Flow Cytometric Analysis of the LE Cell Phenomenon

Böhm, Ingrid

doi:10.1080/08916930310001630325

Cited by 10 publications

(5 citation statements)

References 19 publications

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“…In contrast, the intracellular vesicles contain entire nuclei that had actually been phagocytosed, transferred into the phagolysomes, and partially digested. Phagocytosis has been confirmed by flow cytometry ( 27 , 28 ) and depends on factors in the biological fluids of patients with SLE, in particular autoantibodies recognizing nuclear antigens, histones and DNA, and complement ( 25 ). LE cells have been originally identified in the bone marrow of lupus patients.…”

Section: Neutrophils Recognizing Particulate Substratesmentioning

confidence: 98%

The Neutrophil’s Choice: Phagocytose vs Make Neutrophil Extracellular Traps

et al. 2018

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Neutrophils recognize particulate substrates of microbial or endogenous origin and react by sequestering the cargo via phagocytosis or by releasing neutrophil extracellular traps (NETs) outside the cell, thus modifying and alerting the environment and bystander leukocytes. The signals that determine the choice between phagocytosis and the generation of NETs are still poorly characterized. Neutrophils that had phagocytosed bulky particulate substrates, such as apoptotic cells and activated platelets, appear to be “poised” in an unresponsive state. Environmental conditions, the metabolic, adhesive and activation state of the phagocyte, and the size of and signals associated with the tethered phagocytic cargo influence the choice of the neutrophils, prompting either phagocytic clearance or the generation of NETs. The choice is dichotomic and apparently irreversible. Defects in phagocytosis may foster the intravascular generation of NETs, thus promoting vascular inflammation and morbidities associated with diseases characterized by defective phagocytic clearance, such as systemic lupus erythematosus. There is a strong potential for novel treatments based on new knowledge of the events determining the inflammatory and pro-thrombotic function of inflammatory leukocytes.

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Section: Neutrophils Recognizing Particulate Substratesmentioning

confidence: 98%

The Neutrophil’s Choice: Phagocytose vs Make Neutrophil Extracellular Traps

et al. 2018

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“…Assay for phagocytosis of NC material (PNC assay) and oxidative burst activity. Phagocytosis of NC by PMN was analyzed by flow cytometry, using an assay similar to the one described by Böhm 13 . Propidium iodide-labeled NC (4.5 × 10 5 cells) were incubated with 30 µl undiluted serum at room temperature for 20 min, allowing antibodies to bind to the NC.…”

Section: Preparation Of Phagocytic Cells and Cell Nucleimentioning

confidence: 99%

“…The original diagnostic LE cell test was based on microscopy. In a more recent method, described by Böhm, flow cytometry was used to measure phagocytosis of necrotic cell material by purified PMN in the presence of serum 13 .…”

mentioning

confidence: 99%

Combination of Autoantibodies Against Different Histone Proteins Influences Complement-dependent Phagocytosis of Necrotic Cell Material by Polymorphonuclear Leukocytes in Systemic Lupus Erythematosus

Gullstrand¹,

Lefort²,

Tydén³

et al. 2012

J Rheumatol

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Objective.Polymorphonuclear leukocytes (PMN) with autoantibody-coated engulfed necrotic cell material (NC) are frequently seen in systemic lupus erythematosus (SLE). We evaluated the roles of complement, different antihistone antibodies (anti-H ab), and oxidative burst in the phagocytosis of NC by PMN, as well as association to disease activity and clinical phenotype in SLE.Methods.ELISA and immunoblot were used to measure antibodies to different histone proteins in sera from patients with SLE and complement-deficient individuals. Phagocytosis of NC by PMN and oxidative burst activity was assessed by flow cytometry.Results.A clearly increased phagocytosis of NC was seen in patients with active SLE, which was associated with high levels of anti-H ab concentrations and oxidative burst activity. The complement system contributed to efficient phagocytosis of NC by PMN through activation of the classical pathway, and the phagocytosis was mediated by FcγRIIA, FcγRIIIB, and CR1 in combination. A pattern of high phagocytosis, consumption of classical pathway components, and a broad anti-H ab repertoire was seen particularly in patients with nephritis and serositis. The combination of antibodies to several different histone proteins, often with anti-DNA antibodies, promoted an efficient uptake of NC, whereas antibodies against only histone H1 or a few histones seemed to be of less importance.Conclusion.The distributions of specificities among anti-H ab are of great importance in the complement-dependent phagocytosis of debris from NC in SLE. Measurement of anti-H ab could be useful in monitoring of this disease and contribute to improved understanding of the autoimmune process.

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“…Serum-mediated phagocytosis of NCM by PMNs was performed as described by Böhm [ 29 ], with some modifications [ 30 ]. Briefly, PMNs and peripheral blood mononuclear cells (PBMCs) were isolated from heparinized blood using Polymorphprep™ (Axis-Shield Poc AS, Oslo, Norway) according to the manufacturer’s protocol.…”

Section: Methodsmentioning

confidence: 99%

“…More recently, a flow cytometry–based assay—the phagocytosis of necrotic cells (PNC) assay—was developed as an in vitro assessment and quantification of LE cells in patients with suspected SLE [ 29 ], but its use in clinical practice has never been validated. In a previous study [ 30 ], our group demonstrated that the outcome of the PNC assay is often positive in patients with SLE with increased disease activity.…”

Section: Introductionmentioning

confidence: 99%

The assessment of serum-mediated phagocytosis of necrotic material by polymorphonuclear leukocytes to diagnose and predict the clinical features of systemic lupus erythematosus: an observational longitudinal study

et al. 2016

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BackgroundSerum-mediated phagocytosis of antibody- and complement-opsonized necrotic cell material (NCM) by polymorphonuclear leukocytes can be quantified by using a flow cytometry–based assay. The phagocytosis of necrotic cell material (PNC) assay parallels the well-known lupus erythematosus cell test. In this study, we aimed to investigate the diagnostic accuracy of the assay and the relationship with clinical manifestations and disease activity in systemic lupus erythematosus (SLE).MethodsThe diagnostic accuracy for SLE diagnosis of the PNC assay was studied by cross-sectional assessment of blood samples from 148 healthy control subjects and a multicenter rheumatic group (MRG) of 529 patients with different rheumatic symptoms. A cohort of 69 patients with an established SLE diagnosis (SLE cohort) underwent longitudinal clinical and laboratory follow-up for analysis of the temporal relationships between PNC positivity and specific clinical manifestations.ResultsIn 35 of 529 MRG patients, 13 of whom had SLE, the PNC assay result was positive. Combined positivity of the PNC assay and anti–double-stranded DNA antibodies increased specificity and positive predictive value for SLE diagnosis to 0.99 and 0.67, respectively. In the longitudinal study, 42 of 69 SLE cohort patients had positive results in the PNC assay at least once. PNC assay positivity was associated with current hematological manifestations and could predict mucocutaneous manifestations. When combined with hypocomplementemia, PNC positivity preceded increased Systemic Lupus Erythematosus Disease Activity Index 2000 score, glomerulonephritis, and alopecia.ConclusionsSerum-mediated PNC by polymorphonuclear leukocytes is commonly but not exclusively seen in patients with SLE. The PNC assay may be used in follow-up of patients with SLE and, especially in combination with other routinely assessed laboratory tests, may help to predict flares and different clinical manifestations, including glomerulonephritis. Our results encourage further development of the PNC assay as a complementary laboratory tool in management of patients with SLE.

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Flow Cytometric Analysis of the LE Cell Phenomenon

Cited by 10 publications

References 19 publications

The Neutrophil’s Choice: Phagocytose vs Make Neutrophil Extracellular Traps

The Neutrophil’s Choice: Phagocytose vs Make Neutrophil Extracellular Traps

Combination of Autoantibodies Against Different Histone Proteins Influences Complement-dependent Phagocytosis of Necrotic Cell Material by Polymorphonuclear Leukocytes in Systemic Lupus Erythematosus

The assessment of serum-mediated phagocytosis of necrotic material by polymorphonuclear leukocytes to diagnose and predict the clinical features of systemic lupus erythematosus: an observational longitudinal study

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