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Bioactive murine and human interleukin-12 fusion proteins which retain antitumor activity in vivo

Nature Biotechnology volume 15, pages 35–40 (1997)Cite this article

Abstract

Interleukin-12 (IL-12) is unique amongst cytokines in being a disulfide-linked heterodimer of two separately encoded subunits (p35 and p40). We expressed single chain IL-12 proteins from retroviral constructs in which the two IL-12 subunits were linked by a 6–15 amino acid polypeptide linker, with deletion of the 22 amino acid leader sequence of the trailing subunit. The murine fusion protein IL-12.p40.L.Δp35 containing a (Gly4Ser)3 linker was stably expressed, bioactive in vitro, and had an apparent specific activity comparable to that of native and recombinant IL-12. Western blotting confirmed that murine IL-12.p40.L.Δp35 retained the linking polypeptide sequences. The analogous human IL-12.p40.L.Δp35 fusion protein containing a Gly6Ser linker was bioactive with an apparent specific activity comparable to recombinant human IL-12. In a preexisting CMS-5 tumor model, CMS-5 cells secreting either native or fusion protein forms of IL-12 prolonged survival and led to complete tumor regression.

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Author notes

  1. Graham J. Lieschke

    Present address: Ludwig Institute for Cancer Research, New York Unit at Memorial Sloan Kettering Cancer Center, Box 32, 1275 York Avenue, New York, NY, 10021

  2. Prakash K. Rao & Richard C. Mulligan

    Present address: Howard Hughes Medical Institute, Children's Hospital and Department of Genetics, Harvard Medical School, Boston, MA, 02115

Authors and Affiliations

  1. Whitehead Institute for Biomedical Research, 9 Cambridge Center, Cambridge, MA, 02142

    Graham J. Lieschke, Prakash K. Rao & Richard C. Mulligan

  2. Department of Inflammation/Autoimmune Diseases, Hoffmann-La Roche Inc., 340 Kingsland St., Nutley, NJ, 07110-1199

    Maurice K. Gately

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Correspondence to Richard C. Mulligan.

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Lieschke, G., Rao, P., Gately, M. et al. Bioactive murine and human interleukin-12 fusion proteins which retain antitumor activity in vivo. Nat Biotechnol 15, 35–40 (1997). https://doi.org/10.1038/nbt0197-35

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