{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,1,22]],"date-time":"2026-01-22T10:47:22Z","timestamp":1769078842395,"version":"3.49.0"},"reference-count":33,"publisher":"MDPI AG","issue":"12","license":[{"start":{"date-parts":[[2023,6,19]],"date-time":"2023-06-19T00:00:00Z","timestamp":1687132800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Sarcoidosis is a systemic granulomatous disease mainly affecting the lungs and hilomediastinal lymph nodes. It is characterized by non-caseating epithelioid cell granulomas in lymph nodes and lungs. Our study aimed to evaluate and compare T, B and NK cell subsets in the alveolar compartment, lymph nodes and the bloodstream simultaneously in the same patients to elucidate the immune responses associated with the development and progression of sarcoidosis. A secondary aim was to evaluate the distribution of CD45RA-expressing cells in the different anatomical compartments. Patients suspected to have sarcoidosis and who underwent bronchoscopy with bronchoalveolar lavage (BAL), lung-draining lymph node (LLN) biopsy by EBUS-TBNA and peripheral blood (PB) sampling were included in the study. They were monitored at the Regional Referral Centre of Siena University Hospital and the Respiratory Diseases Unit of Perugia Hospital. Multicolour flow cytometry analysis through FASCLyric was performed to assess T, B and NK cell subsets. Thirty-two patients (median age (IQR) 57 (52\u201358) years) were consecutively and prospectively enrolled. Machine learning analysis created a model which selected CD56dim16bright, CD8, Tfc, Th17, Th12, Tfh17, Tfh2, TcemRA, ThemRA, T na\u00efve, Tc na\u00efve, Breg, CD1d+CD5+, Th-reg, Tfh, Th1 and CD4 cells with an accuracy of 0.9500 (kappa 0.8750). Comparative analysis found 18 cell populations that differed significantly between the three anatomical compartments. The bloodstream was enriched in ThemRA (p = 0.0416), Tfh2 (p = 0.0189), Tfh17 (p = 0.0257), Th2 (p = 0.0212), Th17 (p = 0.0177), Th-na\u00efve (p = 0.0368), CD56dimCD16bright (p &lt; 0.0001), CD8 (p = 0.0319), TcemRA (p &lt; 0.0001) and Tfc cells (p = 0.0004) compared with the alveolar compartment, while Th-reg were lower in PB than BAL (p = 0.0329). The alveolar compartment was enriched in Breg (p = 0.0249) and CD1d+CD5+ (p = 0.0013) with respect to LLN samples and PB. Conversely, Tfh (p = 0.0470), Th1 (p = 0.0322), CD4 (p = 0.0486) and Tc-na\u00efve (p = 0.0009) were more abundant in LLN than in BAL and PB. It has been speculated that changes in the relative contents of PB cells could be related to changes in production and to the selective redistribution of PB cells to granulomatous foci. This study further supports the fact that sarcoidosis is multisystemic in nature. However, the low level of immune cells in peripheral blood of patients with sarcoidosis is concerning. A re-expression of CD45RA on CD4+ and CD8+ cells could result in a reduction in peripheral immune activity. Thus, changes in the spectrum of the bloodstream may reflect both pathogenic and compensatory processes.<\/jats:p>","DOI":"10.3390\/ijms241210344","type":"journal-article","created":{"date-parts":[[2023,6,20]],"date-time":"2023-06-20T01:37:38Z","timestamp":1687225058000},"page":"10344","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["Imbalance of Lymphocyte Subsets and CD45RA-Expressing Cells in Intrathoracic Lymph Nodes, Alveolar Compartment and Bloodstream of Pulmonary Sarcoidosis Patients"],"prefix":"10.3390","volume":"24","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-2368-5722","authenticated-orcid":false,"given":"Miriana","family":"d\u2019Alessandro","sequence":"first","affiliation":[{"name":"Respiratory Diseases Unit, Department of Medical and Surgical Sciences & Neurosciences, University of Siena, 53100 Siena, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1118-3223","authenticated-orcid":false,"given":"Laura","family":"Bergantini","sequence":"additional","affiliation":[{"name":"Respiratory Diseases Unit, Department of Medical and Surgical Sciences & Neurosciences, University of Siena, 53100 Siena, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Sara","family":"Gangi","sequence":"additional","affiliation":[{"name":"Respiratory Diseases Unit, Department of Medical and Surgical Sciences & Neurosciences, University of Siena, 53100 Siena, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8639-2882","authenticated-orcid":false,"given":"Paolo","family":"Cameli","sequence":"additional","affiliation":[{"name":"Respiratory Diseases Unit, Department of Medical and Surgical Sciences & Neurosciences, University of Siena, 53100 Siena, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Martina","family":"Armati","sequence":"additional","affiliation":[{"name":"Respiratory Diseases Unit, Department of Medical and Surgical Sciences & Neurosciences, University of Siena, 53100 Siena, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Matteo","family":"Fanetti","sequence":"additional","affiliation":[{"name":"Respiratory Diseases Unit, Department of Medical and Surgical Sciences & Neurosciences, University of Siena, 53100 Siena, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Fabrizio","family":"Mezzasalma","sequence":"additional","affiliation":[{"name":"Diagnostic and Interventional Bronchoscopy Unit, Cardio-Thoracic and Vascular Department, University Hospital of Siena (Azienda Ospedaliera Universitaria Senese, AOUS), 53100 Siena, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5250-2868","authenticated-orcid":false,"given":"Stefano","family":"Baglioni","sequence":"additional","affiliation":[{"name":"Pneumology Department, Perugia Hospital, 06129 Perugia, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"name":"SARC-SI Study Group","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8351-3703","authenticated-orcid":false,"given":"Elena","family":"Bargagli","sequence":"additional","affiliation":[{"name":"Respiratory Diseases Unit, Department of Medical and Surgical Sciences & Neurosciences, University of Siena, 53100 Siena, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2023,6,19]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"527","DOI":"10.1097\/MCP.0000000000000715","article-title":"Sarcoidosis Epidemiology: Recent Estimates of Incidence, Prevalence and Risk Factors","volume":"26","author":"Arkema","year":"2020","journal-title":"Curr. Opin. Pulm. Med."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"45","DOI":"10.1038\/s41572-019-0096-x","article-title":"Sarcoidosis","volume":"5","author":"Grunewald","year":"2019","journal-title":"Nat. Rev. Dis. Primers"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"1165","DOI":"10.1136\/bmj.2.5261.1165","article-title":"Prognosis of Intrathoracic Sarcoidosis in England. A Review of 136 Cases after Five Years\u2019 Observation","volume":"2","author":"Scadding","year":"1961","journal-title":"Br. Med. J."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"207","DOI":"10.1007\/s10238-020-00608-5","article-title":"BAL Biomarkers\u2019 Panel for Differential Diagnosis of Interstitial Lung Diseases","volume":"20","author":"Carleo","year":"2020","journal-title":"Clin. Exp. Med."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"437","DOI":"10.1007\/s40291-022-00596-0","article-title":"Immune-Checkpoint Expression on CD4, CD8 and NK Cells in Blood, Bronchoalveolar Lavage and Lymph Nodes of Sarcoidosis","volume":"26","author":"Bergantini","year":"2022","journal-title":"Mol. Diagn. Ther."},{"key":"ref_6","doi-asserted-by":"crossref","unstructured":"d\u2019Alessandro, M., Gangi, S., Cavallaro, D., Bergantini, L., Mezzasalma, F., Cattelan, S., Baglioni, S., Abbritti, M., Cameli, P., and Bargagli, E. (2022). CD103 Expression on Regulatory and Follicular T Cells in Lymph Nodes, Bronchoalveolar Lavage Fluid and Peripheral Blood of Sarcoidosis Patients. Life, 12.","DOI":"10.3390\/life12050762"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"e26","DOI":"10.1164\/rccm.202002-0251ST","article-title":"Diagnosis and Detection of Sarcoidosis. An Official American Thoracic Society Clinical Practice Guideline","volume":"201","author":"Crouser","year":"2020","journal-title":"Am. J. Respir. Crit. Care Med."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"244","DOI":"10.1186\/s12931-019-1216-6","article-title":"Aberrant Populations of Circulating T Follicular Helper Cells and Regulatory B Cells Underlying Idiopathic Pulmonary Fibrosis","volume":"20","author":"Asai","year":"2019","journal-title":"Respir. Res."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"215","DOI":"10.1016\/j.jaci.2016.09.042","article-title":"Human Regulatory B Cells Control the TFH Cell Response","volume":"140","author":"Achour","year":"2017","journal-title":"J. Allergy Clin. Immunol."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"476","DOI":"10.1097\/MCP.0000000000000303","article-title":"T-Cell Immunology in Sarcoidosis: Disruption of a Delicate Balance between Helper and Regulatory T-Cells","volume":"22","author":"Broos","year":"2016","journal-title":"Curr. Opin. Pulm. Med."},{"key":"ref_11","doi-asserted-by":"crossref","unstructured":"Kudryavtsev, I., Zinchenko, Y., Starshinova, A., Serebriakova, M., Malkova, A., Akisheva, T., Kudlay, D., Glushkova, A., Yablonskiy, P., and Shoenfeld, Y. (2023). Circulating Regulatory T Cell Subsets in Patients with Sarcoidosis. Diagnostics, 13.","DOI":"10.3390\/diagnostics13081378"},{"key":"ref_12","doi-asserted-by":"crossref","unstructured":"d\u2019Alessandro, M., Bergantini, L., Gangi, S., Conticini, E., Cavallaro, D., Cameli, P., Mezzasalma, F., Cantarini, L., Frediani, B., and Bargagli, E. (2023). Immunological Pathways in Sarcoidosis and Autoimmune Rheumatic Disorders\u2014Similarities and Differences in an Italian Prospective Real-Life Preliminary Study. Biomedicines, 11.","DOI":"10.3390\/biomedicines11061532"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"21463","DOI":"10.3390\/ijms141121463","article-title":"Imbalance between Th17 and Regulatory T-Cells in Sarcoidosis","volume":"14","author":"Huang","year":"2013","journal-title":"Int. J. Mol. Sci."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"1147529","DOI":"10.3389\/fmed.2023.1147529","article-title":"Editorial: Sarcoidosis and Autoimmunity: From Bench to Bedside","volume":"10","year":"2023","journal-title":"Front. Med."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"476","DOI":"10.1016\/j.coi.2012.04.001","article-title":"Properties of End-Stage Human T Cells Defined by CD45RA Re-Expression","volume":"24","author":"Henson","year":"2012","journal-title":"Curr. Opin. Immunol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"315","DOI":"10.4103\/1735-5362.343085","article-title":"Comparison of Follicular T Helper Cells, Monocytes, and T Cells Priming between Newly Diagnosed and Rituximab-Treated MS Patients and Healthy Controls","volume":"17","author":"Yahyazadeh","year":"2022","journal-title":"Res. Pharm. Sci."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"487","DOI":"10.1007\/s10238-019-00578-3","article-title":"NK and NKT-like Cells in Granulomatous and Fibrotic Lung Diseases","volume":"19","author":"Bergantini","year":"2019","journal-title":"Clin. Exp. Med."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"1059","DOI":"10.1038\/s41598-020-57741-0","article-title":"Imbalance in B Cell and T Follicular Helper Cell Subsets in Pulmonary Sarcoidosis","volume":"10","author":"Kudryavtsev","year":"2020","journal-title":"Sci. Rep."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"1080556","DOI":"10.3389\/fimmu.2022.1080556","article-title":"Characterization of Natural Killer and T Cells in Bronchoalveolar Lavage and Peripheral Blood of Sarcoidosis Patients","volume":"13","author":"Bergantini","year":"2022","journal-title":"Front. Immunol."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"307","DOI":"10.1055\/s-0034-1376861","article-title":"Cellular Activation in the Immune Response of Sarcoidosis","volume":"35","author":"Zissel","year":"2014","journal-title":"Semin. Respir. Crit. Care Med."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"e227929","DOI":"10.1136\/bcr-2018-227929","article-title":"Multisystemic Sarcoidosis\u2014Important Lessons Learnt from One of the Great Imitators","volume":"12","author":"Santosa","year":"2019","journal-title":"BMJ Case Reports CP"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"1198","DOI":"10.1164\/rccm.201512-2419ED","article-title":"Sarcoidosis and T-Helper Cells. Th1, Th17, or Th17.1?","volume":"193","author":"Georas","year":"2016","journal-title":"Am. J. Respir. Crit. Care Med."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"144","DOI":"10.1136\/thx.2010.140319","article-title":"Sarcoidosis Is a Th1\/Th17 Multisystem Disorder","volume":"66","author":"Facco","year":"2011","journal-title":"Thorax"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"1701","DOI":"10.1084\/jem.20060772","article-title":"CD127 Expression Inversely Correlates with FoxP3 and Suppressive Function of Human CD4+ T Reg Cells","volume":"203","author":"Liu","year":"2006","journal-title":"J. Exp. Med."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"1176","DOI":"10.5114\/aoms.2018.77772","article-title":"Phenotypic Identification of CD19+CD5+CD1d+ Regulatory B Cells That Produce Interleukin 10 and Transforming Growth Factor \u03921 in Human Peripheral Blood","volume":"15","author":"Karim","year":"2019","journal-title":"Arch. Med. Sci."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"642","DOI":"10.1002\/art.1780310509","article-title":"B Cells Expressing CD5 Are Increased in Sj\u00f6gren\u2019s Syndrome","volume":"31","author":"Tovar","year":"1988","journal-title":"Arthritis Rheum."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"1403","DOI":"10.1164\/rccm.202012-4423OC","article-title":"Follicular Helper\u2013like T Cells in the Lung Highlight a Novel Role of B Cells in Sarcoidosis","volume":"204","author":"Bauer","year":"2021","journal-title":"Am. J. Respir. Crit. Care Med. 204"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"2211","DOI":"10.1681\/ASN.2022030286","article-title":"Effector Memory-Expressing CD45RA (TEMRA) CD8+ T Cells from Kidney Transplant Recipients Exhibit Enhanced Purinergic P2X4 Receptor-Dependent Proinflammatory and Migratory Responses","volume":"33","author":"Tilly","year":"2022","journal-title":"J. Am. Soc. Nephrol."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"104520","DOI":"10.1016\/j.ebiom.2023.104520","article-title":"Expanding Scope of TEMRA in Autoimmunity","volume":"90","author":"Perl","year":"2023","journal-title":"eBioMedicine"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"641013","DOI":"10.3389\/fimmu.2021.641013","article-title":"Frontiers of Autoantibodies in Autoimmune Disorders: Crosstalk Between Tfh\/Tfr and Regulatory B Cells","volume":"12","author":"Ding","year":"2021","journal-title":"Front. Immunol."},{"key":"ref_31","first-page":"149","article-title":"ATS\/ERS\/WASOG Statement on Sarcoidosis. American Thoracic Society\/European Respiratory Society\/World Association of Sarcoidosis and Other Granulomatous Disorders","volume":"16","author":"Hunninghake","year":"1999","journal-title":"Sarcoidosis Vasc. Diffuse Lung Dis."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"735","DOI":"10.1034\/j.1399-3003.1999.14d02.x","article-title":"ATS\/ERS\/WASOG Statement on Sarcoidosis. Sarcoidosis Statement Committee. American Thoracic Society. European Respiratory Society. World Association for Sarcoidosis and Other Granulomatous Disorders","volume":"14","author":"Costabel","year":"1999","journal-title":"Eur. Respir. J."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"53","DOI":"10.1007\/7651_2014_99","article-title":"Peripheral Blood Mononuclear Cells: Isolation, Freezing, Thawing, and Culture","volume":"1304","author":"Riedhammer","year":"2016","journal-title":"Methods Mol. Biol."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/24\/12\/10344\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T19:56:35Z","timestamp":1760126195000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/24\/12\/10344"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,6,19]]},"references-count":33,"journal-issue":{"issue":"12","published-online":{"date-parts":[[2023,6]]}},"alternative-id":["ijms241210344"],"URL":"https:\/\/doi.org\/10.3390\/ijms241210344","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2023,6,19]]}}}