Figure 2.

Influence of human lactoferrin (hLF) on the proinflammatory response of monocyte‐differentiated macrophages (moMϕ). (a) MoMϕ were stimulated with lipopolysaccharide (LPS) or lipoteichoic acid (LTA) for 4 h and tumour necrosis factor (TNF)‐α was measured with enzyme‐linked immunosorbent assay in the cell culture supernatant (n = 5 per group: black bars adults, white bars neonates). (b) Influence of increasing dosage of hLF on adult monocyte‐derived macrophages on the secretion of TNF‐α after 4 h LPS stimulation. Human serum albumin (HSA) was used as protein control (n = 5). (c) In parallel, intracellular (i.c.) TNF‐α was assessed on a single‐cell level (n = 5 per group: black bars adults, white bars neonates) using flow cytometry, and (d) one representative histogram from 10 independent experiments is shown. Bars show mean ± standard deviation. Normal distribution was determined with the Shapiro–Wilk test. Normally distributed data were analysed using one‐way one‐way analysis of variance in accordance with Tukey and non‐normally distributed data were analysed using the Kruskal–Wallis test. *P < 0·05; **P < 0·01.