title: Sickle erythrocytes adhere to polymorphonuclear neutrophils and activate the neutrophil respiratory burst.
abstract: The vasoocclusive process in patients with sickle cell disease (SCD) is complex and involves interactions among sickle erythrocytes (SS-RBC), vascular endothelium, and plasma and cellular components. The role of neutrophils (PMN) in vasoocclusion has not been examined. Patients with SCD appear to have chronically activated PMN. Because the first step in PMN activation is particle recognition, we explored whether normal PMN recognize SS-RBC and whether this recognition results in PMN monolayers, significantly more SS-RBC adhered to the PMN than did normal erythrocytes (AA-RBC; P < .001). Preincubation of erythrocytes with autologous plasma significantly increased the adherence of SS-RBC to PMN but had no effect on AA-RBC (P < .001). When adhesion of density fractionated SS-RBC was performed, dense SS-RBC showed greater adherence to the PMN monolayers than did light SS-RBC (P < .001). To determine mechanisms of this adhesion, IgG and Arg-Gly-Asp-Ser (RGDS) receptor sites on PMN were saturated. IgG inhibited adherence of dense SS-RBC, whereas RGDS inhibited adherence in both fractions, although to a greater extent in the light fraction. We measured SS-RBC activation of PMN by incubating SS-RBC with 2', 7'-Dichloro-fluroescin Diacetate (DCF)-labeled PMN. Incubation of PMN with SS-RBC resulted in a significant increase in fluorescence compared to AA-RBC. We show here that PMN recognize SS-RBC through multiple mechanisms and that this recognition results in activation of PMN. These findings contribute to the understanding of vasoocclusive crisis in patients with SCD and may have therapeutic implications.
authors.affiliation: Department of Pediatrics, Childrens Hospital Los Angeles, CA 90027, USA.
authors.forename: T C
authors.initials: TC
authors.lastname: Hofstra
authors.forename: V K
authors.initials: VK
authors.lastname: Kalra
authors.forename: H J
authors.initials: HJ
authors.lastname: Meiselman
authors.forename: T D
authors.initials: TD
authors.lastname: Coates
mesh_terms: D000595:Amino Acid Sequence; D000755:Anemia, Sickle Cell / Q000097:blood*; D001783:Blood Flow Velocity; D002448:Cell Adhesion / Q000187:drug effects; D004913:Erythrocytes, Abnormal / Q000378:metabolism*; D006451:Hemoglobin, Sickle / Q000737:chemistry; D006801:Humans; D007141:Immunoglobulin Fc Fragments / Q000378:metabolism; D007074:Immunoglobulin G / Q000494:pharmacology; D007249:Inflammation; D008969:Molecular Sequence Data; D009504:Neutrophils / Q000378:metabolism*; D009842:Oligopeptides / Q000494:pharmacology; D011108:Polymers; D017452:Receptors, IgG / Q000502:physiology; D016897:Respiratory Burst
chemical_list: D006451:Hemoglobin, Sickle; D007141:Immunoglobulin Fc Fragments; D007074:Immunoglobulin G; D009842:Oligopeptides; D011108:Polymers; D017452:Receptors, IgG; C058347:glycyl-arginyl-glycyl-glutamyl-seryl-proline; C046617:arginyl-glycyl-aspartyl-serine