{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,7,30]],"date-time":"2026-07-30T14:45:34Z","timestamp":1785422734476,"version":"3.56.0"},"posted":{"date-parts":[[2026,3,2]]},"group-title":"Immunology","reference-count":48,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2026,3,2]],"date-time":"2026-03-02T00:00:00Z","timestamp":1772409600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/www.biorxiv.org\/about\/FAQ#license"}],"funder":[{"award":["5P01AI172523"],"award-info":[{"award-number":["5P01AI172523"]}],"id":[{"id":"https:\/\/ror.org\/043z4tv69","id-type":"ROR","asserted-by":"publisher"}]},{"award":["5U19AI144306"],"award-info":[{"award-number":["5U19AI144306"]}],"id":[{"id":"https:\/\/ror.org\/043z4tv69","id-type":"ROR","asserted-by":"publisher"}]},{"DOI":"10.13039\/100012051","name":"Lupus Research Alliance","doi-asserted-by":"crossref","award":["2022GTSA-1211"],"award-info":[{"award-number":["2022GTSA-1211"]}],"id":[{"id":"10.13039\/100012051","id-type":"DOI","asserted-by":"crossref"}]},{"award":["1068467"],"award-info":[{"award-number":["1068467"]}],"id":[{"id":"https:\/\/ror.org\/03abbex33","id-type":"ROR","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2026,3,2]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                <jats:sec>\n                  <jats:title>Objective<\/jats:title>\n                  <jats:p>To define cellular and molecular mechanisms distinguishing active systemic lupus erythematosus (SLE) from remission by profiling autoreactive antinuclear antigen\u2013 positive (ANA+) and non-autoreactive B cells subsets in three cohorts: active disease (SLE-A), long-term, drug free remission (SLE-R), and healthy controls (HC).<\/jats:p>\n                <\/jats:sec>\n                <jats:sec>\n                  <jats:title>Methods<\/jats:title>\n                  <jats:p>Peripheral blood B cells were phenotyped by flow cytometry, including ANA reactivity. Single-cell RNA sequencing (scRNA-seq) was performed on sorted ANA+ and ANA\u2212 subsets.<\/jats:p>\n                <\/jats:sec>\n                <jats:sec>\n                  <jats:title>Results<\/jats:title>\n                  <jats:p>Seven transcriptomic B cell clusters were resolved: quiescent (Na\u00efve 1, Marginal Zone B cells [MZB], IgG Memory 1) and activated (Age-Associated B cells [ABCs], Na\u00efve 2, IgM Memory, IgG Memory 2). SLE-A showed expansion of activated clusters, MZB contraction, and a higher IgG:IgM B cell ratio. SLE-R exhibited an \u201cimmunological reset,\u201d distinct from healthy homeostasis, with reduced ABCs and IgG Memory 2, persistence of Na\u00efve 2, and partial restoration of MZB and Na\u00efve 1. Interferon-\u03b1 (IFNa) signaling was elevated across clusters in SLE-A (SLE-A &gt; SLE-R &gt; HC), whereas TNF signaling was enriched in activated clusters across cohorts, with minimal differences between SLE-R and SLE-A. IFNa and TNF scores were inversely correlated. B cells predominantly expressed TNFR2, suggesting immunomodulatory TNF effects in remission. ANA+ cells in HC and SLE-R showed enriched Fc\u03b3RIIb inhibitory and IL-4\/STAT6 signaling, suggesting reinstated regulatory control.<\/jats:p>\n                <\/jats:sec>\n                <jats:sec>\n                  <jats:title>Discussion<\/jats:title>\n                  <jats:p>Compared to SLE-A, SLE-R was characterized by partial reversion to HC homeostasis with residual activation. These findings delineate immunologic features of remission and suggest therapeutic opportunities, including modulation of TNFR2, Fc\u03b3RIIb, and IL-4 to help sustain remission.<\/jats:p>\n                <\/jats:sec>\n                <jats:sec>\n                  <jats:title>What is already known on this topic<\/jats:title>\n                  <jats:p>Some patients with SLE achieve complete clinical remission without treatment, referred as \u2018immune reset\u2019; the mechanisms that underlie this state have not been well characterized. Healthy individuals and patients with Systemic Lupus Erythematosus (SLE) normally harbor similar frequencies of autoreactive B cells; the checkpoints that regulate activation of these cells are not fully defined.<\/jats:p>\n                <\/jats:sec>\n                <jats:sec>\n                  <jats:title>What this study adds<\/jats:title>\n                  <jats:p>B cells, stratified by their reactivity to nuclear antigens (ANA), from active SLE (SLE-A), drug-free remission (SLE-R), and healthy controls (HC) were analyzed using single cell sequencing and flow cytometry. We identified B cells states associated with disease activity; SLE-R displayed a distinct profile that differed from SLE-A and HC. TNF signaling was present in activated B cell subsets in SLE-A and SLE-R. This persistence in SLE-R may reflect an immunomodulatory function of TNF on TNFR2, which is expressed on B cells. ANA+ cells in SLE-R and HC were enriched for inhibitory Fc\u03b3RIIb and IL-4\/STAT6 programs.<\/jats:p>\n                <\/jats:sec>\n                <jats:sec>\n                  <jats:title>How this study might affect research, practice or policy<\/jats:title>\n                  <jats:p>The signatures identified help define the \u201cimmunological reset\u201d state in patients with SLE-R. We also identified pathways, such as type I IFN, TNFR2, Fc\u03b3RIIb, IL-4\/STAT6 as potential targets for maintaining remission.<\/jats:p>\n                <\/jats:sec>","DOI":"10.64898\/2026.02.27.708589","type":"posted-content","created":{"date-parts":[[2026,3,2]],"date-time":"2026-03-02T16:50:12Z","timestamp":1772470212000},"source":"Crossref","is-referenced-by-count":0,"title":["Mechanistic insights into B-cell activation and autoreactivity regulation in active SLE and remission"],"prefix":"10.64898","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-7702-8640","authenticated-orcid":false,"given":"Yemil","family":"Atisha-Fregoso","sequence":"first","affiliation":[{"name":"Institute of Molecular Medicine, Feinstein Institutes for Medical Research, Manhasset, NY, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0038-0253","authenticated-orcid":false,"given":"Rita","family":"Pozovskiy","sequence":"additional","affiliation":[{"name":"Institute of Molecular Medicine, Feinstein Institutes for Medical Research, Manhasset, NY, USA"},{"name":"Zucker School of Medicine at Hosftra\/Northwell, NY"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4061-3798","authenticated-orcid":false,"given":"Meggan","family":"Mackay","sequence":"additional","affiliation":[{"name":"Institute of Molecular Medicine, Feinstein Institutes for Medical Research, Manhasset, NY, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9299-0053","authenticated-orcid":false,"given":"Cynthia","family":"Aranow","sequence":"additional","affiliation":[{"name":"Institute of Molecular Medicine, Feinstein Institutes for Medical Research, Manhasset, NY, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3250-3804","authenticated-orcid":false,"given":"Betty","family":"Diamond","sequence":"additional","affiliation":[{"name":"Institute of Molecular Medicine, Feinstein Institutes for Medical Research, Manhasset, NY, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"54368","reference":[{"key":"2026030414300543000_2026.02.27.708589v1.1","doi-asserted-by":"publisher","DOI":"10.1056\/NEJMc2107725"},{"issue":"10","key":"2026030414300543000_2026.02.27.708589v1.2","doi-asserted-by":"crossref","first-page":"565","DOI":"10.1038\/s41584-020-0480-7","article-title":"New insights into the role of antinuclear antibodies in systemic lupus erythematosus","volume":"16","year":"2020","journal-title":"Nature reviews Rheumatology"},{"key":"2026030414300543000_2026.02.27.708589v1.3","doi-asserted-by":"publisher","DOI":"10.1084\/jem.20052033"},{"issue":"9","key":"2026030414300543000_2026.02.27.708589v1.4","first-page":"2210","article-title":"Checkpoints for Autoreactive B Cells in the Peripheral Blood of Lupus Patients Assessed by Flow Cytometry","volume":"68","year":"2016","journal-title":"Arthritis & rheumatology (Hoboken, NJ)"},{"key":"2026030414300543000_2026.02.27.708589v1.5","doi-asserted-by":"publisher","DOI":"10.1016\/j.jaci.2018.10.041"},{"key":"2026030414300543000_2026.02.27.708589v1.6","doi-asserted-by":"crossref","unstructured":"van Vollenhoven RF , Bertsias G , Doria A , Isenberg D , Morand E , Petri MA , Pons-Estel BA , Rahman A , Ugarte-Gil MF , Voskuyl A et al: 2021 DORIS definition of remission in SLE: final recommendations from an international task force. 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