Pre-treatment IL-17 production by blood T cells predicts IVIg response in ICAM-1^-/-NOD mice.

<p>(A) Splenocytes were extracted from placebo (Pla) and IVIg 3x treated mice (n = 5 per group) after 9 weeks of treatment and stimulated in the presence of soluble anti-CD3 (1μg/ml) and anti-CD28 (0.5 μg/ml) antibodies (polyclonal) or mouse sciatic nerve homogenisate (PNS; 50 mg/ml) for 96 hours. Proliferation was measured by Thymidin incorporation in the last 24 hours and calculated as fold non-stimulated wells. (B) Peripheral blood was collected before (left panel) from placebo (Pla) and IVIg 3x treated mice (n = 20 per group), stimulated for 4 hours with PMA/Ionomycin/Golgi Transport Inhibitor and stained for intracellular IL-17. Splenocytes were extracted from treated mice after treatment (right panel) and stained for intracellular IL-17. The proportion of IL-17+ of CD3+ cells before (left panel) and after (right panel) treatment was calculated. (C) Schematic representation of the prognostic analysis. Responders were defined as ≤1 average score increase during treatment. (D) The proportion of IL-17+ CD3+ cells was plotted against the change of score during IVIg 3x treatment. (E) Paraffin sections were generated at different levels of the sciatic nerve at the sciatic notch (proximal, left panel), at mid thigh (mid, middle panel), at the sciatic trifurcation (distal, right panel) were stained for IL-17, IL-17+ cells were counted and the density of IL-17+ cells per area was calculated. ns not significant.