A Reverse Genetics Approach To Study Feline Infectious Peritonitis

Tekes, Gergely; Spies, Danica; Bank-Wolf, Barbara; Thiel, Volker; Thiel, Heinz-Jürgen

doi:10.1128/jvi.00023-12

Cited by 27 publications

(50 citation statements)

References 26 publications

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“…In conclusion, the data obtained in this study support the ''internal mutational theory'' rather than the ''circulating virulent/avirulent FCoV'' hypothesis. However, the significance of specific mutation events in the generation of FIPV needs to be further evaluated using reverse genetic systems for both serotype I and II feline coronaviruses followed by animal experiments (Tekes et al, 2008(Tekes et al, , 2010(Tekes et al, , 2012. .…”

Section: Discussionmentioning

confidence: 99%

Mutations of 3c and spike protein genes correlate with the occurrence of feline infectious peritonitis

Bank-Wolf

Stallkamp

Wiese

et al. 2014

Veterinary Microbiology

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The genes encoding accessory proteins 3a, 3b, 3c, 7a and 7b, the S2 domain of the spike (S) protein gene and the membrane (M) protein gene of feline infectious peritonitis virus (FIPV) and feline enteric coronavirus (FECV) samples were amplified, cloned and sequenced. For this faeces and/or ascites samples from 19 cats suffering from feline infectious peritonitis (FIP) as well as from 20 FECV-infected healthy cats were used. Sequence comparisons revealed that 3c genes of animals with FIP were heavily affected by nucleotide deletions and point mutations compared to animals infected with FECV; these alterations resulted either in early termination or destruction of the translation initiation codon. Two ascites-derived samples of cats with FIP which displayed no alterations of ORF3c harboured mutations in the S2 domain of the S protein gene which resulted in amino acid exchanges or deletions. Moreover, changes in 3c were often accompanied by mutations in S2. In contrast, in samples obtained from faeces of healthy cats, the ORF3c was never affected by such mutations. Similarly ORF3c from faecal samples of the cats with FIP was mostly intact and showed only in a few cases the same mutations found in the respective ascites samples. The genes encoding 3a, 3b, 7a and 7b displayed no mutations linked to the feline coronavirus (FCoV) biotype. The M protein gene was found to be conserved between FECV and FIPV samples. Our findings suggest that mutations of 3c and spike protein genes correlate with the occurrence of FIP.

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Section: Discussionmentioning

confidence: 99%

Mutations of 3c and spike protein genes correlate with the occurrence of feline infectious peritonitis

Bank-Wolf

Stallkamp

Wiese

et al. 2014

Veterinary Microbiology

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“…Whereas serotype II FCoVs replicate well in feline tissue culture cells in vitro, serotype I FCoVs grow poorly, if at all, in cell culture, except for a few cell cultureadapted isolates. Accordingly, in the last decade, most studies on FCoVs were based on serotype II viruses, while the more prevalent serotype I FCoVs were largely neglected (de Haan et al, 2005;Dye and Siddell, 2005;Haijema et al, 2003Haijema et al, , 2004Rottier et al, 2005;Tekes et al, 2012).…”

Section: Fcov Serotypes and Cellular Receptor Usagementioning

confidence: 99%

“…Despite the existence of serotype I and II FIPVs, the characteristics of the disease caused by these serotypes appear to be very similar. The incubation time for naturally occurring FIP cases is difficult to assess, but a number of studies using experimentally infected specific pathogen-free (SPF) cats revealed incubation times of 2-14 days for the wet form and several weeks for the dry form (Kiss et al, 2004;Pedersen and Black, 1983;Pedersen et al, 1981aPedersen et al, , 1984Tekes et al, 2012). Following experimental infections with the prototype serotype II FIPV 79-1146 strain or a recombinant form of this virus, respectively, cats were shown to develop fever after a few days and lost weight rapidly.…”

Section: Feline Infectious Peritonitis Virusmentioning

confidence: 99%

Feline Coronaviruses

Tekes

Thiel

2016

Advances in Virus Research

119

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Feline infectious peritonitis (FIP) belongs to the few animal virus diseases in which, in the course of a generally harmless persistent infection, a virus acquires a small number of mutations that fundamentally change its pathogenicity, invariably resulting in a fatal outcome. The causative agent of this deadly disease, feline infectious peritonitis virus (FIPV), arises from feline enteric coronavirus (FECV). The review summarizes our current knowledge of the genome and proteome of feline coronaviruses (FCoVs), focusing on the viral surface (spike) protein S and the five accessory proteins. We also review the current classification of FCoVs into distinct serotypes and biotypes, cellular receptors of FCoVs and their presumed role in viral virulence, and discuss other aspects of FIPV-induced pathogenesis. Our current knowledge of genetic differences between FECVs and FIPVs has been mainly based on comparative sequence analyses that revealed "discriminatory" mutations that are present in FIPVs but not in FECVs. Most of these mutations result in amino acid substitutions in the S protein and these may have a critical role in the switch from FECV to FIPV. In most cases, the precise roles of these mutations in the molecular pathogenesis of FIP have not been tested experimentally in the natural host, mainly due to the lack of suitable experimental tools including genetically engineered virus mutants. We discuss the recent progress in the development of FCoV reverse genetics systems suitable to generate recombinant field viruses containing appropriate mutations for in vivo studies.

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“…Unfortunately, our mAbs are not well suited to detect the secreted glycosylated form of 7b. To further study the synthesis, intracellular localization and secretion of 7b, we will use our reverse genetics system (Tekes et al, 2008(Tekes et al, , 2012 to obtain recombinant viruses containing the desired changes in the 7b coding sequence. Ultimately, these studies together with the three dimensional structure of 7b will give more insight into the biological role of the 7b protein.…”

Section: Discussionmentioning

confidence: 99%

“…Both cell lines were maintained in Dulbecco's modified Eagle's medium (DMEM) supplemented with 10% fetal calf serum (FCS), penicillin (100 IU ml À1 ) and streptomycin (100 IU ml À1 ) in 5% CO 2 at 37 C. The type-II strain FCoV 79-1146 was kindly provided by R. J. de Groot, University of Utrecht, The Netherlands (GenBank NC_002306). The recombinant type-I FCoV strain recFCoV-DStop-7b was generated by reverse genetics as described previously (Tekes et al, 2012). It comprises the type-I FCoV strain Black with a restored accessory gene 7b (GenBank EU186072).…”

Section: Viruses and Cellsmentioning

confidence: 99%

Characterization of monoclonal antibodies against feline coronavirus accessory protein 7b

Lemmermeyer

Lamp

Schneider

et al. 2016

Veterinary Microbiology

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Feline coronaviruses (FCoVs) encode five accessory proteins termed 3a, 3b, 3c, 7a and 7b of unknown function. These proteins are dispensable for viral replication in vitro but are supposed to play a role in virulence. In the current study, we produced and characterized 7b-specific monoclonal antibodies (mAbs). A recombinant form of the 7b protein was expressed as a fusion protein in Escherichia coli, purified by immobilized metal affinity chromatography and used as immunogen. Two hybridoma lines, 5B6 and 14D8, were isolated that expressed mAbs that recognized 7b proteins of both FCoV serotypes. Using an extensive set of N- and C-terminally truncated 7b proteins expressed in E. coli and a synthetic peptide, the binding sites of mAbs 5B6 and 14D8 were mapped to an 18-residue region that comprises the only potential N-glycosylation site of the FCoV 7b protein. The two mAbs were suitable to detect a 24-kDa protein, which represents the nonglycosylated form of 7b in FCoV-infected cells. We speculate that glycosylation of 7b is part of the viral evasion strategy to prevent an immune response against this antigenic site.

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A Reverse Genetics Approach To Study Feline Infectious Peritonitis

Cited by 27 publications

References 26 publications

Mutations of 3c and spike protein genes correlate with the occurrence of feline infectious peritonitis

Mutations of 3c and spike protein genes correlate with the occurrence of feline infectious peritonitis

Feline Coronaviruses

Characterization of monoclonal antibodies against feline coronavirus accessory protein 7b

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