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Immune responses to citrullinated neoantigens and clinical efficacy of costimulation blockade indicate a general defect in maintaining T cell tolerance in rheumatoid arthritis (RA). To examine whether TCR threshold calibration contributes to disease pathogenesis, signaling in RA T cells was quantified. RA patients had a selective increase in ERK phosphorylation compared with demographically matched controls due to a mechanism distal of Ras activation. Increased ERK responses included naive and memory CD4 and CD8 T cells and did not correlate with disease activity. The augmented ERK activity delayed SHP-1 recruitment to the TCR synapse and sustained TCR-induced Zap70 and NF-κB signaling, facilitating responses to suboptimal stimulation. Increased responsiveness of the ERK pathway was also a characteristic finding in the SKG mouse model of RA where it preceded clinical symptoms. Treatment with subtherapeutic doses of a MEK-1/2 inhibitor delayed arthritis onset and reduced severity, suggesting that increased ERK phosphorylation predisposes for autoimmunity and can be targeted to prevent disease.

In animal models, lymphopenia and the associated function of homeostatic cytokines (HC) set the stage for autoimmunity. This model is relevant for rheumatoid arthritis (RA) which has defects in T cell homeostasis due to a failure in DNA repair and which responds to JAK inhibition. We have previously shown that RA T cells have a hyperreactive ERK response which impairs the induction of anergy. Here, we examined whether HC tune TCR activation thresholds by calibrating the ERK rheostat. RA T cells exhibited increased levels of phosphorylated STAT3 and STAT5 indicating in vivo activity of IL-21/IL6 and IL-7/IL-15. Incubation of healthy donor T cells with IL-7, IL15 or IL21 prior to anti-CD3/CD28 stimulation augmented ERK activation and expression of activation markers (CD69, CD40L) and cytokines (TNFα, IFNγ and IL-17). Preincubation with IL-1β or TNFα did not exert an effect. In DR4 healthy donors, HC priming facilitated responses to self-antigens such as citrullinated vimentin peptide. The effect of HC lasted for less than 3 hours and was sensitive to PI3K inhibition suggesting a non-transcriptional PI3K mediated priming of the ERK pathway. HC dose titration showed a bimodal distribution in pERK consistent with an on-off switch characteristic of a positive feedback loop. Allosteric binding of RAS-GTP to SOS is known to induce such an on-off switch. Indeed, HC incubation increased active RAS. We propose that HC lower the threshold for autoimmune responses by priming of SOS.

Immune responses to citrullinated neoantigens indicate a tolerance defect in rheumatoid arthritis (RA). We examined the hypothesis that abnormal TCR signaling in RA T cells lowers the activation threshold. T cells from 33 seropositive RA patients and 33 controls were stimulated by CD3/CD28 crosslinking or superantigen-coated dendritic cells. Activation-induced signaling was quantified by PhosFlow, Western blotting, and confocal microscopy. Basal as well as post-stimulation p-ERK was higher in naïve, memory and effector CD4 and CD8 T cells from RA patients compared to controls (p