Characterization and reactivity of monoclonal antibodies to the Miller strain of transmissible gastroenteritis virus of swine

Xiao-Ling Zhu From the Veterinary Medical Research Institute, College of Veterinary Medicine, Iowa State University, Ames, Iowa 50011.

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 BVSc, MS
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Prem S. Paul From the Veterinary Medical Research Institute, College of Veterinary Medicine, Iowa State University, Ames, Iowa 50011.

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 BVSc, PhD
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Eric Vaughn From the Veterinary Medical Research Institute, College of Veterinary Medicine, Iowa State University, Ames, Iowa 50011.

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Angeles Morales From the Veterinary Medical Research Institute, College of Veterinary Medicine, Iowa State University, Ames, Iowa 50011.

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 MS

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SUMMARY

Hybridomas secreting monoclonal antibodies (mab) to transmissible gastroenteritis virus (tgev) were produced by fusion of SP2/0 myeloma cells and splenic lymphocytes of BALB/c mice immunized with the virulent cell-passaged Miller strain of tgev. The mab secreted by these hybridomas were partially characterized; 4 of them (MA4, MA5, MH11, MB2) had high-neutralization titer for tgev. The remaining 7 (MC6, MD9, ME5, MG5, MF2, ME9, MG7) did not neutralize tgev at 1:25 dilution. All 4 neutralizing and 2 of the nonneutralizing mab reacted with the E2 protein of tgev in a radioimmunoprecipitation assay. The remaining 5 mab reacted with the E1 protein of tgev. Reactivity of the mab was tested in an indirect immunofluorescent assay with 3 cell culture-adapted strains of tgev (Miller, Purdue, and Illinois) and 13 wild-type isolates of tgev. Neutralizing mab reacted with all 13 wild-type isolates and the 3 cell culture-adapted strains of tgev. In contrast, nonneutralizing mab that reacted with the Miller strain of TGEV varied in their reactivity with the wild-type tgev isolates. Reactivity of neutralizing mab was also tested, using plaque-reduction neutralization assays with Miller, Purdue, and Illinois strains and 5 wild-type isolates. All 4 neutralizing mab neutralized the 8 virus isolates, but the neutralization titer was higher with the homologous virus than with the heterologous virus isolates. However, neutralization titers of the 4 neutralizing mab were 4 to 16 times higher for the homologous Miller strain of tgev than for the heterologous Illinois and Purdue strains, and were 4 to 1,000 times higher than for the wild-type isolates. Extensive antigenic heterogeneity was observed among tgev isolates on epitopes recognized by the nonneutralizing mab directed against either E1 or E2 protein.

SUMMARY

Hybridomas secreting monoclonal antibodies (mab) to transmissible gastroenteritis virus (tgev) were produced by fusion of SP2/0 myeloma cells and splenic lymphocytes of BALB/c mice immunized with the virulent cell-passaged Miller strain of tgev. The mab secreted by these hybridomas were partially characterized; 4 of them (MA4, MA5, MH11, MB2) had high-neutralization titer for tgev. The remaining 7 (MC6, MD9, ME5, MG5, MF2, ME9, MG7) did not neutralize tgev at 1:25 dilution. All 4 neutralizing and 2 of the nonneutralizing mab reacted with the E2 protein of tgev in a radioimmunoprecipitation assay. The remaining 5 mab reacted with the E1 protein of tgev. Reactivity of the mab was tested in an indirect immunofluorescent assay with 3 cell culture-adapted strains of tgev (Miller, Purdue, and Illinois) and 13 wild-type isolates of tgev. Neutralizing mab reacted with all 13 wild-type isolates and the 3 cell culture-adapted strains of tgev. In contrast, nonneutralizing mab that reacted with the Miller strain of TGEV varied in their reactivity with the wild-type tgev isolates. Reactivity of neutralizing mab was also tested, using plaque-reduction neutralization assays with Miller, Purdue, and Illinois strains and 5 wild-type isolates. All 4 neutralizing mab neutralized the 8 virus isolates, but the neutralization titer was higher with the homologous virus than with the heterologous virus isolates. However, neutralization titers of the 4 neutralizing mab were 4 to 16 times higher for the homologous Miller strain of tgev than for the heterologous Illinois and Purdue strains, and were 4 to 1,000 times higher than for the wild-type isolates. Extensive antigenic heterogeneity was observed among tgev isolates on epitopes recognized by the nonneutralizing mab directed against either E1 or E2 protein.

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