Membranoproliferative Glomerulonephritis in a Young Cat

NOTE Pathology

Tomoko ASANO1), Atsushi TSUKAMOTO2), Koichi OHNO2), Kikumi OGIHARA3), Junichi KAMIIE4) and Kinji SHIROTA1,4)*

1)Research Institute of Biosciences, 3)Laboratory of Environmental Pathology and 4)Laboratory of Veterinary Pathology, Azabu University, 1–17–71 Fuchinobe, Sagamihara, Kanagawa 229–8501 and 2)Department of Veterinary Internal Medicine, The University of Tokyo, 1–1–1 Yayoi, Bunkyo 113–8657, Japan

(Received 15 July 2008/Accepted 2 August 2008)

ABSTRACT. A 9-month-old male Japanese domestic cat showed pleural effusion, ascites, azotemia, hypoproteinemia and severe proteinuria. Histopathology of the percutaneous renal biopsy specimen revealed that all glomeruli showed intense mesangial hypercellularity with an increased mesangial matrix and thickening of the capillary walls, resulting in lobular accentuation of the glomerular tufts. Frequent duplication of the capillary walls was also observed. Immunostaining for α-smooth muscle actin distinctly revealed mesangial interposition. Diffuse global and linear deposition of C3 and IgG was observed mostly along the peripheral capillary loops. Electron microscopy con- firmed frequent circumferential mesangial interposition and subendothelial dense-deposits in the glomerulus. The glomerular lesion was consistent with human membranoproliferative glomerulonephritis type I, and might be a rare case that developed at young age. KEY WORDS: feline, glomerulus, pathology. J. Vet. Med. Sci. 70(12): 1373–1375, 2008

Membranoproliferative glomerulonephritis (MPGN) is a The paraffin-embedded sections were made and stained chronic and progressive renal disease characterized by with hematoxylin and eosin (HE) or periodic acid-Schiff intense glomerular hypercellularity and diffuse thickening (PAS). Immunohistochemical staining by the streptoavidin- of the glomerular basement membrane with the appearance biotin peroxidase method (Histofine kit, Nichirei, Tokyo or of “double contours”. According to the World Health Orga- VECTASTAIN ABC kit, Vector Laboratories, Burlingame, nization classifications of human glomerular diseases, CA, U.S.A.) was carried out on the paraffin sections using MPGN can be divided into three subtypes based on morpho- primary antibodies against α-smooth muscle actin (α-SMA, logic appearance [2]. Spontaneous MPGN has been clone1A4, 1:50, DAKO, Glostrup, Denmark), proliferating reported in cats, dogs, horses, pigs, and lambs [7, 9]. Most cell nuclear antigen (PCNA, PC10, 1:100, DAKO), cat IgG of these cases have corresponded to types I or II, and type III (1:200, Kirkegaard & Perry Laboratories, Gaithersburg, MPGN of domestic animals has been reported only in a cat MD, U.S.A.), or cat complement component 3 (C3) (1:200, [6]. There have been no reports on the distinct cases of Biogenesisi, Poole, UK). Antigen retrieval for PCNA was feline MPGN type I. In this report, we describe a young performed by heating sections in 10 mM citrate buffer pH 6 feline case comparable to MPGN type I with typical lobular for 10 min using microwave oven. For IgG and C3 staining, accentuation of the capillary tufs that might be a rare case the sections were pretreated by 0.1% trypsin at 37°C for 30 that developed at a young age. min. Antigen retrieval was not required for α-SMA stain- A 9-month-old male Japanese domestic cat was reported ing. The immunoreaction was visualized by a diaminoben- to have shown signs of anorexia, appendicular edema, and zidine-hydrogen peroxide solution, and the sections were diarrhea 1 month before examination. At the first examina- counterstained with hematoxylin. Electron microscopy was tion, pleural effusion, ascites, and bilateral renal swelling carried out using a formalin-fixed biopsy specimen. were revealed by X-ray examination. The results of a blood Microscopically, there were ten glomeruli in the section test showed non-regenerative anemia (PCV: 20%), thromb- of biopsy specimen. All glomeruli in the specimen showed ocytopenia (26,000/µl), azotemia (blood urea nitrogen: 79 severe mesangial hypercellularity with an increased mesan- mg/dl, creatinine: 1.8 mg/dl), and hypoproteinemia (total gial matrix and prominent thickening of the capillary walls, protein: 3.0 g/dl), indicative of a renal glomerular disorder. resulting in lobular accentuation of the glomerular tufts Proteinuria (1,583 mg/dl) and a high urine protein:creatinine (Fig. 1). Adhesion between glomerular capillary tufs and ratio (25.7) were observed by urinalysis. The laboratory test the Bowman’s capsules often developed. Also, proteina- for feline leukemia virus (FeLV) was positive and that for ceous substances were frequently observed in the Bow- feline infectious peritonitis virus (FIPV) was negative. man’s spaces. PAS and PAM stains revealed frequent A percutaneous renal biopsy was performed to confirm duplication or spliting of the capillary walls. Unusual cellu- diagnosis, and the specimen was fixed in 10% formalin. lar localization was seen in the peripheral capillary walls of the glomeruli, indicating mesangial interposition between *CORRESPONDANCE TO: SHIROTA, K., Laboratory of Veterinary Pathology, Azabu University, 1–17–71 Fuchinobe, Sagamihara, endothelial cells and glomerular basement membrane. Mild Kanagawa 229–8501, Japan. inflammation and fibrosis were seen in the interstitium. e-mail : [email protected] Some proximal tubular epithelium showed swelling due to 1374 T. ASANO ET AL.

Fig. 1. The glomeruli showed intense mesangial hypercellular- Fig. 2. Immunohistochemical staining for α-SMA. Intense ity with an increased mesangial matrix and thickening of the staining was observed along the capillary walls (arrows) and in capillary walls, which often adhered to the Bowman’s capsule the mesangial areas, suggesting mesangial interposition and (large arrow). The capillary tuft structure had changed to activation or phenotypic change of the mesangial cells. Strep- become simple and lobulated. There is a proteinaceous sub- toavidin-biotin peroxidase method. Bar=50 µm. stance in the Bowman’s spaces (small arrows). Mild inflammation and fibrosis were observed in the interstitium. PAS. Bar=100 µm. hydropic degeneration. Immunohistochemically, α-SMA expression was intense in the capillary walls and the mesangial areas, suggesting mesangial interposition and activation or a phenotypic change in the mesangial cells (Fig. 2). In the mesangial areas, PCNA-positive cells were frequently observed. Dif- fuse global and linear deposition of C3 and IgG was observed along the peripheral capillary loops by immuno- histochemistry (Fig. 3). Less intense deposition of these substances was also seen in the mesangium with granular appearance. Electron microscopy revealed prominent mesangial hypercellularity and circumferential mesangial interposition resulting in narrowing of the glomerular capillary lumens Fig. 3. Immunohistochemical staining for C3. Diffuse global (Fig. 4). Further, irregular thickening of the glomerular and linear deposition of C3 was observed along the peripheral basement with frequent wrinkling, and dense-deposits in the capillary loops. Less intense deposition with granular appearance was also seen in the mesangium. Streptoavidin-biotin per- subendothelial (Fig. 5) and occasionally in mesangial areas oxidase method. Bar=50 µm. were detected in the glomerulus. The cat died one week after the biopsy. Necropsy revealed bilateral renal swelling, subcutaneous edema, pleural effusion, ascites, and pulmo- present case, immunostaining for α-SMA clearly proved nary edema. this characteristic lesion. Therefore, α-SMA immunostain- Spontaneous immune complex-mediated glomerulone- ing is preferable as a routine diagnostic method to evaluate phritis has been well-reported in cats and membranous the expansion of the mesangial cells into the glomerular nephropathy is the most frequent type of feline glomerulo- capillary walls. nephritis [1]. On the other hand, the clinical and pathologi- The clinical presentation of this cat was characteristic of cal features of MPGN in cats have not been well so-called “protein-losing nephropathy”, therefore the cat documented. The clinical, pathological, and immunohis- was readily suspected to have a certain glomerular disease. tochemical findings of the glomerulopathy in the present However, the type of glomerular disease could not be con- case were consistent with those of human MPGN type I. firmed without detail histopathological and immunopatho- Circumferential mesangial interposition in the glomeruli is logical examinations on the biopsy specimen. Even though one of characteristic morphological features of MPGN [2], the relationship between the types of feline glomerular dis- however sometimes it may be difficult to detect this glomer- eases and the efficacy of treatments has not been clarified ular lesion by routine HE stain or PAS reaction. In the yet, a renal biopsy is fairly important to evaluate the inten- MEMBRANOPROLIFERATIVE GLOMERULONEPHRITIS 1375

Nephrotic syndrome and macroscopic or microscopic hematuria is often associated with MPGN in human cases. The most characteristic clinical finding in humans is hypocomplementemia, especially with decreased C3 levels. The primary causes of human MPGN have not been elucidated, but nephritic factors are involved in complement activation and hypocomplementemia in this type of glomerular disease [2]. In animal cases of MPGN, congenital or hereditary abnormalities of the complement system have been also suggested to be involved in MPGN of some cases in which the disease develops in early life [3, 5, 7]. However, we could not evaluate complement system in this case and could not trace the family history. In immune-mediated glomerulonephritis in cats, infection with FeLV [1], FIPV [4] or feline immunodeficiency virus [8] is frequently com- plicated, although typical MPGN as seen in the present case has not been reported in these infectious diseases. Fig. 4. Electron micrograph of the glomerulus of a formalin- fixed specimen, showing distinct circumferential mesangial ACKNOWLEGEMENTS. We gratefully thank Dr. Cathy interposition (*) resulting in narrowing of the capillary lumens Brown, College of Veterinary Medicine, University of (c) and wrinkling of the glomerular basement membrane Georgia, and Dr. Johan Hogset Jansen, Norwegian School (arrows). Bar=2 µm. of Veterinary Science for their helpful comments.

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