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genomic organization and expression of the 3' end of the canine and feline enteric coronaviruses.the genomic organization at the 3' end of canine coronavirus (ccv) and feline enteric coronavirus (fecv) was determined by sequence analysis and compared to that of feline infectious peritonitis virus (fipv) and transmissible gastroenteritis virus (tgev) of swine. comparison of the latter two has previously revealed an extra open reading frame (orf) at the 3' end of the fipv genome, lacking in tgev, which is currently designated orf 6b. both ccv and fecv possess 6b-related orfs at the 3' ends of ...19921329312
antigenic variation among transmissible gastroenteritis virus (tgev) and porcine respiratory coronavirus strains detected with monoclonal antibodies to the s protein of tgev.five nonneutralizing monoclonal antibodies (mab) generated to the virulent miller strain of transmissible gastroenteritis virus (tgev) and specific for the s protein were characterized. competition assays between purified and biotinylated mab indicated that mab 75b10 and 8g11 mapped near a new subsite, designated v and 2 mab, 44c11 and 45a8, mapped to a previously designated subsite d. a fifth mab mapped between subsites v and e. these mab were tested with 3 previously characterized mab to subsi ...19921379786
differentiation between transmissible gastroenteritis virus and porcine respiratory coronavirus using a cdna probe.a plasmid, pg3bs, containing a cdna clone from the 5' coding region of the peplomer glycoprotein gene appears to be specific for enteric transmissible gastroenteritis virus (tgev) strains and for live-attenuated tgev vaccines. this cdna probe is used to differentiate porcine respiratory coronavirus (prcv) isolates from tgev field and vaccine strains by a slot blot hybridization assay. probe pg3bs also hybridizes to canine coronavirus (ccv) rna but does not hybridize to antigenically related feli ...19911645595
antigenic analysis of feline coronaviruses with monoclonal antibodies (mabs): preparation of mabs which discriminate between fipv strain 79-1146 and fecv strain 79-1683.we prepared 31 monoclonal antibodies (mabs) against either fipv strain 79-1146 or fecv strain 79-1683, and tested them for reactivity with various coronaviruses by indirect fluorescent antibody assay (ifa). sixteen mabs which reacted with all of the 11 strains of feline coronaviruses, also reacted with canine coronavirus (ccv) and transmissible gastroenteritis virus (tgev). in many of them, the polypeptide specificity was the recognition of transmembrane (e1) protein of the virus. we succeeded i ...19911653482
characterization of monoclonal antibodies against feline infectious peritonitis virus type ii and antigenic relationship between feline, porcine, and canine coronaviruses.seven monoclonal antibodies (mabs) with neutralizing activity against feline infectious peritonitis virus (fipv) strain 79-1149 (type ii) were prepared. when the polypeptide specificity recognized by these monoclonal antibodies (mabs) was investigated by western immunoblotting, all of the mabs reacted with peplomer glycoprotein (s) of the virus. by competitive binding assay these mabs were found to recognize at least 3 different epitopes. the reactivity of these mabs with 6 viruses classified as ...19911706593
differentiation of transmissible gastroenteritis virus from porcine respiratory coronavirus and other antigenically related coronaviruses by using cdna probes specific for the 5' region of the s glycoprotein gene.two cdna clones prepared from the virulent miller strain of transmissible gastroenteritis virus (tgev) were identified, and their nucleotide sequences were determined. the clones were nonoverlapping and located in the 5' region of the s glycoprotein gene. their nucleotide and predicted amino acid sequences were compared with published sequences of the attenuated purdue strain of tgev and feline infectious peritonitis virus (fipv). tgev clone pe21 contained 381 bp of the s glycoprotein gene and h ...19911847152
antigenic structure of transmissible gastroenteritis virus. i. properties of monoclonal antibodies directed against virion proteins.thirty-two hybridoma cell lines producing monoclonal antibodies (mabs) against the three major structural proteins of transmissible gastroenteritis virus (tgev) have been isolated. radioimmunoprecipitation of intracellular viral polypeptides showed that 17 hybridomas recognized both the peplomer protein [e2, 220 x 10(3) mol. wt. (220k)] and a lower mol. wt. species (e'2, 175k), which was characterized as a precursor of e2. six mabs selectively immunoprecipitated the e'2 protein. four hybridomas ...19862418148
comparison of serologic assays for measurement of antibody response to coronavirus in cats.serologic virus neutralization tests, indirect immunofluorescence tests, and elisa, using tissue culture-adapted feline infectious peritonitis virus (fipv) or feline enteric coronavirus (fecv) were compared for their ability to distinguish specific virus exposure in cats. sera of specific-pathogen-free cats inoculated with virulent or modified fipv or fecv were used to compare the sensitivity and specificity of the homologous assays to a heterologous assay that measures antibody reactivity with ...19882851952
porcine epidemic diarrhea virus (cv 777) and feline infectious peritonitis virus (fipv) are antigenically related.using gut sections from pigs infected with porcine epidemic diarrhea virus (strain cv 777) and ascitic fluid from cats which had succumbed to feline infectious peritonitis (fip), a weak cross reaction was found by immunofluorescence. its specificity was confirmed when detergent-treated purified cv 777 showed a prominent reaction with fipv antibodies in elisa; no reaction was obtained with intact virions, which indicated common determinants on an internal component of the particle. antigenic cros ...19883196169
sequence analysis of the 3'-end of the feline coronavirus fipv 79-1146 genome: comparison with the genome of porcine coronavirus tgev reveals large insertions.the genetic information, carried on mrna 6 of feline infectious peritonitis virus (fipv) strain 79-1146, was determined by sequence analysis of cdna clones derived from the 3' end of the fipv genome. two orfs were found, encoding polypeptides of 11k (orf-1) and 22k (orf-2). the fipv sequence was compared to the 3' end sequence of transmissible gastroenteritis virus (tgev). orf-1 has a homologous counterpart (orf-x3) in the tgev genome; both orfs are located at the same position relative to the n ...19883201747
an immunoelectron microscopic and immunofluorescent study on the antigenic relationship between the coronavirus-like agent, cv 777, and several coronaviruses.a possible antigenic relationship between the porcine enteropathogenic coronavirus-like agent (cvla) and 6 known coronaviruses was examined by immunoelectron microscopy (iem) and by immunofluorescence (if). cvla did not show cross reactivity with infectious bronchitis virus, transmissible gastroenteritis virus (tgev), canine coronavirus (ccv) hemagglutinating encephalomyelitis virus (hev), neonatal calf diarrhea coronavirus (ncdcv) or feline infectious peritonitis virus (fipv). antigenic relatio ...19816166280
lesions in the small intestine of newborn pigs inoculated with porcine, feline, and canine coronaviruses.the infectivity and pathogenicity to newborn pigs of antigenically related coronaviruses from pigs (transmissible gastroenteritis virus; tgev), cats (feline infectious peritonitis virus; fipv), and dogs (canine gastroenteritis virus; cgev) were studied by light, scanning electron, and immunofluorescence microscopy. hysterectomy-derived, 12-hour-old pigs were orally given tissue culture or frozen preparations of 6 coronavirus strains (3 porcine, 2 feline, and 1 canine). the pigs were killed at re ...19816168221
antigenic relationships among homologous structural polypeptides of porcine, feline, and canine coronaviruses.transmissible gastroenteritis virus of swine (tgev), feline infectious peritonitis virus (fipv), and canine coronavirus were studied with respect to their serological cross-reactivity in homologous and heterologous virus neutralization, immune precipitation of radiolabeled tgev, electroblotting, and enzyme-linked immunosorbent assay using individual virion polypeptides prepared by polyacrylamide gel electrophoresis. tgev was neutralized by feline anti-fipv serum, and the reaction was potentiated ...19826182101
an enteric coronavirus infection of cats and its relationship to feline infectious peritonitis.an enteric coronavirus that is antigenically closely related to feline infectious peritonitis virus (fipv) is ubiquitous in the cat population. this virus has been designated feline enteric coronavirus to differentiate it from fipv. the virus is shed in the feces by many seropositive cats; in catteries it is a cause of inapparent to mildly severe enteritis in kittens 6 to 12 weeks of age. the virus may produce a more severe enteritis in young specific-pathogen-free kittens. feline enteric corona ...19816267960
macrotiter assay for coronavirus-neutralizing activity in cats using a canine continuous cell line (a-72).a heterologous neutralization assay for feline infectious peritonitis virus serology was developed using a single continuous cell line of canine origin, a-72, which is susceptible to cytopathic infection with both transmissible gastroenteritis virus of pigs and canine coronavirus. of several coronavirus isolates tested, the 1-71 isolate of canine coronavirus demonstrated the most effective neutralization by serum and body fluids of cats with histopathologically confirmed feline infectious perito ...19836319813
genomic relationship of porcine hemagglutinating encephalomyelitis virus to bovine coronavirus and human coronavirus oc43 as studied by the use of bovine coronavirus s gene-specific probes.the genomic relationship of porcine hemagglutinating encephalomyelitis virus (hev) to bovine coronavirus (bcv) and human coronavirus (hcv) strain oc43 was examined by dot blot hybridization assays. two bcv s gene-specific probes were generated by polymerase chain reaction from the avirulent l9-strain of bcv. probes were located in the s1 and the s2 region of the peplomeric (s) glycoprotein gene. the s1 probe (726 bp) hybridized with bcv and hcv-oc43, but not with hev under moderate stringency hy ...19957646353
molecular cloning and sequence determination of the peplomer protein gene of feline infectious peritonitis virus type i.cdna clones spanning the entire region of the peplomer (s) gene of feline infectious peritonitis virus (fipv) type i strain ku-2 were obtained and their complete nucleotide sequences were determined. a long open reading frame (orf) encoding 1464 amino acid residues was found in the gene, which was 12 residues longer than the orf of the fipv type ii strain 79-1146. the sequences of fipv type i and mainly -tpv type ii were compared. the homologies at the n- (amino acid residues 1-693) and c- (resi ...19957733820
further characterization of aminopeptidase-n as a receptor for coronaviruses.we recently reported that porcine aminopeptidase-n (papn) acts as a receptor for transmissible gastroenteritis virus (tgev). in the present work, we addressed the question of whether tgev tropism is determined only by the virus-receptor interaction. to this end, different non-permissive cell lines were transfected with the porcine apn cdna and tested for their susceptibility to tgev infection. the four transfected cell lines shown to express papn at their membrane became sensitive to infection. ...19937911642
nucleotide sequence and expression of the spike (s) gene of canine coronavirus and comparison with the s proteins of feline and porcine coronaviruses.we have cloned, sequenced and expressed the spike (s) gene of canine coronavirus (ccv; strain k378). its deduced amino acid sequence has revealed features in common with other coronavirus s proteins: a stretch of hydrophobic amino acids at the amino terminus (the putative signal sequence), another hydrophobic region at the carboxy terminus (the membrane anchor), heptad repeats preceding the anchor, and a cysteine-rich region located just downstream from it. like other representatives of the same ...19948021609
coronavirus immunogens.coronaviruses (cv) infect a variety of livestock, poultry and companion animals. they belong to at least five antigenic groups. cv cause localized infections of the respiratory and/or intestinal tracts, with the exception of feline infectious peritonitis virus (fipv) and hemagglutinating encephalomyelitis (hev) which cause systemic infections. the enteropathogenic cv infect the villous enterocytes resulting in villous atrophy leading to malabsorptive diarrhea. several cv (bovine cv-bcv, porcine ...19938116187
genomic organization and expression of the 3' end of the canine and feline enteric coronaviruses.the genomic organization at the 3' end of canine coronavirus (ccv) and feline enteric coronavirus (fecv) was determined by sequence analysis and compared to that of feline infectious peritonitis virus (fipv) and transmissible gastroenteritis virus (tgev) of swine. comparison of the latter two has previously revealed an extra open reading frame (orf) at the 3' end of the fipv genome, lacking in tgev, now designated orf 6b. both ccv and fecv possess 6b-related orfs. the ccv orf 6b is colinear with ...19938209715
cloning, sequencing and expression of the s protein gene from two geographically distinct strains of canine coronavirus.the gene encoding the spike (s) protein from two geographically distinct strains (american and british) of canine coronavirus (ccv) was cloned and sequenced. the nucleotide sequence revealed open reading frames of 1443 or 1453 amino acids, respectively. structural features include an n-terminal hydrophobic signal sequence, a hydrophilic cysteine-rich cluster near the c-terminus, two heptad repeats and 29 or 33 potential n-glycosylation sites. pairwise comparisons of s amino acid sequences from t ...19958607285
multiple receptor-dependent steps determine the species specificity of hcv-229e infection.human coronavirus (hcv)-229e causes disease only in humans and grows in human cells and in cells of other species that express recombinant human aminopeptidase n (hapn), the receptor for hcv-229e. we compared the species specificity of hcv-229e infection with the species specificity of virus binding using immunofluorescence, assay of virus yields, fluorescence activated cell sorting and a monoclonal antibody directed against hapn that blocks infection. we found that hcv-229e binds to intestinal ...19958830504
comparison of the amino acid sequence and phylogenetic analysis of the peplomer, integral membrane and nucleocapsid proteins of feline, canine and porcine coronaviruses.complete nucleotide sequences were determined by cdna cloning of peplomer (s), integral membrane (m) and nucleocapsid (n) genes of feline infectious peritonitis virus (fipv) type i strain ku-2, ucd1 and black, and feline enteric coronavirus (fecv) type ii strain 79-1683. only m and n genes were analyzed in strain ku-2 and strain 79-1683 which still had unknown nucleotide sequences. deduced amino acid sequences of s, m and n proteins were compared in a total of 7 strains of coronaviruses, which i ...19968839428
feline aminopeptidase n serves as a receptor for feline, canine, porcine, and human coronaviruses in serogroup i.two members of coronavirus serogroup i, human respiratory coronavirus hcv-229e and porcine transmissible gastroenteritis virus (tgev), use aminopeptidase n (apn) as their cellular receptors. these viruses show marked species specificity in receptor utilization, as hcv-229e can utilize human but not porcine apn, while tgev can utilize porcine but not human apn. to determine whether feline apn could serve as a receptor for two feline coronaviruses in serogroup i, feline infectious peritonitis viru ...19968970993
characterization of determinants involved in the feline infectious peritonitis virus receptor function of feline aminopeptidase n.feline aminopeptidase n (fapn) is a major cell surface receptor for feline infectious peritonitis virus (fipv), transmissible gastroenteritis virus (tgev), human coronavirus 229e (hcv 229e) and canine coronavirus (ccv). by using chimeric molecules assembled from porcine, human and feline apn we have analysed the determinants involved in the coronavirus receptor function of fapn. our results show that amino acids 670-840 of fapn are critically involved in its fipv and tgev receptor function where ...19989634079
differences in virus receptor for type i and type ii feline infectious peritonitis virus.feline infectious peritonitis viruses (fipvs) are classified into type i and type ii serogroups. here, we report that feline aminopeptidase n (apn), a cell-surface metalloprotease on the intestinal, lung and kidney epithelial cells, is a receptor for type ii fipv but not for type i fipv. a monoclonal antibody (mab) r-g-4, which blocks infection of felis catus whole fetus (fcwf-4) cells by type ii fipv, was obtained by immunizing mice with fcwf-4 cells which are highly susceptible to fipv. this m ...19989645192
molecular analysis of the coronavirus-receptor function of aminopeptidase n.aminopeptidase n (apn) is a major cell surface for coronaviruses of the serogroup i. by using chimeric apn proteins assembled from human, porcine and feline apn we have identified determinants which are critically involved in the coronavirus-apn interaction. our results indicate that human coronavirus 229e (hcv 229e) is distinct from the other serogroup i coronaviruses in that determinants located within the n-terminal parts of the human and feline apn proteins mediate the infection of hcv 229e, ...19989782265
a novel internal open reading frame product expressed from a polycistronic mrna of porcine epidemic diarrhoea virus may not contribute to virus attenuation.cell-culture-adapted (ca) porcine epidemic diarrhoea virus (pedv) contains three internal open reading frames (i orf) within the nucleocapsid protein gene and lacks the downstream counterpart of porcine transmissible gastroenteritis virus orf7 or feline infectious peritonitis virus orf6a. to confirm whether such features also exist in wild-type (wt) pedv, the 3' 1800 nucleotides of its genome were sequenced and were found to be identical to those of ca virus. the coding potential of i-1 orf was ...199910466791
the s gene of canine coronavirus, strain ucd-1, is more closely related to the s gene of transmissible gastroenteritis virus than to that of feline infectious peritonitis virus.to gain insight into the genetic relationships among six canine coronavirus (ccv) strains, the variable region of the spike (s) protein gene was sequenced. the ccv strains were: two atcc reference strains, the insavc-1 vaccine strain, the national veterinary services laboratories (ames, ia) challenge strain, and two california field isolates (ucd-1 and ucd-2) from the 1970s. all six strains, downstream of the nucleocapsid (n) protein gene, had sufficient size for an orf 7b, and thus, none were t ...199910475084
feline and canine coronaviruses are released from the basolateral side of polarized epithelial llc-pk1 cells expressing the recombinant feline aminopeptidase-n cdna.in this study feline (fecv and fipv) and canine (ccov) coronavirus entry into and release from polarized porcine epithelial llc-pk1 cells, stably expressing the recombinant feline aminopeptidase-n cdna, were investigated. virus entry appeared to occur preferentially through the apical membrane, similar to the entry of the related porcine coronavirus transmissible gastroenteritis virus (tgev) into these cells. however, whereas tgev is released apically, feline and canine coronaviruses were found ...200111402864
conservation of substrate specificities among coronavirus main proteases.the key enzyme in coronavirus replicase polyprotein processing is the coronavirus main protease, 3cl(pro). the substrate specificities of five coronavirus main proteases, including the prototypic enzymes from the coronavirus groups i, ii and iii, were characterized. recombinant main proteases of human coronavirus (hcov), transmissible gastroenteritis virus (tgev), feline infectious peritonitis virus, avian infectious bronchitis virus and mouse hepatitis virus (mhv) were tested in peptide-based t ...200211842254
mosaic evolution of the severe acute respiratory syndrome coronavirus.severe acute respiratory syndrome (sars) is a deadly form of pneumonia caused by a novel coronavirus, a viral family responsible for mild respiratory tract infections in a wide variety of animals including humans, pigs, cows, mice, cats, and birds. analyses to date have been unable to identify the precise origin of the sars coronavirus. we used bayesian, neighbor-joining, and split decomposition phylogenetic techniques on the sars virus replicase, surface spike, matrix, and nucleocapsid proteins ...200414671089
two-way antigenic cross-reactivity between severe acute respiratory syndrome coronavirus (sars-cov) and group 1 animal covs is mediated through an antigenic site in the n-terminal region of the sars-cov nucleoprotein.in 2002, severe acute respiratory syndrome-associated coronavirus (sars-cov) emerged in humans, causing a global epidemic. by phylogenetic analysis, sars-cov is distinct from known covs and most closely related to group 2 covs. however, no antigenic cross-reactivity between sars-cov and known covs was conclusively and consistently demonstrated except for group 1 animal covs. we analyzed this cross-reactivity by an enzyme-linked immunosorbent assay (elisa) and western blot analysis using specific ...200717913799
coronavirus m proteins accumulate in the golgi complex beyond the site of virion budding.the prevailing hypothesis is that the intracellular site of budding of coronaviruses is determined by the localization of its membrane protein m (previously called e1). we tested this by analyzing the site of budding of four different coronaviruses in relation to the intracellular localization of their m proteins. mouse hepatitis virus (mhv) and infectious bronchitis virus (ibv) grown in sac(-) cells, and feline infectious peritonitis virus (fipv) and transmissible gastroenteritis virus (tgev) g ...19948083990
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