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J Exp Med. Sep 1, 1989; 170(3): 827–845.
PMCID: PMC2189443

Identification and purification of natural killer cell stimulatory factor (NKSF), a cytokine with multiple biologic effects on human lymphocytes

Abstract

We have identified and purified a novel cytokine, NK cell stimulatory factor (NKSF), from the cell-free supernatant fluid of the phorbol diester-induced EBV-transformed human B lymphoblastoid cell line RPMI 8866. NKSF activity is mostly associated to a 70-kD anionic glycoprotein. The purified 70-kD protein, isolated from an SDS-PAGE gel, yields upon reduction two small species of molecular masses of 40 and 35 kD, suggesting that this cytokine is a heterodimer. When added to human PBL, purified NKSF preparations induce IFN-gamma production and synergize with rIL-2 in this activity, augment the NK cell-mediated cytotoxicity of PBL preparations against both NK-sensitive and NK- resistant target cell lines, and enhance the mitogenic response of T cells to mitogenic lectins and phorbol diesters. The three activities remain associated through different purification steps resulting in a 9,200-fold purification, and purified NKSF mediates the three biological activities at concentrations in the range of 0.1-10 pM. These data strongly suggest that the same molecule mediates these three activities, although the presence of traces of contaminant peptides even in the most purified NKSF preparations does not allow us to exclude the possibility that distinct biologically active molecules have been co-purified. The absence of other known cytokines in the purified NKSF preparations, the unusual molecular conformation of NKSF, the high specific activity of the purified protein, and the spectrum of biological activities distinguish NKSF from other previously described cytokines.

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Selected References

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  • Hercend T, Meuer S, Reinherz EL, Schlossman SF, Ritz J. Generation of a cloned NK cell line derived from the "null cell" fraction of human peripheral blood. J Immunol. 1982 Sep;129(3):1299–1305. [PubMed]
  • van de Griend RJ, van Krimpen BA, Ronteltap CP, Bolhuis RL. Rapidly expanded activated human killer cell clones have strong antitumor cell activity and have the surface phenotype of either T gamma, T-non-gamma, or null cells. J Immunol. 1984 Jun;132(6):3185–3191. [PubMed]
  • London L, Perussia B, Trinchieri G. Induction of proliferation in vitro of resting human natural killer cells: expression of surface activation antigens. J Immunol. 1985 Feb;134(2):718–727. [PubMed]
  • London L, Perussia B, Trinchieri G. Induction of proliferation in vitro of resting human natural killer cells: IL 2 induces into cell cycle most peripheral blood NK cells, but only a minor subset of low density T cells. J Immunol. 1986 Dec 15;137(12):3845–3854. [PubMed]
  • Phillips JH, Le AM, Lanier LL. Natural killer cells activated in a human mixed lymphocyte response culture identified by expression of Leu-11 and class II histocompatibility antigens. J Exp Med. 1984 Apr 1;159(4):993–1008. [PMC free article] [PubMed]
  • Perussia B, Ramoni C, Anegon I, Cuturi MC, Faust J, Trinchieri G. Preferential proliferation of natural killer cells among peripheral blood mononuclear cells cocultured with B lymphoblastoid cell lines. Nat Immun Cell Growth Regul. 1987;6(4):171–188. [PubMed]
  • Itoh K, Shiiba K, Shimizu Y, Suzuki R, Kumagai K. Generation of activated killer (AK) cells by recombinant interleukin 2 (rIL 2) in collaboration with interferon-gamma (IFN-gamma). J Immunol. 1985 May;134(5):3124–3129. [PubMed]
  • Landolfo S, Cofano F, Giovarelli M, Prat M, Cavallo G, Forni G. Inhibition of interferon-gamma may suppress allograft reactivity by T lymphocytes in vitro and in vivo. Science. 1985 Jul 12;229(4709):176–179. [PubMed]
  • Reem GH, Cook LA, Henriksen DM, Vilcek J. Gamma interferon induction in human thymocytes activated by lectins and B cell lines. Infect Immun. 1982 Jul;37(1):216–221. [PMC free article] [PubMed]
  • Michael A, Hackett JJ, Bennett M, Kumar V, Yuan D. Regulation of B lymphocytes by natural killer cells. Role of IFN-gamma. J Immunol. 1989 Feb 15;142(4):1095–1101. [PubMed]
  • Scala G, Kuang YD, Hall RE, Muchmore AV, Oppenheim JJ. Accessory cell function of human B cells. I. Production of both interleukin 1-like activity and an interleukin 1 inhibitory factor by an EBV-transformed human B cell line. J Exp Med. 1984 Jun 1;159(6):1637–1652. [PMC free article] [PubMed]
  • Klein G, Vilcek J. Attempts to induce interferon production by IdUrd induction and EBV superinfection in human lymphoma lines and their hybrids. J Gen Virol. 1980 Jan;46(1):111–117. [PubMed]
  • Yamamoto RS, Johnson DL, Masunaka IK, Granger GA. Phorbol myristate acetate induction of lymphotoxins from continuous human B lymphoid cell lines in vitro. J Biol Response Mod. 1984;3(1):76–87. [PubMed]
  • Ambrus JL, Jr, Jurgensen CH, Brown EJ, Fauci AS. Purification to homogeneity of a high molecular weight human B cell growth factor; demonstration of specific binding to activated B cells; and development of a monoclonal antibody to the factor. J Exp Med. 1985 Oct 1;162(4):1319–1335. [PMC free article] [PubMed]
  • Ythier A, Delmon L, Reinherz E, Nowill A, Moingeon P, Mishal Z, Bohuon C, Hercend T. Proliferative responses of circulating human NK cells: delineation of a unique pathway involving both direct and helper signals. Eur J Immunol. 1985 Dec;15(12):1209–1215. [PubMed]
  • Jensen JR, Sand TT, Spiegelberg HL. Generation of IgE-binding and IgG-binding factors from human lymphoblastoid cell lines. Immunology. 1984 Sep;53(1):1–8. [PMC free article] [PubMed]
  • Perussia B, Mangoni L, Engers HD, Trinchieri G. Interferon production by human and murine lymphocytes in response to alloantigens. J Immunol. 1980 Oct;125(4):1589–1595. [PubMed]
  • Cuturi MC, Murphy M, Costa-Giomi MP, Weinmann R, Perussia B, Trinchieri G. Independent regulation of tumor necrosis factor and lymphotoxin production by human peripheral blood lymphocytes. J Exp Med. 1987 Jun 1;165(6):1581–1594. [PMC free article] [PubMed]
  • Elias JA, Trinchieri G, Beck JM, Simon PL, Sehgal PB, May LT, Kern JA. A synergistic interaction of IL-6 and IL-1 mediates the thymocyte-stimulating activity produced by recombinant IL-1-stimulated fibroblasts. J Immunol. 1989 Jan 15;142(2):509–514. [PubMed]
  • Muraguchi A, Kishimoto T, Miki Y, Kuritani T, Kaieda T, Yoshizaki K, Yamamura Y. T cell-replacing factor- (TRF) induced IgG secretion in a human B blastoid cell line and demonstration of acceptors for TRF. J Immunol. 1981 Aug;127(2):412–416. [PubMed]
  • Cuturi MC, Anegón I, Sherman F, Loudon R, Clark SC, Perussia B, Trinchieri G. Production of hematopoietic colony-stimulating factors by human natural killer cells. J Exp Med. 1989 Feb 1;169(2):569–583. [PMC free article] [PubMed]
  • Trinchieri G, De Marchi M, Mayr W, Savi M, Ceppellini R. Lymphocyte antibody lymphocytolytic interaction (LALI) with special emphasis on HL-A. Transplant Proc. 1973 Dec;5(4):1631–1649. [PubMed]
  • Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. [PubMed]
  • Trinchieri G, Kobayashi M, Rosen M, Loudon R, Murphy M, Perussia B. Tumor necrosis factor and lymphotoxin induce differentiation of human myeloid cell lines in synergy with immune interferon. J Exp Med. 1986 Oct 1;164(4):1206–1225. [PMC free article] [PubMed]
  • Trinchieri G, Santoli D. Anti-viral activity induced by culturing lymphocytes with tumor-derived or virus-transformed cells. Enhancement of human natural killer cell activity by interferon and antagonistic inhibition of susceptibility of target cells to lysis. J Exp Med. 1978 May 1;147(5):1314–1333. [PMC free article] [PubMed]
  • Trinchieri G, Matsumoto-Kobayashi M, Clark SC, Seehra J, London L, Perussia B. Response of resting human peripheral blood natural killer cells to interleukin 2. J Exp Med. 1984 Oct 1;160(4):1147–1169. [PMC free article] [PubMed]
  • Wilson AB, Harris JM, Coombs RR. Interleukin-2-induced production of interferon-gamma by resting human T cells and large granular lymphocytes: requirement for accessory cell factors, including interleukin-1. Cell Immunol. 1988 Apr 15;113(1):130–142. [PubMed]
  • Garman RD, Jacobs KA, Clark SC, Raulet DH. B-cell-stimulatory factor 2 (beta 2 interferon) functions as a second signal for interleukin 2 production by mature murine T cells. Proc Natl Acad Sci U S A. 1987 Nov;84(21):7629–7633. [PMC free article] [PubMed]
  • Antonelli G, Dianzani F, Van Damme J, Amicucci P, De Marco F, Cefaro A. A macrophage-derived factor different from interleukin 1 and able to induce interferon-gamma and lymphoproliferation in resting T lymphocytes. Cell Immunol. 1988 May;113(2):376–386. [PubMed]
  • Antonelli G, Blalock JE, Dianzani F. Generation of a soluble IFN-gamma inducer by oxidation of galactose residues on macrophages. Cell Immunol. 1985 Sep;94(2):440–446. [PubMed]
  • Ostensen ME, Thiele DL, Lipsky PE. Tumor necrosis factor-alpha enhances cytolytic activity of human natural killer cells. J Immunol. 1987 Jun 15;138(12):4185–4191. [PubMed]
  • Henney CS, Kuribayashi K, Kern DE, Gillis S. Interleukin-2 augments natural killer cell activity. Nature. 1981 May 28;291(5813):335–338. [PubMed]
  • Ortaldo JR, Mason AT, Gerard JP, Henderson LE, Farrar W, Hopkins RF, 3rd, Herberman RB, Rabin H. Effects of natural and recombinant IL 2 on regulation of IFN gamma production and natural killer activity: lack of involvement of the Tac antigen for these immunoregulatory effects. J Immunol. 1984 Aug;133(2):779–783. [PubMed]
  • Kehrl JH, Dukovich M, Whalen G, Katz P, Fauci AS, Greene WC. Novel interleukin 2 (IL-2) receptor appears to mediate IL-2-induced activation of natural killer cells. J Clin Invest. 1988 Jan;81(1):200–205. [PMC free article] [PubMed]
  • Mond JJ, Brunswick M. A role for IFN-gamma and NK cells in immune responses to T cell-regulated antigens types 1 and 2. Immunol Rev. 1987 Oct;99:105–118. [PubMed]
  • Grimm EA, Mazumder A, Zhang HZ, Rosenberg SA. Lymphokine-activated killer cell phenomenon. Lysis of natural killer-resistant fresh solid tumor cells by interleukin 2-activated autologous human peripheral blood lymphocytes. J Exp Med. 1982 Jun 1;155(6):1823–1841. [PMC free article] [PubMed]
  • Rosenberg S. Lymphokine-activated killer cells: a new approach to immunotherapy of cancer. J Natl Cancer Inst. 1985 Oct;75(4):595–603. [PubMed]

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