human-neutrophil-flow-chamber-adhesion-assay

Human Neutrophil Flow Chamber Adhesion Assay

Neutrophil firm adhesion to endothelial cells plays a critical role in inflammation in both health and disease. The process of neutrophil firm adhesion ...

University of Alabama at Birmingham

We report eight microsatellite markers developed for Scirpus mariqueter Wang et Tang using 5'-anchored PCR. They were tested using 30 S. mariqueter individuals. The observed and expected heterozygosity levels ranged from 0.0690 to 0.8426 and from 0.0678 to 0.5249, respectively. Cross-amplification was carried out in a congener, S. planiculmis F. Schmidt. All these markers will be very helpful in population genetics study of S. mariqueter and S. planiculmis.

Development of microsatellites for Scirpus mariqueter Wang et Tang (Cyperaceae) and cross-species amplification in Scirpus planiculmis F. Schmidt.

Granulomatosis with polyangiitis (Wegener's) is a rare autoimmune neutrophil-mediated vasculitis that can cause renal disease and mucosal manifestations. Antineutrophil cytoplasmic antibodies (ANCA) are present in many patients, vary in level over time, and induce neutrophil activation through engagement with Fc receptors (FcRs). Given roles for FcRs in ANCA-mediated neutrophil activation and IgA antibodies in mucosal immunity, we hypothesized that FcR genetics and previously unappreciated IgA ANCA affect clinical presentation. We assembled a total of 673 patients and 413 controls from two multicenter cohorts, performed ELISA and immunofluorescence assays to determine IgA and IgG ANCA positivity, and used Illumina, TaqMan, or Pyrosequencing to genotype eight haplotype-tagging SNPs in the IgA FcR (FCAR) and to determine NA1/NA2 genotype of FCGR3B, the most prevalent neutrophil IgG FcR. We evaluated neutrophil activation by measuring degranulation marker CD11b with flow cytometry or neutrophil extracellcular trap formation with confocal microscopy. Functional polymorphisms in FCGR3B and FCAR differed between patient groups stratified by renal involvement. IgA ANCA were found in ∼30% of patients and were less common in patients with severe renal disease. Neutrophil stimulation by IgA or IgG ANCA led to degranulation and neutrophil extracellcular trap formation in a FcR allele-specific manner (IgA:FCAR P = 0.008; IgG:FCGR3B P = 0.003). When stimulated with IgA and IgG ANCA together, IgG ANCA induced neutrophil activation was reduced (P = 0.0001). FcR genotypes, IgA ANCA, and IgG ANCA are potential prognostic and therapeutic targets for understanding the pathogenesis and presentation of granulomatosis with polyangiitis (Wegener's).

IgA and IgG antineutrophil cytoplasmic antibody engagement of Fc receptor genetic variants influences granulomatosis with polyangiitis.

Multiple studies have demonstrated that single-nucleotide polymorphisms (SNPs) in the ITGAM locus (including the nonsynonymous SNPs rs1143679, rs1143678, and rs1143683) are associated with systemic lupus erythematosus (SLE). ITGAM encodes the protein CD11b, a subunit of the β2 integrin Mac-1. The purpose of this study was to determine the effects of ITGAM genetic variation on the biologic functions of neutrophil Mac-1.

Article	Year
Development of microsatellites for Scirpus mariqueter Wang et Tang (Cyperaceae) and cross-species amplification in Scirpus planiculmis F. Schmidt. Molecular ecology resources\| PubMed ID: 21564653	2009
IgA and IgG antineutrophil cytoplasmic antibody engagement of Fc receptor genetic variants influences granulomatosis with polyangiitis. Proceedings of the National Academy of Sciences of the United States of America\| PubMed ID: 22147912	2011
Multiple lupus-associated ITGAM variants alter Mac-1 functions on neutrophils. Arthritis and rheumatism\| PubMed ID: 23918739	2013

Yebin Zhou

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