Figure 9.
Evidence for immunological imprinting after bivalent mRNA vaccine boosting in the LO chip. (A) Experimental setup: LO chips were perfused for 6 days with BSA, a monovalent mRNA vaccine encoding the Wuhan spike (mRNA-1273, Moderna), or a bivalent mRNA vaccine encoding the Wuhan and Omicron BA.1 spikes (mRNA-1273.214, Moderna). (B) Representative analysis of CD27 and CD38 expression in CD19+ B cells (left) and frequency of CD27hiCD38hi PB among B cells (right; n = 7). (C) Representative analysis of CD19+ B cells binding to the RBD of the Wuhan strain (top row) or the BA.1 strain (bottom row) (left) and frequency of Wuhan (top) and BA.1 (bottom) RBD-specific cells among B cells (right; n = 7). (D) Positive correlation between the frequency of Wuhan RBD-specific B cells and the frequency of BA.1 RBD-specific B cells. Data were analyzed by simple linear regression. The correlation coefficient R and the associated P value are reported. (E) S-Flow assay analysis showing the MFI of spike-specific IgG associated bound to spike-expressing 293T cells (n = 7). (F) Positive correlation between spike-specific IgG levels and the frequency of Wuhan (left) or BA.1 (right) RBD-specific B cells; Data were analyzed by simple linear regression. The correlation coefficient R and the associated P value are reported. (G) The neutralization capacity of antibodies produced in the LO chip was assessed by the S-Fuse assay for n = 7 donors. The threshold for the ID50 was set as 14 based on negative pre-pandemic controls (dashed line). ID50 are reported for neutralization of the ancestral-like D614G strain and the Omicron BA.1 strain, after stimulation with BSA, mRNA-1273, and mRNA-1273.214. (H) The dose response of D614G neutralization (top) and BA.1 neutralization (bottom) is reported in function of chip extract dilution for three representative donors to illustrate individual variability. (B, C, E, and G) Each color represents an independent donor. Differences were evaluated with a Wilcoxon matched pairs test; *P < 0.05; ns, not significant.

Evidence for immunological imprinting after bivalent mRNA vaccine boosting in the LO chip. (A) Experimental setup: LO chips were perfused for 6 days with BSA, a monovalent mRNA vaccine encoding the Wuhan spike (mRNA-1273, Moderna), or a bivalent mRNA vaccine encoding the Wuhan and Omicron BA.1 spikes (mRNA-1273.214, Moderna). (B) Representative analysis of CD27 and CD38 expression in CD19+ B cells (left) and frequency of CD27hiCD38hi PB among B cells (right; n = 7). (C) Representative analysis of CD19+ B cells binding to the RBD of the Wuhan strain (top row) or the BA.1 strain (bottom row) (left) and frequency of Wuhan (top) and BA.1 (bottom) RBD-specific cells among B cells (right; n = 7). (D) Positive correlation between the frequency of Wuhan RBD-specific B cells and the frequency of BA.1 RBD-specific B cells. Data were analyzed by simple linear regression. The correlation coefficient R and the associated P value are reported. (E) S-Flow assay analysis showing the MFI of spike-specific IgG associated bound to spike-expressing 293T cells (n = 7). (F) Positive correlation between spike-specific IgG levels and the frequency of Wuhan (left) or BA.1 (right) RBD-specific B cells; Data were analyzed by simple linear regression. The correlation coefficient R and the associated P value are reported. (G) The neutralization capacity of antibodies produced in the LO chip was assessed by the S-Fuse assay for n = 7 donors. The threshold for the ID50 was set as 14 based on negative pre-pandemic controls (dashed line). ID50 are reported for neutralization of the ancestral-like D614G strain and the Omicron BA.1 strain, after stimulation with BSA, mRNA-1273, and mRNA-1273.214. (H) The dose response of D614G neutralization (top) and BA.1 neutralization (bottom) is reported in function of chip extract dilution for three representative donors to illustrate individual variability. (B, C, E, and G) Each color represents an independent donor. Differences were evaluated with a Wilcoxon matched pairs test; *P < 0.05; ns, not significant.

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