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. 2024 Mar 17;16(3):462.
doi: 10.3390/v16030462.

Serologic, Virologic and Pathologic Features of Cats with Naturally Occurring Feline Infectious Peritonitis Enrolled in Antiviral Clinical Trials

Affiliations

Serologic, Virologic and Pathologic Features of Cats with Naturally Occurring Feline Infectious Peritonitis Enrolled in Antiviral Clinical Trials

Brian G Murphy et al. Viruses. .

Abstract

Feline infectious peritonitis (FIP) is a multisystemic, generally lethal immuno-inflammatory disease of domestic cats caused by an infection with a genetic variant of feline coronavirus, referred to as the FIP virus (FIPV). We leveraged data from four different antiviral clinical trials performed at the University of California, Davis. Collectively, a total of 60 client-owned domestic cats, each with a confirmed diagnosis of naturally occurring FIP, were treated with a variety of antiviral compounds. The tested therapies included the antiviral compounds GS-441524, remdesivir, molnupiravir and allogeneic feline mesenchymal stem/stroma cell transfusions. Four client-owned cats with FIP did not meet the inclusion criteria for the trials and were not treated with antiviral therapies; these cats were included in the data set as untreated FIP control cats. ELISA and Western blot assays were performed using feline serum/plasma or ascites effusions obtained from a subset of the FIP cats. Normalized tissue/effusion viral loads were determined in 34 cats by a quantitative RT-PCR of nucleic acids isolated from either effusions or abdominal lymph node tissue. Twenty-one cats were PCR "serotyped" (genotyped) and had the S1/S2 region of the coronaviral spike gene amplified, cloned and sequenced from effusions or abdominal lymph node tissue. In total, 3 untreated control cats and 14 (23.3%) of the 60 antiviral-treated cats died or were euthanized during (13) or after the completion of (1) antiviral treatment. Of these 17 cats, 13 had complete necropsies performed (10 cats treated with antivirals and 3 untreated control cats). We found that anticoronaviral serologic responses were persistent and robust throughout the treatment period, primarily the IgG isotype, and focused on the viral structural Nucleocapsid and Membrane proteins. Coronavirus serologic patterns were similar for the effusions and serum/plasma of cats with FIP and in cats entering remission or that died. Viral RNA was readily detectable in the majority of the cats in either abdominal lymph node tissue or ascites effusions, and all of the viral isolates were determined to be serotype I FIPV. Viral nucleic acids in cats treated with antiviral compounds became undetectable in ascites or abdominal lymph node tissue by 11 days post-treatment using a sensitive quantitative RT-PCR assay. The most common pathologic lesions identified in the necropsied cats were hepatitis, abdominal effusion (ascites), serositis, pancreatitis, lymphadenitis, icterus and perivasculitis. In cats treated with antiviral compounds, gross and histological lesions characteristic of FIP persisted for several weeks, while the viral antigen became progressively less detectable.

Keywords: FIP; antiviral compound; cat; feline coronavirus; serology.

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Conflict of interest statement

The authors declare no conflicts of interest.

Figures

Figure 1
Figure 1
ELISA results for plasma and effusions derived from cats with FIP. (a) ELISA absorbance values for paired feline plasma from week 0 (dark blue bars) and week 11 (light blue bars). Data are presented as the mean (or median) of technical triplicates and standard deviation (or standard error of the median). The grey bar represents plasma from a cat experimentally infected with FIPV (positive control) and the light grey bar represents plasma from a specific-pathogen-free cat (negative control) [19]. (b) ELISA absorbance values for cats with FIP that died or were euthanized (red bars) and cats that successfully entered remission (blue bars). Controls are as for (a). (c) For cats that successfully entered remission, the anti-FCoV response is depicted for each patient using samples derived from ascites effusions (dark blue bars) or plasma (light blue bars). Controls are as for (a). (d) For cats that successfully entered remission, the isotype-specific anti-FCoV response is indicated for IgM (light blue bars), IgA (med blue) and IgG (dark blue). SPF control plasma serves as the negative control.
Figure 2
Figure 2
Western blot (immunoblot) assays demonstrating anti-FCoV responses from cats with FIP. FCoV N protein (~43 kDa), M protein (~25 kDa) and S2 (~90 kDa) are labeled with the corresponding letter. (a) Western blot assay demonstrating total Ig binding to FCoV proteins for study cats. Cats that successfully entered remission are indicated by the blue bar (lanes 3–6), and cats that died or were euthanized are indicated by the red bar (lanes 7–11). Lane 1—molecular weight markers, lane 2—SPF negative control plasma, lane 3—cat 14, lane 4—cat 15, lane 5—cat 16, lane 6—cat 17, lane 7—cat 9, lane 8—cat 12, lane 9—cat 10, lane 10—cat 7, lane 11—cat 11. (b) Western blot assay demonstrating Ig subtype binding for a subset of study cats. Lane 1—cat 18, lane 2—cat 14, lane 4—cat 19, lane 5—SPF feline plasma (negative control).
Figure 3
Figure 3
Frequency of pathologic lesions identified in the necropsied cats with FIP.
Figure 4
Figure 4
Select gross lesions identified in the necropsied cats with FIP. (a) Open abdomen demonstrating icterus, abdominal effusion, fibrinous serositis and hepatic capsulitis (wet/effusive FIP, 4 days of antiviral (AV) treatment, cat 8). (b) Open abdomen demonstrating granulomatous mesenteric lymphadenitis/lymphadenomegaly (dry/granulomatous FIP, untreated cat 3). (c) Open abdomen and thorax demonstrating abdominal effusion, icterus and serositis (wet/effusive FIP, 3 days of AV treatment, cat 6). (d) Bilateral kidneys demonstrating granulomatous nephritis and capsulitis (dry/granulomatous FIP, 6 days of AV treatment, cat 10). (e) Spleen demonstrating splenic capsulitis (4 days of antiviral treatment, cat 8). (f) Liver demonstrating hepatic capsulitis (1 day of antiviral treatment, cat 4).
Figure 5
Figure 5
Select histological lesions identified in the necropsied cats with FIP. (a) Section of brainstem demonstrating periventriculitis and encephalitis. The inset image demonstrates higher magnification of an IHC for FCoV antigen (+++ viral antigen, untreated cat 3). (b) Mesenteric lymph node with necrotizing lymphadenitis lesion. IHC for FCoV demonstrated abundant viral antigen (IHC not shown, untreated cat 3). (c) Kidney demonstrating necrotizing interstitial nephritis (6 days of antiviral treatment, cat 5). (d) Pancreas demonstrating necrotizing pancreatitis and steatitis (5 days of antiviral treatment, cat 9).
Figure 6
Figure 6
Select histological lesions identified in the necropsied cats with FIP. (a,b) Kidney demonstrating interstitial nephritis lesion, HE-stained section: (a). FCoV IHC of renal section demonstrating abundant (+++) viral antigen; (b) untreated cat 2. (c,d) Liver demonstrating fibrinonecrotizing hepatitis, HE-stained section: (c) FCoV IHC of same hepatic section demonstrating minimal (+) viral antigen; (d) inset—higher magnification of same section demonstrating single positive macrophage (7 days of antiviral treatment, cat 11). (e,f) Seven days of antiviral therapy, cat 11. Serosa and mesentery of gut demonstrating fibrinous phlebitis and perivasculitis lesion, HE-stained section (e). FCoV IHC of same section demonstrating no (0) viral antigen (f).
Figure 7
Figure 7
FCoV nucleic acid quantification in lymph nodes and effusions in response to antiviral therapy. Numbers along the “undetected” dotted line refer to the number of cases with no FCoV RNA detected at that specific time point.
Figure 8
Figure 8
S1/S2 nucleotide sequences for the FCoV spike gene. Sequences derived from study cats that successfully entered remission are listed in the upper block, while those derived from cats that died or were euthanized are listed in the lower block. Individual cat numbers are listed on the left, and 1–3 sequences are depicted per animal as a, b or c. The hypervariable region (yellow) flanks the S1/S2 sequence (green). Primer sites are indicated by blue lines. Asterisks (*) indicate conservation of the nucleotide sequence at that locus.
Figure 9
Figure 9
S1/S2 amino acid sequences for the FCoV spike gene. (a) Sequences are organized as in Figure 8. (b) A schematic of the FCoV S protein demonstrating the S1 and S2 portions and the cleavage sites and locus 23,531. (c) The relative frequency of the amino acids at each locus are depicted in this WebLogo 3.1 analysis (http://weblogo.threeplusone.com/create.cgi (accessed on 1 June 2023)). The S1/S2 site is boxed in red.

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