Evidence of conserved epitopes in variable region of VP8* subunit of VP4 protein of rotaviruses of P[8]-1 and P[8]-3 lineages.

Although antibody responses to the human rotavirus VP4 protein have been reported, few studies have analyzed the specificity of these responses to the VP8* subunit. This study investigated antibody responses generated against the variable region of the VP4 protein (VP8* subunit) in children infected with rotavirus genotype P[8]. Recombinant VP8* subunit (rVP8*) and truncations corresponding aa 1-102
(peptide A) and 84-180 (peptide B) of rotavirus strains P[8]-1 and P[8]-3 lineages were expressed in Escherichia coli and examined for antibody reactivity using ELISA and Western blot assays. Sera from infected children had IgG antibodies that reacted with full-length rVP8*, peptide A and B of both lineages, with stronger reactivity observed against peptide B. In addition, anti-strain Wa (P[8]-1) and anti-rVP8* (P[8]-3) rabbit polyclonal antiserum reacted against peptide B sequences of both lineages. These data indicate that the VP8* variable region of rotavirus belonging to P[8]-1 and P[8]-3 lineages have conserved epitopes recognized by antibodies elicited during natural infections.

Heterogeneity of VP4 neutralization epitopes among serotype P1A human rotavirus strains.

We have used serotype-specific VP4 and VP7 neutralizing monoclonal antibodies (Nt-MAbs), as well as subgroup (SG)-specific MAbs, to characterize by enzyme immunoassay rotavirus strains isolated from diarrheic infants in the city of Monterrey, Mexico, from July 1993 to March 1994. Of a total of 465 children studied, 140 were rotavirus positive, including 3 patients infected with non-group A rotaviruses. The SG and VP7 (G) serotype specificities could be determined for 118 (84%) of the 140 rotavirus-positive stool specimens; 4 rotavirus strains were serotype G1 and SGII; 1 strain was serotype G2 and SGI+II; 112 strains were serotype G3 and SGII; 1 strain was serotype G3 and SGI; and none of the strains was serotype G4. Fifty-eight specimens, representing the 13 different group A rotavirus electropherotypes detected, were chosen for VP4 (P) serotyping. Of these, 48 (83%) strains reacted with the P1A serotype-specific Nt-MAb 1A10. None of the strains reacted with the serotype P2-specific Nt-MAbs tested. Not all viruses that reacted with Nt-MAb 1A10 were recognized by Nt-MAbs 2A3 and 2G1, which also recognize P1A strains, indicating heterogeneity of neutralization epitopes among serotype P1A human rotaviruses. This heterogeneity could be relevant for the specificity of the VP4-mediated neutralizing antibody immune response and indicates the need for antigenic characterization, in addition to genomic typing, of the VP4 proteins of circulating human rotavirus field strains.