{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,5]],"date-time":"2026-09-05T06:10:59Z","timestamp":1788588659032,"version":"build-2803163510"},"reference-count":75,"publisher":"National Academy of Sciences","issue":"35","license":[{"start":{"date-parts":[[2026,8,25]],"date-time":"2026-08-25T00:00:00Z","timestamp":1787616000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001659","name":"Deutsche Forschungsgemeinschaft","doi-asserted-by":"publisher","award":["450648163"],"award-info":[{"award-number":["450648163"]}],"id":[{"id":"10.13039\/501100001659","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001659","name":"Deutsche Forschungsgemeinschaft","doi-asserted-by":"publisher","award":["INST 161\/778-1 FuGG"],"award-info":[{"award-number":["INST 161\/778-1 FuGG"]}],"id":[{"id":"10.13039\/501100001659","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100000781","name":"EC | European Research Council","doi-asserted-by":"publisher","award":["101141396"],"award-info":[{"award-number":["101141396"]}],"id":[{"id":"10.13039\/501100000781","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001659","name":"Deutsche Forschungsgemeinschaft","doi-asserted-by":"publisher","award":["TA157\/12-1"],"award-info":[{"award-number":["TA157\/12-1"]}],"id":[{"id":"10.13039\/501100001659","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["www.pnas.org"],"crossmark-restriction":true},"short-container-title":["Proc. Natl. Acad. Sci. U.S.A."],"published-print":{"date-parts":[[2026,9,1]]},"abstract":"<jats:p>Major histocompatibility complex class I (MHC I) molecules present antigenic peptides to cytotoxic T cells, a process central to immune surveillance. However, the nanoscale spatial organization of peptide\u2013MHC I (pMHC I) on human dendritic cells (DCs), key initiators of cytotoxic T cell responses, remains largely unexplored. Here, we combine high-affinity soluble T cell receptors with DNA-based point accumulation for imaging in nanoscale topography (DNA-PAINT) to quantitatively map and count defined pMHC I complexes at single-molecule resolution on HLA-A*02:01-expressing cells and primary human monocyte-derived DCs. We found no evidence for higher-order pMHC I nanoclusters under conditions of extracellular peptide exchange or physiological intracellular loading. Instead, detected signals correspond to individual pMHC I complexes. Notably, DC differentiation and activation modulate pMHC I surface abundance and spatial compartmentalization. These findings refine current models of antigen presentation by emphasizing regulation through surface density and spatial distribution, and establish a quantitative framework for epitope-specific, single-molecule quantification of antigen presentation in human immune cells.<\/jats:p>","DOI":"10.1073\/pnas.2615838123","type":"journal-article","created":{"date-parts":[[2026,8,25]],"date-time":"2026-08-25T16:36:09Z","timestamp":1787675769000},"update-policy":"https:\/\/doi.org\/10.1073\/pnas.cm10313","source":"Crossref","is-referenced-by-count":1,"title":["Quantitative nanoscale imaging shows peptide\u2013MHC I complexes are monomeric and spatially regulated in human dendritic cells"],"prefix":"10.1073","volume":"123","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-4443-9632","authenticated-orcid":false,"given":"Olivia","family":"Jacobs","sequence":"first","affiliation":[{"id":[{"id":"https:\/\/ror.org\/04cvxnb49","id-type":"ROR","asserted-by":"publisher"}],"name":"Institute of Biochemistry, Biocenter, Goethe University Frankfurt","place":["Frankfurt, Germany"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5615-4170","authenticated-orcid":false,"given":"Tanja","family":"Menche","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/04cvxnb49","id-type":"ROR","asserted-by":"publisher"}],"name":"Institute of Physical and Theoretical Chemistry, Goethe University Frankfurt","place":["Frankfurt, Germany"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0000-8857-871X","authenticated-orcid":false,"given":"Cindy","family":"H\u00f6per","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/04cvxnb49","id-type":"ROR","asserted-by":"publisher"}],"name":"Institute of Biochemistry, Biocenter, Goethe University Frankfurt","place":["Frankfurt, Germany"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0000-8811-0148","authenticated-orcid":false,"given":"Fr\u00e9d\u00e9ric","family":"Gerhards","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/04cvxnb49","id-type":"ROR","asserted-by":"publisher"}],"name":"Institute of Biochemistry, Biocenter, Goethe University Frankfurt","place":["Frankfurt, Germany"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ivica","family":"Fucek","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/023b0x485","id-type":"ROR","asserted-by":"publisher"}],"name":"TRON gGmbH-Translational Oncology, Medical Center, Johannes Gutenberg University Mainz","place":["Mainz, Germany"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Fulvia","family":"Vascotto","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/023b0x485","id-type":"ROR","asserted-by":"publisher"}],"name":"TRON gGmbH-Translational Oncology, Medical Center, Johannes Gutenberg University Mainz","place":["Mainz, Germany"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0504-1824","authenticated-orcid":false,"given":"Marina S.","family":"Dietz","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/04cvxnb49","id-type":"ROR","asserted-by":"publisher"}],"name":"Institute of Physical and Theoretical Chemistry, Goethe University Frankfurt","place":["Frankfurt, Germany"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mike","family":"Heilemann","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/04cvxnb49","id-type":"ROR","asserted-by":"publisher"}],"name":"Institute of Physical and Theoretical Chemistry, Goethe University Frankfurt","place":["Frankfurt, Germany"]},{"id":[{"id":"https:\/\/ror.org\/04cvxnb49","id-type":"ROR","asserted-by":"publisher"}],"name":"Cluster of Excellence SubCellular Architecture of Life (SCALE), Goethe University Frankfurt","place":["Frankfurt, Germany"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0403-2160","authenticated-orcid":false,"given":"Robert","family":"Tamp\u00e9","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/04cvxnb49","id-type":"ROR","asserted-by":"publisher"}],"name":"Institute of Biochemistry, Biocenter, Goethe University Frankfurt","place":["Frankfurt, Germany"]},{"id":[{"id":"https:\/\/ror.org\/04cvxnb49","id-type":"ROR","asserted-by":"publisher"}],"name":"Cluster of Excellence SubCellular Architecture of Life (SCALE), Goethe University Frankfurt","place":["Frankfurt, Germany"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"341","published-online":{"date-parts":[[2026,8,25]]},"reference":[{"key":"e_1_3_4_1_2","doi-asserted-by":"publisher","DOI":"10.1146\/annurev-immunol-032712-095910"},{"key":"e_1_3_4_2_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.it.2016.08.010"},{"key":"e_1_3_4_3_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.coi.2021.02.004"},{"key":"e_1_3_4_4_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-022-32841-9"},{"key":"e_1_3_4_5_2","doi-asserted-by":"publisher","DOI":"10.1126\/sciadv.aea7735"},{"key":"e_1_3_4_6_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-022-32384-z"},{"key":"e_1_3_4_7_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.smim.2023.101713"},{"key":"e_1_3_4_8_2","doi-asserted-by":"publisher","DOI":"10.1146\/annurev-immunol-041015-055523"},{"key":"e_1_3_4_9_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41590-021-00903-7"},{"key":"e_1_3_4_10_2","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.271.24.13993"},{"key":"e_1_3_4_11_2","doi-asserted-by":"publisher","DOI":"10.1083\/jcb.140.1.71"},{"key":"e_1_3_4_12_2","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.166.2.800"},{"key":"e_1_3_4_13_2","doi-asserted-by":"publisher","DOI":"10.1016\/1074-7613(94)90040-X"},{"key":"e_1_3_4_14_2","doi-asserted-by":"publisher","DOI":"10.1002\/cpim.115"},{"key":"e_1_3_4_15_2","doi-asserted-by":"publisher","DOI":"10.1038\/ni1559"},{"key":"e_1_3_4_16_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.coi.2007.04.013"},{"key":"e_1_3_4_17_2","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.166.11.6625"},{"key":"e_1_3_4_18_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.2122188119"},{"key":"e_1_3_4_19_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1208696109"},{"key":"e_1_3_4_20_2","doi-asserted-by":"publisher","DOI":"10.1021\/acs.chemrev.6b00667"},{"key":"e_1_3_4_21_2","doi-asserted-by":"publisher","DOI":"10.1039\/C9NR06364A"},{"key":"e_1_3_4_22_2","doi-asserted-by":"publisher","DOI":"10.1038\/nprot.2017.024"},{"key":"e_1_3_4_23_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-021-21142-2"},{"key":"e_1_3_4_24_2","doi-asserted-by":"publisher","DOI":"10.3390\/biom11101503"},{"key":"e_1_3_4_25_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.celrep.2017.12.045"},{"key":"e_1_3_4_26_2","doi-asserted-by":"publisher","DOI":"10.1021\/nl103427w"},{"key":"e_1_3_4_27_2","doi-asserted-by":"publisher","DOI":"10.1117\/1.NPh.6.3.035008"},{"key":"e_1_3_4_28_2","doi-asserted-by":"crossref","unstructured":"E. Betzig Imaging Intracellular Fluorescent Proteins at Nanometer Resolution. Science 313 1642\u20131645 (2006).","DOI":"10.1126\/science.1127344"},{"key":"e_1_3_4_29_2","doi-asserted-by":"publisher","DOI":"10.1002\/anie.200802376"},{"key":"e_1_3_4_30_2","doi-asserted-by":"publisher","DOI":"10.1038\/nmeth.3804"},{"key":"e_1_3_4_31_2","doi-asserted-by":"publisher","DOI":"10.1146\/annurev-biophys-102521-112912"},{"key":"e_1_3_4_32_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-019-12301-7"},{"key":"e_1_3_4_33_2","doi-asserted-by":"publisher","DOI":"10.1002\/smtd.202401799"},{"key":"e_1_3_4_34_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.2321600121"},{"key":"e_1_3_4_35_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41571-018-0142-8"},{"key":"e_1_3_4_36_2","doi-asserted-by":"publisher","DOI":"10.1093\/protein\/gzm033"},{"key":"e_1_3_4_37_2","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M113.464560"},{"key":"e_1_3_4_38_2","doi-asserted-by":"publisher","DOI":"10.1038\/35008103"},{"key":"e_1_3_4_39_2","doi-asserted-by":"publisher","DOI":"10.1002\/ctm2.1384"},{"key":"e_1_3_4_40_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41586-025-10004-2"},{"key":"e_1_3_4_41_2","doi-asserted-by":"publisher","DOI":"10.1186\/s12943-021-01335-5"},{"key":"e_1_3_4_42_2","doi-asserted-by":"publisher","DOI":"10.1038\/nrd4278"},{"key":"e_1_3_4_43_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.aar7112"},{"key":"e_1_3_4_44_2","doi-asserted-by":"publisher","DOI":"10.1038\/mt.2008.200"},{"key":"e_1_3_4_45_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.2219790120"},{"key":"e_1_3_4_46_2","doi-asserted-by":"publisher","DOI":"10.3389\/fimmu.2019.00061"},{"key":"e_1_3_4_47_2","doi-asserted-by":"publisher","DOI":"10.1007\/s00018-022-04472-2"},{"key":"e_1_3_4_48_2","doi-asserted-by":"publisher","DOI":"10.1182\/bloodadvances.2018027268"},{"key":"e_1_3_4_49_2","doi-asserted-by":"publisher","DOI":"10.1126\/sciadv.aeb0821"},{"key":"e_1_3_4_50_2","doi-asserted-by":"publisher","DOI":"10.3389\/fimmu.2022.983000"},{"key":"e_1_3_4_51_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-024-50774-3"},{"key":"e_1_3_4_52_2","doi-asserted-by":"publisher","DOI":"10.1158\/2326-6066.CIR-24-0320"},{"key":"e_1_3_4_53_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.coi.2019.12.003"},{"key":"e_1_3_4_54_2","doi-asserted-by":"publisher","DOI":"10.3389\/fimmu.2025.1563789"},{"key":"e_1_3_4_55_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.molimm.2008.06.009"},{"key":"e_1_3_4_56_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.2013554117"},{"key":"e_1_3_4_57_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41592-024-02242-5"},{"key":"e_1_3_4_58_2","doi-asserted-by":"publisher","DOI":"10.1158\/1535-7163.MCT-21-0115"},{"key":"e_1_3_4_59_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41573-023-00809-z"},{"key":"e_1_3_4_60_2","doi-asserted-by":"publisher","DOI":"10.1039\/C6SC05420J"},{"key":"e_1_3_4_61_2","doi-asserted-by":"publisher","DOI":"10.1002\/eji.202048599"},{"key":"e_1_3_4_62_2","doi-asserted-by":"crossref","unstructured":"M. -L. I. E. Harwardt Single-molecule super-resolution microscopy reveals heteromeric complexes of MET and EGFR upon Ligand Activation. Int. J. Mol. Sci. 21 2803 (2020).","DOI":"10.3390\/ijms21082803"},{"key":"e_1_3_4_63_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41586-021-03275-y"},{"key":"e_1_3_4_64_2","doi-asserted-by":"publisher","DOI":"10.1182\/blood-2006-04-015024"},{"key":"e_1_3_4_65_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.ymthe.2018.12.011"},{"key":"e_1_3_4_66_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41592-020-0869-x"},{"key":"e_1_3_4_67_2","article-title":"Computer control of microscopes using microManager","volume":"14","author":"Edelstein A.","year":"2010","unstructured":"A. Edelstein, N. Amodaj, K. Hoover, R. Vale, N. Stuurman, Computer control of microscopes using microManager. Curr. Protoc. Mol. Biol. Chapter 14, Unit14.20 (2010).","journal-title":"Curr. Protoc. Mol. Biol."},{"key":"e_1_3_4_68_2","doi-asserted-by":"publisher","DOI":"10.1038\/nmeth.2019"},{"key":"e_1_3_4_69_2","doi-asserted-by":"publisher","DOI":"10.1038\/nmeth.1449"},{"key":"e_1_3_4_70_2","doi-asserted-by":"publisher","DOI":"10.1364\/OE.22.015982"},{"key":"e_1_3_4_71_2","unstructured":"M. Ester H. Kriegel J. Sander X. Xu \u201cA density-based algorithm for discovering clusters in large spatial databases with noise\u201d in KDD\u201996 (AAI Press 1996) pp. 226\u2013231."},{"key":"e_1_3_4_72_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41586-023-05925-9"},{"key":"e_1_3_4_73_2","doi-asserted-by":"publisher","DOI":"10.1007\/s00418-014-1192-3"},{"key":"e_1_3_4_74_2","doi-asserted-by":"crossref","unstructured":"T. Menche M. S. Dietz M. Heilemann Data from \u201cQuantitative nanoscale imaging of peptide\u2013MHC I complexes in HAP1 cells and human dendritic cells.\u201d BioImage Archive. https:\/\/doi.org\/10.6019\/S-BIAD2942. Deposited 5 May 2026.","DOI":"10.6019\/S-BIAD2942"},{"key":"e_1_3_4_75_2","doi-asserted-by":"crossref","unstructured":"O. Jacobs . Data from \u201cQuantitative nanoscale imaging shows peptide\u2013MHC I complexes are monomeric and spatially regulated in human dendritic cells.\u201d Goethe University Data Repository. https:\/\/doi.org\/10.25716\/gude.0pc9-s4ze. Deposited 13 July 2026.","DOI":"10.1073\/pnas.2615838123"}],"container-title":["Proceedings of the National Academy of Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/pnas.org\/doi\/pdf\/10.1073\/pnas.2615838123","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,9,4]],"date-time":"2026-09-04T18:30:55Z","timestamp":1788546655000},"score":1,"resource":{"primary":{"URL":"https:\/\/pnas.org\/doi\/10.1073\/pnas.2615838123"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,8,25]]},"references-count":75,"journal-issue":{"issue":"35","published-print":{"date-parts":[[2026,9,1]]}},"alternative-id":["10.1073\/pnas.2615838123"],"URL":"https:\/\/doi.org\/10.1073\/pnas.2615838123","relation":{},"ISSN":["0027-8424","1091-6490"],"issn-type":[{"value":"0027-8424","type":"print"},{"value":"1091-6490","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,8,25]]},"assertion":[{"value":"2026-05-05","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2026-07-30","order":2,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2026-08-25","order":3,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}],"article-number":"e2615838123"}}