SEARCH · PubMed Health

Results for “Transmissible gastroenteritis”

Explore indexed PubMed citations for clinical trials, systematic reviews and public health research. Read source abstracts and follow each citation to its original PubMed record.

Quote a phrase for an exact phrase match. Source license links do not imply unrestricted reuse.

At least 415 records · Page 23Linked to original sources

PubMedDate not supplied

[Isolation of a concentrated purified preparation of the transmissible gastroenteritis virus of swine and a study of its antigenic properties].

Pretreatment of the transmissible gastroenteritis virus of swine (TGE) (coronavirus) with trypsin increased the yield of the infectious virus approximately 100-fold when propagated in roller cultures of SPEV cells, the time required for the development of marked CPE being twice shorter than without treatment. The method of concentration and purification of TGE virus is described which yields a preparative amount of highly purified virions retaining their structure. Such preparations were used for immunization of guinea pigs. It is concluded that purified preparations of TGE virus possess marked antigenic potency and may be used for generation of highly active and specific hyperimmune sera.

Animals↗

PubMed1994

Immunogenicity of the S protein of transmissible gastroenteritis virus expressed in baculovirus.

Seven fragments of the spike (S) gene cDNA of transmissible gastroenteritis virus (TGEV), as well as the full length cDNA, were cloned and expressed in baculovirus vectors. Piglets were immunized with cells infected with the recombinant viruses. Each of the recombinants induced TGEV-specific antibodies detected in a fixed cell enzyme immunoassay. The amino terminal half of the S protein, containing all four major antigenic sites (A, B, C and D), and encoded by a 2.2 kb fragment of the S gene, induced virus neutralizing (VN) antibody titers comparable with those induced by the complete S protein. Recombinant proteins lacking the A antigenic site, or with a deletion including the putative receptor binding sites and the D antigenic site, were not capable of inducing levels of VN antibodies similar to those induced by the whole S protein.

Animals↗

PubMed2001

Sialic acid binding activity of transmissible gastroenteritis coronavirus affects sedimentation behavior of virions and solubilized glycoproteins.

The sedimentation behavior of transmissible gastroenteritis coronavirus (TGEV) was analyzed. Upon sucrose gradient centrifugation, the major virus band was found at a density of 1.20 to 1.22 g/cm(3). This high density was observed only when TGEV with a functional sialic acid binding activity was analyzed. Mutants of TGEV that lacked sialic acid binding activity due to a point mutation in the sialic acid binding site of the S protein were mainly recovered at a lower-density position on the sucrose gradient (1.18 to 1.19 g/cm(3)). Neuraminidase treatment of purified virions resulted in a shift of the sedimentation value from the higher to the lower density. These results suggest that binding of sialoglycoproteins to the virion surface is responsible for the sedimentation behavior of TGEV. When purified virions were treated with octylglucoside to solubilize viral glycoproteins, ultracentrifugation resulted in sedimentation of the S protein of TGEV. However, when neuraminidase-treated virions or mutants with a defective sialic acid binding activity were analyzed, the S protein remained in the supernatant rather than in the pellet fraction. These results indicate that the interaction of the surface protein S with sialoglycoconjugates is maintained after solubilization of this viral glycoprotein by detergent treatment.

Animals↗

PubMed1998

Progress towards the construction of a transmissible gastroenteritis coronavirus self-replicating RNA using a two-layer expression system.

Three transmissible gastroenteritis coronavirus (TGEV) defective interfering RNAs of 21, 10.6 and 9.7 kb (DI-A, DI-B and DI-C, respectively) were isolated. Dilution experiments showed that the largest DI RNA, DI-A, is a self-replicating RNA (replicon), and thus codes for a functional RNA polymerase and all the necessary replication signals. In order to engineer a cDNA encoding the RNA replicon a strategy based on the cloning of DI-C cDNA, followed by the insertion of the sequences required to complete the DI-A sequence has been developed. A cDNA complementary to DI-C RNA was cloned under the control of the CMV promoter (pDI-C-CMV) and rescued with a helper virus. In the ORF 1a of polymerase gene pDI-C-CMV contained a 10 kb deletion and in ORF 1b a 1.1 kb deletion. The consensus sequence corresponding to the deleted regions was cloned, and the deletions in pDI-C-CMV were replaced to yield a complete cDNA clone of DI-A, pDI-A-21-CMV, containing a full-length TGEV polymerase, driven by a CMV promoter. Expression of a functional TGEV polymerase is being investigated.

Animals↗

PubMed2002

Generation of a replication-competent, propagation-deficient virus vector based on the transmissible gastroenteritis coronavirus genome.

Replication-competent propagation-deficient virus vectors based on the transmissible gastroenteritis coronavirus (TGEV) genome that are deficient in the essential E gene have been developed by complementation within E(+) packaging cell lines. Cell lines expressing the TGEV E protein were established using the noncytopathic Sindbis virus replicon pSINrep21. In addition, cell lines stably expressing the E gene under the CMV promoter have been developed. The Sindbis replicon vector and the ectopic TGEV E protein did not interfere with the rescue of infectious TGEV from full-length cDNA. Recombinant TGEV deficient in the nonessential 3a and 3b genes and the essential E gene (rTGEV-Delta3abDeltaE) was successfully rescued in these cell lines. rTGEV-Delta3abDeltaE reached high titers (10(7) PFU/ml) in baby hamster kidney cells expressing porcine aminopeptidase N (BHK-pAPN), the cellular receptor for TGEV, using Sindbis replicon and reached titers up to 5 x 10(5) PFU/ml in cells stably expressing E protein under the control of the CMV promoter. The virus titers were proportional to the E protein expression level. The rTGEV-Delta3abDeltaE virions produced in the packaging cell line showed the same morphology and stability under different pHs and temperatures as virus derived from the full-length rTGEV genome, although a delay in virus assembly was observed by electron microscopy and virus titration in the complementation system in relation to the wild-type virus. These viruses were stably grown for >10 passages in the E(+) packaging cell lines. The availability of packaging cell lines will significantly facilitate the production of safe TGEV-derived vectors for vaccination and possibly gene therapy.

Animals↗

PubMed1981

[Morphology of coronaviruses-- electron microscopic demonstration by negative contrast technic of the transmissible gastroenteritis virus of swine].

This contribution to the morphology of corona virus is based on the use of electron microscopy in a negative contrast investigation of the virus of transmissible gastroenteritis of swine. Viruses of transmissible gastro-enteritis are spherical or, in rare cases, pleomorphous particles of typical corona virus structure. The viruses investigated by the authors were 132 nm in diameter. The surface projections, consisting of a flattened knob and conical shaft, are 18 nm in length. Internal bodies, 50 +/- 2 nm in diameter, occur as "filled" or "empty" particles. The thickness of all internal bodies and virus membranes is 8 +/- 1 nm. Differences were found to exist between virus of transmissible gastro-enteritis and particles similar to corona virus and isolated from faecal samples. A first comparison had been made for that purpose. Those differences were relating to shape and size of individual particles and to morphological setup of surface projections.

Animals↗

PubMed2001

In situ hybridization for the detection of transmissible gastroenteritis virus in pigs and comparison with other methods.

Archived formalin-fixed, paraffin-embedded tissues from 25 pigs naturally infected with transmissible gastroenteritis virus (TGEV) were examined by in situ hybridization for TGEV nucleic acid using a nonradioactive digoxigenin-labeled cDNA probe that targeted the nucleocapsid sequence of TGEV strains. The results of in situ hybridization for the detection of TGEV were compared with virus isolation (VI), a fluorescent antibody test (FAT), and transmission electron microscopy (TEM). VI, FAT, and TEM were tested over a course of time before the in situ hybridization was performed. Positive hybridization signals were detected in duodenal, jejunal, and ileal enterocytes from 21 pigs. Hybridization signals were confined to the cytoplasm. Intestinal specimens from 25 piglets were evaluated by 4 tests. Twenty-one of 25 were positive by in situ hybridization. Of these 21 samples, 5 (24%) were positive for TGEV by all 4 tests, 15 (71%) were positive by FAT, 14 (67%) were positive by VI, and 6 (29%) were positive by TEM. In situ hybridization for the detection of TGEV in formalin-fixed, paraffin-embedded tissues provides a rapid means of confirmation of a histopathological diagnosis of TGEV without virus isolation, or when only formalin-fixed intestinal specimens were available.

Animals↗

PubMed1978

[Action of actidion on the replication of the transmissible gastroenteritis virus].

Explored was the possibility of inhibiting the replication of the virus of the transmissive gastroenteritis by means of actidione. It was found that in certain concentrations the antibiotic completely inhibited the replication of this virus. In individual strains the process of replication was not influenced by the various concentrations applied. It was demonstrated that when the antibiotic was added to the infected cell cultures during individual phases its varying concentrations influenced the virus replication. When actidione was added to the cells at the time of virus adsorption it did not inhibit the synthesis of the virus. Similar were the results when the cells were cultivated 24 hours with specified concentrations of actidione prior to their infection with the virus.

Adsorption↗

PubMed2002

Stabilization of a full-length infectious cDNA clone of transmissible gastroenteritis coronavirus by insertion of an intron.

The stable propagation of a full-length transmissible gastroenteritis coronavirus (TGEV) cDNA in Escherichia coli cells as a bacterial artificial chromosome has been considerably improved by the insertion of an intron to disrupt a toxic region identified in the viral genome. The viral RNA was expressed in the cell nucleus under the control of the cytomegalovirus promoter and the intron was efficiently removed during translocation of this RNA to the cytoplasm. The insertion in two different positions allowed stable plasmid amplification for at least 200 generations. Infectious TGEV was efficiently recovered from cells transfected with the modified cDNAs.

Animals↗

PubMed1986

Characterisation of the effector cells in antibody-dependent and spontaneous cell-mediated cytotoxicity in swine against target cells infected with transmissible gastroenteritis virus.

When porcine peripheral blood leucocytes were fractionated, lymphocytes were the most active effectors in both antibody-dependent cell-mediated cytotoxicity (ADCC) and spontaneous cell-mediated cytotoxicity (SCMC), although both polymorphs and macrophages showed some activity in ADCC. Adsorption of lymphocytes to antibody-sensitised or unsensitized PK-15-transmissible gastroenteritis (TGE) cells caused similar reductions in ADCC and SCMC effector activities. Over 60 per cent of the target-binding lymphocytes were non-specific esterase positive large lymphocytes, which did not form erythrocyte (E)-rosettes, and about 30 per cent were non-specific esterase positive medium sized lymphocytes, which formed low avidity E-rosettes. The remainder were non-specific esterase negative small lymphocytes, some of which formed high avidity E-rosettes. None of the eluted lymphocytes stained for surface immunoglobulin and all formed low avidity erythrocyte-antibody rosettes. Porcine killer and natural killer cells resembled in many respects those described in humans and rodents.

Animals↗

PubMed1987

Transmissible gastroenteritis (TGE) of swine: in vitro virus attachment and effects of polyanions and polycations.

Four transmissible gastroenteritis virus (TGEV) strains (Purdue-115, D-52, 188-SG and Gep-II) and two cell lines (swine testis-ST and pig kidney-RPD) were used to study virus attachment and cell susceptibility. Virus attachment was partially thermodependent and the rate varied, depending on the strain. Identical TGEV inocula produced a higher plaque number by plaque assay in the swine testis cell line (ST) than in the pig kidney cell line (RPD) but [3H]uridine-labelled virus was found associated equally well with both cell lines. A field TGEV strain (Gep-II), which was unable to multiply in cell cultures, appeared able to inhibit the attachment of radiolabelled cell-passaged virus. Therefore, the susceptibility to TGEV infection was apparently not determined at the virus-to-cell attachment stage. The attachment sites on the cell surface were specific, however, differences in TGEV attachment determinant between strains were not observed. Attachment of all the virus strains tested was enhanced by DEAE-dextran and inhibited by dextran sulfate, poly-L-lysine (PLL), poly-L-alpha-ornithine (PLO) and protamine sulfate.

Animals↗

PubMed1999

Replication and packaging of transmissible gastroenteritis coronavirus-derived synthetic minigenomes.

The sequences involved in the replication and packaging of transmissible gastroenteritis virus (TGEV) RNA have been studied. The structure of a TGEV defective interfering RNA of 9.7 kb (DI-C) was described previously (A. Mendez, C. Smerdou, A. Izeta, F. Gebauer, and L. Enjuanes, Virology 217: 495-507, 1996), and a cDNA with the information to encode DI-C RNA was cloned under the control of the T7 promoter. The molecularly cloned DI-C RNA was replicated in trans upon transfection of helper virus-infected cells and inhibited 20-fold the replication of the parental genome. A collection of 14 DI-C RNA deletion mutants (TGEV minigenomes) was synthetically generated and tested for their ability to be replicated and packaged. The smallest minigenome (M33) that was replicated by the helper virus and efficiently packaged was 3.3 kb. A minigenome of 2.1 kb (M21) was also replicated, but it was packaged with much lower efficiency than the M33 minigenome, suggesting that it had lost either the sequences containing the main packaging signal or the required secondary structure in the packaging signal due to alteration of the flanking sequences. The low packaging efficiency of the M21 minigenome was not due to minimum size restrictions. The sequences essential for minigenome replication by the helper virus were reduced to 1,348 nt and 492 nt at the 5' and 3' ends, respectively. The TGEV-derived RNA minigenomes were successfully expressed following a two-step amplification system that couples pol II-driven transcription in the nucleus to replication supported by helper virus in the cytoplasm, without any obvious splicing. This system and the use of the reporter gene beta-glucuronidase (GUS) allowed minigenome detection at passage zero, making it possible to distinguish replication efficiency from packaging capability. The synthetic minigenomes have been used to design a helper-dependent expression system that produces around 1.0 microgram/10(6) cells of GUS.

Animals↗

PubMed1994

Simulation of the economic impact of transmissible gastroenteritis on commercial pig production in Australia.

A simulation model (AUSPIG) was used to predict the effect of an increase in piglet deaths, and a reduction in growth rate and an increase in feed conversion ratio of grower pigs on the profitability of two herds representative of the Australian pig industry caused by the introduction of transmissible gastroenteritis virus (TGE) into those herds. For each herd, mortality rates for piglets under 1 week of age of 50% and 95% were assumed to represent a 'moderate' and a 'severe' outbreak, respectively. A reduction in net revenue of 70% was predicted to occur in the 6 months after a 'moderate' outbreak of TGE (100% for a 'severe' outbreak). This represents a total loss of between $260 and $330 per breeding sow in the 12 months after infection with the TGE virus. The likely financial impact of an outbreak of TGE on an Australian piggery is substantial and should be considered when addressing quarantine issues.

Animals↗

PubMed1990

Antigenic homology among coronaviruses related to transmissible gastroenteritis virus.

The antigenic homology of 26 coronavirus isolates, of which 22 were antigenically related to transmissible gastroenteritis virus (TGEV), was determined with 42 monoclonal antibodies. Type, group, and interspecies specific epitopes were defined. Two group specific MAbs distinguished the enteric TGEV isolates from the respiratory variants. An antigenic subsite involved in neutralization was conserved in porcine, feline, and canine coronavirus. The classification of the human coronavirus 229E in a taxonomic cluster distinct from TGEV group is suggested.

Animals↗

PubMed1990

Sequence analysis of the leader RNA of two porcine coronaviruses: transmissible gastroenteritis virus and porcine respiratory coronavirus.

The leader RNA sequence was determined for two pig coronaviruses, transmissible gastroenteritis virus (TGEV), and porcine respiratory coronavirus (PRCV). Primer extension, of a synthetic oligonucleotide complementary to the 5' end of the nucleoprotein gene of TGEV was used to produce a single-stranded DNA copy of the leader RNA from the nucleoprotein mRNA species from TGEV and PRCV, the sequences of which were determined by Maxam and Gilbert cleavage. Northern blot analysis, using a synthetic oligonucleotide complementary to the leader RNA, showed that the leader RNA sequence was present on all of the subgenomic mRNA species. The porcine coronavirus leader RNA sequences were compared to each other and to published coronavirus leader RNA sequences. Sequence homologies and secondary structure similarities were identified that may play a role in the biological function of these RNA sequences.

Amino Acid Sequence↗

PubMed1988

Sequence of the nucleoprotein gene from a virulent British field isolate of transmissible gastroenteritis virus and its expression in Saccharomyces cerevisiae.

Subgenomic mRNA from a virulent isolate of porcine transmissible gastroenteritis virus (TGEV) was used to produce cDNA which was sequenced. Two non-overlapping open reading frames (ORFs) were identified. The largest, encoding a polypeptide of 382 amino acids (relative molecular mass (Mr) 43,483), was shown to be the viral nucleoprotein gene. The second ORF, found 3' to the larger ORF, encodes a polypeptide of 78 amino acids (Mr 9068) which has yet to be assigned to a viral product. The nucleoprotein gene was expressed in yeast cells under the control of two types of yeast promoters: the constitutive PGK promoter, and the inducible GAL1 promoter. Yeast cells containing recombinant plasmids, with the nucleoprotein gene in the correct orientation, produced a polypeptide of Mr 47,000, identical to the viral product, that reacted with a specific monoclonal antibody.

Amino Acid Sequence↗

PubMed1985

Enzymatic and acidic sensitivity profiles of selected virulent and attenuated transmissible gastroenteritis viruses of swine.

This study identifies the in vitro differences (markers) between virulent and attenuated transmissible gastroenteritis (TGE) viruses. Exposure of virulent Miller strain and attenuated Purdue strain TGE viruses to a spectrum of acidities indicated that the Miller strain was more stable at pH 2. Acidities at or above pH 3 did not reduce viral infectivity of either strain. When virulent and attenuated viruses were exposed to gastric fluids of either fed or fasted swine, there was a similar degree of sensitivity. Carboxypeptidase B, alpha-amylase, and alkaline phosphatase present in porcine small intestinal fluids did not cause a significant difference in sensitivity between virulent and attenuated virus isolates. The digestive enzymes: trypsin, alpha-chymotrypsin, pancreatin, peptidase, and carboxypeptidase A did not (or only slightly) inactivate virulent Miller strain TGE virus, but greatly reduced infectivity of attenuated viruses (Purdue strain and TGE vaccine virus isolates). The attenuated strains were significantly more sensitive to small intestinal fluids from both fasted and fed adult swine. Differential sensitivities between virulent and attenuated TGE viruses to digestive fluids from stomach and small intestine further substantiate the notion of differential susceptibility to small intestinal proteases as a correlate of viral virulence.

Alkaline Phosphatase↗

PubMed1989

Induction of transmissible gastroenteritis coronavirus-neutralizing antibodies in vitro by virus-specific T helper cell hybridomas.

Three transmissible gastroenteritis (TGE) virus-specific T helper (Th) cell hybridomas have been generated from virus-primed BALB/c mice, by fusion with the thymoma BW5147. The hybridomas responded to purified u.v.-inactivated TGE virus with interleukin production and growth inhibition. TGE virus recognition by the hybridomas was restricted by the major histocompatibility complex: only splenocytes from syngeneic or semi-syngeneic mice were able to recognize the antigen. The three hybridomas were Thy 1.2+, but did not express detectable levels of Lyt 1 or Lyt 2 antigens by fluorescent cell sorting analysis. Only one hybridoma (T.1J.B5) expressed the L3T4 marker. These hybridomas had helper activity, as they were able to reconstitute in vitro the synthesis of TGE virus-specific antibodies by Th cell-depleted spleen cells from immune BALB/c mice. The antibodies that they induced specifically neutralized by 10(3)- to 10(4)-fold the infectivity of TGE virus, ruling out the possibility of inhibition of virus replication by interferon. These hybridomas could be very useful for identifying antigenic domains in TGE virus recognized by Th cells, which cooperate with B cells in the synthesis of neutralizing antibodies.

Animals↗