Interleukin-23 promotes Th17 differentiation by inhibiting T-bet and FoxP3 and is required for elevation of interleukin-22, but not interleukin-21, in autoimmune experimental arthritis.

OBJECTIVE: To examine the role of interleukin-23 (IL-23) in subgroup polarization of IL-17A-positive and/or interferon-gamma (IFNgamma)-positive T cells in autoimmune disease-prone DBA/1 mice with and without collagen-induced arthritis. METHODS: A magnetic-activated cell sorting system was used to isolate CD4+ T cells from the spleen of naive and type II collagen (CII)-immunized DBA/1 mice. These CD4+ T cells were stimulated in vitro under Th0, Th1, or different Th17 culture conditions. Intracellular staining for IL-17A and IFNgamma was evaluated by flow cytometry. In addition, Th17 cytokines and T helper-specific transcription factors were analyzed by enzyme-linked immunosorbent assay and/or quantitative polymerase chain reaction. RESULTS: In CD4+ T cells from naive DBA/1 mice, IL-23 alone hardly induced retinoic acid-related orphan receptor gammat (RORgammat), Th17 polarization, and Th17 cytokines, but it inhibited T-bet expression. In contrast, transforming growth factor beta1 (TGFbeta1)/IL-6 was a potent inducer of RORgammat, RORalpha, IL-17A, IL-17F, IL-21, and FoxP3 in these cells. In contrast to TGFbeta1/IL-6, IL-23 was critical for the induction of IL-22 in CD4+ T cells from both naive and CII-immunized DBA/1 mice. Consistent with these findings, IL-23 showed a more pronounced induction of the IL-17A+IFNgamma- subset in CD4+ T cells from CII-immunized mice. However, in CD4+ T cells from naive mice, IL-23 significantly increased the TGFbeta1/IL-6-induced Th17 polarization, including elevated levels of IL-17A and IL-17F and decreased expression of T-bet and FoxP3. Of note, the IL-23-induced increase in IL-17A and IL-17F levels was prevented in T-bet-deficient mice. CONCLUSION: IL-23 promotes Th17 differentiation by inhibiting T-bet and FoxP3 and is required for elevation of IL-22, but not IL-21, levels in autoimmune arthritis. These data indicate different mechanisms for IL-23 and TGFbeta1/IL-6 at the transcription factor level during Th17 differentiation in autoimmune experimental arthritis.

Congenital deficiency of thromboxane and prostacyclin.

Animal work suggests that with certain doses of aspirin the antithrombotic effect exerted via the inhibition of the proaggregatory platelet thromboxane A2 (TXA2) may be neutralised by the concomitant vascular reduction of the antiaggregatory prostacyclin (PGI2). Such a situation might result not only in therapeutic ineffectiveness but also in a thrombotic tendency. A patient with a bleeding disorder characterised by a mildly prolonged bleeding time and defective platelet-release reaction due to a congenital deficiency of cyclo-oxygenase provided an opportunity for studying this problem. Her platelets did not aggregate with arachidonic acid, but they did so with a synthetic endoperoxide analogue. Thrombin added to her platelet-rich plasma and whole blood did not generate thromboxane B2 (TXB2). Washed platelets, when incubated with 14C-arachidonic acid, did not produce the cyclo-oxygenase metabolites. A biopsy specimen of her vein did not generate PGI2, as measured both by platelet-aggregation inhibition and radioimmunoassay of 6-keto-PGF1 alpha. Clinically, the patient had a mild bleeding tendency but no thrombotic problems. The findings suggest that in man aspirin therapy, even at doses which inhibit PGI2 formation, would only impair haemostasis mildly without producing a thrombotic tendency.