TNF-alpha-induced transendothelial neutrophil migration is IL-8 dependent

Am J Physiol. 1994 Mar;266(3 Pt 1):L238-45. doi: 10.1152/ajplung.1994.266.3.L238.


The early phases of airway inflammation include complex interactions between leukocytes, vascular endothelium, and inflammatory cytokines. Therefore, we examined tumor necrosis factor-alpha (TNF-alpha)-induced neutrophil migration through polycarbonate filters and human umbilical vein endothelial (HUVE) cells cultured as monolayers on these filters. TNF-alpha induced both dose- and time-dependent migration of neutrophils across both barriers. At 10(-11)-10(-9) M TNF-alpha, neutrophil migration across HUVE monolayers was more than twofold greater than that observed across naked filters. Modified checkerboard experiments indicated that neutrophils crossed naked filters as a chemokinetic rather than chemotactic response. Supernatants of TNF-alpha (10(-9) M)-stimulated HUVE monolayers induced threefold greater migration of neutrophils across naked filters than 10(-9) M TNF-alpha itself, suggesting release of soluble chemotactic factor(s). Pretreatment of HUVE monolayers with actinomycin D inhibited both TNF-alpha-induced production of soluble chemotactic factors and transendothelial neutrophil migration by > 90%. Supernatants from TNF-alpha-treated HUVE cells contained significant concentrations of interleukin 8 (IL-8), and coincubation of these supernatants with anti-IL-8 decreased supernatant-induced chemotaxis. Finally, coincubation of TNF-alpha with anti-IL-8 during transmigration experiments nearly completely inhibited the increase in neutrophil migration measured across HUVE monolayers. In contrast, WEB 2086, a platelet-activating factor receptor antagonist, had no effect. Therefore, endothelial cells greatly facilitate TNF-alpha-induced transcellular migration of neutrophils. This facilitation is dependent on TNF-alpha-stimulated production of IL-8. These data further support the active role of vascular endothelium in recruiting leukocytes to sites of inflammation.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Antibodies / immunology
  • Cell Movement / drug effects
  • Cells, Cultured
  • Chemotaxis, Leukocyte
  • Dose-Response Relationship, Drug
  • Endothelium, Vascular / cytology
  • Endothelium, Vascular / metabolism
  • Humans
  • Interleukin-8 / pharmacology*
  • Kinetics
  • Neutrophils / drug effects*
  • Neutrophils / metabolism
  • Solubility
  • Tumor Necrosis Factor-alpha / immunology
  • Tumor Necrosis Factor-alpha / pharmacology*
  • Umbilical Veins / cytology
  • Umbilical Veins / metabolism


  • Antibodies
  • Interleukin-8
  • Tumor Necrosis Factor-alpha