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Research ArticleArticle

Characterization of Interleukin-1α Binding to Mouse Brain Endothelial Cells

William A. Banks
Journal of Pharmacology and Experimental Therapeutics November 1999, 291 (2) 665-670;
William A. Banks
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Abstract

In vivo studies have shown that interleukin (IL)-1α binds to and is transported across brain endothelial cells, whereas in vitro studies have shown that brain endothelial cells respond to IL and contain mRNA for the IL-type 1 receptor. However, these binding sites have yet to be characterized. Herein, we used murine brain microvessels to characterize the binding of IL labeled with 125I. Binding was temperature- and time-dependent with maximal binding after 4 h of incubation at 37°C. The amount of radioactivity determined by HPLC to represent intact 125I-labeled murine IL-1α at 4 h was ∼100% in the incubation fluid and 80 to 90% for radioactive material recovered from the incubated cells.Bmax was 0.955 fmol and theKd was 292 pM for human 125I-IL and binding was displaced by interleukin-1β and interleukin-1 receptor antagonist but not by tumor necrosis factor α. Binding was dependent on magnesium and glucose. Incubation with antibodies showed that the binding site was not identical with the IL-type 1 receptor but closely resembled the blood-brain barrier transporter. These results show that murine brain endothelial cells have specific binding sites for IL and that these sites more closely resemble the transporter than the type 1 receptor.

Footnotes

  • Send reprint requests to: William A. Banks, VAMC, 915 N. Grand Blvd., St. Louis, MO 63106. E-mail: bankswa{at}slu.edu

  • ↵1 Supported by Veterans Affairs merit review and RO1 MH54979.

  • Abbreviations:
    IL
    interleukin
    CNS
    central nervous system
    BBB
    blood-brain barrier
    I-hIL,125I-labeled human IL-1α
    I-mIL, 125I-labeled murine IL-1α
    %TB
    percent of total binding
    IL-1ra
    IL-1 receptor antagonist
    TNF-α
    tumor necrosis factor α
    R-B
    receptor blocking
    R-NB
    receptor nonblocking
    I-B
    IL blocking
    I-NB
    IL nonblocking
    • Received March 30, 1999.
    • Accepted July 12, 1999.
  • The American Society for Pharmacology and Experimental Therapeutics
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Journal of Pharmacology and Experimental Therapeutics: 291 (2)
Journal of Pharmacology and Experimental Therapeutics
Vol. 291, Issue 2
1 Nov 1999
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Research ArticleArticle

Characterization of Interleukin-1α Binding to Mouse Brain Endothelial Cells

William A. Banks
Journal of Pharmacology and Experimental Therapeutics November 1, 1999, 291 (2) 665-670;

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Research ArticleArticle

Characterization of Interleukin-1α Binding to Mouse Brain Endothelial Cells

William A. Banks
Journal of Pharmacology and Experimental Therapeutics November 1, 1999, 291 (2) 665-670;
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