Figure 3.

Resting and primed neutrophils, and their contents, differentially affect T cells. T cells and neutrophils were isolated from peripheral blood of healthy human donors using negative magnetic separation, and were cultured together at a 5:1 neutrophil: T cell ratio. Activation of (A) CD4+ and (B) CD8+ T cells following 24 hours co-culture with neutrophils was assessed via quantification of PD1 expression by flow cytometry. Proliferation of (C) CD4+ and (D) CD8+ T cells was assessed following 72 hours co-culture with contents released from neutrophils during lysing. (E) Following 24 hours culture of T cells with released contents, culture supernatant was collected and TNF measured by ELISA. In all cases black symbols = early activating T cells, blue symbols = late activating T cells. N values of individual donors, each of which was plated separately: (A) resting 10 early 10 late; contents 10 early 7 late; CB FMLF 14 early 9 late; TNF 5 early 4 late; LPS 4 early 4 late; NETotic 10 early 6 late. (B) resting 10 early 9 late; contents 10 early 7 late; CB FMLF 10 early 11 late; TNF 4 early 4 late; LPS 4 early 4 late; NETotic 10 early 6 late. (C) 3 early 5 late; (D) 3 early 5 late; (E) 7 early 3 late. Data between T cells exposed to neutrophils and control T cells were analyzed by paired t tests on raw data before conversion to percentages. CB/FMLF = primed with cytochalasin B and N-Formylmethionyl-leucyl-phenylalanine.