, the movement of bound MoAb, presumably together with corresponding antigenic molecule along the glomerular epithelial cell surface followed by endocytosis into the epithelial cell, seems to be closely related to the induction of proteinuria.

DOI： 10.1111/j.1365-2249.1995.tb08361.x

Visceral epithelial cells in rat glomerular cell culture Reviewed

E. Yaoita, T. Yamamoto, N. Takashima, K. Kawasaki, H. Kawachi, F. Shimizu, I. Kihara

European Journal of Cell Biology 67 ( 2 ) 136 - 144 1995

Language：English Publishing type：Research paper (scientific journal) Publisher：Elsevier GmbH

The purpose of this study was to identify and characterize visceral epithelial cells (VECs) of renal glomeruli in culture. Such cells have been described variably as regular, polygonal cells showing a high rate of replication and a cobblestone-like appearance at confluency and also as irregular, arborized and often multinucleated cells showing a very limited proliferative capacity. We examined early outgrowths from the glomeruli by immunofluorescence microscopy using antibodies specific for VECs (anti podocalyxin(1A), anti-pp44 and 5-1-6), for endothelial cells (anti-von Willebrand factor (vWF) and RECA-1) and for mesangial cells (anti-Thy-1). 1A and anti-pp44 reacted with several types of irregular, arborized cells, but never with regular, polygonal cells. 5-1-6 did not react with any of the cells. Neither the 1A- nor anti-pp44-positive cells (1A/pp44(+) cells) stained with anti-vWF, RECA-1 or anti-Thy-1. However, all the 1A/pp44(+) cells expressed desmin and vimentin but not cytokeratin. These results show that the 1A/pp44(+) cells are derived from VECs, supporting the idea that most polygonal cells in glomerular cultures are of parietal epithelial origin.

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