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International Immunology, Vol. 2, No. 2, pp. 181-187,February 1990
© 1990 Japanese Society for Immunology

Characterization of the murine interleukin 5 receptor by using a monoclonal antibody

Naoto Yamaguchi, Yasumichi Hitoshi, Seiji Mita, Yukio Hosoya, Yushiyuki Murata, Yuji Kikuchi, Akira Tominaga and Kiyoshi Takatsu

Department of Biology, Institute for Medical Immunology, Kumamoto University Medical School 2-2-1, Honjo, Kumamoto 860, Japan

Correspondence to: Correspondence to. N. Yamaguchi, as above

Murine interleukin 5 (IL-5), a lymphokine produced by helper T cells, is involved in the regulation of growth and differentiation of B cells and other hematopoletic cells. The receptor for IL-5 has been Identified as two cross-linked complexes on T88-M cells (a murine IL-5-dependent early B cell line). In this study the IL-5 receptor was directly characterized by utilizing an immobilized IL-5 column and a rat monoclonal antibody, designated H7, directed against the IL-5 receptor. H7 completely inhibited specific binding of 35S-labeled IL-5 to T88-M cells, and bound to IL-5-responsive cells, e.g. T88-M, BCL1-B20 (a chronic B-cell leukemia), and MOPC104E (a myeloma), whereas H7 did not bind to IL-5-non-responsive cells, e.g. X5563 (a myeloma), FDC-P1 (an IL-3-dependent line), and MTH (an IL-2-dependent CTLL). H7 could barely bind to T88-M cells in the presence of IL-5, and immunoprecipitated a major band with an Mr of {small tilde}60 kd from the extract of surface-radioiodinated T88-M cells. The precipitation of this 60 kd molecule was inhibited by the addition of IL-5. Analysis with immobilized IL-5 also revealed that a 60 kd molecule bound specifically to IL-5-coupled beads compared with control beads. Furthermore, no additional molecule with a higher Mr that was recognized by H7 appeared under non-reducing, compared with reducing, conditions. The 60 kd molecule recognized by H7 could be digested with N-glycanase to yield a protein band of {small tilde}55 kd. These results suggest that the 60 kd glycoprotein defined by H7 represents at least one peptide-chain of the murine IL-5 receptor.

Keywords: lymphokine, early B-cell line, immunoprecipitation, glycoprotein

Received 10 October 1989, accepted 17 November 1989.


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