Figure 5.

Modulation of cytokine production and lytic and apoptotic molecules expression in innate immune cells by multiple doses of the BNT162b1 vaccine. (A) INF-γ and TNF- producing monocytes; (B) Gate strategy used to identify cytokine-producing monocytes. The CD14+ vs. the SS dot plot (left) allows the discrimination of monocytes (Gate CD14); cytokine-producing monocytes were identified in the CD14 vs. TNF (upper right) and the CD14 vs. IFN-γ (lower right) dot plots. (C) INF-γ and TNF- producing NK cells; (D) Gate strategy used to identify NK cells producing cytokines. Double negative cells (Gate CD3-CD19-) were analyzed within a CD56 vs. CD16 dot plot (left) leading to the identification of NK cells Cytokine-producing NK cells were selected in the TNF vs. IFN-γ dot plot (right); (E) CD107a+/perforin+ and CD107a+/granzymes+ NK cells. (F) Gate strategy used to identify NK cells expressing perforin and granzymes. Double negative cells (Gate CD3-CD19-) were analyzed within a CD56 vs. CD16 dot plot (left) leading to the identification of granzymes- and perforin-producing NK cells in the granzyme vs. CD107 (right) and in the perforin vs. CD107 (right) dot plots. PBMC were stimulated with the SARS-CoV-2 spike protein. Individuals who had never been SARS-CoV-2-infected and received three doses of the BNT162b1 mRNA vaccine were analyzed at different time points: baseline (immediately before the first inoculation (white triangle) (T0), 7 (T1) and 21 (T2) days after initial inoculation, one (T3), three (T4) and six (T5) months after the first vaccine booster (grey triangle), and, finally, ten days after the second vaccine booster (black triangle) (T6). Median values are shown. Boxes stretch from the 25th to the 75th percentile. Lines across the boxes indicate the median values. Lines stretching from the boxes indicate extreme values. Statistical significance is shown, **p<0,05.