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J. Biol. Chem., Vol. 266, Issue 22, 14147-14150, Aug, 1991
EV Granowitz, BD Clark, J Mancilla and CA Dinarello
The interleukin-1 receptor antagonist (IL-1ra) inhibits the binding of
interleukin-1 (IL-1) to T-cell lines possessing the type I IL-1 receptor;
evidence has been published (Carter, D. B., Deibel, M. R. J., Dunn, C. J.,
Tomich, C. S., Laborde, A. L., Slightom, J. L., Berger, A. E., Bienkowski,
M. J., Sun, F. F., McEwan, R. N., Harris, P. K. W., Yem, A. W., Waszak, G.
A., Chosay, J. G., Sieu, L. C., Hardee, M. M., Zurcher-Neely, H. A.,
Reardon, I. M., Heinrickson, R. L., Truesdell, S. E., Shelly, J. A.,
Eessalu, T. E., Taylor, B. M., and Tracey, D. E. (1990) Nature 344,
633-638; Hannum, C. H., Wilcox, C. J., Arend, W. P., Joslin, F. G., Dripps,
D. J., Heimdal, P. L., Armes, L. G., Sommer, A., Eisenberg, S. P., and
Thompson, R. C. (1990) Nature 343, 336-340) that IL-Ira does not bind to
the type II IL-1 receptor (IL-1RtII). In this study we examined the ability
of human recombinant IL-1ra to block the binding of IL-1 to the IL-1RtII on
human polymorphonuclear leukocytes (PMN) and Raji human B-lymphoma cells.
The binding of 125I-IL-1 beta to PMN was competively inhibited by IL-1ra.
IL-1 beta was more potent in inhibiting the binding of 125I-IL-1 beta than
IL-1ra. Incubating PMN with 125I-IL-1ra in the presence of increasing
concentrations of IL-1 beta or IL-1ra showed that IL-1 beta was an
approximately 40-fold more potent inhibitor of binding of 125I-IL-1ra than
unlabeled IL-1ra. The IL-1ra was approximately 500-fold less potent in
inhibiting the binding of 125I-IL-1 alpha than IL-1 alpha. IL-1ra was also
able to competitively inhibit binding of 125I-IL-1 beta to Raji cells. PMN
or Raji cells were also incubated with 125I-IL-1 in the absence or presence
of IL-1 or IL-1ra. After cross-linking of IL-1 to cells followed by
specific immunoprecipitation, sodium dodecyl sulfate- polyacrylamide gel
electrophoresis showed a band at 85 kDa corresponding to the 68-kDa
IL-1RtII. However, in the presence of an excess of either unlabeled IL-1 or
IL-1ra, the 85-kDa IL-1.IL-1RtII complex was not present. These findings
demonstrate that the IL-1ra recognizes and blocks IL-1 binding to the
IL-1RtII.
Interleukin-1 receptor antagonist competitively inhibits the binding of interleukin-1 to the type II interleukin-1 receptor
Department of Medicine, New England Medical Center Hospitals, Boston, Massachusetts 02111.
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