Journal: medRxiv

Article Title: Immunological and Antigenic Signatures Associated with Chronic Illnesses after COVID-19 Vaccination

doi: 10.1101/2025.02.18.25322379

Figure Lengend Snippet: A. Proportion of each group (PVS: n = 42, control: n = 22, pre-pandemic healthy control: n = 3448) seropositive for each of 31 common pathogen panels as determined by SERA, grouped by pathogen-type. Statistical significance determined by Fisher’s exact test corrected with FDR (Benjamini Hochberg). Star indicates panels for which pre-pandemic healthy controls were not analyzed. B. Heatmap showing supervised clustering of SERA-determined seropositivity to EBV, CMV, HSV-1, and HSV-2 across samples. Clusters were named for their herpesvirus dominance and are labeled accordingly. C. Herpes seropositivity composition for each cohort. Significance of relative enrichment for each cluster was calculated using Chi-square test of observed composition vs. expected composition. D, E Plasma reactivity to EBV gp42 protein measured by ELISA shown by cohort, PVS and Control (D) and by groups Control-I, Control+I, PVS-I, and PVS+I (E) . F. SERA-derived z scores for the gp42 motif [VI]XLPHW among EBV-seropositive individuals only, plotted by cohort, n = 20 (Control), n = 38 (PVS) (F) and group, n = 11 (Control-I), n = 9 (Control+I), n = 12 (PVS-I), n = 26 (PVS+I) (G) . The dashed line represents the z -score threshold for epitope positivity defined by SERA. H. Three-dimensional mapping of the PVS-enriched linear peptide sequence [VI]XLPHW (purple) onto EBV gp42 (blue) in a complex with gH (light grey) and gL (dark grey) (PDB: 5T1D). I. Relationship between EBV gp42 [VI]XLPHW SERA z score and plasma concentration of anti-gp42 IgG. J. SERA-derived z scores for the gp350 motif KXRX[RQ]WXF among EBV-seropositive individuals only, plotted by cohort. The dashed line represents the z -score threshold for epitope positivity defined by SERA. K. The relationship between plasma concentration of IgG against EBV gp42 and the percentage of TNFα CD8+ T cells (of total CD8+ T cells). For all box plots, the central lines indicate the group median values, the top and bottom lines indicate the 75th and 25th percentiles, respectively, the whiskers represent 1.5× the interquartile range. Each dot represents one individual. Statistical significance of the difference in median values was determined using Kruskal–Wallis tests with Post hoc Dunn’s test and Bonferroni–Holm’s method to adjust for multiple comparisons. Correlation was assessed using Spearman’s correlation. The black line shows linear regression, and shading shows the 95% CIs.

Article Snippet: Samples and a serial dilution curve of monoclonal antibody against EBV gH/gL/gp42 (MyBioSource #MBS430548) were diluted in 1% Omniblock non-fat dry milk in PBS and added to the plate to incubate for 1 hour at RT.

Techniques: Control, Labeling, Clinical Proteomics, Enzyme-linked Immunosorbent Assay, Derivative Assay, Sequencing, Concentration Assay