Figure 4

Progranulin (PGRN) and/or its fragments GRNs augment the IL-6 production by human peripheral blood mononuclear cells (PBMCs) via TLR9 signaling. (A) PBMCs from healthy control individuals were incubated for 24 hours in serum-free medium with or without 10 nM CpG-B, 100 ng/ml poly (I:C), 60 pg/ml LPS, or 1 µg/ml imiquimod and purified PGRN (250 ng/ml). The supernatant IL-6 levels were measured with ELISA. The data are representative of three independent experiments (average and SEM). (B, C) The binding of anti-PGRN Ab to PGRN was verified by immunoprecipitation and immunoblot with anti-PGRN Ab. In C, lupus sera were partly treated for 1 hour with 6,000 U/ml DNase I before stimulation. The PBMCs were then incubated in serum-free medium or were stimulated with 10% serum from systemic lupus erythematosus (SLE) patients in the presence of anti-PGRN Ab or control Ab for 24 hours. The supernatant IL-6 levels were measured with ELISA. (D) The serum PGRN levels correlated significantly with the serum IL-6 levels in SLE patients. *P < 0.05.