Antigenic Structure of Transmissible Gastroenteritis Virus. I. Properties of Monoclonal Antibodies Directed against Virion Proteins

Laude, Hubert; Chapsal, Jean-Michel; Gelfi, Jacqueline; Labiau, Suzanne; Grosclaude, Jeanne

doi:10.1099/0022-1317-67-1-119

Cited by 106 publications

(116 citation statements)

References 25 publications

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“…We have recently reported the derivation of MAbs directed against the three major structural polypeptides of TGEV and confirmed that all essential neutralization-mediating determinants were carried by the 220K glycoprotein E2 (Laude et al, 1986). As was observed in the case of MHV4 (Wege et al, 1984), all the 17 MAbs which recognized E2 also immunoprecipitated its intracellular precursor (E'2, 175K), whereas six other MAbs selectively recognized the 175K species.…”

confidence: 89%

“…The preparation of MAbs, as well as their characterization with respect to polypeptide specificity (E2 and/or E'2) and neutralizing ability have recently been published (Laude et al, 1986). Immunoglobufins (Igs) from ascites fluids were purified by gel permeation as already described (Laude et al, 1986), and stored at -20°C.…”

Section: Methodsmentioning

confidence: 99%

“…The Purdue strain of TGEV was propagated in the pig kidney cell line PD5. A stock of 10 mg of purified virion was prepared as described elsewhere (Laude et al, 1986), divided into aliquots, stored at -70 °C and used throughout the experiments.…”

Section: Methodsmentioning

confidence: 99%

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Antigenic Structure of Transmissible Gastroenteritis Virus. II. Domains in the Peplomer Glycoprotein

Delmas¹,

Gelfi²,

Laude³

1986

Journal of General Virology

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105

118

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SUMMARYThe antigenic structure of the peplomer glycoprotein E2 of the porcine transmissible gastroenteritis coronavirus (TGEV) was explored using a panel of 23 hybridoma antibodies (MAbs). The topography of the epitopes was established by means of a competition radioimmunoassay. Four main antigenic sites, termed A, B, C and D, were thus clearly delineated. Most of the neutralization-mediating determinants were found to cluster in the A-B area, which has been shown to be highly conserved among TGEV strains. Cooperative enhancement of binding to sites B and D was observed following attachment of MAbs relevant to site A. Additional epitopes were identified on E2 by MAbs that selectively recognized its intracellular precursor.

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confidence: 89%

Section: Methodsmentioning

confidence: 99%

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Antigenic Structure of Transmissible Gastroenteritis Virus. II. Domains in the Peplomer Glycoprotein

Delmas¹,

Gelfi²,

Laude³

1986

Journal of General Virology

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105

118

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“…An independent group of recent isolates from TGE field outbreaks (S387, T232, T507, T517, T988, U328 and CC717) was different from the historical Miller and Purdue types. Although only one serotype of TGEV was described [22], it has been suggested that antigenic variation of TGEV may occur in nature [17,25,44,52]. Delmas et al [6] reported that four antigenic sites (A, B, C and D) of TGEV are located on the N-terminal half of the S glycoprotein.…”

Section: Discussionmentioning

confidence: 99%

“…Site C was composed of largely conformationindependent epitopes and showed slight variation among TGEV strains [6]. Epitopes involved in the antigenic variation of TGEV strains have been found outside the major neutralization domain and induced non-neutralizing antibodies [25,41]. However, these MAbs bound to wide range of TGEV strains.…”

Section: Discussionmentioning

confidence: 99%

Field Isolates of Transmissible Gastroenteritis Virus Differ at the Molecular Level from the Miller and Purdue Virulent and Attenuated Strains and from Porcine Respiratory Coronaviruses.

1998

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ABSTRACT. The diversity in selected regions of the transmissible gastroenteritis virus (TGEV) and porcine respiratory coronavirus (PRCV) genomes was analyzed among known TGEV and PRCV strains and field isolates. The N-terminal half of the spike (S) glycoprotein gene and open reading frames (ORF) 3, 3-1 and 4 were amplified by reverse transcriptase reaction and polymerase chain reaction (RT/PCR), and analyzed using restriction fragment length polymorphism (RFLP) patterns of the amplified DNA. Reference TGEV strains (Miller and Purdue) and a PRCV strain (ISU-1), and TGEV and PRCV field isolates were analyzed. Based on the size of the ORF 3, 3-1 and 4 RT/PCR products, TGEV and PRCV strains could be quickly and easily differentiated into three groups designated TGEV Miller, Purdue types and PRCV. By RFLP analysis of the N-terminal region of the S glycoprotein gene and ORFs 3, 3-1 and 4, TGEV and PRCV strains were differentiated into five groups using the restriction enzyme Sau3AI. Sequence analysis of a PCR product in the ORFs 3, 3-1 and 4 from virulent and attenuated Miller strains demonstrated additional differences in that region which have been correlated with a change in virulence of TGEV isolates. -KEY WORDS: coronavirus, PRCV, RT/PCR-RFLP analysis, TGEV.

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Interferon‐gamma gene and protein are spontaneously expressed by the porcine trophectoderm early in gestation

Lefèvre

Martinat-Botté

Guillomot³

et al. 1990

Eur J Immunol

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The nature and the source of the antiviral activity found in the reproductive tract of pregnant gilts early in gestation were analyzed. Two antigenically distinct antiviral activities were found in uterine flushings and in supernatants of conceptus-conditioned culture medium between days 12 and 20 of gestation, using Madin Darby bovine kidney cells and vesicular stomatitis virus as a challenge in the antiviral bioassay. One component was antigenically identified as interferon-gamma (IFN-gamma). Northern blot analysis of conceptus poly(A)+ RNA with a human IFN-gamma cDNA probe revealed two mRNA of 1.3 and 1.4 kb. In addition, immunoprecipitation of metabolically labeled conceptus secretory proteins with an antiserum raised against purified porcine rIFN-gamma resulted in four bands in sodium dodecyl sulfate-polyacrylamide gel electrophoresis, with molecular mass 18.5 to 24.5 kDa. Pre-electrophoresis incubation of the immunoprecipitate with glycopeptidase F, which removes N-linked carbohydrates, yielded a single band of 16.5 kDa. Finally, staining of ultrathin sections by indirect immunofluorescence using the same antiserum to rIFN-gamma revealed that all cells of extra-embryonic trophectoderm contained intensely fluorescent granules in their apical cytoplasm. Neither endoderm nor embryonic cells stained positive. These results clearly show that IFN-gamma, known so far as a T or NK cell-derived lymphokine, is spontaneously and intensively secreted by the porcine trophectoderm, an embryonic tissue not related to the hematopoietic lineage. They also suggest that the implanting conceptus, at least in the porcine species, could play an active role in immune interactions with the mother.

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Antigenic Structure of Transmissible Gastroenteritis Virus. I. Properties of Monoclonal Antibodies Directed against Virion Proteins

Cited by 106 publications

References 25 publications

Antigenic Structure of Transmissible Gastroenteritis Virus. II. Domains in the Peplomer Glycoprotein

Antigenic Structure of Transmissible Gastroenteritis Virus. II. Domains in the Peplomer Glycoprotein

Field Isolates of Transmissible Gastroenteritis Virus Differ at the Molecular Level from the Miller and Purdue Virulent and Attenuated Strains and from Porcine Respiratory Coronaviruses.

Interferon‐gamma gene and protein are spontaneously expressed by the porcine trophectoderm early in gestation

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