Initial interaction of rotavirus strains with N-acetylneuraminic (sialic) acid residues on the cell surface correlates with VP4 genotype, not species of origin

Abstract

We examined 41 human and animal rotavirus strains representative of all known P genotypes for their dependency on cellular N-acetylneuraminic (sialic) acid (SA) residues for infectivity. Our results showed that all rotaviruses studied, whether of animal or human origin, belonging to P genotypes [1], [2], [3], and [7] depended on SA residues on the cell surface for efficient infectivity but that all human and animal rotavirus strains representative of the remaining known P genotypes were SA independent. The SA residue requirement for efficient infectivity did not change for reassortant rotavirus strains with altered VP4-VP7 combinations. The initial interaction of rotavirus strains with SA residues on the cell surface correlated with VP4 genotype specificity, not with species of origin or VP7 G serotype specificity (P = 0.001; r2 = 1.00, Pearson's correlation coefficient). In addition to being a requirement for infectivity, the presence of SA residues on the cell surface is a requirement for efficient growth in cell culture; recognition of the association of specific P genotypes with the binding of rotavirus to SA residues will facilitate our understanding of the molecular basis of the early events of rotavirus-cell interactions in cell culture models and of pathogenicity in vivo.

FIG. 1.

Effects of treatment of MA104 cells with neuraminidase (from A. ureafaciens) on the infectivities of P[1] and P[2] bovine and simian rotavirus strains reported previously (A); P[3] human, simian, canine, and feline rotavirus strains (B); P[7] porcine rotavirus strains (C); P[5] bovine and porcine rotavirus strains (D); P[4], P[6], and P[8] human rotavirus strains (E); P[9], P[10], P[11], and P[14] feline, human, and bovine rotavirus strains (F); P[12], P[13], and P[15] equine, porcine, and ovine rotavirus strains (G); P[16], P[17], P[18], and P[19] murine, avian (chicken), equine, and porcine rotavirus strains (H); P11[14] human-human reassortant and P5B[3] RRV-human rotavirus reassortant strains (I); and P7[5] UK-human rotavirus reassortant candidate vaccine strains (J). Viral infectivity was expressed as a percentage reflective of the number of FFU in neuraminidase-treated cells relative to the number observed in control (TNC buffer-treated) cells. Values shown are the arithmetic means of results from at least three replicate experiments. Error bars represent 1 standard error of the mean. The genotype and serotype are given in parentheses for each strain. Abbreviations: Si, simian; Eq, equine; Po, porcine; Ca, canine; Fe, feline; Bo, bovine; Ov, ovine; Mu, murine; Hu, human.

FIG. 1.

Effects of treatment of MA104 cells with neuraminidase (from A. ureafaciens) on the infectivities of P[1] and P[2] bovine and simian rotavirus strains reported previously (A); P[3] human, simian, canine, and feline rotavirus strains (B); P[7] porcine rotavirus strains (C); P[5] bovine and porcine rotavirus strains (D); P[4], P[6], and P[8] human rotavirus strains (E); P[9], P[10], P[11], and P[14] feline, human, and bovine rotavirus strains (F); P[12], P[13], and P[15] equine, porcine, and ovine rotavirus strains (G); P[16], P[17], P[18], and P[19] murine, avian (chicken), equine, and porcine rotavirus strains (H); P11[14] human-human reassortant and P5B[3] RRV-human rotavirus reassortant strains (I); and P7[5] UK-human rotavirus reassortant candidate vaccine strains (J). Viral infectivity was expressed as a percentage reflective of the number of FFU in neuraminidase-treated cells relative to the number observed in control (TNC buffer-treated) cells. Values shown are the arithmetic means of results from at least three replicate experiments. Error bars represent 1 standard error of the mean. The genotype and serotype are given in parentheses for each strain. Abbreviations: Si, simian; Eq, equine; Po, porcine; Ca, canine; Fe, feline; Bo, bovine; Ov, ovine; Mu, murine; Hu, human.

FIG. 1.

Effects of treatment of MA104 cells with neuraminidase (from A. ureafaciens) on the infectivities of P[1] and P[2] bovine and simian rotavirus strains reported previously (A); P[3] human, simian, canine, and feline rotavirus strains (B); P[7] porcine rotavirus strains (C); P[5] bovine and porcine rotavirus strains (D); P[4], P[6], and P[8] human rotavirus strains (E); P[9], P[10], P[11], and P[14] feline, human, and bovine rotavirus strains (F); P[12], P[13], and P[15] equine, porcine, and ovine rotavirus strains (G); P[16], P[17], P[18], and P[19] murine, avian (chicken), equine, and porcine rotavirus strains (H); P11[14] human-human reassortant and P5B[3] RRV-human rotavirus reassortant strains (I); and P7[5] UK-human rotavirus reassortant candidate vaccine strains (J). Viral infectivity was expressed as a percentage reflective of the number of FFU in neuraminidase-treated cells relative to the number observed in control (TNC buffer-treated) cells. Values shown are the arithmetic means of results from at least three replicate experiments. Error bars represent 1 standard error of the mean. The genotype and serotype are given in parentheses for each strain. Abbreviations: Si, simian; Eq, equine; Po, porcine; Ca, canine; Fe, feline; Bo, bovine; Ov, ovine; Mu, murine; Hu, human.

FIG. 2.

Phylogenetic relationships among the VP8* cleavage products of the VP4 spike proteins of 52 animal and human rotavirus strains representative of the 20 known P genotypes, for which we know if the presence of SA residues is required for efficient adsorption to and infectivity of MA104 cells. The tree shows an analysis based on amino acid sequences deduced from the gene encoding the VP8* cleavage product of VP4 and was constructed by using the PAUP program (Wisconsin Package, version 9.0). A consensus tree of 95% majority rule is presented. The tree is rooted with the VP8* amino acid sequence of the group A avian (turkey) rotavirus strain Ty-1. The vertical distances are arbitrary. VP8* amino acid sequence data were obtained from the following rotavirus strains, with the GenBank accession numbers in parentheses: SA11 4F (X57319), NCDV (VPXRT2), RF (U65924), BRV033 (U62155), SA11 Cl3 (M23188), RRV (M18736), CU-1 (D13401), K9 (D13400), Cat97 (D13402), HCR3a (L19712), DS-1 (P11196), L26 (M58292), UK (P12474), WC3 (AY050271), B-641 (M63267), 678 (D32054), 4S (L10358), Gottfried (M33516), ST3 (L33895), 1076 (P11198), M37 (L20877), OSU (X13190), TFR-41 (L07889), CRW-8 (L07888), H-1 (D16341), YM (P25174), MO (AB008278), YO (AB008279), Wa (L34161), KU (M21014), K8 (D90260), Cat2 (D13403), 69M (M60600), B223 (M92986), I321 (L07657), H-2 (D13397), FI-14 (D13398), FI-23 (D13342), A46 (AY050274), Mc35 (D140032), HAL1166 (L20875), PA169 (L20874), ALA (U62149), C-11 (U62150), Lp14 (L11599), EW (U08429), EB (U08419), EC (U08421), Ty-1 (L41493), L338 (D13399), 4F (L10359), and EHP (U08424). On the figure, the genotype and serotype for each strain are given in parentheses. Abbreviations: Si, simian; La, lapine; Eq, equine; Po, porcine; Ca, canine; Fe, feline; Bo, bovine; Ov, ovine; Mu, murine; Hu, human.