Fig. 3
Variable effects of HMOs identified from field studies on rotavirus infectivity and VP8* binding. Of the HMOs identified as associated with rotavirus infections in the field studies, only 2’FL and LNFP I but not 6’SL and LNFP II enhance G10P[11] infectivity in vitro (a). LNT was included as a positive control while 3’SL that was not associated with rotavirus infections in the field studies was included as a negative control. All bars represent the mean percentage of infectivity with error bars denoting standard error of the mean from a minimum of three independent experiments. No HMO treatment was considered 100% infectivity. The mean baseline titer of G10P[11] used in these experiments (100% infectivity) was 8.8 × 105 fluorescent focus units (FFU) per ml. p value <0.05 (analysis of variance with Dunnett’s post hoc test) was considered statistically significant (*p < 0.05, **p < 0.001). Nuclear magnetic resonance (NMR) analysis shows differences in binding of HMOs to P[11] VP8* (b, c). The highest binding was seen with LNT (b) while no binding was seen with 6’SL (c). The binding curves of NMR titrations of LNT to P[11] VP8* for nine representative NMR peaks of changes in proton chemical shifts are shown in different colors (b). The binding curves of NMR titrations of LNFP1, 2’FL, and 6’SL to P[11] VP8* for the same NMR peaks (greatest chemical shift change) are shown in red, blue, and black, respectively (c)
