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Generation and characterization of monoclonal antibodies specific for PR8 influenza A virus-infected cells
Abstract
The majority of the MAbs induced by cells six hours after infection reacted with infected cells only, while most of the MAbs induced by cells twenty-four hours after infection recognized both infected cells and virus. Antigens recognized by the 6-hour MAbs are present two to four hours after infection, and plateau around six hours after infection. Antigens recognized by the 24-hour antibodies appear between six and twelve hours after infection and reach a peak around 12 to 24 hours after infection. Significantly, antigens recognized by both sets of antibodies persist for the duration of the experiment, which means that the immune response to viral surface antigens becomes dominant later in infection, despite the persistence of "early immunogens" at the cell surface. Western blot analysis showed that the 6-hour MAbs reacted either with a 56-kD viral nucleoprotein or with a 27-kD matrix protein, and the 24-hour MAbs were either NP-specific or HA-specific. All the 6-hour MAbs bound to PR8-infected 3T3 cells (H-2$\sp{\rm d}$) and PR8-infected 2F6 cells (H-2$\sp{\rm b}$), and two of them also reacted with PR8-infected L929 cells (H-2$\sp{\rm k}$). These results suggest that, either these MAbs do not recognize NP peptide-MHC complex, or, if they do, all of them cross-react with infected or uninfected cells of H-2$\sp{\rm b}$ haplotype and 2 of them also recognize cells with an H-2$\sp{\rm k}$ haplotype. A modified peptide (NP147-158R$\sp-$) corresponding to the NP epitope of the K$\sp{\rm d}$-restricted CTL bound to H-2$\sp{\rm d}$, H-2$\sp{\rm b}$, and H-2$\sp{\rm k}$ cells. Two of the 6-hour MAbs reacted with peptide-pulsed cells of both H-2$\sp{\rm d}$ and H-2$\sp{\rm b}$ haplotypes, while one reacted with cells of all three haplotypes by ELISA. This peptide inhibited binding of two 6-hour MAbs by 63% and 43% respectively to PR8-infected cells. These findings suggest that murine B cells might recognize a processed viral peptide that also is the viral epitope of the K$\sp{\rm d}$ restricted CTL on PR8-infected BALB/c spleen cells (H-2$\sp{\rm d}$).
Subject Area
Microbiology|Immunology
Recommended Citation
Lin, Boh Chang, "Generation and characterization of monoclonal antibodies specific for PR8 influenza A virus-infected cells" (1992). ETD collection for University of Nebraska-Lincoln. AAI9233407.
https://digitalcommons.unl.edu/dissertations/AAI9233407