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

By Michiko Kobayashi, L. Fitz, M. Ryan, R. Hewick, S. Clark, Susan Chan, R. Loudon, Frederick Sherman, I. B. Perussiaj, Giorgio TRINCHIERIt. 1989

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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Citations in the corpus

IL-17 family cytokines and the expanding diversity of effector T cell lineages. (2007, background): “IL-12, a heterodimeric cytokine that had been defined as a potent inducer of IFN-γ production by NK cells (16), was soon identified as the Th1-inducing cofactor responsible for Th1 development (17, 18).”
Interleukin 12: still a promising candidate for tumor immunotherapy? (2014, background): “Based on its actions, IL-12 was initially designated as “cytotoxic lymphocyte maturation factor” [1] and “natural killer cell stimulatory factor” [2].”
Interleukin-12 and the regulation of innate resistance and adaptive immunity (2003, background): “cgi?/&Q=il12a%20or%20il12b&ORG=Hs">IL-12 was discovered and recognized to have a central role in T H 1 response”
Human Dendritic Cells Activate Resting Natural Killer (NK) Cells and Are Recognized via the NKp30 Receptor by Activated NK Cells (2002, background): “The costimulatory molecule B7–1/CD80 (23–26) and several cytokines (IL-12 [27–29]; IL-15 [30–33]; and IFN / [34–38]) have been implicated in NK activation, and early during infection, DCs can act as a major source of these stimuli (39, 40).”
Antitumor and antimetastatic activity of interleukin 12 against murine tumors (1993, background): “Among its properties are the ability to act as a NK cell and T cell growth factor (5-7), to enhance NK/LAK cell cytolytic activity (1, 7-9), to augment cytolytic T cell responses (9), and to induce secretion of cytokines, particularly IFN-3,, from T and NK cells (1, 10).”
Therapeutic targeting of the IL-12/23 pathways: generation and characterization of ustekinumab (2011, background): “and drives cell-mediated immunity by activating natural killer cells, inducing lymphokine-activated killer cells, and triggering IFN-γ production and T-cell proliferation by T H 1 cell”
Inflammatory and immune pathways in the pathogenesis of periodontal disease (2014, background): “It has the capacity to enhance T-cell and natural killer cell proliferation after activation by other stimuli (121, 189).”
New IL-12-family members: IL-23 and IL-27, cytokines with divergent functions (2005, background): “IL-12 was identified in 1989, as a soluble factor that could stimulate natural killer (NK) cells to produce IFN-&gamma”
Interleukin-23 rather than interleukin-12 is the critical cytokine for autoimmune inflammation of the brain (2003, background): “Analyses in vitro have defined IL-12 as an important factor for the differentiation of naive T cells into T-helper type 1 CD4 p lymphocytes secreting interferon-γ (ref”
Interleukins (from IL-1 to IL-38), interferons, transforming growth factor β, and TNF-α: Receptors, functions, and roles in diseases. (2016, background): “IL-12 The bioactive form of IL-12 (IL-12p70), first described as NKstimulating factor, is a heterodimer that consists of 2 subunits: a 35-kDa light chain (p35) and a 40-kDa heavy chain (p40).(114) It is produced by activated monocytes, macrophages, neutrophils, microglia, and DCs.”