GM-CSF-dependent CD301b+ lung dendritic cells confer tolerance to inhaled allergens

GM-CSF-dependent CD301b+ lung dendritic cells confer tolerance to inhaled allergens | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Article GM-CSF-dependent CD301b + lung dendritic cells confer tolerance to inhaled allergens Hideki Nakano, Christina Wilkinson, Keiko Nakano, Sara Grimm, and 6 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-4414130/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 29 Sep, 2025 Read the published version in Nature Communications → Version 1 posted You are reading this latest preprint version Abstract The severity of allergic asthma is driven by the balance between allergen-specific T regulatory (Treg) and T helper (Th)2 cells. However, it is unclear whether specific subsets of conventional dendritic cells (cDCs) promote the differentiation of these two T cell lineaeges. We have identified a subset of lung resident type 2 cDCs (cDC2s) that display high levels of CD301b and have potent Treg-inducing activity ex vivo. Single cell RNA sequencing and adoptive transfer experiments show that during allergic sensitization, many CD301b+ cDC2s transition in a stepwise manner to CD200+ cDC2s that selectively promote Th2 differentiation. GM-CSF augments the development and maintenance of CD301b+ cDC2s in vivo, and also selectively expands Treg-inducing CD301b+ cDC2s derived from bone marrow. Upon their adoptive transfer to recipient mice, lung-derived CD301b+ cDC2s confer immunological tolerance to inhaled allergens. Thus, GM-CSF maintains lung homeostasis by increasing numbers of Treg-inducing CD301b+ cDC2s. Biological sciences/Immunology/Antigen processing and presentation/Immune tolerance Health sciences/Diseases/Immunological disorders/Inflammatory diseases/Asthma Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 Figure 7 Full Text Additional Declarations There is NO Competing Interest. Supplementary Files ExtDataFigures.pdf Cite Share Download PDF Status: Published Journal Publication published 29 Sep, 2025 Read the published version in Nature Communications → Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. 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Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-4414130","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":303821217,"identity":"fa56f525-9b9f-4bf6-9cf3-aa3a1aabc8e7","order_by":0,"name":"Hideki Nakano","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA40lEQVRIiWNgGAWjYFACNhBxQI6BGcLiMSBWizHpWhIboCwGglrM2Y8lfi6ouJO+nZ352MMfDHdkzBnYLz7mwaPFsiftsPSMM89ydzazpRvzMDzjsWzgKTbGp8XgQHqDNG/b4dwNh3nMpBkYDvMYHOBJk5yBT8v5582/gVrSDQ7zf5P8QZSWG2nHQLYkGBzmYZPgAWthPybxAa+WZ2nWPGcOG244zGYmzQPUCETMBni1nE8zvs1TcVje4PzhZ5I/Kg7bGxxvf/ggAY8WdBOAmJmY2EQD7A9I1jIKRsEoGAXDGgAAgsdLt+acN80AAAAASUVORK5CYII=","orcid":"https://orcid.org/0000-0002-6343-4709","institution":"National Institute of Environmental Health Sciences","correspondingAuthor":true,"prefix":"","firstName":"Hideki","middleName":"","lastName":"Nakano","suffix":""},{"id":303821218,"identity":"ee2a51bb-938c-4535-941a-8ffb8c3e70d4","order_by":1,"name":"Christina Wilkinson","email":"","orcid":"","institution":"National Institute of Environmental Health Sciences","correspondingAuthor":false,"prefix":"","firstName":"Christina","middleName":"","lastName":"Wilkinson","suffix":""},{"id":303821219,"identity":"e4d3b5a7-eed9-4711-b91b-5587415e20d5","order_by":2,"name":"Keiko Nakano","email":"","orcid":"https://orcid.org/0000-0003-2735-4119","institution":"NIEHS","correspondingAuthor":false,"prefix":"","firstName":"Keiko","middleName":"","lastName":"Nakano","suffix":""},{"id":303821220,"identity":"7b524e2c-0bff-4b70-85ae-4fd0057acbf7","order_by":3,"name":"Sara Grimm","email":"","orcid":"https://orcid.org/0000-0003-3050-1909","institution":"NIH/NIEHS","correspondingAuthor":false,"prefix":"","firstName":"Sara","middleName":"","lastName":"Grimm","suffix":""},{"id":303821221,"identity":"40000fc4-4f3f-4e06-867a-a2830f769385","order_by":4,"name":"Gregory Whitehead","email":"","orcid":"","institution":"NIEHS","correspondingAuthor":false,"prefix":"","firstName":"Gregory","middleName":"","lastName":"Whitehead","suffix":""},{"id":303821222,"identity":"2cb4c8c6-9b89-4d49-b257-658adacc02a1","order_by":5,"name":"Yukitomo Arao","email":"","orcid":"","institution":"NIH/NIEHS","correspondingAuthor":false,"prefix":"","firstName":"Yukitomo","middleName":"","lastName":"Arao","suffix":""},{"id":303821223,"identity":"84152032-7b61-4d98-96db-091c3a4b7e75","order_by":6,"name":"Perry Blackshear","email":"","orcid":"https://orcid.org/0000-0002-9561-8529","institution":"NIH","correspondingAuthor":false,"prefix":"","firstName":"Perry","middleName":"","lastName":"Blackshear","suffix":""},{"id":303821224,"identity":"45e17dbc-b694-4f21-8f32-ac28f015f37b","order_by":7,"name":"Peer Karmaus","email":"","orcid":"","institution":"National Institute of Environmental Health Sciences","correspondingAuthor":false,"prefix":"","firstName":"Peer","middleName":"","lastName":"Karmaus","suffix":""},{"id":303821225,"identity":"0c6aa2d4-6a01-424e-b231-435ed81bcb68","order_by":8,"name":"Michael Fessler","email":"","orcid":"https://orcid.org/0000-0002-8262-8613","institution":"National Institute of Environmental Health Sciencess","correspondingAuthor":false,"prefix":"","firstName":"Michael","middleName":"","lastName":"Fessler","suffix":""},{"id":303821226,"identity":"cf7a1e95-f01e-4758-ad44-75cc9ed88eaa","order_by":9,"name":"Donald Cook","email":"","orcid":"","institution":"NIEHS","correspondingAuthor":false,"prefix":"","firstName":"Donald","middleName":"","lastName":"Cook","suffix":""}],"badges":[],"createdAt":"2024-05-13 15:11:57","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-4414130/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-4414130/v1","draftVersion":[],"editorialEvents":[{"content":"https://doi.org/10.1038/s41467-025-63547-3","type":"published","date":"2025-09-29T04:00:00+00:00"}],"editorialNote":"","failedWorkflow":false,"files":[{"id":57665187,"identity":"80cb9b31-71d8-40f0-887a-a3ccf1135962","added_by":"auto","created_at":"2024-06-04 04:49:54","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":511865,"visible":true,"origin":"","legend":"\u003cp\u003eCD301b+ lung cDC2s promote Treg differentiation and induce immunological tolerance. a, Timelines for mouse models of asthma. To induce asthma, mice were sensitized with OVA/HDE twice by o.p. and challenged once with OVA aerosol. Bronchoalveolar lavage fluid (BALF) was harvested 48 h post- challenge. Assays of tolerance assays were similar, except that mice were also exposed to OVA alone by o.p. aspiration on day -7. b, Cell numbers for the indicated leukocytes in BALF, as determined by differential microscopy (n=8-12). c, Activation of CD4+ T cells (left), representative cytograms of CD4+CD44hi T cells (middle) and compiled data of Tregs (right) are shown. Naïve CD4+ T cells from Foxp3eGFP OT-II mice were cocultured for 5 days with indicated cDC2 subsets isolated from C57BL/6 mice that received OVA by o.p. aspiration (n=4). Gating strategy for purified cDC subsets and CD4+ T cell analysis (CD4+CD3e+MHCII–Live/Dead–) is shown in Extended Data Fig. 1a and b. d, Activation of CD4+ T cells (left) and Treg induction in CD44+ CD4+ T cells (right) by distinct cDC subsets isolated from mice that received OVA/HDE by o.p. aspiration (n=5). e, Timeline for mouse model of asthma to test tolerogenic function of cDC2 subsets. CD301b+ or CD301b– cDC2s were purified and incubated with OVA323-339 peptides, then adoptively transferred by o.p. to C57BL/6 mice on day 0. After OVA/HDE sensitization and OVA challenge, cells in BALF were analyzed. f, Cell numbers of the indicated leukocytes in BALF. (n=6-16). (b, c, d) Data were analyzed by one-way ANOVA with Tukey’s multiple comparison test. (f) Data were analyzed by one-way ANOVA with LSD test. Data are presented as mean values ± SEM. P values are indicated above the graphs. (b, f) Each dot represents individual mouse. Combined results from two independent experiments are shown. (c, d) Each dot represents separately cultured CD4+ T cells. Representative results from 2 independent experiments are shown.\u003c/p\u003e","description":"","filename":"MainFigure1.png","url":"https://assets-eu.researchsquare.com/files/rs-4414130/v1/7c3994a48594c05cdcc15fa3.png"},{"id":57665188,"identity":"3e7cbefb-c36b-4e7d-90a1-aea1778e419b","added_by":"auto","created_at":"2024-06-04 04:49:54","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":851984,"visible":true,"origin":"","legend":"\u003cp\u003eCD301b+ lung resident cDC2s give rise to CD200+ migratory cDC2s. a, Timeline for lung cDC2 isolation following OVA/HDE sensitization. Lung cells were harvested at 0 h (steady state), 6h, and 18h post-sensitization, and purified cDC2 were analyzed by CITE-Seq. b, UMAP plots showing cDC2 clusters identified in scRNA-Seq (left panel). Clusters are denoted by collection timepoints (right panel). c, UMAPs showing expression of subset-marker genes (top) and proteins (bottom). d, Time course of each cluster’s abundance in cDC2s at steady state and after allergic sensitization. e, RNA velocity analysis of cDC2s at each time point and combination. Maturation stages inferred by scVelocity latent time are indicated by different colors. f, g, The abundance of lung cDCs at steady state and 16 h after sensitization with OVA/HDE were calculated based on total cell counts and flow cytometric analysis. Each dot represents an individual mouse. Data were analyzed by two-tailed t-test (n=3) and presented as mean values ± SEM. P values are indicated above the graphs. h, Adoptive transfer of purified CD301b+ lung cDC2s from C57BL/6 mice (CD45.2) to CD45.1 recipient mice. Representative cytograms of purified donor cells and recipient lung cDC2s post-transfer are shown. i, Lung cDC2s at 1 day and 3 days post-transfer were analyzed by flow cytometry. Cytogram showing the phenotype of donor CD301b+ cDC2s-derived cells at the indicated time points. (a, f, g, h) Gating strategies depicted in Extended Data Figures 3a-c. (f, g, h, i) Representative results from 2 independent experiments are shown.\u003c/p\u003e","description":"","filename":"MainFigure2.png","url":"https://assets-eu.researchsquare.com/files/rs-4414130/v1/339b9b970abe02a09827a21e.png"},{"id":57665651,"identity":"beadf6f3-21c0-4dce-a47b-91f1e299d470","added_by":"auto","created_at":"2024-06-04 04:57:54","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":272379,"visible":true,"origin":"","legend":"\u003cp\u003eTregs can be induced in the lung without cDC migration. a,b, Migration of cDC2 subsets. All C57BL/6 mice received PKH26 dye and some mice received OVA or OVA/HDE by o.p. aspiration. The frequency of each subset in PKH+ migratory cDC2s in mLNs of the recipient mice were evaluated by flow cytometry (n=5-6). Gating strategy depicted 710 in Extended Data Figure 4a. Representative results of 2 independent experiments are shown. c, Treg generation in the lung of WT and Ccr7–/– mice. All mice received naïve CD4+ T cells isolated from CD45.1 OT-II mice by intravenous injection, and OVA by o.p. aspiration. The phenotype of CD45.1+ donor-derived CD4+ T cells was analyzed by flow cytometric analysis of surface proteins and intracellular Foxp3 (n=10-11). Gating strategy depicted in Extended Data Figure 4b. d, Timeline for mouse model of asthma to test tolerance induction. Some WT and Ccr7–/– mice received OVA by o.p. aspiration and were sensitized with HDE/OVA. After OVA aerosol challenge, cells in BAL fluid were analyzed. e, Cell numbers of the indicated leukocytes in BALF (n=4-6). (a, b, c, e) Each dot represents an individual mouse. Data are presented as mean values ± SEM. (a, b, e) Data were analyzed by one-way ANOVA with Fisher’s LSD test. (c) Data were analyzed by Kruskal-Wallis test. P values are indicated above the graphs.\u003c/p\u003e","description":"","filename":"MainFigure3.png","url":"https://assets-eu.researchsquare.com/files/rs-4414130/v1/d62bdebcbb1b931e9fe87ec8.png"},{"id":57665190,"identity":"49f9656a-7954-4c16-8b5d-e800bf04ddab","added_by":"auto","created_at":"2024-06-04 04:49:54","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":879198,"visible":true,"origin":"","legend":"\u003cp\u003eGM-CSF promotes the development of lung CD301b+ cDC2s. a, Surface display level of CSF2Ra on lung cDC2 subsets at steady state and 16 h after OVA/HDE inhalation were analyzed by flow cytometry. b, GM-CSF level in lung homogenates was measured by ELISA (n=5-7). c-f, The frequencies of cDC1s and cDC2 subsets in mouse lungs were analyzed by flow cytometry at steady state (c-e) or after OVA/HDE inhalation (f). (e, f) Representative cytograms (left panels) and compiled data (right panels) of Ly6C– cDC2s are shown (n=6-8). Gating strategy depicted in Extended Data Fig. 6a. (g) Representative image of an alveolar duct in a PCLS of a mouse lung analyzed by a laser-scanning microscope. Individual staining for CD11c (red), CD301b (green), and E-cadherin (blue) are shown, as well as a merged image. Arrows indicate CD11c+CD301b+ cells. AD indicates alveolar duct. Scale bar represents 50 μm. (a-f) Data are presented as mean values ± SEM. Each dot represents an individual 733 mouse. Data were analyzed by one-way ANOVA with Tukey’s multiple comparison test. Combined results from two independent experiments are shown.\u003c/p\u003e","description":"","filename":"MainFigure4.png","url":"https://assets-eu.researchsquare.com/files/rs-4414130/v1/de74334d42f19b182368e3d7.png"},{"id":57665194,"identity":"3c7795b0-fea4-4c94-b1e6-d4098b45cc45","added_by":"auto","created_at":"2024-06-04 04:49:54","extension":"png","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":456989,"visible":true,"origin":"","legend":"\u003cp\u003eGM-CSF signal is required for CD301b+ cDC2 development. a-d, Frequencies of cDC1s and cDC2 subsets in mouse lungs at steady state (a-c) and after OVA/HDE inhalation (d). Each dot represents an individual mouse. Representative results from 2 independent experiments are shown. (c, d) Representative cytograms (left panels) and compiled data (right panels) of Ly6C– cDC2s are shown. e, Treg induction by lung cDC2s. Flow cytometric analysis of CD25+ and Foxp3+ Tregs after culture of naïve CD4+ T cells from Foxp3eGFP OT-II mice with or without total lung cDC2s (n=8). Combined results from two independent experiments are shown. f, Effector T cell responses induced by lung cDC2s. Proliferation and cytokine production by T cells, as determined by cell count and ELISA, respectively, after the culture of naïve CD4+ T cells from OT-II mice with or without total lung cDC2s (n=3). Representative results from two independent experiments are shown. . (a-f) Data are presented as mean values ± SEM. P values are indicated above the graphs. (a-d) Data were analyzed by ordinary one-way ANOVA with Tukey’s multiple comparison test. (e-f) Each dot represents separately cultured CD4+ T cells. Data were analyzed by one-way ANOVA with Fisher’s LSD test. Comparison between Csf2rbfx and Csf2rbΔDC cDC2s are shown.\u003c/p\u003e","description":"","filename":"MainFigure5.png","url":"https://assets-eu.researchsquare.com/files/rs-4414130/v1/b172e82e6b45ed147788bcd4.png"},{"id":57665652,"identity":"110bbfb2-a50c-409a-beb2-e0d8d0403999","added_by":"auto","created_at":"2024-06-04 04:57:54","extension":"png","order_by":6,"title":"Figure 6","display":"","copyAsset":false,"role":"figure","size":334145,"visible":true,"origin":"","legend":"\u003cp\u003eCostimulatory molecule levels on cDC2 subsets and the role of TGF-b on Treg induction. a, Surface display levels of MHC-II (I-A) and costimulatory molecules on cDC2 subsets at steady state (top) and 16 h after OVA/HDE inhalation (bottom) were analyzed by flow cytometry. Gating strategy depicted in Extended Data Figure 6b. Data 756 were analyzed by one-way ANOVA with Tukey’s multiple comparison test (n=9). Each dot represents an individual mouse. Combined results from two independent experiments are shown. b, UMAP of lung cDC2 scRNA759 analysis depicting cells expressing Tgfb1, Furin and Nrros (left), and percentage of cells expressing Tgfb1, Furin and Nrros in each cluster (right). c, Effect of the TGF-b receptor inhibitor, SB43142, on Treg induction by lung cDC2s. CD25 on and Foxp3GFP in CD4+ T cells were analyzed by flow cytometry 5 days after coculture with total lung cDC2s. Gating strategy is depicted in Extended Data Fig. 7a. Each dot represents a separate culture of CD4+ T cells. Data were analyzed by one-way ANOVA with Dunnett’s multiple comparison test (a), or with Tukey’s multiple comparison test (c). Data are presented as mean values ± SEM. P values are indicated above the graphs.\u003c/p\u003e","description":"","filename":"MainFigure6.png","url":"https://assets-eu.researchsquare.com/files/rs-4414130/v1/e9308855b7ae72f2561edfda.png"},{"id":57665192,"identity":"bff09595-259b-48db-a2b1-bdc150546d9d","added_by":"auto","created_at":"2024-06-04 04:49:54","extension":"png","order_by":7,"title":"Figure 7","display":"","copyAsset":false,"role":"figure","size":366320,"visible":true,"origin":"","legend":"\u003cp\u003eCD301b+ BMDC2s induce Tregs in vitro. a, BMDC2s displaying CD301b and CD200 on their surface. BMDC2s were generated by in vitro culture with FLT3L for 6 days, and further cultured with or without GM-CSF. Representative cytograms (left panels) and compiled data (right panels) of flow cytometric analysis are shown. The gating strategy for BMDC2s (CD172a+CD11c+MHCII+CD24–Ly6C–CD88–Live/Dead–) is shown in Extended Data Fig. 7d. Data were analyzed by two-way ANOVA with Dunnett’s multiple comparison test. b, c, Induction of effector CD4+ T cells and Tregs by BMDC2 subsets. Cytokine production from OT-II CD4+ T cells following culture with the indicated BMDC2 subsets was analyzed by ELISA (n=3). Representative data from two independent experiments shown (b). Tregs in CD4+ T cells from Foxp3eGFP OT-II mice were analyzed by flow cytometry after 5 days culture with BMDC2 subsets (n=11). Combined results from two independent experiments shown (c). Gating strategies are depicted in Extended Data Fig. 7e and f. Data were analyzed by 779 one-way ANOVA with Tukey’s multiple comparison test. (a-c) Each dot represents separately cultured cells. Data are presented as mean values ± SEM. P values are indicated above the graphs.\u003c/p\u003e","description":"","filename":"MainFigure7.png","url":"https://assets-eu.researchsquare.com/files/rs-4414130/v1/98270cf87aa6cecb5eec37ea.png"},{"id":92475858,"identity":"5fbbd6e2-407f-4b86-9693-47c493957f0e","added_by":"auto","created_at":"2025-09-30 07:18:08","extension":"pdf","order_by":1,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":2902408,"visible":true,"origin":"","legend":"","description":"","filename":"Maintext.pdf","url":"https://assets-eu.researchsquare.com/files/rs-4414130/v1_covered_c5259c55-5638-42aa-b657-992d7b1d9c04.pdf"},{"id":57665193,"identity":"a0a20e36-3168-406b-b18b-cda3c9a8c6e7","added_by":"auto","created_at":"2024-06-04 04:49:54","extension":"pdf","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":13821533,"visible":true,"origin":"","legend":"","description":"","filename":"ExtDataFigures.pdf","url":"https://assets-eu.researchsquare.com/files/rs-4414130/v1/c990e51919d099336c28dc62.pdf"}],"financialInterests":"There is \u003cb\u003eNO\u003c/b\u003e Competing Interest.","formattedTitle":"\u003cp\u003eGM-CSF-dependent CD301b\u003csup\u003e+\u003c/sup\u003e lung dendritic cells confer tolerance to inhaled allergens\u003c/p\u003e","fulltext":[],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":false,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":true,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":true,"isAuthorSuppliedPdf":true,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":true,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"nature-portfolio","isNatureJournal":true,"hasQc":false,"allowDirectSubmit":false,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"","title":"Nature Portfolio","twitterHandle":"","acdcEnabled":false,"dfaEnabled":false,"editorialSystem":"ejp","reportingPortfolio":"","inReviewEnabled":true,"inReviewRevisionsEnabled":false},"keywords":"","lastPublishedDoi":"10.21203/rs.3.rs-4414130/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-4414130/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"The severity of allergic asthma is driven by the balance between allergen-specific T regulatory (Treg) and T helper (Th)2 cells. However, it is unclear whether specific subsets of conventional dendritic cells (cDCs) promote the differentiation of these two T cell lineaeges. We have identified a subset of lung resident type 2 cDCs (cDC2s) that display high levels of CD301b and have potent Treg-inducing activity ex vivo. Single cell RNA sequencing and adoptive transfer experiments show that during allergic sensitization, many CD301b+ cDC2s transition in a stepwise manner to CD200+ cDC2s that selectively promote Th2 differentiation. GM-CSF augments the development and maintenance of CD301b+ cDC2s in vivo, and also selectively expands Treg-inducing CD301b+ cDC2s derived from bone marrow. Upon their adoptive transfer to recipient mice, lung-derived CD301b+ cDC2s confer immunological tolerance to inhaled allergens. Thus, GM-CSF maintains lung homeostasis by increasing numbers of Treg-inducing CD301b+ cDC2s.","manuscriptTitle":"GM-CSF-dependent CD301b+ lung dendritic cells confer tolerance to inhaled allergens","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2024-06-04 04:49:49","doi":"10.21203/rs.3.rs-4414130/v1","editorialEvents":[],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"nature-communications","isNatureJournal":true,"hasQc":false,"allowDirectSubmit":false,"externalIdentity":"NCOMMS","sideBox":"Learn more about [Nature Communications](http://www.nature.com/ncomms/)","snPcode":"","submissionUrl":"https://mts-ncomms.nature.com/","title":"Nature Communications","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"ejp","reportingPortfolio":"Nature Communications","inReviewEnabled":true,"inReviewRevisionsEnabled":false}}],"origin":"","ownerIdentity":"6620d456-1373-477e-964d-c44417da41cc","owner":[],"postedDate":"June 4th, 2024","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"published-in-journal","subjectAreas":[{"id":32784794,"name":"Biological sciences/Immunology/Antigen processing and presentation/Immune tolerance"},{"id":32784795,"name":"Health sciences/Diseases/Immunological disorders/Inflammatory diseases/Asthma"}],"tags":[],"updatedAt":"2025-09-30T07:17:55+00:00","versionOfRecord":{"articleIdentity":"rs-4414130","link":"https://doi.org/10.1038/s41467-025-63547-3","journal":{"identity":"nature-communications","isVorOnly":false,"title":"Nature Communications"},"publishedOn":"2025-09-29 04:00:00","publishedOnDateReadable":"September 29th, 2025"},"versionCreatedAt":"2024-06-04 04:49:49","video":"","vorDoi":"10.1038/s41467-025-63547-3","vorDoiUrl":"https://doi.org/10.1038/s41467-025-63547-3","workflowStages":[]},"version":"v1","identity":"rs-4414130","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-4414130","identity":"rs-4414130","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}