POS0048 LIPOPOLYSACCHARIDE ACTIVATED ENTHESEAL MYELOID CELL INHIBITION WITH UPADACITINIB PARADOXICALLY INCREASES MYELOID PRO-INFLAMMATORY CYTOKINES INCLUDING TNF AND IL-23 BY RESTRAINING AN IL-10 NEGATIVE FEEDBACK-BUT T-CELL TNFΑ AND IL-17 IS STRONGLY BLOCKED
Bibliographic record
Abstract
Background Upadacitinib is effective in spondyloarthritis (SpA) associated arthropathy and is licensed for ankylosing spondylitis and psoriatic arthritis. Collectively SpA is linked to enthesitis with both myeloid lineage and T-lineage cells contributing to the IL-23/IL-17 and TNF axes that underscore disease. Objectives To investigate the effect of upadacitinib on an in vitro human enthesis model by focusing separately on the IL-23/IL-17 axis component myeloid and T-cells to better elucidate mechanisms of action in vivo. Methods Normal spinous process enthesis (n=26) was obtained from patients undergoing spinal decompression or surgery for scoliosis correction as previously described [1]. Enthesis cells were subsequently isolated by mechanical digestion. Entheseal myeloid cells were stimulated with LPS with and without upadacitinib. IL-23, TNFα and other proinflammatory cytokines in the supernatants were quantified using ELISA. IL-23 levels were measured after myeloid cell stimulation with LPS in the presence of upadacitinib and IL-10. IL-23 levels were also measured after stimulation with LPS in the presence of an IL-10Rα blocking antibody. Entheseal cells were stimulated with anti-CD3 with or without LPS, Th17-driving cytokines (IL-23 and IL-1β) and upadacitinib. Supernatants were measured by ELISA and cytokine-positive cell populations assessed by intracellular flow cytometry. Results Unexpectedly, upadacitinib significantly increased myeloid cell secretion of IL-23, TNFα and other proinflammatory cytokines after stimulation with LPS (Figure 1) (LPS = 51.2 pg/ml vs LPS + 1 µM Upa = 1048 pg/ml IL-23, n= 11). Addition of recombinant IL-10 attenuated such IL-23. Blockade of the IL-10 receptor with a blocking antibody increased IL-23 secretion from myeloid cells after stimulation with LPS (100 ng/ml IL-10 = 2.79 mean fold increase in IL-23, n=5). Despite this effect, upadacitinib strongly blocked TNFα and IL-17 from Th17 and Tc17 entheseal derived cells stimulated with exogenous IL-23 (138.19 pg/ml vs 14.2 pg/ml IL-17A after addition of 1 µM Upa to Th17 driving conditions, n = 5). Furthermore, addition of upadacitinib to LPS + anti-CD3 activated entheseal cells strongly downregulated T-cell derived IL-17 and TNFα despite a simultaneous increase in myeloid IL-23. Conclusion LPS activated myeloid cells, predominantly monocyte lineage further increase key cytokines IL-23 and TNFα post upadacitinib. This involves IL-10 pathway inhibition. However, in this system T-cell derived cytokine production is completely blocked despite elevation of myeloid IL-23 and TNFα. These unexpected findings highlight the pivotal role of T-cell derived cytokines in SpA given the known efficacy of JAK inhibition and these findings merit consideration in other domains such as intestinal inflammation that may have a greater innate immune driven component. Reference [1]Bridgewood, C., et al., Identification of myeloid cells in the human enthesis as the main source of local IL-23 production. Annals of the rheumatic diseases, 2019. 78(7): p. 929-933. Acknowledgements The study is funded by an Abbvie research grant. Disclosure of Interests Sami Giryes: None declared, Tom Macleod: None declared, Chi Wong: None declared, Mark Harland: None declared, Nicole McDermott: None declared, Charlie Bridgewood: None declared, Abhay S Rao: None declared, Almas Khan: None declared, Peter Loughenbury: None declared, Dennis McGonagle Grant/research support from: The study is funded by an Abbvie research grant.
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How this classification was reachedexpand
Full frame machine prediction
Teacher imitationNot calibrated prevalence, not ground truth. Human validation pending. The Gemma side is a direct model label for every work in the frame, read from the title-only record. The Codex side is a classifier learned from the 10,348 direct Codex labels and calibrated to design-weighted sample rates; fields without enough sample support carry no Codex call. Candidate is the union of the two sides; consensus is their intersection. These outputs are machine_predicted_unvalidated and are not human labels.
Distilled classifier scores by category (both heads)
| Category | Codex | Gemma |
|---|---|---|
| Metaresearch | 0.000 | 0.000 |
| Meta-epidemiology (narrow) | 0.000 | 0.000 |
| Meta-epidemiology (broad) | 0.001 | 0.000 |
| Bibliometrics | 0.000 | 0.000 |
| Science and technology studies | 0.000 | 0.000 |
| Scholarly communication | 0.000 | 0.000 |
| Open science | 0.000 | 0.000 |
| Research integrity | 0.000 | 0.002 |
| Insufficient payload (model declined to judge) | 0.008 | 0.002 |
Machine scores (provisional)
The two teacher heads of the student model, read on this work. A score orders the frame for review; it never asserts a category, and the validation status ships verbatim with every row.
Baseline scores from an immature model (maturity gate not passed, 7 training rounds). Scores rank; they never assert a category.
score_only:v0-immature-baseline · verbatim from the scoring run: score_only means the number may rank works, and no category label ships from itClassification
machine, unvalidatedMachine predicted; a candidate call from one source (direct Gemma or distilled Codex), not a consensus.
How this classification was reached, model by model and score by score, is at the end of the page under "How this classification was reached".