title: L-selectin expression on neutrophils from allergic patients.
abstract: BACKGROUND
  L-selectin (CD62L) is an adhesion molecule involved in leucocyte attachment to endothelium at sites of inflammation, and it has been demonstrated that L-selectin is rapidly shed after neutrophil activation. Recently, it has been reported that there is increasing evidence of neutrophil participation in asthma and the allergic process.
  
  
  OBJECTIVE
  The present study was designed to determine whether an IgE-dependent mechanism can modulate L-selectin expression on the surface of neutrophils. Moreover, we analyse the potential implication of intracellular signal-transduction pathways and whether specific immunotherapy (IT), glucocorticoids and antihistamines might regulate this process.
  
  
  METHODS
  Peripheral blood neutrophils from three groups of donors (asthmatic group without IT treatment, IT-treated asthmatic group and healthy group) were used. Cells were challenged in vitro with the specific allergen that produced clinical symptoms in asthmatic patients and also with the allergen to which the patients were not sensitive. Neutrophils from healthy donors were also challenged with allergens. Expression of CD62L on the neutrophil surface was analysed by flow cytometry, and soluble CD62L (sCD62L) in culture supernatant by ELISA. In an attempt to discover which IgE receptor is involved, we also challenged the neutrophils with monoclonal antibody to FcepsilonRI, FcepsilonRII (CD23) and galectin-3 receptors.
  
  
  RESULTS
  When neutrophils from allergic patients were challenged with specific allergens that produce clinical allergy symptoms, L-selectin was down-regulated from the surface of those cells, accompanied by a concomitant up-regulation of soluble L-selectin in the supernatant. The challenge with antibodies against FCepsilonRI, FCepsilonRII (CD23) and galectin-3, induces down-modulation of L-selectin on the surface of the neutrophils in all three cases. Calphostin C, wortmannin and manoalide attenuated CD62L down-regulation, suggesting the potential implication of protein kinase C, phosphatidylinositol 3-kinase and phospholipase A(2) in the process. IT and glucocorticoids modulated allergen-dependent CD62L down-regulation, whereas antihistamines (terfenadine, loratadine and cetirizine) or nedocromil sodium did not affect the shedding of L-selectin.
  
  
  CONCLUSIONS
  We present evidence that the neutrophil surface expression of CD62L can be modulated by an allergen-dependent mechanism. The modulation of CD62L expression can be induced through the three receptors of IgE. This process can be affected by IT.
authors.affiliation: Servicio de Inmunología y Alergia, Hospital Universitario Virgen Macarena, Asunción 27, 41011 Seville, Spain. fmonteseirinm@meditix.es
authors.forename: J
authors.initials: J
authors.lastname: Monteseirín
authors.forename: P
authors.initials: P
authors.lastname: Chacón
authors.forename: A
authors.initials: A
authors.lastname: Vega
authors.forename: H
authors.initials: H
authors.lastname: Sánchez-Monteseirín
authors.forename: J A
authors.initials: JA
authors.lastname: Asturias
authors.forename: A
authors.initials: A
authors.lastname: Martínez
authors.forename: P
authors.initials: P
authors.lastname: Guardia
authors.forename: R
authors.initials: R
authors.lastname: Pérez-Cano
authors.forename: J
authors.initials: J
authors.lastname: Conde
mesh_terms: D000293:Adolescent; D000328:Adult; D000485:Allergens / Q000494:pharmacology; D000704:Analysis of Variance; D016022:Case-Control Studies; D002478:Cells, Cultured; D017077:Culture Media, Conditioned / Q000737:chemistry; D004305:Dose-Response Relationship, Drug; D015536:Down-Regulation; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D005260:Female; D005434:Flow Cytometry; D006801:Humans; D006967:Hypersensitivity / Q000276:immunology / Q000378:metabolism* / Q000628:therapy; D007073:Immunoglobulin E / Q000276:immunology*; D007167:Immunotherapy; D019041:L-Selectin / Q000032:analysis*; D008297:Male; D008875:Middle Aged; D009504:Neutrophils / Q000737:chemistry*; D013997:Time Factors
chemical_list: D000485:Allergens; D017077:Culture Media, Conditioned; D019041:L-Selectin; D007073:Immunoglobulin E