Eosinophils accumulate adjacent to epithelial cells in the mucosa of patients with eosinophilic esophagitis (EoE), yet the bidirectional communication between these cells is not well understood. Herein, we investigated the crosstalk between human eosinophils and esophageal epithelial cells. We report that blood-derived eosinophils have prolonged survival when cocultured with epithelial cells; 96 ± 1% and 30 ± 6% viability was observed after 7 and 14 days of coculture, respectively, compared with 1 ± 0% and 0 ± 0% of monoculture. In the presence of IL-13 and epithelial cells, eosinophils had greater survival (68 ± 1%) at 14 days compared with cocultures lacking IL-13. Prolonged eosinophil viability did not require cellular contact and was observed when eosinophils were cultured in conditioned media from esophageal epithelial cells; neutralizing GM-CSF attenuated eosinophil survival. The majority of eosinophil transcripts (58%) were dysregulated in cocultured eosinophils compared with freshly isolated cells. Analysis of epithelial cell transcripts indicated that exposure to eosinophils induced differential expression of a subset of genes that were part of the EoE esophageal transcriptome. Collectively, these results uncover a network of crosstalk between eosinophils and esophageal epithelial cells involving epithelial mediated eosinophil survival and reciprocal changes in cellular transcripts, events likely to occur in EoE.
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Zuo, L. et al. IL-13 induces esophageal remodeling and gene expression by an eosinophil-independent, IL-13R alpha 2-inhibited pathway. J. Immunol. 185, 660–669 (2010).
Ben Baruch-Morgenstern, N. et al. Paired Ig-like receptor B inhibits IL-13-driven eosinophil accumulation and activation in the esophagus. J. Immunol. 197, 707–714 (2016).
Rothenberg, M. E. et al. Intravenous anti-IL-13 mAb QAX576 for the treatment of eosinophilic esophagitis. J. Allergy Clin. Immunol. 135, 500–507 (2015).
Hirano, I. et al. RPC4046, a monoclonal antibody against IL13, reduces histologic and endoscopic activity in patients with eosinophilic esophagitis. Gastroenterology 156, 592–603 e510 (2019).
Kc, K., Rothenberg, M. E. & Sherrill, J. D. In vitro model for studying esophageal epithelial differentiation and allergic inflammatory responses identifies keratin involvement in eosinophilic esophagitis. PLoS One 10, e0127755 (2015).
Rochman, M. et al. Profound loss of esophageal tissue differentiation in patients with eosinophilic esophagitis. J. Allergy Clin. Immunol. 140, 738–749 e733 (2017).
Kottyan, L. C. et al. Genome-wide association analysis of eosinophilic esophagitis provides insight into the tissue specificity of this allergic disease. Nat. Genet. 46, 895–900 (2014).
Davis, B. P. et al. Eosinophilic esophagitis-linked calpain 14 is an IL-13-induced protease that mediates esophageal epithelial barrier impairment. JCI Insight 1, e86355 (2016).
Litosh, V. A. et al. Calpain-14 and its association with eosinophilic esophagitis. J. Allergy Clin. Immunol. 139, 1762–1771 e1767 (2017).
Masterson, J. C. et al. Eosinophil-mediated signalling attenuates inflammatory responses in experimental colitis. Gut 64, 1236–1247 (2015).
Sugawara, R. et al. Small intestinal eosinophils regulate Th17 cells by producing IL-1 receptor antagonist. J. Exp. Med. 213, 555–567 (2016).
Weller, P. F. & Spencer, L. A. Functions of tissue-resident eosinophils. Nat. Rev. Immunol. 17, 746–760 (2017).
Rozenberg, P. et al. CD300f:IL-5 cross-talk inhibits adipose tissue eosinophil homing and subsequent IL-4 production. Sci. Rep. 7, 5922 (2017).
Lee, J. J., Jacobsen, E. A., McGarry, M. P., Schleimer, R. P. & Lee, N. A. Eosinophils in health and disease: the LIAR hypothesis. Clin. Exp. Allergy 40, 563–575 (2010).
Jacobsen, E. A. et al. Eosinophil activities modulate the immune/inflammatory character of allergic respiratory responses in mice. Allergy 69, 315–327 (2014).
Shah, K., Ignacio, A., McCoy, K. D. & Harris, N. L. The emerging roles of eosinophils in mucosal homeostasis. Mucosal Immunol. 13, 574–583 (2020).
Simon, H. U., Yousefi, S., Dibbert, B., Levi-Schaffer, F. & Blaser, K. Anti-apoptotic signals of granulocyte-macrophage colony-stimulating factor are transduced via Jak2 tyrosine kinase in eosinophils. Eur. J. Immunol. 27, 3536–3539 (1997).
Nutku-Bilir, E., Hudson, S. A. & Bochner, B. S. Interleukin-5 priming of human eosinophils alters siglec-8 mediated apoptosis pathways. Am. J. Respir. Cell Mol. Biol. 38, 121–124 (2008).
Johansson, M. W. et al. Anti-IL-5 attenuates activation and surface density of beta(2) -integrins on circulating eosinophils after segmental antigen challenge. Clin. Exp. Allergy 43, 292–303 (2013).
Fulkerson, P. C., Schollaert, K. L., Bouffi, C. & Rothenberg, M. E. IL-5 triggers a cooperative cytokine network that promotes eosinophil precursor maturation. J. Immunol. 193, 4043–4052 (2014).
Han, S. T. & Mosher, D. F. IL-5 induces suspended eosinophils to undergo unique global reorganization associated with priming. Am. J. Respir. Cell Mol. Biol. 50, 654–664 (2014).
Egea, L., Hirata, Y. & Kagnoff, M. F. GM-CSF: a role in immune and inflammatory reactions in the intestine. Expert Rev. Gastroenterol. Hepatol. 4, 723–731 (2010).
Sherrill, J. D. et al. Desmoglein-1 regulates esophageal epithelial barrier function and immune responses in eosinophilic esophagitis. Mucosal Immunol. 7, 718–729 (2014).
Dunn, J. L. M. et al. Esophageal type 2 cytokine expression heterogeneity in eosinophilic esophagitis in a multisite cohort. J. Allergy Clin. Immunol. 145, 1629–1640 e1624 (2020).
Shimizu, S. et al. Eosinophil-epithelial cell interactions stimulate the production of MUC5AC mucin and profibrotic cytokines involved in airway tissue remodeling. Am. J. Rhinol. Allergy 28, 103–109 (2014).
Kottyan, L. C. & Rothenberg, M. E. Genetics of eosinophilic esophagitis. Mucosal Immunol. 10, 580–588 (2017).
Namjou, B. et al. Phenome-wide association study (PheWAS) in EMR-linked pediatric cohorts, genetically links PLCL1 to speech language development and IL5-IL13 to EOsinophilic Esophagitis. Front. Genet. 5, 401 (2014).
Barnig, C. et al. Circulating human eosinophils share a similar transcriptional profile in asthma and other hypereosinophilic disorders. PLoS One 10, e0141740 (2015).
Ding, J. et al. Systematic comparison of single-cell and single-nucleus RNA-sequencing methods. Nat. Biotechnol. 38, 737–746 (2020).
Andreev, D. et al. Regulatory eosinophils induce the resolution of experimental arthritis and appear in remission state of human rheumatoid arthritis. Ann. Rheum. Dis. 80, 451–468 (2020).
Luttmann, W. et al. Activation of human eosinophils by IL-13. Induction of CD69 surface antigen, its relationship to messenger RNA expression, and promotion of cellular viability. J. Immunol. 157, 1678–1683 (1996).
Le-Carlson M, Seki S, Abarbanel D, Quiros A, Cox K, Nadeau KC. Markers of antigen presentation and activation on eosinophils and T cells in the esophageal tissue of patients with eosinophilic esophagitis. J Pediatr Gastroenterol Nutr. 56, 257–62. https://doi.org/10.1097/MPG.0b013e3182758d49 (2013).
Bullock, J. Z. et al. Interplay of adaptive th2 immunity with eotaxin-3/c-C chemokine receptor 3 in eosinophilic esophagitis. J. Pediatr. Gastroenterol. Nutr. 45, 22–31 (2007).
Bhattacharya, B. et al. Increased expression of eotaxin-3 distinguishes between eosinophilic esophagitis and gastroesophageal reflux disease. Hum. Pathol. 38, 1744–1753 (2007).
Johnsson, M. et al. Distinctive blood eosinophilic phenotypes and cytokine patterns in eosinophilic esophagitis, inflammatory bowel disease and airway allergy. J. Innate Immun. 3, 594–604 (2011).
Penido, C. et al. LPS induces eosinophil migration via CCR3 signaling through a mechanism independent of RANTES and Eotaxin. Am. J. Respir. Cell Mol. Biol. 25, 707–716 (2001).
Fulkerson, P. C. et al. A central regulatory role for eosinophils and the eotaxin/CCR3 axis in chronic experimental allergic airway inflammation. Proc. Natl Acad. Sci. USA 103, 16418–16423 (2006).
Trautmann, A. et al. T cells and eosinophils cooperate in the induction of bronchial epithelial cell apoptosis in asthma. J. Allergy Clin. Immunol. 109, 329–337 (2002).
Pope, S. M., Zimmermann, N., Stringer, K. F., Karow, M. L. & Rothenberg, M. E. The eotaxin chemokines and CCR3 are fundamental regulators of allergen-induced pulmonary eosinophilia. J. Immunol. 175, 5341–5350 (2005).
Johansson, M. W., Kelly, E. A., Nguyen, C. L., Jarjour, N. N. & Bochner, B. S. Characterization of Siglec-8 expression on lavage cells after segmental lung allergen challenge. Int Arch. Allergy Immunol. 177, 16–28 (2018).
Fulkerson, P. C. & Rothenberg, M. E. Eosinophil development, disease involvement, and therapeutic suppression. Adv. Immunol. 138, 1–34 (2018).
Straumann, A. et al. Anti-interleukin-5 antibody treatment (mepolizumab) in active eosinophilic oesophagitis: a randomised, placebo-controlled, double-blind trial. Gut 59, 21–30 (2010).
Assa’ad, A. H. et al. An antibody against IL-5 reduces numbers of esophageal intraepithelial eosinophils in children with eosinophilic esophagitis. Gastroenterology 141, 1593–1604 (2011).
Otani, I. M. et al. Anti-IL-5 therapy reduces mast cell and IL-9 cell numbers in pediatric patients with eosinophilic esophagitis. J. Allergy Clin. Immunol. 131, 1576–1582 (2013).
Nelson, R. K. et al. Human eosinophils express a distinct gene expression program in response to IL-3 compared with common beta-chain cytokines IL-5 and GM-CSF. J. Immunol. 203, 329–337 (2019).
Cherry, W. B., Yoon, J., Bartemes, K. R., Iijima, K. & Kita, H. A novel IL-1 family cytokine, IL-33, potently activates human eosinophils. J. Allergy Clin. Immunol. 121, 1484–1490 (2008).
Matsumoto, K. et al. CD44 and CD69 represent different types of cell-surface activation markers for human eosinophils. Am. J. Respir. Cell Mol. Biol. 18, 860–866 (1998).
Barthel, S. R., Jarjour, N. N., Mosher, D. F. & Johansson, M. W. Dissection of the hyperadhesive phenotype of airway eosinophils in asthma. Am. J. Respir. Cell Mol. Biol. 35, 378–386 (2006).
Reichman, H. et al. Activated eosinophils exert antitumorigenic activities in colorectal cancer. Cancer Immunol. Res. 7, 388–400 (2019).
Wen, T. et al. Single-cell RNA sequencing identifies inflammatory tissue T cells in eosinophilic esophagitis. J. Clin. Investig. 130, 2014–2028 (2019).
Kodavanti, U. P., Jaskot, R. H., Bonner, J., Badgett, A. & Dreher, K. L. Eosinophilic lung inflammation in particulate-induced lung injury: technical consideration in isolating RNA for gene expression studies. Exp. Lung Res. 22, 541–554 (1996).
Toni, L. S. et al. Optimization of phenol-chloroform RNA extraction. MethodsX 5, 599–608 (2018).
Bouffi, C. et al. Transcription factor repertoire of homeostatic eosinophilopoiesis. J. Immunol. 195, 2683–2695 (2015).
Vallabh, S., Kartashov, A. V. & Barski, A. Analysis of ChIP-seq and RNA-seq data with BioWardrobe. Methods Mol. Biol. 1783, 343–360 (2018).
This study was supported by the NIH R01 A1045898. All flow cytometric data were acquired using equipment maintained by the Research Flow Cytometry Core in the Division of Rheumatology at Cincinnati Children’s Hospital Medical Center, and this work was supported in part by the Digestive Health Center (NIDDK P30 DK078392). The authors would like to thank Shawna Hottinger for editorial support.
M.E.R. is a consultant for Pulm One, Spoon Guru, ClostraBio, Serpin Pharm, Allakos, Celgene, Astra Zeneca, Arena Pharmaceuticals, Glaxo Smith Kline, Guidepoint, and Suvretta Capital Management and has an equity interest in the first five listed and royalties from reslizumab (Teva Pharmaceuticals), PEESSv2 (Mapi Research Trust) and UpToDate. M.E.R. is an inventor of patents owned by Cincinnati Children’s Hospital.
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Dunn, J.L.M., Caldwell, J.M., Ballaban, A. et al. Bidirectional crosstalk between eosinophils and esophageal epithelial cells regulates inflammatory and remodeling processes. Mucosal Immunol (2021). https://doi.org/10.1038/s41385-021-00400-y