Increased expression and phosphorylation of the two S100A9 isoforms in mononuclear cells from patients with systemic lupus erythematosus: A proteomic signature for circulating low-density granulocytes

Pavón, Esther J.; García‐Rodríguez, Sonia; Zumaquero, Esther; Perandrés-López, Rubén; Rosal-Vela, Antonio; Lario, Antonio; Longobardo, Victoria; Carrascal, Montserrat; Abián, Joaquín; Callejas‐Rubio, José Luis; Ortego-Centeno, Norbérto; Zubiaur, Mercedes; Sancho, Jaime

doi:10.1016/j.jprot.2011.12.020

Cited by 21 publications

(18 citation statements)

References 43 publications

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“…3B) highlighted that spots 1A and 11 corresponded to phosphorylated proteins and spots 2 and 1B to non-phosphorylated proteins. This result is in agreement with the identification of the S100A9 protein species recently obtained by Pavon et al on peripheral blood mononuclear cells from adults [18]. These authors reported a decrease in pI of 0.16 units between the non-phosphorylated long and short S100A9, due to the loss of Lys 4 in the short-form, and an identical pI decrease in phosphorylated long and short S100A9 with respect to the corresponding non-phosphorylated forms.…”

Section: S100 Familysupporting

confidence: 92%

Proteomic characterization of the acid-insoluble fraction of whole saliva from preterm human newborns

Arba

Iavarone

Vincenzoni

et al. 2016

Journal of Proteomics

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Section: S100 Familysupporting

confidence: 92%

Proteomic characterization of the acid-insoluble fraction of whole saliva from preterm human newborns

Arba

Iavarone

Vincenzoni

et al. 2016

Journal of Proteomics

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“…In support of the gene array analysis, high levels of calprotectin (S100A8/S100A9) were recently reported in a proteomic analysis of SLE PBMCs, in direct correlation with the presence of LDGs. These observations suggest a proteomic signature for circulating LDGs in lupus patients that requires further investigation [26]. Overall, based on the gene expression profile and the ultrastructural analysis of LDGs, it is possible that they represent a more immature granulocyte subset prematurely released from the marrow due to yet uncharacterized stimuli.…”

Section: Gene Array Analysis Of Ldgsmentioning

confidence: 99%

Low-density granulocytes: a distinct class of neutrophils in systemic autoimmunity

2013

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Recent studies have renewed the interest on the potential role that neutrophils play in the development of systemic lupus erythematosus (SLE) and other autoimmune conditions. A distinct subset of proinflammatory, low-density granulocytes (LDGs) isolated from the peripheral blood mononuclear cell fractions of patients with SLE has been described. While the origin and role of LDGs needs to be fully characterized, there is evidence that these cells may contribute to lupus pathogenesis and to the development of end-organ damage through heightened proinflammatory responses, altered phagocytic capacity, enhanced ability to synthesize type I Interferons and to kill endothelial cells. Furthermore, these cells readily form neutrophil extracellular traps (NETs), a phenomenon that may promote autoantigen externalization and organ damage. This review examines the biology and potential origin of LDGs, describes the ultrastructural characteristics of these cells, and discusses their putative pathogenic role in systemic autoimmune diseases.

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“…Now the research extends beyond rheumatism to diseases such as human immunodeficiency virus (HIV) infection, severe infections, tumors, and neonatal disorders where a significantly increased number of LDGs in PBMCs have been reported (Morisaki et al, 1992;Rodriguez et al, 2009;Lin et al, 2011;Cloke et al, 2012;Hoffmann et al, 2013;Ssemaganda et al, 2014). In the classical autoimmune disease of SLE, the LDG percentage in PBMCs was determined for 35 patients and 31 healthy controls, and the results indicated that the average percentage was 15% in SLE patents and 2% in healthy controls (Pavón et al, 2012). In another study, the LDG average percentage was 18% in 65 patients with SLE and 5% in 22 healthy controls (Denny et al, 2010).…”

Section: Discussionmentioning

confidence: 99%

“…Fluorescein isothiocyanate (FITC)-labeled anti-CD15 (Biolegend, Catalog: 301904) and phycoerythrin (PE)-labeled anti-CD14 (Biolegend, Catalog: 325606) antibodies were used for flow (Biolegend, Catalog: 401605, 400111) were used to calculate the amount of nonspecific staining. Lymphocytes, monocytes, and granulocytes were gated according to their forward and scatter characteristics as previously described (Denny et al, 2010;Pavón et al, 2012). Two color immunofluorescence analyses were performed on a Beckman Coulter Epics XL flow cytometer (Beckman Coulter, Inc., CA, USA).…”

Section: Pbmc Isolation and Ldg Measurementmentioning

confidence: 99%

“…Low-density granulocytes (LDGs) have been observed in peripheral blood mononuclear cells (PBMCs) of patients with systemic lupus erythematosus (SLE), a typical autoimmune disease (Denny et al, 2010;Pavón et al, 2012). Similar to mature autologous and healthy control neutrophils, LDGs express highly the neutrophil surface markers CD15, CD10, CD11b, CD11c, and CD16, but unlike neutrophils, their nuclear shape is of an immature phenotype (Denny et al, 2010).…”

Section: Introductionmentioning

confidence: 99%

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A simple method for removing low-density granulocytes to purify T lymphocytes from peripheral blood mononuclear cells

Zhang

Song

Shu

et al. 2017

J. Zhejiang Univ. Sci. B

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Abstract:Objective: Low-density granulocytes (LDGs) can form neutrophil extracellular traps (NETs) spontaneously and excessively. When peripheral blood mononuclear cells (PBMCs) are used for studying T lymphocytes, LDGs contained in the PBMCs may decrease the threshold of activating T lymphocytes by forming NETs. This study focused on the profiles of LDGs in common autoimmune diseases and methods for removing LDGs from PBMCs. Methods: The percentages of LDGs in PBMCs from 55 patients with dermatomyositis (DM), 15 with polymyositis (PM), 42 with rheumatoid arthritis (RA), 25 with systemic lupus erythematosus (SLE), and 19 healthy controls were determined by flow cytometry. Three methods of removing LDGs were explored and compared. After removal, PBMCs from six patients with positive T-SPOT.TB were tested again to find out if LDGs contained in the PBMCs could influence T lymphocyte reactions. Results: Significantly higher LDG percentages were found in PBMCs from patients with DM ((8.41±10.87)%, P<0.0001), PM ((8.41±10.39)%, P<0.0001), RA ((4.05±6.97)%, P=0.0249), and SLE ((7.53±11.52)%, P=0.0006), compared with the controls ((1.28±0.73)%). The T-SPOT.TB values significantly decreased after LDGs were removed. Increasing relative centrifugal force (RCF) within a limited range can decrease the LDG percentage from an initial high level, but not markedly increase the LDG clearance rate. Compared with the whole blood sediment method, the PBMC adherence method can significantly remove LDGs yet scarcely influence the T lymphocyte percentage in PBMCs. Conclusions: The LDG percentage in PBMCs is significantly increased in patients with SLE, DM, PM, and RA. The influence of LDGs on T lymphocytes cannot be ignored in PBMC cultures. The adherence method is a simple and easy-to-use method for removing LDGs and purifying T lymphocytes from PBMCs.

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Increased expression and phosphorylation of the two S100A9 isoforms in mononuclear cells from patients with systemic lupus erythematosus: A proteomic signature for circulating low-density granulocytes

Cited by 21 publications

References 43 publications

Proteomic characterization of the acid-insoluble fraction of whole saliva from preterm human newborns

Proteomic characterization of the acid-insoluble fraction of whole saliva from preterm human newborns

Low-density granulocytes: a distinct class of neutrophils in systemic autoimmunity

A simple method for removing low-density granulocytes to purify T lymphocytes from peripheral blood mononuclear cells

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