Fig. 6

Evaluation of NOS2 antigen expression at baseline and 4 weeks post-treatment with anti-TNF-α antibody (cA2). (A) Immunoblot analysis of N0S2 antigen expression in blood mononuclear cells isolated from healthy control subjects (C1–C7), patients with RA (P1–P9) at baseline measurements (Pre) and 4 weeks after a single infusion of 5 mg/kg, 10 mg/kg, or 20 mg/kg of cA2, or a placebo (Post). Positive controls for NOS2 antigen expression included stimulated human adenocarcinoma cells (DLD-1, designated D1), and stimulated mouse macrophage cells (RAW 264.7, designated R2). All immunoblotting was performed concomitantly with exactly the same exposure time so that comparisons between subjects were valid. (B) Relative optical density expressed as NOS2/actin in arbitrary units. Solid lines represent RA patients infused with of 5 mg/kg (1), 10 mg/kg(s), or 20 mg/kg(t) of cA2, whereas dashed lines represent RA patients infused with a placebo(n). The open squares represent the means ± SEM of the pre- and post-treatment values. The asterisk denotes statistical significance of p < 0.03 for differences between the levels of NOS2/actin before and 4 weeks after treatment with cA2 (n = 7; P1, P3, P4, P5, P7, P8, and P9). [Reproduced with permission (131).]