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Preprint WatchModerateOctober 10th, 2026

Direct visualization of lysosome-mediated intracellular transport and membrane anchoring of TNF-α in human T cells

Burgess, A.; Gunasekara, H.; Xie, J.; Hu, Y. S.

Lysosome-mediated intracellular transport and membrane anchoring govern TNF-alpha release from human T cells, with activation increasing vesicle motility in a microtubule-dependent manner

Moderate contradiction

1 prior failure

Two documented clinical failures match this mechanism, or a single Phase 3 failure is on record.

This preprint uses single-particle tracking and structured illumination microscopy in Jurkat T cells to show that TNF-alpha vesicles travel with lysosomes and that T cell activation increases vesicle motility, placing trafficking machinery upstream of TNF release at the cellular source. Claidex holds one TNF failure, inb03-soluble-tnf-covid19-ards-futility, recorded as an efficacy failure in which neutralising soluble TNF did not change outcomes. A trafficking-level control point does not rescue that result, and it sharpens the question the failure raised, which is whether the relevant pool of TNF in a given indication is the soluble ligand, the membrane-anchored form, or the secretion rate itself. Programs targeting TNF biology should state which pool they intend to modulate before selecting an indication.

Abstract excerpt

Tumor necrosis factor-alpha (TNF-) represents a major pro-inflammatory cytokine associated with chronic inflammation. Enhanced understanding of intracellular trafficking of TNF- enables the identification of drug targets that modulate TNF- secretion at the cellular source. However, conventional cytokine assays provide only bulk measurements of secreted TNF- or static measurements in fixed cells and therefore cannot provide mechanistic information. Here, we report a fluorescence imaging platform to visualize and quantify intracellular TNF- trafficking in human T cells. Using a Jurkat T cell line stably expressing TNF- fused to mStayGold as a model system, we revealed through single-particle tracking (SPT) that T cell activation increased the motility of TNF- vesicles and that microtubule perturbation disrupted this increased motility. 3D structured illumination microscopy uncovered spatial associations between TNF- vesicles and lysosomes, and SPT further demonstrated their coordinated transport. Using total internal reflection fluorescence microscopy, we captured the sub-second splitting of single TNF- vesicles into multiple puncta that subsequently separated, suggesting delivery of TNF- from the vesicle to the plasma membrane. Confocal-based super-resolution microscopy further resolved TNF- vesicles fusing with the plasma membrane and transmembrane TNF-. Together, these imaging capabilities for tracking and super-resolving the intracellular transport and membrane delivery of TNF- in live T cells enable mechanistic studies of TNF- trafficking and release.

Matching Claidex post-mortems

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This is an automated contradiction flag, not an editorial judgment on the preprint's quality. Flags identify where the preclinical literature and the clinical failure record diverge.