Monoclonal antipeptide antibodies recognize epitopes upon VP4 and VP7 of simian rotavirus SA11 in infected MA104 cells

Abstract

To study morphogenetic events of rotavirus SA11-infected MA104 cells with strictly defined reagents we produced monoclonal antibodies against synthetic peptides from both outer capsid proteins VP4 (aa residues 228-241: QNTRNIVPVSIVSR) and VP7 (aa residues 319-326: SAAFYYRV) of simian rotavirus SA11. Two of the selected monoclonal antibodies proved to be reactive with determinants of SA11-infected MA104 rhesus monkey kidney cells, with purified SA11 as well as with the particular peptides used for immunization. The anti-VP4 antibody had a demonstrable neutralizing titer of 200 (50% focus reduction) whereas the anti-VP7 MuMAb revealed no detectable neutralizing activity. In peptide-inhibition experiments, the corresponding peptide inhibited its MuMAb whereas the noncorresponding peptide had no effect on antibody binding to intracellular viral antigen. Localization of VP7 was preceded by VP4 as shown by immunofluorescence microscopy.

Studies on the single-shelled rotavirus receptor with a synthetic peptide derived from the cytoplasmic domain of NS28

“…In immunofluorescence the antipeptide serum demonstrates Fig. 2A) resembling that previously obtained with VP4-or VPT-specific monoclonal antibodies [4]. Preincubation of the diluted antipeptide serum with 20 gg of peptide per ml resulted in a complete inhibition of immunofluorescence (Fig.…”

Rotavirus Hospital Surveillance in the Era of Immunization~!2009-10-15~!2009-12-30~!2010-03-05~!

“…In line with the finding that the vaccine remains effective is the fact that the now circulating G3P [8] strains in our area remain relatively stable at the gene level (at least for the genes and segments sequenced). Some of the amino acid differences detected fall within regions of VP7 (amino acid 87-101 and 208-221) [16][17][18][19] and VP4 (amino acid 180, 183, 228-241) [20,21] that have been suggested as important for immune protection. Yet the circulating strains do not seem to be evading vaccine-induced immunity.…”

The humoral immune response of children and infants to an RSV infection: its maturation and association with illness

“…The applied microneutralization assay has been described for rotaviruses in detail previously (Hansen et al, 1992). Biefly, 100 /LI of the virus suspension containing approximately 200 focus-forming units were mixed with 100 /LI of the appropriate antibody dilution and incubated for 90 min at 37°C.…”