In vitro Stimulation of Presensitized Mouse Spleen Cells with Poliovirus Type 1, Mahoney, and Enhancement of Poliovirus-specific Hybridomas

Antigenic Structure of Picornaviruses

“…While the region corresponding to the immunodominant site of type 3 was not strongly immunogenic in type 1 in any of the studies reviewed above, DERNICK and co-workers have isolated a large number of antibodies specific for type 1 poliovirus which select for mutations within this locus (UHLIG et al 1983;WIEGERS and DERNICK 1987;WIEGERS et al 1986WIEGERS et al , 1988. They have also demonstrated the involvement of the loop composed of residues 141 ~ 152 ofVPl in this site (WIEGERS et al 1989).…”

“…They have also demonstrated the involvement of the loop composed of residues 141 ~ 152 ofVPl in this site (WIEGERS et al 1989). In some instances the antibodies were generated by immunisation with purified viral proteins and the splenocytes stimulated in vitro with infectious virus before fusion (WIEGERS et al 1986) but this was not always the case (UHLIG et al 1983). The success of this group in generating antibodies against this site of type 1 poliovirus by conventional means remains unexplained (WIEGERS et al 1988).…”

“…Monoclonal antibodies isolated by another group prepared by normal immunisation schedules (UHUG et al 1983) or in vivo priming followed by in vitro stimulation (WIEGERS et al 1986) were shown to recognise linear epitopes within site 1 of poliovirus type 1 (WIEGERS and DERNICK 1987;WIEGERS et al 1988). The methods included isolation of mutants, binding of antibodies to isolated proteins and competition with a peptide with the sequence of VP 1 from residues 93 to 104.…”

Molecular basis for linkage of a continuous and discontinuous neutralization epitope on the structural polypeptide VP2 of poliovirus type 1

“…At 2 to 6 months after boosting, the animals were sacrificed and the spleens were removed. In vitro cultivation of splenocytes was done in the presence of 5 ,ug of poliovirus per ml as described previously (39). Fusion of spleen cells after conventional immunization of BALB/c mice or after in vitro stimulation of primed spleen cells was done as described previously (18,39).…”

“…In vitro cultivation of splenocytes was done in the presence of 5 ,ug of poliovirus per ml as described previously (39). Fusion of spleen cells after conventional immunization of BALB/c mice or after in vitro stimulation of primed spleen cells was done as described previously (18,39). Hybridoma supernatants were initially screened in a solid-phase enzyme-linked immunosorbent assay with poliovirus type 1 (Mahoney) per ml as the antigen bound either directly to microdilution plates or by polyclonal rabbit anti-poliovirus immunoglobulins.…”