The Immunoregulatory Architecture of the Adult Oral Cavity
Cite
Matuck, B.F., Huynh, K., Pereira, D., Zhang, X., Kunz, M., Kumar, N., Easter, Q.T., Fernandes, A., Ghodke, A., Predeus, A. and Szabo, L., 2024. The Immunoregulatory Architecture of the Adult Oral Cavity. bioRxiv. https://doi.org/10.1101/2024.12.01.626279
Abstract
The oral cavity is a critical barrier with immunosurveillance capabilities. A detailed understanding of its cellular, molecular, and spatial architecture is essential for advancing precision medicine across aerodigestive tissues. Here, we present the first integrated atlas of human adult oral and craniofacial tissues, derived from single-cell RNA sequencing of ∼250,000 cells from 70 samples across 13 niches, including salivary glands and oral mucosae. Complementary spatial atlases using a 40-plex protein and 300-plex transcriptomics panels were developed, covering >2 million cells. Using our new AstroSuite package, we identified 8 tissue cellular neighborhoods (TCNs) and 17 multicellular interaction modules (MCIMs) across gland and mucosae in health. Diverse fibrovascular TCNs emerged as centralized hubs, coordinating immune cell migration and activation. Mucosal hubs within stroma displayed higher immune cell density, diversity, and activation than glands, underscoring differences in functional architecture. In keratinized and nonkeratinized mucosae, common fibrovascular hubs, and MCIMs revealed a specialized biogeography of immunoregulation. Under chronic inflammation, mucosal TCNs were fragmented, and MCIMs involving fibroblasts and immune cells were reorganized and expanded. Focused analysis of tertiary lymphoid structures (TLS) and fibroblasts identified TLS-associated MCIMs and drug candidates targeting these spatial interactions, offering a foundational framework for broadly dissecting mucosal immunity to guide precision medicine approaches.
Datasets
1. Oral and Craniofacial Atlas
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Epithelial cells62303 cells Lymphoid cells49863 cells Neural Crest-derived9664 cells Williams et al.72687 cells Tabula Sapiens35234 cells Pagella et al.28775 cells Costa-da-Silva et al.24010 cells Krivanek et al.22530 cells Opasawatchai et al.2024 cells Kretzschmar et al.835 cells Gingival Mucosa54436 cells Minor Salivary Gland31473 cells Parotid Salivary Gland30983 cells Submandibular Gland14143 cells Sublingual Gland5180 cells Tabula Sapiens_TSP1423171 cells Chen_parotid gland 113265 cells Tabula Sapiens_TSP712063 cells Costa-da-Silva_HV210842 cells Horeth_Submandibular Gland 128952 cells Costa-da-Silva_HV16023 cells Horeth_Submandibular Gland 155191 cells Costa-da-Silva_HV44345 cells Caetano_Healthy23059 cells Costa-da-Silva_HV32800 cells Opasawatchai_Donor C1445 cells Opasawatchai_Donor A579 cells Caetano_Healthy1389 cells https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8359928/72687 cells https://www.science.org/stoken/author-tokens/ST-495/full35234 cells https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8099559/28775 cells https://www.sciencedirect.com/science/article/pii/S2589004221015625?via%3Dihub24010 cells https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7511944/22530 cells https://journals.sagepub.com/doi/10.1177/00220345221147908?url_ver=Z39.88-2003&rfr_id=ori:rid:crossref.org&rfr_dat=cr_pub%20%200pubmed14143 cells https://www.nature.com/articles/s41591-021-01296-813639 cells https://journals.sagepub.com/doi/full/10.1177/0022034522107606913265 cells https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9827523/8807 cells https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7781605/3448 cells https://www.frontiersin.org/articles/10.3389/fdmed.2021.806294/full2024 cells National Institute of Dental and Craniofacial Research (NIDCR)112546 cells Chan-Zuckerberg BioHub35234 cells University of Zurich26411 cells Medical University of Vienna22530 cells University at Buffalo14143 cells Sichuan University13265 cells University of Pennsylvania8807 cells University Medical Center Groningen4496 cells King's College London3448 cells University of Zurich (UZH)2364 cells Mahidol University2024 cells MSNZ University of Würzburg835 cells site_1_Switzerland28775 cells site_1_Austria22530 cells site_1_Netherlands4496 cells site_1_Thailand2024 cells parotid gland 113265 cells Submandibular Gland 12 scRNA8952 cells Submandibular Gland 15 scRNA5191 cells not applicable210869 cells surgical donor210869 cells postmortem donor35234 cells surgical resection42102 cells Labial minor salivary gland24010 cells Parotid salivary gland17718 cells Submandibular gland14143 cells Lower pole of the parotid salivary gland 13265 cells Posterior tongue: circumvallate papilla anterior to sulcus terminalis9900 cells Rugous Area anterior Hard palate8807 cells Mandibular labial minor salivary glands7463 cells Anterior tongue: tongue dorsum 6932 cells Palatal side, interdental papilla #14 distal/15 mesial 6176 cells Sublingual gland5180 cells Buccal gingival margin UR3/UL33059 cells Mandibular labial mucosa3045 cells Apical two-thirds of the roots of extracted third molars2364 cells Tooh number 18 (upper maxillary right third molar)2024 cells Buccal gingival margin UR6
389 cells STAR v2.7.10a_alpha_220818 / STARsolo246103 cells American Indian1991 cells epithelial cell62303 cells endothelial cell of vascular tree43549 cells minor salivary gland31473 cells mucosa of dorsum of tongue16832 cells submandibular gland14143 cells sublingual gland5180 cells Hispanic or Latin American2002 cells Native American1991 cells Author cell type
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Vascular endothelial cells23920 cells Lymphatic endothelial cells2278 cells Monocytes/Macrophages1925 cells Natural Killer cells1604 cells Dendritic cells1497 cells Langerhans cells618 cells Ductal epithelial cells265 cells Myoepithelial cells26 cells Williams et al.72100 cells Kretzschmar et al.831 cells Gingival Mucosa48259 cells Caetano_Healthy23059 cells Caetano_Healthy1389 cells https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8359928/72100 cells https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9827523/8432 cells https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7781605/3448 cells National Institute of Dental and Craniofacial Research (NIDCR)74289 cells University of Pennsylvania8432 cells King's College London3448 cells MSNZ University of Würzburg831 cells not applicable87000 cells surgical donor87000 cells surgical resection4279 cells Rugous Area anterior Hard palate8432 cells Buccal gingival margin UR3/UL33059 cells Mandibular labial mucosa3020 cells Buccal gingival margin UR6
389 cells STAR v2.7.10a_alpha_220818 / STARsolo87000 cells American Indian1991 cells blood vessel endothelial cell23920 cells CD4-positive helper T cell7644 cells CD8-positive, alpha-beta T cell5367 cells endothelial cell of lymphatic vessel2278 cells natural killer cell1604 cells acinar cell of salivary gland558 cells salivary gland glandular cell265 cells adult skeletal muscle myoblast51 cells myoepithelial cell26 cells Hispanic or Latin American2002 cells Native American1991 cells Author cell type
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Vascular endothelial cells10658 cells Monocytes/Macrophages1555 cells Natural Killer cells217 cells Peripherial neurons51 cells Pagella et al.26373 cells Krivanek et al.22530 cells Opasawatchai et al.2024 cells Opasawatchai_Donor C1445 cells Opasawatchai_Donor A579 cells https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8099559/26373 cells https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7511944/22530 cells https://www.frontiersin.org/articles/10.3389/fdmed.2021.806294/full2024 cells University of Zurich26373 cells Medical University of Vienna22530 cells Mahidol University2024 cells site_1_Switzerland26373 cells site_1_Austria22530 cells site_1_Thailand2024 cells not applicable50927 cells surgical donor50927 cells surgical resection24554 cells Tooh number 18 (upper maxillary right third molar)2024 cells STAR v2.7.10a_alpha_220818 / STARsolo50927 cells blood vessel endothelial cell10658 cells CD4-positive helper T cell1060 cells CD8-positive, alpha-beta T cell1053 cells natural killer cell217 cells peripheral nervous system neuron51 cells Author cell type
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Ductal epithelial cells3846 cells Vascular endothelial cells2205 cells Monocytes/Macrophages1092 cells Myoepithelial cells220 cells Natural Killer cells155 cells Lymphatic endothelial cells81 cells Costa-da-Silva et al.24008 cells Minor Salivary Gland31468 cells Submandibular Gland14142 cells Parotid Salivary Gland4630 cells Costa-da-Silva_HV210842 cells Horeth_Submandibular Gland 128952 cells Costa-da-Silva_HV16023 cells Horeth_Submandibular Gland 155190 cells Costa-da-Silva_HV44345 cells Costa-da-Silva_HV32798 cells Tabula Sapiens_TSP14104 cells Tabula Sapiens_TSP742 cells https://www.sciencedirect.com/science/article/pii/S2589004221015625?via%3Dihub24008 cells https://journals.sagepub.com/doi/10.1177/00220345221147908?url_ver=Z39.88-2003&rfr_id=ori:rid:crossref.org&rfr_dat=cr_pub%20%200pubmed14142 cells https://www.nature.com/articles/s41591-021-01296-87460 cells https://www.science.org/stoken/author-tokens/ST-495/full146 cells National Institute of Dental and Craniofacial Research (NIDCR)31468 cells University at Buffalo14142 cells University Medical Center Groningen4484 cells Chan-Zuckerberg BioHub146 cells site_1_Netherlands4484 cells Submandibular Gland 12 scRNA8952 cells Submandibular Gland 15 scRNA5190 cells not applicable50094 cells surgical donor50094 cells postmortem donor146 cells Labial minor salivary gland24008 cells Submandibular gland14142 cells Mandibular labial minor salivary glands7460 cells Parotid salivary gland4630 cells STAR v2.7.10a_alpha_220818 / STARsolo50240 cells acinar cell of salivary gland29997 cells salivary gland glandular cell3846 cells blood vessel endothelial cell2205 cells CD8-positive, alpha-beta T cell2063 cells CD4-positive helper T cell657 cells myoepithelial cell220 cells natural killer cell155 cells endothelial cell of lymphatic vessel81 cells adult skeletal muscle myoblast56 cells minor salivary gland31468 cells submandibular gland14142 cells 5. Minor Salivary Gland Atlas
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Ductal epithelial cells2361 cells Vascular endothelial cells1513 cells Myoepithelial cells431 cells Monocytes/Macrophages416 cells Lymphatic endothelial cells78 cells Natural Killer cells31 cells Costa-da-Silva et al.24008 cells Minor Salivary Gland31468 cells Costa-da-Silva_HV210842 cells Costa-da-Silva_HV16023 cells Costa-da-Silva_HV44345 cells Costa-da-Silva_HV32798 cells https://www.sciencedirect.com/science/article/pii/S2589004221015625?via%3Dihub24008 cells https://www.nature.com/articles/s41591-021-01296-87460 cells National Institute of Dental and Craniofacial Research (NIDCR)31468 cells not applicable31468 cells surgical donor31468 cells Labial minor salivary gland24008 cells Mandibular labial minor salivary glands7460 cells STAR v2.7.10a_alpha_220818 / STARsolo31468 cells acinar cell of salivary gland18786 cells salivary gland glandular cell2361 cells blood vessel endothelial cell1513 cells CD8-positive, alpha-beta T cell1025 cells myoepithelial cell431 cells CD4-positive helper T cell300 cells endothelial cell of lymphatic vessel78 cells adult skeletal muscle myoblast47 cells natural killer cell31 cells minor salivary gland31468 cells Author cell type
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Vascular endothelial cells9788 cells Lymphatic endothelial cells877 cells Monocytes/Macrophages699 cells Natural Killer cells603 cells Langerhans cells365 cells Ductal epithelial cells176 cells Myoepithelial cells22 cells Williams et al.27289 cells https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8359928/27289 cells National Institute of Dental and Craniofacial Research (NIDCR)29478 cells not applicable29478 cells surgical donor29478 cells Mandibular labial mucosa2189 cells STAR v2.7.10a_alpha_220818 / STARsolo29478 cells blood vessel endothelial cell9788 cells CD4-positive helper T cell2167 cells CD8-positive, alpha-beta T cell1932 cells endothelial cell of lymphatic vessel877 cells natural killer cell603 cells acinar cell of salivary gland451 cells salivary gland glandular cell176 cells myoepithelial cell22 cells 7. Major Salivary Gland Atlas
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Ductal epithelial cells1485 cells Vascular endothelial cells741 cells Monocytes/Macrophages676 cells Myoepithelial cells381 cells Natural Killer cells124 cells Submandibular Gland14142 cells Parotid Salivary Gland4630 cells Horeth_Submandibular Gland 128952 cells Horeth_Submandibular Gland 155190 cells Tabula Sapiens_TSP14104 cells Tabula Sapiens_TSP742 cells https://journals.sagepub.com/doi/10.1177/00220345221147908?url_ver=Z39.88-2003&rfr_id=ori:rid:crossref.org&rfr_dat=cr_pub%20%200pubmed14142 cells https://www.science.org/stoken/author-tokens/ST-495/full146 cells University at Buffalo14142 cells University Medical Center Groningen4484 cells Chan-Zuckerberg BioHub146 cells site_1_Netherlands4484 cells Submandibular Gland 12 scRNA8952 cells Submandibular Gland 15 scRNA5190 cells not applicable18626 cells surgical donor18626 cells postmortem donor146 cells Submandibular gland14142 cells Parotid salivary gland4630 cells STAR v2.7.10a_alpha_220818 / STARsolo18772 cells acinar cell of salivary gland11211 cells salivary gland glandular cell1485 cells CD8-positive, alpha-beta T cell1038 cells blood vessel endothelial cell741 cells myoepithelial cell381 cells CD4-positive helper T cell357 cells natural killer cell124 cells submandibular gland14142 cells Author cell type
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Vascular endothelial cells1057 cells Monocytes/Macrophages203 cells Lymphatic endothelial cells190 cells Natural Killer cells154 cells Kretzschmar et al.831 cells https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9827523/8432 cells University of Pennsylvania8432 cells MSNZ University of Würzburg831 cells surgical resection831 cells Rugous Area anterior Hard palate8432 cells Mandibular labial mucosa831 cells STAR v2.7.10a_alpha_220818 / STARsolo9263 cells blood vessel endothelial cell1057 cells CD4-positive helper T cell858 cells CD8-positive, alpha-beta T cell412 cells endothelial cell of lymphatic vessel190 cells natural killer cell154 cells Author cell type
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Innate lymphoid cells2149 cells Dendritic cells1928 cells Monocytes/Macrophages1878 cells Williams et al.17354 cells Kretzschmar et al.104 cells Gingival Mucosa15490 cells Caetano_Healthy2524 cells Caetano_Healthy1228 cells https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8359928/17354 cells https://www.nature.com/articles/s41591-021-01296-82979 cells https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9827523/1875 cells https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7781605/752 cells National Institute of Dental and Craniofacial Research (NIDCR)20333 cells University of Pennsylvania1875 cells King's College London752 cells MSNZ University of Würzburg104 cells not applicable23064 cells surgical donor23064 cells surgical resection856 cells Palatal side, interdental papilla #14 distal/15 mesial 2979 cells Rugous Area anterior Hard palate1875 cells Buccal gingival margin UR3/UL3524 cells Buccal gingival margin UR6
228 cells Mandibular labial mucosa104 cells STAR v2.7.10a_alpha_220818 / STARsolo23064 cells CD8-positive, alpha-beta T cell6617 cells CD4-positive helper T cell6151 cells innate lymphoid cell2149 cells Hispanic or Latin American500 cells Author cell type
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Monocytes/Macrophages832 cells Innate lymphoid cells146 cells Costa-da-Silva et al.2388 cells Minor Salivary Gland3764 cells Parotid Salivary Gland1832 cells Submandibular Gland766 cells Costa-da-Silva_HV11179 cells Costa-da-Silva_HV4527 cells Horeth_Submandibular Gland 12399 cells Costa-da-Silva_HV2384 cells Horeth_Submandibular Gland 15367 cells Costa-da-Silva_HV3298 cells https://www.sciencedirect.com/science/article/pii/S2589004221015625?via%3Dihub2388 cells https://www.nature.com/articles/s41591-021-01296-81376 cells https://journals.sagepub.com/doi/10.1177/00220345221147908?url_ver=Z39.88-2003&rfr_id=ori:rid:crossref.org&rfr_dat=cr_pub%20%200pubmed766 cells National Institute of Dental and Craniofacial Research (NIDCR)3764 cells University Medical Center Groningen1832 cells University at Buffalo766 cells site_1_Netherlands1832 cells Submandibular Gland 12 scRNA399 cells Submandibular Gland 15 scRNA367 cells Labial minor salivary gland2388 cells Parotid salivary gland1832 cells Mandibular labial minor salivary glands1376 cells Submandibular gland766 cells STAR v2.7.10a_alpha_220818 / STARsolo6362 cells CD8-positive, alpha-beta T cell1325 cells CD4-positive helper T cell587 cells innate lymphoid cell146 cells minor salivary gland3764 cells submandibular gland766 cells Alias names
Human Oral and Craniofacial Cell Atlas