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 cells62,303 cells Lymphoid cells49,863 cells Myeloid cells19,492 cells Neural Crest-derived9,664 cells Williams et al.72,687 cells Tabula Sapiens35,234 cells Pagella et al.28,775 cells Costa-da-Silva et al.24,010 cells Krivanek et al.22,530 cells Horeth et al.14,143 cells Pringle et al.4,496 cells Caetano et al.3,448 cells Opasawatchai et al.2,024 cells Kretzschmar et al.835 cells Gingival Mucosa54,436 cells Minor Salivary Gland31,473 cells Parotid Salivary Gland30,983 cells Buccal Mucosa27,875 cells Dorsal Tongue16,832 cells Submandibular Gland14,143 cells Sublingual Gland5,180 cells Tabula Sapiens_TSP1423,171 cells Chen_parotid gland 113,265 cells Tabula Sapiens_TSP712,063 cells Costa-da-Silva_HV210,842 cells Horeth_Submandibular Gland 128,952 cells Williams_BM1688,738 cells Williams_BM1578,734 cells Ko_replicate 28,233 cells Williams_GM2387,436 cells Williams_GM1486,461 cells Costa-da-Silva_HV16,023 cells Williams_GM2835,974 cells Williams_GM2425,487 cells Horeth_Submandibular Gland 155,191 cells Williams_GM1834,564 cells Costa-da-Silva_HV44,345 cells Williams_GM2413,796 cells Williams_BM1693,512 cells Caetano_Healthy23,059 cells Williams_BM1583,032 cells Williams_GM2892,916 cells Costa-da-Silva_HV32,800 cells Williams_GM1432,559 cells Williams_GM1362,002 cells Williams_GM1841,991 cells Williams_BM1561,904 cells Williams_GM1441,626 cells Williams_BM1651,475 cells Opasawatchai_Donor C1,445 cells Pagella_Perio41,074 cells Opasawatchai_Donor A579 cells Caetano_Healthy1389 cells https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8359928/72,687 cells https://www.science.org/stoken/author-tokens/ST-495/full35,234 cells https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8099559/28,775 cells https://www.sciencedirect.com/science/article/pii/S2589004221015625?via%3Dihub24,010 cells https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7511944/22,530 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%200pubmed14,143 cells https://www.nature.com/articles/s41591-021-01296-813,639 cells https://journals.sagepub.com/doi/full/10.1177/0022034522107606913,265 cells https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9827523/8,807 cells https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7781605/3,448 cells https://www.frontiersin.org/articles/10.3389/fdmed.2021.806294/full2,024 cells National Institute of Dental and Craniofacial Research (NIDCR)112,546 cells Chan-Zuckerberg BioHub35,234 cells University of Zurich26,411 cells Medical University of Vienna22,530 cells University at Buffalo14,143 cells Sichuan University13,265 cells University of Pennsylvania8,807 cells University Medical Center Groningen4,496 cells King's College London3,448 cells University of Zurich (UZH)2,364 cells Mahidol University2,024 cells MSNZ University of Würzburg835 cells site_1_Switzerland28,775 cells site_1_Austria22,530 cells site_1_Netherlands4,496 cells site_1_Thailand2,024 cells parotid gland 113,265 cells Submandibular Gland 12 scRNA8,952 cells Submandibular Gland 15 scRNA5,191 cells not applicable210,869 cells surgical donor210,869 cells postmortem donor35,234 cells surgical resection42,102 cells Labial minor salivary gland24,010 cells Parotid salivary gland17,718 cells Submandibular gland14,143 cells Lower pole of the parotid salivary gland 13,265 cells Posterior tongue: circumvallate papilla anterior to sulcus terminalis9,900 cells Rugous Area anterior Hard palate8,807 cells Mandibular labial minor salivary glands7,463 cells Anterior tongue: tongue dorsum 6,932 cells Palatal side, interdental papilla #14 distal/15 mesial 6,176 cells Sublingual gland5,180 cells Buccal gingival margin UR3/UL33,059 cells Mandibular labial mucosa3,045 cells Apical two-thirds of the roots of extracted third molars2,364 cells Tooh number 18 (upper maxillary right third molar)2,024 cells Buccal gingival margin UR6
389 cells frozen at -80C6,706 cells STAR v2.7.10a_alpha_220818 / STARsolo246,103 cells American Indian1,991 cells epithelial cell62,303 cells endothelial cell of vascular tree43,549 cells minor salivary gland31,473 cells parotid gland30,983 cells buccal mucosa27,875 cells mucosa of dorsum of tongue16,832 cells submandibular gland14,143 cells sublingual gland5,180 cells Hispanic or Latin American2,002 cells Native American1,991 cells 
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Vascular endothelial cells23,920 cells Lymphatic endothelial cells2,278 cells Monocytes/Macrophages1,925 cells Natural Killer cells1,604 cells Dendritic cells1,497 cells Langerhans cells618 cells Ductal epithelial cells265 cells Myoepithelial cells26 cells Williams et al.72,100 cells Caetano et al.3,448 cells Warner, et al.2,189 cells Kretzschmar et al.831 cells Gingival Mucosa48,259 cells Buccal Mucosa27,289 cells Williams_BM1578,611 cells Williams_BM1688,456 cells Ko_replicate 27,863 cells Williams_GM2387,435 cells Williams_GM1486,461 cells Williams_GM2835,974 cells Williams_GM2425,487 cells Williams_GM1834,564 cells Williams_GM2413,796 cells Williams_BM1693,425 cells Caetano_Healthy23,059 cells Williams_BM1583,012 cells Williams_GM2892,916 cells Williams_GM1432,559 cells Williams_GM1362,002 cells Williams_GM1841,991 cells Williams_BM1561,870 cells Williams_GM1441,626 cells Williams_BM1651,455 cells Caetano_Healthy1389 cells https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8359928/72,100 cells https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9827523/8,432 cells https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7781605/3,448 cells National Institute of Dental and Craniofacial Research (NIDCR)74,289 cells University of Pennsylvania8,432 cells King's College London3,448 cells MSNZ University of Würzburg831 cells not applicable87,000 cells surgical donor87,000 cells surgical resection4,279 cells Rugous Area anterior Hard palate8,432 cells Buccal gingival margin UR3/UL33,059 cells Mandibular labial mucosa3,020 cells Buccal gingival margin UR6
389 cells frozen at -80C2,189 cells STAR v2.7.10a_alpha_220818 / STARsolo87,000 cells American Indian1,991 cells blood vessel endothelial cell23,920 cells CD4-positive helper T cell7,644 cells CD8-positive, alpha-beta T cell5,367 cells endothelial cell of lymphatic vessel2,278 cells natural killer cell1,604 cells dendritic cell1,497 cells acinar cell of salivary gland558 cells salivary gland glandular cell265 cells adult skeletal muscle myoblast51 cells myoepithelial cell26 cells buccal mucosa27,289 cells Hispanic or Latin American2,002 cells Native American1,991 cells 
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Vascular endothelial cells10,658 cells Monocytes/Macrophages1,555 cells Natural Killer cells217 cells Peripherial neurons51 cells Pagella et al.26,373 cells Krivanek et al.22,530 cells Opasawatchai et al.2,024 cells Opasawatchai_Donor C1,445 cells Opasawatchai_Donor A579 cells https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8099559/26,373 cells https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7511944/22,530 cells https://www.frontiersin.org/articles/10.3389/fdmed.2021.806294/full2,024 cells University of Zurich26,373 cells Medical University of Vienna22,530 cells Mahidol University2,024 cells site_1_Switzerland26,373 cells site_1_Austria22,530 cells site_1_Thailand2,024 cells not applicable50,927 cells surgical donor50,927 cells surgical resection24,554 cells Tooh number 18 (upper maxillary right third molar)2,024 cells STAR v2.7.10a_alpha_220818 / STARsolo50,927 cells blood vessel endothelial cell10,658 cells CD4-positive helper T cell1,060 cells CD8-positive, alpha-beta T cell1,053 cells natural killer cell217 cells peripheral nervous system neuron51 cells 
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Ductal epithelial cells3,846 cells Vascular endothelial cells2,205 cells Monocytes/Macrophages1,092 cells Myoepithelial cells220 cells Natural Killer cells155 cells Lymphatic endothelial cells81 cells Costa-da-Silva et al.24,008 cells Horeth et al.14,142 cells Pringle et al.4,484 cells Minor Salivary Gland31,468 cells Submandibular Gland14,142 cells Parotid Salivary Gland4,630 cells Costa-da-Silva_HV210,842 cells Horeth_Submandibular Gland 128,952 cells Costa-da-Silva_HV16,023 cells Horeth_Submandibular Gland 155,190 cells Costa-da-Silva_HV44,345 cells Costa-da-Silva_HV32,798 cells Tabula Sapiens_TSP14104 cells Tabula Sapiens_TSP742 cells https://www.sciencedirect.com/science/article/pii/S2589004221015625?via%3Dihub24,008 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%200pubmed14,142 cells https://www.nature.com/articles/s41591-021-01296-87,460 cells https://www.science.org/stoken/author-tokens/ST-495/full146 cells National Institute of Dental and Craniofacial Research (NIDCR)31,468 cells University at Buffalo14,142 cells University Medical Center Groningen4,484 cells Chan-Zuckerberg BioHub146 cells site_1_Netherlands4,484 cells Submandibular Gland 12 scRNA8,952 cells Submandibular Gland 15 scRNA5,190 cells not applicable50,094 cells surgical donor50,094 cells postmortem donor146 cells Labial minor salivary gland24,008 cells Submandibular gland14,142 cells Mandibular labial minor salivary glands7,460 cells Parotid salivary gland4,630 cells frozen at -80C4,484 cells STAR v2.7.10a_alpha_220818 / STARsolo50,240 cells acinar cell of salivary gland29,997 cells salivary gland glandular cell3,846 cells blood vessel endothelial cell2,205 cells CD8-positive, alpha-beta T cell2,063 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 gland31,468 cells submandibular gland14,142 cells 
5. Minor Salivary Gland Atlas
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Ductal epithelial cells2,361 cells Vascular endothelial cells1,513 cells Myoepithelial cells431 cells Monocytes/Macrophages416 cells Lymphatic endothelial cells78 cells Natural Killer cells31 cells Costa-da-Silva et al.24,008 cells Minor Salivary Gland31,468 cells Costa-da-Silva_HV210,842 cells Costa-da-Silva_HV16,023 cells Costa-da-Silva_HV44,345 cells Costa-da-Silva_HV32,798 cells https://www.sciencedirect.com/science/article/pii/S2589004221015625?via%3Dihub24,008 cells https://www.nature.com/articles/s41591-021-01296-87,460 cells National Institute of Dental and Craniofacial Research (NIDCR)31,468 cells not applicable31,468 cells surgical donor31,468 cells Labial minor salivary gland24,008 cells Mandibular labial minor salivary glands7,460 cells STAR v2.7.10a_alpha_220818 / STARsolo31,468 cells acinar cell of salivary gland18,786 cells salivary gland glandular cell2,361 cells blood vessel endothelial cell1,513 cells CD8-positive, alpha-beta T cell1,025 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 gland31,468 cells 
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Vascular endothelial cells9,788 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.27,289 cells Warner, et al.2,189 cells Buccal Mucosa27,289 cells Williams_BM1578,611 cells Williams_BM1688,456 cells Williams_BM1693,425 cells Williams_BM1583,012 cells Williams_BM1561,870 cells Williams_BM1651,455 cells https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8359928/27,289 cells National Institute of Dental and Craniofacial Research (NIDCR)29,478 cells not applicable29,478 cells surgical donor29,478 cells Mandibular labial mucosa2,189 cells frozen at -80C2,189 cells STAR v2.7.10a_alpha_220818 / STARsolo29,478 cells blood vessel endothelial cell9,788 cells CD4-positive helper T cell2,167 cells CD8-positive, alpha-beta T cell1,932 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 buccal mucosa27,289 cells 
7. Major Salivary Gland Atlas
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Ductal epithelial cells1,485 cells Vascular endothelial cells741 cells Monocytes/Macrophages676 cells Myoepithelial cells381 cells Natural Killer cells124 cells Horeth et al.14,142 cells Pringle et al.4,484 cells Submandibular Gland14,142 cells Parotid Salivary Gland4,630 cells Horeth_Submandibular Gland 128,952 cells Horeth_Submandibular Gland 155,190 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%200pubmed14,142 cells https://www.science.org/stoken/author-tokens/ST-495/full146 cells University at Buffalo14,142 cells University Medical Center Groningen4,484 cells Chan-Zuckerberg BioHub146 cells site_1_Netherlands4,484 cells Submandibular Gland 12 scRNA8,952 cells Submandibular Gland 15 scRNA5,190 cells not applicable18,626 cells surgical donor18,626 cells postmortem donor146 cells Submandibular gland14,142 cells Parotid salivary gland4,630 cells frozen at -80C4,484 cells STAR v2.7.10a_alpha_220818 / STARsolo18,772 cells acinar cell of salivary gland11,211 cells salivary gland glandular cell1,485 cells CD8-positive, alpha-beta T cell1,038 cells blood vessel endothelial cell741 cells myoepithelial cell381 cells CD4-positive helper T cell357 cells natural killer cell124 cells submandibular gland14,142 cells 
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Vascular endothelial cells1,057 cells Monocytes/Macrophages203 cells Lymphatic endothelial cells190 cells Natural Killer cells154 cells Kretzschmar et al.831 cells Ko_replicate 27,863 cells https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9827523/8,432 cells University of Pennsylvania8,432 cells MSNZ University of Würzburg831 cells not applicable9,263 cells surgical donor9,263 cells surgical resection831 cells Rugous Area anterior Hard palate8,432 cells Mandibular labial mucosa831 cells STAR v2.7.10a_alpha_220818 / STARsolo9,263 cells blood vessel endothelial cell1,057 cells CD4-positive helper T cell858 cells CD8-positive, alpha-beta T cell412 cells endothelial cell of lymphatic vessel190 cells natural killer cell154 cells 
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Innate lymphoid cells2,149 cells Dendritic cells1,928 cells Monocytes/Macrophages1,878 cells Williams et al.17,354 cells Kretzschmar et al.104 cells Gingival Mucosa15,490 cells Williams_GM2382,905 cells Williams_GM2832,657 cells Williams_BM1572,119 cells Ko_replicate 21,763 cells Williams_GM2421,274 cells Williams_BM1581,196 cells Williams_BM1681,018 cells Williams_GM1831,012 cells Caetano_Healthy2524 cells Caetano_Healthy1228 cells https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8359928/17,354 cells https://www.nature.com/articles/s41591-021-01296-82,979 cells https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9827523/1,875 cells https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7781605/752 cells National Institute of Dental and Craniofacial Research (NIDCR)20,333 cells University of Pennsylvania1,875 cells King's College London752 cells MSNZ University of Würzburg104 cells not applicable23,064 cells surgical donor23,064 cells surgical resection856 cells Palatal side, interdental papilla #14 distal/15 mesial 2,979 cells Rugous Area anterior Hard palate1,875 cells Buccal gingival margin UR3/UL3524 cells Buccal gingival margin UR6
228 cells Mandibular labial mucosa104 cells STAR v2.7.10a_alpha_220818 / STARsolo23,064 cells CD8-positive, alpha-beta T cell6,617 cells CD4-positive helper T cell6,151 cells innate lymphoid cell2,149 cells dendritic cell1,928 cells Hispanic or Latin American500 cells 
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Monocytes/Macrophages832 cells Innate lymphoid cells146 cells Costa-da-Silva et al.2,388 cells Pringle et al.1,832 cells Minor Salivary Gland3,764 cells Parotid Salivary Gland1,832 cells Submandibular Gland766 cells Costa-da-Silva_HV11,179 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%3Dihub2,388 cells https://www.nature.com/articles/s41591-021-01296-81,376 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)3,764 cells University Medical Center Groningen1,832 cells University at Buffalo766 cells site_1_Netherlands1,832 cells Submandibular Gland 12 scRNA399 cells Submandibular Gland 15 scRNA367 cells not applicable6,362 cells surgical donor6,362 cells Labial minor salivary gland2,388 cells Parotid salivary gland1,832 cells Mandibular labial minor salivary glands1,376 cells Submandibular gland766 cells frozen at -80C1,832 cells STAR v2.7.10a_alpha_220818 / STARsolo6,362 cells CD8-positive, alpha-beta T cell1,325 cells CD4-positive helper T cell587 cells innate lymphoid cell146 cells minor salivary gland3,764 cells submandibular gland766 cells 
Alias names
Human Oral and Craniofacial Cell Atlas