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Blood, Vol. 109, Issue 1, 155-158, January 1, 2007
A role for IL-1 receptor antagonist or other cytokines in the acute therapeutic effects of IVIg?
Blood Crow et al.
109: 155
Supplemental materials for: Crow et al, Vol 109, Issue 1, 155-158
Files in this Data Supplement:
- Figure S1. A monoclonal antibody that mimics the action of anti-D in the amelioration of murine ITP induces IL-1Ra production in vivo yet ameliorates ITP in IL-1 receptor–deficient mice (PDF, 14.5 KB) -
(A) C57BL/6 mice were injected intravenously with a therapeutic amount of the anti-RBC antibody TER-119 (50 µg), and each mouse was bled at the indicated time points on the x-axis. Sera were assessed for IL-1Ra by ELISA (R&D Systems, Minneapolis, MN). Values are expressed in pg IL-1Ra/mL on the y-axis. Data are expressed as mean ± SEM; n = 4 mice from 2 separate experiments. (B) C57BL/6 mice (wild-type) and IL-1R–deficient mice were injected on days 0 to 3 with antiplatelet antibody (anti–integrin  IIb antibody; clone MWReg30; PharMingen, Mississauga, ON, Canada) to induce thrombocytopenia. On day 2, mice received 50 µg TER-119 (arrow). Mice were bled daily for platelet enumeration by flow cytometry. Filled triangle indicates C57BL/6 mice; open triangle, IL-1R–deficient mice (n = 6 mice per group).
- Figure S2. IVIg is not dependent on nitric oxide for its acute therapeutic effect in ITP (PDF, 13.6 KB) -
Inducible nitric oxide synthase (iNOS) was depleted according to the method of Bang et al.1 Briefly, mice were injected intraperitoneally with 100 mg/kg of aminoguanidine (AMG) in dH2O daily beginning on day –1 and continuing to day 3 (A). The drinking water was dosed with AMG to a final concentration of 1.0% (wt/vol) to maintain constant in vivo levels of AMG. To inhibit iNOS, eNOS, and nNOS, mice were injected intraperitoneally with100 mg/kg of N(omega)-Nitro-L-arginine methyl ester hydrochloride (L-NAME) in dH2O daily beginning on day –1 and continuing to day 3 (B). To maintain in vivo levels, L-NAME was added to drinking water at a final concentration of 1.0% (wt/vol). All mice were rendered thrombocytopenic by daily injection of antiplatelet antibody (days 0-3). On day 2, mice received 50 mg IVIg (arrow). Mice were bled daily for platelet enumeration by flow cytometry. The x-axis denotes length of experiment, in days. The y-axis denotes mouse platelet count; n = 8 mice per group.
REFERENCE
1. Bang A, Speck ER, Blanchette VS, Freedman J, Semple JW. Recipient humoral immunity against leukoreduced allogeneic platelets is suppressed by aminoguanidine, a selective inhibitor of inducible nitric oxide synthase. Blood. 1996;88:2959-2966.
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