Fig. 2
Development of TNFRI peptides with anti-TNF activity. a Schematic representation of TNFRI and mutants. WT wild type, CRD cysteine-rich domain, His histidine, linker restriction enzyme linker, M mutant. b p38 activation in cells transfected with WT or mutant TNFRI. 70Z/3 pro B cells transfected with WT TNFRI or truncated TNFRI mutants, M1 or M4, were treated with either HBSS or TNF70–80 (10 μM) for 5 min and p38 activity assayed. Results are mean ± s.e.m. of three separate experiments. Significance of difference between control and TNF70–80 (one-tailed one sample t-test): *p < 0.05, **p < 0.01. There was no difference between WT and M4 (p > 0.05, n = 3 experiments, two-tailed Mann–Whitney test). c Generation and schematic representation of TNFRI-derived peptides. The boldfaced text represents residues in the natural TNFRI sequence. Leu-Lys-Pro was introduced to generate a restriction site for coupling to the His-tag. d ∆HM4 (Leu Lys Pro Gly Thr Thr) inhibited the TNF70–80-induced CL production in neutrophils. Results are mean ± s.e.m. of four separate experiments. Significance of difference between TNF70–80 and TNF70–80 + ∆HM4 (Kruskal–Wallis test, followed by Dunn’s multiple comparison test): *p < 0.05. e Kinetics of TNF70–80-induced CL production in the presence of ∆HM4 peptide
