Table 1.
Characteristics of study participants.
Fig 1.
Cytokine secretion in spontaneous and Bacille Calmette-Guerin (BCG) stimulated Whole Blood Assay (WBA) and peripheral blood mononuclear cells (PBMCs) short-term culture supernatants.
The shaded area compares secretion of WBA and PBMCs (n = 20) as the mean level of individual cytokine in BCG stimulated cultures (12 hrs.) shown on the primary axis (left side). The line graph compares the spontaneous secretion of the cytokine to be read on the secondary axis (right side). The “4-sided star” indicates a significant p-value for spontaneous cytokine secretion, whereas “asterisks” shows the difference in BCG stimulated cytokines in WBA and PBMCs. Wilcoxon Sign Rank test was applied for the significant difference in the cytokine level. p < 0.05 was considered significant.
Fig 2.
Expression of innate and adaptive genes in response to Bacille Calmette-Guerin (BCG).
The scatter plot shows the fold changes in the expression of Tumor Necrosis Factor (TNFα), Granzyme A (GZMA), and Butyrophilin (BTN3A2) genes in response to live BCG stimulation of PBMCs. The fold changes are shown as relative expression compared to non-stimulated media control (10% FBS) using 2x10^6 cells between 12–24 hrs. Wilcoxon Sign Rank test were applied for the comparison of Scar positive and negative subjects.
Fig 3.
Assay dependent clustering pattern of BCG stimulated cytokines.
The orange dots indicate cytokines analyzed through PBMCs while the red dot indicates cytokines analyzed through WBA. The PCA separated data into two major components. All variables explained 80% of the variability in the model.
Fig 4.
Cytokine correlation index in WBA and PBMCs cultures.
Correlation plot showing a correlation coefficient of cytokines using the intensity of colors at the scale of -1.0 to +1.0 in WBA (A) and PBMCs assay (B).
Fig 5.
Comparative analysis of Bacille Calmette-Guerin (BCG) and Lipopolysaccharide (LPS) stimulated cytokine secretion in Whole Blood Assay (WBA) cell culture.
Comparison of cytokine secretion in response to BCG and LPS stimulation in WBA cell cultures at 12 and 48 hrs. respectively (n = 20). Horizontal bars indicate mean levels with standard error bars around the mean. Student t-tests were carried out to assess the significance of differences between BCG and LPS.
Fig 6.
Assessment of cross-reactive IgG antibodies to COVID-19 Receptor-Binding Domain (RBD).
Results are expressed as units /ml. Antibody assessment was carried out in plasma and BCG stimulated supernatants. All samples were collected before the COVID-19 pandemic. Mann-Whitney U tests were carried out to determine the significance of the difference.
Fig 7.
Relationship of IgG anti-RBD with BCG and LPS driven cytokines in Whole Blood Assay (WBA).
Correlation plot of cytokines derived from WBA (BCG and LPS) and IgG. Corrplot showing a correlation coefficient of cytokines using the intensity of colors at the scale of -1.0 to +1.0 in WBA. All significant correlation at the level of p = 0.01 is shown on the plot as circles and p > 0.01 are shown as blank.