TNF-α producing monocyte subsets in QFT negative CCs and PTB patients according to helminth species.

The frequency of monocytes within each subset of monocytes (classical, intermediate, and non-classical monocytes) in helminth negative (Helm-) and helminth positive (as indicated) after 6h ex vivo stimulation of PBMCs with medium (Unstim.) or PPD, where TNF positivity was analyzed by flow cytometry following intracellular staining with anti-human TNFα PE. (A) Intracellular PE-isotype stained PBMCs were used to set the TNF+ gates for the monocyte subsets, gated as in Fig 2A. Gating for PPD-stimulated PBMC from patients with TB is shown, and the numbers in gates are the mean values for each group. (B) Data is presented as the mean frequency ± SEM of TNF-α producing monocytes within each monocyte subset. n = 18/10/25 for Helm-/Ascaris+/Schistosoma+ CCs, and n = 35/6/14/14 for Helm-/Ascaris+/Schistosoma+/hookworm+ PTB patients. Statistical analysis was performed separately for CCs and PTB groups, using one-way ANOVA followed by Tukey’s multiple comparison test. Additionally to the differences within the PTB groups, there was a significant increase in % TNF+ intermediate monocytes in S. mansoni+/PTB compared to S. mansoni+/CCs, both in unstimulated and PPD-stimulated PBMCs (p < 0.05). *, p < 0.05; **, p < 0.01.