{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,23]],"date-time":"2026-06-23T17:08:13Z","timestamp":1782234493454,"version":"3.54.5"},"reference-count":39,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2025,2,26]],"date-time":"2025-02-26T00:00:00Z","timestamp":1740528000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Immunol."],"abstract":"<jats:sec><jats:title>Introduction<\/jats:title><jats:p>The molecular pathogenesis of ocular adnexal mucosa-associated lymphoid tissue (MALT) lymphoma and IgG4-related ophthalmic disease (IgG4-ROD) remains incompletely understood. Differentiating between the two diseases is vital given that the diagnostic evaluation and treatment approaches can vary significantly; this difficulty in distinction is exacerbated by the absence of specific biomarkers. This study aimed to investigate the differences between these two diseases based on their cellular composition, transcriptional heterogeneity, and the immune microenvironment using single-cell RNA transcriptional sequencing (scRNA-seq) technology.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>We collected orbital lacrimal gland region tissue samples from three patients with MALT lymphoma and another three with IgG4-ROD and performed single-cell sequencing experiments. Subsequently, we conducted bioinformatics analyses, including cell subpopulation segmentation and inter-group comparison, tumor cell identification, functional enrichment analysis, and pseudotime trajectory analysis. Furthermore, we analyzed the cellular communication between tumor B-cell and T-cell subsets within the immune microenvironment of MALT lymphoma tissues. We performed immunofluorescence assays to verify the co-expression of receptor-ligand pairs.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>A total of six major cell subpopulations were identified, with B-cells and T-cells being the predominant cell types. All B-cell subpopulations in MALT lymphomas are malignant, exhibiting significant intratumoral and intertumoral heterogeneity. Reclustering of the T-cell subpopulation identified five major T-cell subpopulations. Pseudotime analysis revealed that CD4+ naive T-cells in MALT lymphoma patients were highly likely to differentiate into follicular helper T-cells, whereas, in IgG4-ROD patients, CD4+ naive T-cells were highly likely to differentiate into regulatory T-cells. Intercellular communication analysis revealed that the CD27-CD70 immune checkpoint receptor\u2212ligand pair and CXCL13-CXCR5 chemokine receptor\u2212ligand pair were significantly upregulated between malignant B-cells and T-cells subpopulations.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusion<\/jats:title><jats:p>This study is the first to conduct a comparative single-cell transcriptome sequencing analysis of ocular adnexal MALT lymphoma and IgG4-ROD. Our results reveal the cellular composition, key pathways, and critical immune microenvironment implicated in the development of these two diseases. These findings provide important insights into the pathogenesis of these two diseases and highlight the differences between them.<\/jats:p><\/jats:sec>","DOI":"10.3389\/fimmu.2025.1508559","type":"journal-article","created":{"date-parts":[[2025,2,26]],"date-time":"2025-02-26T09:06:16Z","timestamp":1740560776000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":6,"title":["Single-cell RNA-sequencing reveals cellular heterogeneity and immune microenvironment characteristics between ocular adnexal mucosa-associated lymphoid lymphoma and IgG4\u2212related ophthalmic disease"],"prefix":"10.3389","volume":"16","author":[{"given":"Yu","family":"Yang","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yujiao","family":"Wang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xuelian","family":"Jin","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Weimin","family":"He","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2025,2,26]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","DOI":"10.1097\/IOP.0b013e31821cc4bb","article-title":"Primary, unilateral ocular adnexal lymphoma: disease progression and long-term survival","volume":"27","author":"Rootman","year":"2011","journal-title":"Ophthalmic Plast reconstructive Surg"},{"key":"B2","doi-asserted-by":"publisher","first-page":"45","DOI":"10.1016\/j.survophthal.2018.08.002","article-title":"Orbital lymphoma","volume":"64","author":"Olsen","year":"2019","journal-title":"Survey Ophthalmol"},{"key":"B3","doi-asserted-by":"publisher","first-page":"2538","DOI":"10.3390\/cancers12092538","article-title":"Orbital and eyelid B-cell lymphoma: A multicenter retrospective study","volume":"12","author":"Savino","year":"2020","journal-title":"Cancers"},{"key":"B4","doi-asserted-by":"publisher","DOI":"10.1016\/j.beha.2016.11.002","article-title":"Ocular adnexal marginal zone lymphoma: Clinical presentation, pathogenesis, diagnosis, prognosis, and treatment","volume":"30","author":"Sassone","year":"2017","journal-title":"Best Pract Res Clin haematology"},{"key":"B5","doi-asserted-by":"publisher","DOI":"10.1007\/s10147-019-01481-3","article-title":"Treatment-associated outcomes of patients with primary ocular adnexal MALT lymphoma after accurate diagnosis","volume":"24","author":"Masuda","year":"2019","journal-title":"Int J Clin Oncol"},{"key":"B6","doi-asserted-by":"publisher","DOI":"10.1136\/bjo-2022-321656","article-title":"Predicting the risk of distant and local recurrence for patients with ocular adnexal extranodal marginal zone lymphoma: a matched case-control study","volume":"107","author":"Shi","year":"2023","journal-title":"Br J Ophthalmol"},{"key":"B7","doi-asserted-by":"publisher","DOI":"10.1002\/cam4.v12.3","article-title":"Long-term survival outcomes of patients with primary ocular adnexal MALT lymphoma: A large single-center cohort study","volume":"12","author":"Liang","year":"2023","journal-title":"Cancer Med"},{"key":"B8","doi-asserted-by":"publisher","DOI":"10.1002\/ijc.v154.6","article-title":"Transformation to diffuse large B-cell lymphoma and its impact on survival in patients with marginal zone lymphoma: A population-based study","volume":"154","author":"Du","year":"2024","journal-title":"Int J Cancer"},{"key":"B9","doi-asserted-by":"publisher","first-page":"176","DOI":"10.3390\/cancers14010176","article-title":"Recent advances in the genetic of MALT lymphomas","volume":"14","author":"Rodr\u00edguez-Sevilla","year":"2021","journal-title":"Cancers"},{"key":"B10","doi-asserted-by":"publisher","first-page":"1264","DOI":"10.3390\/cancers14051264","article-title":"The biology of ocular adnexal marginal zone lymphomas","volume":"14","author":"Johansson","year":"2022","journal-title":"Cancers"},{"key":"B11","doi-asserted-by":"publisher","first-page":"3","DOI":"10.12701\/yujm.2021.01263","article-title":"Ocular adnexal mucosa-associated lymphoid tissue lymphoma: a narrative review","volume":"39","author":"Chung","year":"2022","journal-title":"J Yeungnam Med Sci"},{"key":"B12","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0131458","article-title":"Clinicopathological analysis of ocular adnexal extranodal marginal zone B-cell lymphoma with igG4-positive cells","volume":"10","author":"Lee","year":"2015","journal-title":"PloS One"},{"key":"B13","doi-asserted-by":"publisher","DOI":"10.1097\/IOP.0000000000000965","article-title":"Immunoglobulin G4 (IgG4)-positive ocular adnexal mucosa-associated lymphoid tissue lymphoma and idiopathic orbital inflammation","volume":"34","author":"Sohn","year":"2018","journal-title":"Ophthalmic Plast reconstructive Surg"},{"key":"B14","doi-asserted-by":"publisher","DOI":"10.2147\/OTT.S242852","article-title":"The expression of igG and igG4 in orbital MALT lymphoma: the similarities and differences of igG4-related diseases","volume":"13","author":"Li","year":"2020","journal-title":"OncoTargets Ther"},{"key":"B15","doi-asserted-by":"publisher","first-page":"9","DOI":"10.4103\/tjo.tjo_12_17","article-title":"Immunoglobulin G4-related ophthalmic disease","volume":"8","author":"Yu","year":"2018","journal-title":"Taiwan J Ophthalmol"},{"key":"B16","doi-asserted-by":"publisher","first-page":"1660","DOI":"10.3390\/life12101660","article-title":"Novel insights into pathophysiology of orbital inflammatory diseases and progression to orbital lymphoma by pathway enrichment analysis","volume":"12","author":"Al-Ghazzawi","year":"2022","journal-title":"Life (Basel Switzerland)"},{"key":"B17","doi-asserted-by":"publisher","DOI":"10.3960\/jslrt.52.51","article-title":"Ocular adnexal IgG4-producing mucosa-associated lymphoid tissue lymphoma mimicking IgG4-related disease","volume":"52","author":"Sato","year":"2012","journal-title":"J Clin Exp hematopathology: JCEH"},{"key":"B18","doi-asserted-by":"publisher","DOI":"10.1007\/s10384-017-0513-1","article-title":"Immunophenotypic profiles for distinguishing orbital mucosa-associated lymphoid tissue lymphoma from benign lymphoproliferative tumors","volume":"61","author":"Ueda","year":"2017","journal-title":"Japanese J Ophthalmol"},{"key":"B19","doi-asserted-by":"publisher","DOI":"10.21873\/anticanres.11913","article-title":"Usefulness of flow cytometry in diagnosis of igG4-related ophthalmic disease and extranodal marginal zone B-cell lymphoma of the ocular adnexa","volume":"37","author":"Kase","year":"2017","journal-title":"Anticancer Res"},{"key":"B20","doi-asserted-by":"publisher","first-page":"3458","DOI":"10.3390\/jcm9113458","article-title":"Comprehensive gene analysis of igG4-related ophthalmic disease using RNA sequencing","volume":"9","author":"Asakage","year":"2020","journal-title":"J Clin Med"},{"key":"B21","doi-asserted-by":"publisher","first-page":"2530","DOI":"10.3390\/jcm9082530","article-title":"Distinctive tissue and serum microRNA profile of igG4-related ophthalmic disease and MALT lymphoma","volume":"9","author":"Nezu","year":"2020","journal-title":"J Clin Med"},{"key":"B22","doi-asserted-by":"publisher","DOI":"10.1038\/s41581-018-0021-7","article-title":"Single-cell RNA sequencing for the study of development, physiology and disease","volume":"14","author":"Potter","year":"2018","journal-title":"Nat Rev Nephrol"},{"key":"B23","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1007\/s10384-014-0352-2","article-title":"Diagnostic criteria for IgG4-related ophthalmic disease [J","volume":"59","author":"Goto","year":"2015","journal-title":"Japanese J Ophthalmol"},{"key":"B24","doi-asserted-by":"publisher","DOI":"10.1111\/j.1365-2141.2011.08841.x","article-title":"Genomic analysis of non-splenic marginal zone lymphomas (MZL) indicates similarities between nodal and extranodal MZL and supports their derivation from memory B-cells","volume":"155","author":"Novak","year":"2011","journal-title":"Br J haematology"},{"key":"B25","doi-asserted-by":"publisher","DOI":"10.1097\/IOP.0000000000000363","article-title":"IgG4-related ophthalmic disease. Part I: background and pathology","volume":"31","author":"Mcnab","year":"2015","journal-title":"Ophthalmic Plast reconstructive Surg"},{"key":"B26","doi-asserted-by":"publisher","DOI":"10.1007\/s13277-014-1999-5","article-title":"Alterations of circulating follicular helper T cells and interleukin 21 in diffuse large B-cell lymphoma","volume":"35","author":"Cha","year":"2014","journal-title":"Tumour biology: J Int Soc Oncodevelopmental Biol Med"},{"key":"B27","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0190468","article-title":"Circulating Tfh1 (cTfh1) cell numbers and PD1 expression are elevated in low-grade B-cell non-Hodgkin's lymphoma and cTfh gene expression is perturbed in marginal zone lymphoma","volume":"13","author":"Byford","year":"2018","journal-title":"PloS One"},{"key":"B28","doi-asserted-by":"publisher","DOI":"10.1056\/NEJMoa1606774","article-title":"Pembrolizumab versus chemotherapy for PD-L1-positive non-small-cell lung cancer","volume":"375","author":"Reck","year":"2016","journal-title":"New Engl J Med"},{"key":"B29","doi-asserted-by":"publisher","DOI":"10.2217\/imt-2016-0025","article-title":"PD-L1 expression and outcome of advanced melanoma patients treated with anti-PD-1\/PD-L1 agents: a meta-analysis","volume":"8","author":"Abdel-Rahman","year":"2016","journal-title":"Immunotherapy"},{"key":"B30","doi-asserted-by":"publisher","DOI":"10.17305\/bjbms.2019.4204","article-title":"Targeted immunotherapy with a checkpoint inhibitor in combination with chemotherapy: A new clinical paradigm in the treatment of triple-negative breast cancer","volume":"19","author":"Cyprian","year":"2019","journal-title":"Bosnian J basic Med Sci"},{"key":"B31","doi-asserted-by":"publisher","DOI":"10.1001\/jamaoncol.2019.3343","article-title":"Assessment of combined nivolumab and bevacizumab in relapsed ovarian cancer: A phase 2 clinical trial","volume":"5","author":"Liu","year":"2019","journal-title":"JAMA Oncol"},{"key":"B32","doi-asserted-by":"publisher","DOI":"10.1016\/S1470-2045(16)30167-X","article-title":"Nivolumab for classical Hodgkin's lymphoma after failure of both autologous stem-cell transplantation and brentuximab vedotin: a multicentre, multicohort, single-arm phase 2 trial","volume":"17","author":"Younes","year":"2016","journal-title":"Lancet Oncol"},{"key":"B33","doi-asserted-by":"publisher","DOI":"10.1002\/med.21765","article-title":"Understanding the tumor microenvironment for effective immunotherapy","volume":"41","author":"Sadeghi Rad","year":"2021","journal-title":"Medicinal Res Rev"},{"key":"B34","doi-asserted-by":"publisher","first-page":"bbac234","DOI":"10.1093\/bib\/bbac234","article-title":"Cell-cell communication inference and analysis in the tumour microenvironments from single-cell transcriptomics: data resources and computational strategies","volume":"23","author":"Peng","year":"2022","journal-title":"Briefings Bioinf"},{"key":"B35","doi-asserted-by":"publisher","DOI":"10.1182\/blood.2020010527","article-title":"SOX11, CD70, and Treg cells configure the tumor-immune microenvironment of aggressive mantle cell lymphoma","volume":"138","author":"Balsas","year":"2021","journal-title":"Blood"},{"key":"B36","doi-asserted-by":"publisher","first-page":"110713","DOI":"10.1016\/j.celrep.2022.110713","article-title":"A single-cell atlas of diffuse large B cell lymphoma","volume":"39","author":"Ye","year":"2022","journal-title":"Cell Rep"},{"key":"B37","doi-asserted-by":"publisher","DOI":"10.1002\/cncr.v128.5","article-title":"Cusatuzumab for treatment of CD70-positive relapsed or refractory cutaneous T-cell lymphoma","volume":"128","author":"Leupin","year":"2022","journal-title":"Cancer"},{"key":"B38","doi-asserted-by":"publisher","DOI":"10.1016\/j.ajo.2005.02.026","article-title":"B-Cells in ocular adnexal lymphoproliferative lesions express B-cell attracting chemokine 1 (CXCL13)","volume":"140","author":"Falkenhagen","year":"2005","journal-title":"Am J Ophthalmol"},{"key":"B39","doi-asserted-by":"publisher","first-page":"10","DOI":"10.1186\/s40164-015-0004-3","article-title":"Immunohistochemical analysis indicates that the anatomical location of B-cell non-Hodgkin's lymphoma is determined by differentially expressed chemokine receptors, sphingosine-1-phosphate receptors and integrins","volume":"4","author":"Middle","year":"2015","journal-title":"Exp Hematol Oncol"}],"container-title":["Frontiers in Immunology"],"original-title":[],"link":[{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fimmu.2025.1508559\/full","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,2,26]],"date-time":"2025-02-26T09:06:18Z","timestamp":1740560778000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fimmu.2025.1508559\/full"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,2,26]]},"references-count":39,"alternative-id":["10.3389\/fimmu.2025.1508559"],"URL":"https:\/\/doi.org\/10.3389\/fimmu.2025.1508559","relation":{},"ISSN":["1664-3224"],"issn-type":[{"value":"1664-3224","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,2,26]]},"article-number":"1508559"}}