Preparation and characterization of polyclonal and monoclonal antibodies against human interleukin 1α (IL 1α)

T. Kasahara, N. Mukaida, H. Shinomiya, M. Imai, Koji Matsushima, H. Wakasugi, K. Nakano

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Abstract

Polyclonal and monoclonal anti-human IL 1α antibodies (Ab) have been established. These Ab neutralized human recombinant IL 1α (rIL 1α) activity effectively, but did not interfere with human rIL 1β, murine rIL 1α, or human rIL 2 activity. Fifty percent of rIL 1α activity (25 U/ml, or 2.5 ng/ml) was neutralized by less than 0.06 μg/ml of rabbit anti-IL 1α Ab (R-38.3G) and by less than 0.13 μg/ml of monoclonal Ab (clone 28(3B1)), respectively. In other experiments, 10 μg/ml of rabbit anti-IL 1α Ab could effectively neutralize 50% of 2000 U of rIL 1α activity, and the same amount of monoclonal Ab neutralized 50% of 500 U/ml of rIL 1α activity. Not only IL 1α activity in the thymocyte costimulator assay, but also IL 1-dependent IL 2 production by a human leukemic cell line, HSB.2 subclone, were blocked by these polyclonal or monoclonal Ab. In addition, pI 4.9 IL 1 activity produced by the myelomonocytic cell line THP-1 and by the Epstein-Barr virus-transformed B cell lines, were neutralized by these Ab, suggesting that these cell lines also produce IL 1α. The specificity of these polyclonal and monoclonal Ab was further confirmed by immunochemical method (Western blotting), in which anti-IL 1α Ab reacted with rIL 1α in a specific manner. Furthermore, an enzyme-linked immunosorbent assay system has been developed that can detect low levels of IL 1α activity (<0.3 ng/ml or <3 U/ml), which is still less sensitive than thymocyte comitogenic assay and considerably less sensitive than the D10 assay. Finally, anti-IL 1α Ab-conjugated affinity columns were prepared, by which IL 1α activity, but not IL 1β activity, was specifically adsorbed and eluted effectively.

Original languageEnglish
Pages (from-to)1804-1812
Number of pages9
JournalJournal of Immunology
Volume138
Issue number6
Publication statusPublished - 19 May 1987

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    Kasahara, T., Mukaida, N., Shinomiya, H., Imai, M., Matsushima, K., Wakasugi, H., & Nakano, K. (1987). Preparation and characterization of polyclonal and monoclonal antibodies against human interleukin 1α (IL 1α). Journal of Immunology, 138(6), 1804-1812.