Fig 1.
Box and whisker plots of major humoral immune response outcomes pre-vaccination (baseline) and post-vaccination (Day 3, Day 28, and Day 75).
The top (bottom) of the box indicate the 75th (25th) percentiles, respectively, while the bold line within the box indicates the median. The ‘whiskers’ extend up to 1.5 times the interquartile range above or below the 75th or 25th percentiles respectively. Beyond that point, individual points are plotted. A. B cell ELISPOT counts representing the influenza A/H1N1-specific IgG-producing memory-like B cell response plotted for each timepoint as spot forming units (SFUs) per 2x105 PBMCs. B. HAI antibody titers (presented as the inverse of the greatest serum dilution that still gave a positive HAI result) are plotted over time. C. Viral neutralization antibody (VNA) titers (presented as the inverse of the greatest serum dilution that still gave a positive VN result) are plotted over time. D. CD20+ CD27+memory B cells (% of B cells) over time. Parts of Fig. 1(Fig. 1A) have been published in Viral Immunology [18,19].
Table 1.
Influence of age and immunosenescence on immune response variables after influenza A/H1N1vaccination.
Fig 2.
Correlations between markers of immunosenescence (and/or immune outcomes) and influenza A/H1N1 vaccine-induced immune responses.
A. The panel illustrates the positive correlation between early change in TERT expression at Day 3 relative to baseline on x-axis and influenza-specific B cell ELISPOT response change on y-axis (Day 28 compared to baseline). B and C. The panels illustrate the positive correlation between TREC levels on x-axis and influenza-specific B cell ELISPOT response on y-axis (Day 0 or Day 3). D, E and F. The panels illustrate the positive correlation between CD28 expression (MFI) on CD4+ T cells on x-axis (or change in expression at Day 3 relative to baseline, panel F) and influenza-specific B cell ELISPOT response on y-axis (Day 75 or increase at Day 28 compared to baseline for panel F). G and H. The panels illustrate the positive correlation between CD28 expression (MFI) on CD8+ T cells on x-axis (Day 0 or Day 3) and influenza-specific B cell ELISPOT response at Day 75 on y-axis. I. The panel illustrates the positive correlation between early change in influenza-specific B cell ELISPOT response (Day 3 relative to baseline, on x-axis) and influenza-specific HAI titer increase (Day 28 compared to baseline, on y-axis). The values on the x-axis for panels A, B, C, and F, y-axis for panels D, E, G, and H, and both x- and y-axes for panel I are displayed on the cube root scale for ease of visualization, but labeled on the raw scale. Influenza A/H1N1-specific memory-like IgG B cell ELISPOT response is presented as median SFUs per 2 x 105 cells; TERT expression is presented as TERT relative abundance value (relative expression in mRNA) divided by the GAPDH abundance value (ratio); TREC levels are presented as TREC versus CCR5 copies (ratio) in genomic DNA; HAI represents the influenza A/H1N1-specific hemagglutination inhibition titer as the reciprocal of the highest serum dilution that completely inhibits hemagglutination. The expression of CD28 on CD4+ and/or CD8+ T cells is measured by the mean fluorescence intensity/MFI using flow cytometry. “rs” indicates Spearman’s correlation coefficient.
Table 2.
Correlation between overall and influenza-specific humoral immune response variables following influenza vaccination.
Fig 3.
Multivariable associations of early variables with peak influenza/H1N1 vaccine-induced immune responses.
Standardized linear regression coefficients from the elastic net penalized linear regression models for association of early (Day 3) variables with peak (Day 28) influenza-specific humoral immune response outcomes (HAI, VNA, B cell ELISPOT). Standardized linear regression coefficients for early variables associated with: A. Day 28 HAI response; B. Day 28 VNA response; and C. Day 28 B cell ELISPOT response. The model with the minimum cross validated mean squared error is presented.