title: Intercellular Adhesion Molecule-1 and Vascular Cell Adhesion Molecule Are Induced by Ionizing Radiation on Lymphatic Endothelium.
abstract: The goal of this study was to assess the effects of ionizing radiation on the expression of the integrin ligands ICAM-1 and VCAM that control leucocyte transit by lymphatic endothelial cells.
  
  
  
  Confluent monolayers of primary human lymphatic endothelial cells (LEC) were irradiated with single dose of 2, 5, 10 or 20 Gy, with 6 MeV-x-rays using a Linear-Accelerator. ICAM-1 and VCAM expression was determined by flow cytometry. Human tissue specimens received a single dose of 20 Gy with 15 MeV-x-rays. MC38, B16-OVA or B16-VEGF-C tumors grown in C57BL/6 mice were irradiated with single dose of 20Gy using a Linear-Accelerator fitted with a 10mm Radiosurgery collimator. Clinical samples were obtained from patients previous and 4 weeks after complete standard radiotherapy. ICAM-1 and VCAM expression was detected in all tissue specimens by confocal microscopy. To understand the role of TGFβ in this process anti-TGFβ blocking mAb were injected i.p. 30min before radiotherapy. Cell adhesion to irradiated LEC was analyzed in adhesion experiments performed in the presence or in the absence of anti- TGFβ and /or anti-ICAM1 blocking mAb.
  
  
  
  We demonstrate that lymphatic endothelial cells in tumor samples experience induction of surface ICAM-1 and VCAM when exposed to ionizing radiation in a dose- and time-dependent manner. These effects can be recapitulated in cultured LEC, and are in part mediated by TGFβ. These data are consistent with increases in ICAM-1 and VCAM expression on LYVE-1+ endothelial cells in freshly explanted human tumor tissue and in mouse transplanted tumors after radiotherapy. Finally, ICAM-1 and VCAM expression accounts for enhanced adherence of human T lymphocytes to irradiated LEC.
  
  
  
  Our results show induction of ICAM-1 and VCAM on LVs in irradiated lesions and offer a starting point for elucidating the biological and therapeutic implications of targeting leukocyte traffic in combination to immunotherapy.
authors.affiliation: Division of Immunology and Immunotherapy, Center for Applied Medical Research, University of Navarra, Pamplona, Spain; Radiation Oncology, University Clinic, University of Navarra, Pamplona, Spain. Electronic address: mrruiz@unav.es.
authors.forename: María E
authors.initials: ME
authors.lastname: Rodriguez-Ruiz
authors.affiliation: Division of Immunology and Immunotherapy, Center for Applied Medical Research, University of Navarra, Pamplona, Spain.
authors.forename: Saray
authors.initials: S
authors.lastname: Garasa
authors.affiliation: Division of Immunology and Immunotherapy, Center for Applied Medical Research, University of Navarra, Pamplona, Spain.
authors.forename: Inmaculada
authors.initials: I
authors.lastname: Rodriguez
authors.affiliation: Radiation Oncology, University Clinic, University of Navarra, Pamplona, Spain.
authors.forename: Jose Luis
authors.initials: JL
authors.lastname: Solorzano
authors.affiliation: Radiation Oncology, University Clinic, University of Navarra, Pamplona, Spain.
authors.forename: Benigno
authors.initials: B
authors.lastname: Barbes
authors.affiliation: Division of Immunology and Immunotherapy, Center for Applied Medical Research, University of Navarra, Pamplona, Spain; Department of Biochemistry and Genetics, University of Navarra, Pamplona, Spain.
authors.forename: Alba
authors.initials: A
authors.lastname: Yanguas
authors.affiliation: Division of Immunology and Immunotherapy, Center for Applied Medical Research, University of Navarra, Pamplona, Spain.
authors.forename: Alvaro
authors.initials: A
authors.lastname: Teijeira
authors.affiliation: Division of Immunology and Immunotherapy, Center for Applied Medical Research, University of Navarra, Pamplona, Spain.
authors.forename: Iñaki
authors.initials: I
authors.lastname: Etxeberria
authors.affiliation: Radiation Oncology, University Clinic, University of Navarra, Pamplona, Spain.
authors.forename: José Javier
authors.initials: JJ
authors.lastname: Aristu
authors.affiliation: Pharmaceutical Immunology, Institute of Pharmaceutical Sciences, ETH Zurich, Zurich, Switzerland.
authors.forename: Cornelia
authors.initials: C
authors.lastname: Halin
authors.affiliation: Division of Immunology and Immunotherapy, Center for Applied Medical Research, University of Navarra, Pamplona, Spain; Radiation Oncology, University Clinic, University of Navarra, Pamplona, Spain.
authors.forename: Ignacio
authors.initials: I
authors.lastname: Melero
authors.affiliation: Division of Immunology and Immunotherapy, Center for Applied Medical Research, University of Navarra, Pamplona, Spain; Department of Biochemistry and Genetics, University of Navarra, Pamplona, Spain.
authors.forename: Ana
authors.initials: A
authors.lastname: Rouzaut
mesh_terms: D000818:Animals; D002448:Cell Adhesion; D045744:Cell Line, Tumor; D002465:Cell Movement; D004307:Dose-Response Relationship, Radiation; D004729:Endothelium, Lymphatic / Q000187:drug effects / Q000378:metabolism / Q000528:radiation effects*; D005434:Flow Cytometry; D015200:Fluoroimmunoassay; D006801:Humans; D018799:Intercellular Adhesion Molecule-1 / Q000378:metabolism*; D051379:Mice; D008810:Mice, Inbred C57BL; D009369:Neoplasms / Q000378:metabolism / Q000532:radiotherapy; D010315:Particle Accelerators / Q000295:instrumentation; D011829:Radiation Dosage; D011897:Random Allocation; D013601:T-Lymphocytes / Q000502:physiology; D013997:Time Factors; D016212:Transforming Growth Factor beta / Q000037:antagonists & inhibitors / Q000378:metabolism / Q000494:pharmacology; D019010:Vascular Cell Adhesion Molecule-1 / Q000378:metabolism*
chemical_list: D016212:Transforming Growth Factor beta; D019010:Vascular Cell Adhesion Molecule-1; D018799:Intercellular Adhesion Molecule-1