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Interleukin-6 is antiproliferative to a mouse hybridoma cell line and promotive for its antibody productivity

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Abstract

Monoclonal antibody production by hybridoma cells at moderately slowed growth states would be favorable for commercial scale production since cells can devote their resources to performing the differentiated function, immunoglobulin production. We found that a purified recombinant human interleukin-6, which had been reported to support or stimulate proliferation of B cell hybridoma/plasmacytoma cells, suppressed growth of a hybridoma cell line in serum-free medium. In the presence of the interleukin, the growth-suppressed cells were viable for remarkably long periods in batch culture, and after removal of the interleukin from the culture medium, they started to proliferate at their normal growth rate. As the concentration of the interleukin increased in the culture, the growth rate decreased and the specific antibody productivity (antibody production rate per cell) increased to 5-fold of control at 10 U ml−1 (2 ng ml−1) of the interleukin.

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Abbreviations

IL-1,2, and 6:

interleukin-1, 2 and 6

rhIL-6:

recombinant human interleukin-6

MCAb:

monoclonal antibody

TNP:

trinitrophenyl

unit (U) of interleukin-6:

A unit (U) is equivalent to the amounts of IL-6 which gives one-half maximal IgM secretion by SKW6-CL4 cells (1U ml−1=200 pg ml−1)

References

  • Andus, T, Geiger, T, Hirano, T, Northoff, H, Ganter, U, Bauer, J, Kishimoto, T and Heinrich, PC (1987) Recombinant human B cell stimulatory factor 2 (BSF-2, IFN-β2) regulates β-fibrinogen and albumin mRNA levels in FAO-9 cells. FEBS Lett. 221: 18–22.

    Google Scholar 

  • Dean, RC Jr, Karkare, SB, Ray, NG, Runstadler, PW Jr and Venkatasubramanian, K (1988) Large scale culture of hybridoma and mammalian cells in fluidized bed bioreactors. In: Moo-Young, M (ed.), Bioreactor Immobilized Enzymes and Cells: Fundamentals and Applications (pp. 125–142). Elsevier Applied Science Publishers, New York.

    Google Scholar 

  • Engvall, E (1980) Enzyme immunoassay ELISA and EMIT. In: Van Vunakis, H and Langone, JJ (eds) Methods in Enzymology. Vol. 70 (pp. 419–439). Academic Press, New York.

    Google Scholar 

  • Flickinger MC and Bibila TA (1991) Modeling and experimental evidence of the effect of intracellular energy availability on monoclonal antibody secretion rate. Presented at 4th Annual Meeting of Japanese Association for Animal Cell Technology (JAACT '91), 13–15 November 1991, Fukuoka, Japan.

  • Garatun-Tjeldstø, O, Pryme, IF, Weltman, JK and Dowben, RM (1976) Synthesis and secretion of light-chain immunoglobulin in two successive cycles of synchronized plasmacytoma cells. J. Cell. Biol. 68: 232–239.

    Google Scholar 

  • Garman, RD, Jacobs, KA, Clark, SC and Raulet, DH (1987) B cell stimulatory factor 2 (β2 interferon) functions as a second signal for interleukin-2 production by mature murine T cells. Proc. Natl. Acad. Sci. USA 84: 7629–7633.

    Google Scholar 

  • Gauldie, J, Richards, C, Harmish, D, Lansdorp, P and Baumann, H (1987) Interferon β2/B cell stimulatory factor type 2 shares identity with monocyte-derived hepatocyte-stimulating factor and regulates the major acute phase protein response in liver cells. Proc. Natl. Acad. Sci. USA 84: 7251–7255.

    Google Scholar 

  • Hirano, T, Taga, T, Nakano, N, Yasukawa, K Kashiwamura, S, Shimizu, K, Nakajima, K, Pyun, KH and Kishimoto, T (1985) Purification to homogeneity and characterization of human B cell differentiation factor (BCDF or BSFp-2). Proc. Natl. Acad. Sci. USA 82: 5490–5494.

    Google Scholar 

  • Ikebuchi, K, Wong, GG, Clark, SC, Ihle, JN, Hirai, Y and Ogawa, M (1987) Interleukin-6 enhancement of interleukin-3-dependent proliferation of multipotential hemopoietic progenitors. Proc. Natl. Acad. Sci. USA 84: 9035–9039.

    Google Scholar 

  • Kawano, M, Hirano, T, Matsuda, T, Taga, T, Horii, Y, Iwato, K, Asaoku, H, Tang, B, Tanabe, O, Tanaka, H, Kuramoto, A and Kishimoto, T (1988) Autocrine generation and requirement of BSF-2/IL-6 for human multiple myelomas. Nature 332: 83–85.

    Google Scholar 

  • Lotz, M, Jirik, F, Kabouridis, P, Tsoukas, C, Hirano, T, Kishimoto, T and Carson, DA (1988) B cell stimulating factor 2/interleukin-6 is a co-stimulant for human thymocytes and T lymphocytes. J. Exp. Med. 167: 1253–1258.

    Google Scholar 

  • Miller, WM, Blanch, HW and Wilke, CR (1988) A kinetic analysis of hybridoma growth and metabolism in batch and continuous suspension culture: effect of nutrient concentration, dilution rate and pH. Biotechnol. Bioeng. 32: 947–965.

    Google Scholar 

  • Miyaura, C, Onozaki, K, Akiyama, Y, Taniyama, T, Hirano, T, Kishimoto, T and Suda, T (1988) Recombinant human interleukin-6 (B cell stimulatory factor 2) is a potent inducer of differentiation of mouse myeloid leukemia cells (MI). FEBS Lett. 234: 17–21.

    Google Scholar 

  • Muraguchi, A, Hirano, T, Tang, B, Matsuda, T, Horii, Y, Nakajima, K and Kishimoto, T (1988) The essential role of B cell stimulatory factor 2 (BSF-2/IL-6) for the terminal differentiation of B cells. J. Exp. Med. 167: 332–344.

    Google Scholar 

  • Nordan, RP and Potter, M (1986) A macrophage-derived factor required by plasmacytomas for survival and proliferation in vitro. Science 233: 566–569.

    Google Scholar 

  • Okada, M, Kitahara, M, Kishimoto, S, Matsuda, T, Hirano, T and Kishimoto, T (1988) IL-6/BSF-2 functions as a killer helper factor in the in vitro induction of cytotoxic T cells. J. Immunol. 141: 1543–1549.

    Google Scholar 

  • Onozaki, K, Akiyama, Y, Okano, A, Hirano, T, Kishimoto, T, Hashimoto, T, Yoshizawa, K and Taniyama, T (1989) Synergistic regulatory effects of interleukin-6 and interleukin-1 on the growth and differentiation of human and mouse myeloid leukemic cell lines. Cancer Res. 49: 3602–3607.

    Google Scholar 

  • Reuveny, S, Velez, D, Miller, L and Macmillan, JD (1986) Comparison of cell propagation methods for their effect on monoclonal antibody yield in fermentors. J. Immunol. Methods 86: 61–69.

    Google Scholar 

  • Suzuki, E, Ollis, DF (1990) Enhanced antibody production at slowed growth rates: experimental demonstration and a simple structured model. Biotechnol. Prog. 6: 231–236.

    Google Scholar 

  • Suzuki E, Takahashi K and Ollis DF (1991) A simple structured model for antibody production rate: cell cycle model combined with mRNA balance. Proceedings 4th Annual Meeting of Japanese Association for Animal Cell Technology (JAACT '91) Fukuoka Japan, in printing.

  • Takai, Y, Wong, GG, Clark, SC, Burakoff, SJ and Hermann, SH (1988) B cell stimulatory factor 2 is involved the differentiation of cytotoxic T lymphocytes. J. Immunol. 140: 508–512.

    Google Scholar 

  • Tohyama, N, Karasuyama, H and Tada, T (1990) Growth autonomy and tumorigenicity of interleukin-6-dependent B cells transfected with interleukin-6 cDNA. J. Exp. Med. 171: 389–400.

    Google Scholar 

  • Van Damme, J, Opdenakker, G, Simpson, RJ, Rubira, MR, Cayphas, S, Vink, A, Billiau, A and Van Snick, J (1987) Identification of the human 26 kD protein, interferon β2 (IFN-β2), as a B cell hybridoma/plasmacytoma growth factor induced by interleukin-1 and tumor necrosis factor. J. Exp. Med. 165: 914–919.

    Google Scholar 

  • Van Snick, J, Cayphas, S, Szikora, J-P, Renauld, J-C, Roost, EV, Boon, T and Simpson, RJ (1988) cDNA cloning of murine interleukin-HP1: homology with human interleukin-6. Eur. J. Immunol. 18: 193–197.

    Google Scholar 

  • Van Snick, J, Cayphas, S, Vink, A, Uyttenhove, C, Coulie, PG, Rubira, MR and Simpson, RJ (1986) Purification and NH2-terminal amino acid sequence of a T cell-derived lymphokine with growth factor activity for B cell hybridomas. Proc. Natl. Acad. Sci. USA 83: 9679–9683.

    Google Scholar 

  • Van Snick, J, Vink, A, Cayphas, S and Uyttenhove, C (1987) Interleukin-HP1, a T cell-derived hybridoma growth factor that supports the in vitro growth of murine plasmacytomas. J. Exp. Med. 165: 641–649.

    Google Scholar 

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Makishima, F., Terada, S., Mikami, T. et al. Interleukin-6 is antiproliferative to a mouse hybridoma cell line and promotive for its antibody productivity. Cytotechnology 10, 15–23 (1992). https://doi.org/10.1007/BF00376096

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