title: Eosinophil transmigration across VCAM-1-expressing endothelial cells is upregulated by antigen-stimulated mononuclear cells.
abstract: BACKGROUND
  Adhesion to and transmigration across endothelial cells expressing vascular cell adhesion molecule 1 (VCAM-1) may be key steps in the development of selective eosinophil accumulation at the allergic inflammation sites. There is evidence that cytokines/chemokines produced by CD4+ T cells play a prominent role in these processes.
  
  
  OBJECTIVE
  The objective of this study was to evaluate whether eosinophil migration across human pulmonary microvascular endothelial cells (HPMEC) expressing VCAM-1 is modulated by the supernatants of antigen-stimulated mononuclear cells obtained from atopic asthmatics.
  
  
  METHODS
  Peripheral blood mononuclear cells (PBMC) were isolated from Dermatophagoides farinae (Df)-sensitive asthmatic subjects and cultured for 96 h at 37 degrees C in the presence or absence of 1 microg/ml Df antigen. Eosinophils were isolated from blood of healthy subjects and placed on the HPMEC monolayers cultured on a transwell filter (3-microm pore size) stimulated with IL-4 plus TNF-alpha (both at 100 pM, 24 h) The supernatants of PBMC were then applied to the lower compartment and the transmigration of eosinophils was examined.
  
  
  RESULTS
  The supernatants of PBMC stimulated with Df significantly enhanced the eosinophil transmigration across VCAM-1-expressing HPMEC (% migration: 7.6 +/- 0.6 by the supernatants of PBMC cultured without Df vs. 12.3 +/- 1.2 by the PBMC cultured with Df, p < 0.01, n = 8). The enhanced migration, but not spontaneous migration, was blocked by the anti-alpha4 integrin antibody. Moreover, the enhanced transmigration was blocked by anti-CCR3 antibody.
  
  
  CONCLUSIONS
  The antigen-stimulated PBMC from atopic asthmatics produce an activity to induce eosinophil migration across VCAM-1-expressing endothelial cells. This activity appears to involve CCR3 as an essential molecule.
authors.affiliation: Pulmonary Division, Second Department of Internal Medicine, Saitama Medical School, Iruma-gun, Saitama, Japan. favre4mn@saitama-med.ac.jp
authors.forename: M
authors.initials: M
authors.lastname: Nagata
authors.forename: H
authors.initials: H
authors.lastname: Yamamoto
authors.forename: K
authors.initials: K
authors.lastname: Tabe
authors.forename: Y
authors.initials: Y
authors.lastname: Sakamoto
mesh_terms: D000906:Antibodies / Q000494:pharmacology; D015703:Antigens, CD / Q000276:immunology / Q000502:physiology; D039741:Antigens, Dermatophagoides; D001249:Asthma / Q000276:immunology*; D002478:Cells, Cultured; D002634:Chemotaxis, Leukocyte; D017077:Culture Media, Conditioned / Q000494:pharmacology; D004730:Endothelium, Vascular / Q000378:metabolism*; D004804:Eosinophils / Q000276:immunology*; D006023:Glycoproteins / Q000276:immunology; D006801:Humans; D039441:Integrin alpha4; D007963:Leukocytes, Mononuclear / Q000276:immunology*; D054397:Receptors, CCR3; D019707:Receptors, Chemokine / Q000276:immunology / Q000502:physiology; D019010:Vascular Cell Adhesion Molecule-1 / Q000378:metabolism*
chemical_list: D000906:Antibodies; D015703:Antigens, CD; D039741:Antigens, Dermatophagoides; C515350:CCR3 protein, human; D017077:Culture Media, Conditioned; D006023:Glycoproteins; D054397:Receptors, CCR3; D019707:Receptors, Chemokine; D019010:Vascular Cell Adhesion Molecule-1; D039441:Integrin alpha4