Jo-1 autoantigen-specific B cells are skewed towards distinct functional B cell subsets in anti-synthetase syndrome patients

Young-Glazer, Jennifer J.; Cisneros, Alberto; Wilfong, Erin M.; Smith, Scott A.; Crofford, Leslie J.; Bonami, Rachel H.

doi:10.1186/s13075-020-02412-8

Cited by 10 publications

(15 citation statements)

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“…As shown in Figure 2A , specific B cell subsets were purified using flow cytometry sorting. The focus of this manuscript is on B cells derived from autoimmune donors; we presented detailed immunophenotyping to compare B cell subset percentages between patients with several of the autoimmune diseases highlighted in this manuscript and healthy controls in separate manuscripts ( 36 , 41 ). Purified B cell subsets (e.g.…”

Section: Resultsmentioning

confidence: 99%

“…We previously identified ASBCs using fluorescently-labeled autoantigen ( 36 ). This detection method could however miss B cells that downregulate surface BCR because of chronic antigen stimulation and/or immune tolerance mechanisms such as anergy.…”

Section: Resultsmentioning

confidence: 99%

“…Flow cytometry detection of ASBCs can be challenging due to low frequency of target cells. However, we recently identified a readily detectable population of Jo-1-binding B cells in the peripheral blood of Jo-1 autoantibody-positive anti-histidyl tRNA synthetase syndrome patients using flow cytometry staining with labeled autoantigen ( 36 ). This created an opportunity to directly examine expansion of ASBCs following our in vitro stimulation protocol.…”

Section: Resultsmentioning

confidence: 99%

“…Data were analyzed using FlowJo software (Tree Star, Inc.). Jo-1-binding B cells were identified by flow cytometry using GST-tagged Jo-1 antigen we previously cloned and recombinantly expressed, followed by detection with anti-GST secondary antibody (Abcam), as previously described ( 36 ).…”

Section: Methodsmentioning

confidence: 99%

“…For example, whereas naïve B cells proliferate in response to BCR stimulation, anergic (B ND ) and CD21 lo B cells do not (30,31). B ND and CD21 lo subsets may serve as reservoirs for autoreactive B cells in several autoimmune diseases, including type 1 diabetes, Sjögren's syndrome, anti-histidyl tRNA synthetase syndrome, and systemic sclerosis (32)(33)(34)(35)(36). We sought to develop high-throughput stimulation and screening methods to identify ASBCs among total PBMCs using ELISA detection of BCRs secreted as antibody.…”

Section: Introductionmentioning

confidence: 99%

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High-Throughput Detection of Autoantigen-Specific B Cells Among Distinct Functional Subsets in Autoimmune Donors

et al. 2021

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Antigen-specific B cells (ASBCs) can drive autoimmune disease by presenting autoantigen to cognate T cells to drive their activation, proliferation, and effector cell differentiation and/or by differentiating into autoantibody-secreting cells. Autoantibodies are frequently used to predict risk and diagnose several autoimmune diseases. ASBCs can drive type 1 diabetes even when immune tolerance mechanisms block their differentiation into antibody-secreting cells. Furthermore, anti-histidyl tRNA synthetase syndrome patients have expanded IgM+ Jo-1-binding B cells, which clinically diagnostic IgG Jo-1 autoantibodies may not fully reflect. Given the potential disconnect between the pathologic function of ASBCs and autoantibody secretion, direct study of ASBCs is a necessary step towards developing better therapies for autoimmune diseases, which often have no available cure. We therefore developed a high-throughput screening pipeline to 1) phenotypically identify specific B cell subsets, 2) expand them in vitro, 3) drive them to secrete BCRs as antibody, and 4) identify wells enriched for ASBCs through ELISA detection of antibody. We tested the capacity of several B cell subset(s) to differentiate into antibody-secreting cells following this robust stimulation. IgM+ and/or IgD+, CD27- memory, memory, switched memory, and BND B cells secreted B cell receptor (BCR) as antibody following in vitro stimulation, whereas few plasmablasts responded. Bimodal responses were observed across autoimmune donors for IgM+ CD21lo and IgM- CD21lo B cells, consistent with documented heterogeneity within the CD21lo subset. Using this approach, we detected insulin-binding B cell bias towards CD27- memory and CD27+ memory subsets in pre-symptomatic type 1 diabetes donors. We took advantage of routine detection of Jo-1-binding B cells in Jo-1+ anti-histidyl tRNA synthetase syndrome patients to show that Jo-1-binding B cells and total B cells expanded 20-30-fold using this culture system. Overall, these studies highlight technology that is amenable to small numbers of cryopreserved peripheral blood mononuclear cells that enables interrogation of phenotypic and repertoire attributes of ASBCs derived from autoimmune patients.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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High-Throughput Detection of Autoantigen-Specific B Cells Among Distinct Functional Subsets in Autoimmune Donors

et al. 2021

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Association of Juvenile Dermatomyositis Disease Activity With the Expansion of Blood Memory B and T Cell Subsets Lacking Follicular Markers

Gofshteyn

Mansfield

Spitznagle

et al. 2023

Arthritis & Rheumatology

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Objective. This study was undertaken to identify blood markers of juvenile dermatomyositis (DM) disease activity (DA), which are needed to improve disease management.Methods. The study comprised a total of 123 juvenile DM patients and 53 healthy controls. Results of laboratory tests (aldolase, creatinine kinase, lactate dehydrogenase [LDH], aspartate aminotransferase) and clinical measures of DA in patients with juvenile DM, including the Manual Muscle Testing in 8 muscles (MMT-8), Childhood Myositis Assessment Scale (CMAS), and disease activity scores (DAS) (total DAS for juvenile DM, the muscle DAS, and the skin DAS), were recorded when available. Surface phenotype of peripheral blood mononuclear cells was assessed using flow cytometry. Whole blood transcriptional profiles were studied using either RNA-sequencing or microarrays. Differential gene expression was determined using DESeq and compared by pathway and gene ontology analyses.Results. Conventional memory (CD27+IgD-) B cells expressing low CXCR5 levels (CXCR5 low/-CM B cells) were significantly increased in frequency and absolute numbers in 2 independent cohorts of juvenile DM patients compared with healthy controls. The frequency of CD4+ Th2 memory cells (CD45RA-CXCR5-CCR6-CXCR3-) was also increased in juvenile DM, especially in patients who were within <1 year from diagnosis. The frequency of CXCR5 low/-CM B cells correlated with serum aldolase levels and with a blood interferon-stimulated gene transcriptional signature. Furthermore, both the frequency and absolute numbers of CXCR5 low/-CM B cells correlated with clinical and laboratory measures of muscle DA (MMT-8, CMAS, aldolase, and LDH).Conclusion. These findings suggest that both CM B cells lacking the CXCR5 follicular marker and CXCR5-Th2 cells represent potential biomarkers of DA in juvenile DM and may contribute to its pathogenesis.

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Anti‐PL‐12 anti‐synthetase syndrome manifesting with multiple digital ischemia: Case report & review of the literature

Doumeth,

Petrinec,

Chaudhary

et al. 2024

Clinical Case Reports

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Key Clinical MessageAcute digital ischemia is a rare manifestation of anti‐synthetase syndrome in the absence of Raynaud's phenomenon. A high index of suspicion may result in early diagnosis and better clinical outcomes.AbstractA 61‐year‐old male patient was admitted to the hospital for worsening arthralgias with morning stiffness lasting hours, as well as left sided headaches, and jaw pain while eating. He had significant weight loss and subjective fever at home. Multiple fingers and toes were noted to be ischemic. His laboratory workup was pertinent for significantly elevated inflammatory markers, and mild Creatinine kinase elevation. Chest imaging and later lung biopsy were notable for organizing pneumonia. Conventional angiogram showed evidence of significant digital disease without collaterals. Subsequent autoimmune screening tests with extended myositis‐specific and myositis‐associated panels revealed a strongly positive anti‐PL‐12 antibody and moderately positive anti‐ SSA‐52KD IgG ab. After ruling out infectious etiologies and malignancy, anti‐synthetase syndrome (ASyS) diagnosis was considered in the presence of ischemic digits, organizing pneumonia, polyarthralgia, constitutional symptoms, increased inflammatory markers and positive antibodies. The patient was treated with high dose prednisone and mycophenolate mofetil along with amlodipine and sildenafil for digital vasodilation. Acute digital ischemia may be the first manifestation of ASyS with ILD. A high index of suspicion is warranted for early diagnosis and better outcomes.

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Jo-1 autoantigen-specific B cells are skewed towards distinct functional B cell subsets in anti-synthetase syndrome patients

Cited by 10 publications

References 50 publications

High-Throughput Detection of Autoantigen-Specific B Cells Among Distinct Functional Subsets in Autoimmune Donors

High-Throughput Detection of Autoantigen-Specific B Cells Among Distinct Functional Subsets in Autoimmune Donors

Association of Juvenile Dermatomyositis Disease Activity With the Expansion of Blood Memory B and T Cell Subsets Lacking Follicular Markers

Anti‐PL‐12 anti‐synthetase syndrome manifesting with multiple digital ischemia: Case report & review of the literature

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