Macrophage-specific NF-kappa B activation dynamics can segregate inflammatory bowel disease patients

Papoutsopoulou, S.; Burkitt, M. D.; Bergey, F.; England, H.; Hough, R.; Schmidt, L.; Spiller, D. G.; White, M. H.; Paszek, P.; Jackson, D. A.; Martins Dos Santos, V. A.; Sellge, G.; Pritchard, D. M.; Campbell, B. J.; Muller, W.; Probert, C.

doi:10.1101/535096

bioRxiv · 10.1101/535096

Macrophage-specific NF-kappa B activation dynamics can segregate inflammatory bowel disease patients

Papoutsopoulou, S.·Burkitt, M. D.·Bergey, F.·England, H.·Hough, R.·Schmidt, L.·Spiller, D. G.·White, M. H.·Paszek, P.·Jackson, D. A.·Martins Dos Santos, V. A.·Sellge, G.·Pritchard, D. M.·Campbell, B. J.·Muller, W.·Probert, C.

View original source ↗Export metadata ↓

Abstract

The heterogeneous nature of inflammatory bowel disease (IBD) presents challenges, particularly when choosing therapy. Activation of the NF-{kappa}B transcription factor is a highly-regulated, dynamic event in IBD pathogenesis. We expressed the human NF-{kappa}B/p65 subunit in blood-derived macrophages, using lentivirus. Confocal imaging of p65 activation revealed that a higher proportion of macrophages from Crohns patients responded to lipid-A compared to controls. In contrast, cells from ulcerative colitis (UC) patients exhibited a shorter duration of p65 nuclear localisation compared to healthy controls and Crohns donors. Using a similar lentivirus approach, NF-{kappa}B-regulated luciferase was expressed in patient macrophages, isolated from frozen peripheral blood mononuclear cell samples. Following activation, samples could be segregated into three clusters based on the NF-{kappa}B-regulated luciferase response. The majority of UC samples appeared in hypo-responsive cluster 1, with Crohns patients representing the majority of hyper-responsive cluster 3. A positive correlation was seen between NF-{kappa}B-induced luciferase activity and cytokine levels released to medium from stimulated macrophages, but not in serum or biopsy. Analysis of macrophage cytokine responses and patient metadata revealed a strong correlation between Crohns patients who smoked and hyper-activation of p65. These in vitro dynamic assays of NF-{kappa}B activation in blood-derived macrophages segregate IBD patients into groups with different phenotypes and therefore may help determine response to therapy. Significance statementThis manuscript describes two dynamic assays of NF-{kappa}B activation in blood-derived macrophages that can segregate IBD patients into groups with different phenotypes. For the first time we introduce the use of dynamic measurements of a transcription factor activation as a method to stratify patients and we are confident that our approach will lead in future to early patient stratification and prediction of treatment outcome.

Source connections

Published article ↗

Explore related subjects

immunology Preprint

Keep this discovery

Explore connections, maps & timelines

BibTeX RIS

Papoutsopoulou, S., Burkitt, M. D., Bergey, F., England, H., Hough, R., Schmidt, L., Spiller, D. G., White, M. H., Paszek, P., Jackson, D. A., Martins Dos Santos, V. A., Sellge, G., Pritchard, D. M., Campbell, B. J., Muller, W., Probert, C.. 2019-01-30. Macrophage-specific NF-kappa B activation dynamics can segregate inflammatory bowel disease patients. https://doi.org/10.1101/535096

Cite the original work for its findings. Save a collection to share your selection of sources.

KEEP EXPLORING

Related preprints

bioRxiv2026

γδ T cell-derived IL-17A aggravates experimental epidermolysis bullosa acquisita in response to IL-1β

Epidermolysis bullosa acquisita (EBA) is a mucocutaneous autoimmune blistering disorder mediated by autoantibodies against type VII collagen and subsequent splitting of the dermal-epidermal junction. Interleukin (IL)-17A promotes the production of chemokines and cytokines involved in neutrophil recruitment and activation, suggesting that IL-17A may contribute to EBA pathogenesis. We aimed to clarify the role of IL-17A in experimental EBA. Using an antibody transfer-induced EBA mouse model, we demonstrate that IL-17A promotes disease development, as pharmacological blockade of IL-17A with a neutralizing antibody ameliorated skin lesions. Gamma-delta ({gamma}{delta}) T cells were identified as the predominant cellular source of IL-17A, supported by the attenuated disease phenotype observed in {gamma}{delta}TCR-deficient mice. In addition, CD4+ T cells and neutrophils contributed to IL-17A production in experimental EBA. Mechanistically, IL-1-beta, rather than IL-23, was identified as a critical mediator of IL-17A production by {gamma}{delta} T cells in vitro, consistent with our in vivo findings. Whereas EBA severity was not reduced in IL-23p19-deficient mice, disease was markedly attenuated in IL-1R-deficient animals. Together, our study provides the first evidence that {gamma}{delta} T cell-derived IL-17A plays a pivotal role in experimental EBA and identifies a previously unrecognized IL-1{beta}/{gamma}{delta} T cell/IL-17A axis that expands current understanding of EBA immunopathogenesis.

immunology↗

bioRxiv2026

A mammalian fibroblast zoo reveals barriers to cross-species virus replication and differential regulation of antiviral immunity

Virus zoonosis represents a continual threat to humans. Viruses emerge in humans directly or indirectly through interactions with mammals. However, our understanding of virus-host susceptibility across the diversity of mammals is limited and experimental model systems are restricted to a very small set of model organisms. Mammals are highly diverse and different habitats, lifestyles, diets, and exposures to different pathogens have driven substantial genetic diversity. We generated a library of primary fibroblasts across 86 mammalian species and used these cells to build an atlas of virus-host susceptibility across a panel of arboviruses that infect diverse mammals including humans. Perturbing the innate immune system revealed some virus-host pairs where restriction is controlled by the immune system and others where there is an incompatibility with host factors. Transcriptomic profiling of the library in response to virus sensing mimicked by poly(I:C) or antiviral signaling with type I interferon reveals a core set of conserved genes that respond as well as substantial species- and order-specific responses. Together these data provide a comparative atlas of innate immune responses across mammals and offer a window into how these systems have evolved across animals.

immunology↗

bioRxiv2026

A reporter mouse model to map RIG-I expression and function in vivo

The cytosolic RNA sensor RIG-I is a critical component of the innate immune response to RNA viruses, yet few tools exist to track RIG-I in vivo. Herein, we report the development of a RIG-I reporter mouse, termed RIG-ImScar, that enables fluorescence imaging and tractable immunoprecipitation of RIG-I in vivo. These mice revealed enrichment of RIG-I expression in circulatory, lymphoid, and mucosal tissues and broad expression across immune cell types. RIG-I itself is upregulated in response to IFN. To distinguish RIG-I sentinel cells, which are programmed to express RIG-I at steady state, from IFN-responder cells, we crossed RIG-ImScar mice with Mx1GFP mice. Blocking IFNAR with a monoclonal antibody showed that basal RIG-I expression in immune cells was largely independent of type I IFN signaling, although Ly6Chi monocytes required IFNAR for RIG-I expression. To probe functional responses in vivo, we intranasally administered a RIG-I-specific agonist, SLR14. Mapping RIG-I expression across the lung identified alveolar macrophages as a critical RIG-I sensor cell population, and depletion of alveolar macrophages, but not neutrophils, slightly impaired lung IFN responses. Together, these findings support the use of the RIG-ImScar model to probe RIG-I dynamics in vivo and predict functional sensor cell populations across tissues.

immunology↗