Cellular composition, coronavirus antigen expression and production of specific antibodies in lesions in feline infectious peritonitis A. Kipar*, S. Bellmann, J. Kremendahl, K. Ko hler, M. Reinacher Institut fu r Veterina r-Pathologie, Universita t Leipzig, Margarete Blank Strasse 4, 04103 Leipzig, Germany Abstract Twenty-three cats with spontaneous feline infectious peritonitis (FIP) were examined by light microscopy including immunohistology and histochemistry in order to determine the cellular composition and the expression of viral antigen in lesions in FIP. Furthermore, the presence of plasma-cells producing coronavirus-specific antibodies was evaluated in situ. Macrophages and neutrophils were demonstrated by an antibody against calprotectin (leukocyte protein L1, myeloid/ histiocyte antigen), neutrophils were recognized due to their chloroacetate esterase activity, and B- and T-lymphocytes were identified by antibodies against the CD3 antigen and the CD45R antigen, respectively. Expression of viral antigen was immunohistologically demonstrated by a monoclonal antibody (mAb) against coronavirus while coronavirus-specific antibodies in situ were identified by the application of feline coronavirus prior to the coronavirus antibody. Lesions were classified as diffuse alterations at serosal surfaces, granulomas with areas of necrosis, granulomas without extended necrosis, focal and perivascular lymphoplasmocytic infiltrates, and granulomatous- necrotizing vasculitis. Diffuse alterations on serosal surfaces were represented either by activated mesothelial cells with single coronavirus antigen-bearing macrophages or by layers of precipitated exudate containing single to numerous granulomas with areas of necrosis. In liver and spleen, the exudate was often underlaid by a small band of subcapsular B-cells with an occasional plasma-cell producing coronavirus-specific antibodies. In other locations, a variably broad band of B-cells and plasma-cells, often infiltrating between underlying muscle fibers, separated the exudate from the unaltered tissue. Some of these plasma-cells were positive for coronavirus-specific antibodies. In granulomas with areas of necrosis, the central necrosis was surrounded by macrophages usually expressing considerable amounts of viral antigen. Few B-cells and plasma-cells were found in the periphery. In granulomas without extended necrosis, the number of macrophages were lower. Only few macrophages expressing low amounts of viral antigen were present. B-cells and plasma-cells formed a broad rim. Few plasma-cells stained positive for coronavirus-specific antibodies. In both types of granulomas, few neutrophils were found between macrophages. Few T-cells were seen scattered throughout the lesions. Focal and perivascular lymphoplasmocytic infiltrates were mainly Veterinary Immunology and Immunopathology 65 (1998) 243257 * Corresponding author. Tel. +49 341 9738278; fax: +49 341 9738299; e-mail: kiparrz.uni-leipzig.de 0165-2427/98/$ see front matter # 1998 Elsevier Science B.V. All rights reserved. PII: S0165-2427(98)00158-5 seen in omentum and leptomeninx. B-cells were the predominant cells; some plasma-cells were positive for coronavirus-specific antibodies. Viral antigen was not readily detected in these alterations. Granulomatous-necrotizing vasculitis was occasionally found in kidneys and leptomeninx. It was dominated by macrophages which often stained strongly positive for coronavirus antigen. Different types of alteration were often seen in the same animal and even the same tissue. There was no obvious correlation between the cats age, gross pathological changes, and the histological types of alteration. Single plasma-cells positive for coronavirus-specific antibodies were found around blood vessels distant from inflammatory alterations, within the lung parenchyma, as infiltrating cells in the mucosa of the small intestine, and in spleen and mesenteric lymph node. Results show that alterations in FIP are heterogeneous concerning cellular composition and expression of viral antigen. The dominance of B-cells in part of the lesions together with the presence of plasma-cells positive for coronavirus-specific antibodies indicate that these cells may play a role in the maintenance of inflammatory processes in FIP. # 1998 Elsevier Science B.V. All rights reserved. Keywords: Feline infectious peritonitis; Immunohistology; Coronavirus antigen; Coronavirus-specifi c anti- bodies; B-cells; Plasma-cells 1. Introduction The histopathological findings in spontaneous and experimental feline infectious peritonitis (FIP) have been described in several previous reports on effusive and non- effusive FIP (Wolfe and Griesemer, 1966; Montali and Strandberg, 1972; Ward et al., 1974; Hayashi et al., 1977, 1980). Although usually characteristic, both macroscopic and histological alterations were not always indicative of FIP. Therefore, differential diagnoses can sometimes not be readily excluded by routi