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Antibodies to natural killer stimulatory factorRelated Patent Categories: Drug, Bio-affecting And Body Treating Compositions, LymphokineAntibodies to natural killer stimulatory factor description/claimsThe Patent Description & Claims data below is from USPTO Patent Application 20070104680, Antibodies to natural killer stimulatory factor. Brief Patent Description - Full Patent Description - Patent Application Claims
[0001] This is a division of application Ser. No. 09/325,958, filed Jun. 4, 1999; which was a division of application Ser. No. 08/858,000, filed May 16,1997, now abandoned; which was a continuation of application Ser. No. 08/403,013, filed Mar. 13, 1995 and issued as U.S. Patent No. 5,648,467; which was a division of application Serial No. 07/584,941 filed Sep. 18, 1990 and issued as U.S. Pat. No. 5,457,038, which was a continuation-in-part of Ser. No. 07/307,817, filed Feb. 7, 1989, now abandoned; which was a continuation-in-part of Ser. No. 07/269,945, filed Nov. 10, 1988, now abandoned, all of which are incorporated herein by reference. [0002] The present invention relates to a novel cytokine that stimulates the function of natural killer cells and other cells of the immune system, and to processes for obtaining the factor in homogeneous form and producing it by recombinant genetic engineering techniques. BACKGROUND OF THE INVENTION [0003] Natural killer (NK) cells are a subset of lymphocytes active in the immune system and representing an average 15% of mononuclear cells in human peripheral blood [G. Trinchieri and B. Perussia, Lab. Invest., 50: 489 (1984)]. Among the surface markers used to identify human NK cells is a receptor binding with low affinity to the Fc fragment of IgG antibodies, such as Fc-gamma receptor III or CD16 antigen [B. Perussia et al, J. Immunol., 133:180 (1984)]. NK cells have been demonstrated to play an important role in vivo in the defense against tumors, tumor metastases, virus infection, and to regulate normal and malignant hematopoiesis. [0004] A growing family of regulatory proteins that deliver signals between cells of the immune system has been identified. These regulatory molecules are known as cytokines. Many of the cytokines have been found to control the growth, development and biological activities of cells of the hematopoietic and immune systems. These regulatory molecules include all of the colony-stimulating factors (GM-CSr, G-CSF, M-CSF, and multi CSF or interleukin-3), the interleukins (I-1 through IL-11), the interferons (alpha, beta and gamma), the tumor necrosis factors (alpha and beta) and leukemia inhibitory factor (LIF). These cytokines exhibit a wide range of biologic activities with target cells from bone marrow, peripheral blood, fetal liver, and other lymphoid or hematopoietic organs. See, e.g., G. Wong and S. Clark, Immunology Today, 2(5):137 (1988). [0005] The biochemical and biological identification and characterization of certain cytokines was hampered by the small quantities of the naturally occurring factors available from natural sources, e.g., blood and urine. Many of the cytokines have recently been molecularly cloned, heterologously expressed and purified to homogeneity. [D. Metcalf, "The Molecular Biology and Functions of the Granulocyte-Macrophage Colony Stimulating Factors," Blood, 67(2):257-267 (1986).] Among these cytokines are gamma interferon, human and murine GM-CSF, human G-CSF, human CSF-1 and human and murine IL-3. Several of these purified factors have been found to demonstrate regulatory effects on the hematopoietic and immune systems in vivo, including GM-CSF, G-CSF, IL-3 and IL-2. [0006] There remains a need in the art for additional proteins purified from their natural sources or otherwise produced in homogeneous form, which are capable of stimulating or enhancing immune responsiveness and are suitable for pharmaceutical use. BRIEF SUMMARY OF THE INVENTION [0007] In one aspect the present invention provides a novel human natural killer stimulatory factor called NKSF, which is substantially free from other mammalian proteins. Active NKSF has an apparent molecular weight of approximately 70-80 kD. Pure preparations of NKSF reveal the presence of two polypeptides, subunits of approximately 40 kD and 30 kD, which, when associated, yield active NKSF. It is presently speculated that NKSF is a heterodimer formed by association of both the larger and smaller subunits through one or more disulfide bonds. This apparent heterodimeric structure can be generated by association of the two individual subunits. [0008] The active, approximately 70-80 kD, NKSF is further characterized by containing all or a portion of the amino acid sequences of FIG. 1 (SEQ ID NO:4) and/or FIG. 2 (SEQ ID NO:6). Additionally, one or more of nine sequences of amino acids is present in the primary sequence of either the larger or smaller of the NKSF subunits. These nine amino acid fragments are listed and discussed in detail below (SEQ ID NOS:7-15). [0009] The larger subunit polypeptide of NKSF is characterized by having an apparent molecular weight of 40 kD. This subunit is further characterized by having the same or substantially the same amino acid sequence as described in FIG. 1 (SEQ ID NO:4), containing the N-terminal sequence: [0010] Ile-Trp-Glu-Leu-Lys-Lys-Asp-Val-Tyr-Val-Val-Glu-Leu-Asp-Trp-Tyr-Pro- -Asp-Ala-Pro-Gly-Glu-Met (SEQ ID NO: 1). This N-terminal amino acid sequence corresponds to amino acids #23-45 of FIG. 1. This polypeptide is further characterized by containing six of the nine amino acid fragments. [0011] The smaller polypeptide subunit of NKSF is characterized bv an apparent molecular weight of approximately 30-35 kD. Two cDNA sequences have been identified for the smaller subunit. The shorter of the two sequences is substantially contained within the longer sequence in plasmid p35nksf14-1-1, illustrated in FIG. 2. The smaller subunit is further characterized by having the same or substantially the same amino acid sequence as described in FIG. 2 (SEQ ID NO:6), containing the following N-terminal sequence: [0012] Arg-Asn-Leu-Pro-Val-Ala-Thr-Pro-Asp-Pro-Gly-Met-Phe-Pro (SEQ ID NO:2). This fragment corresponds to underlined amino acids #57-70 of the p35nksf14-1-1 clone. [0013] This smaller polypeptide is further characterized by containing three of the nine fragments of amino acids identified by underlining in FIG. 2 (SEQ ID NOS: 10-12). [0014] NKSF displays biological activity in inducing the production of gamma interferon in vitro in human peripheral blood lymphocytes (PBLs). In homogeneous form, NKSF is characterized by a specific activity of greater than 1.times.10.sup.7 dilution units per milligram in the gamma interferon induction assay, described in detail below. [0015] In addition to the induction of gamma interferon in PBLs, NKSF demonstrates the following biological activities: [0016] (1) biological activity in a granulocyte-macrophage colony stimulating factor (GM-CSF) inducing assay with PBLs; [0017] (2) biological activity in activating Natural Killer (NK) cells to kill leukemia and tumor-derived cells; [0018] (3) biological activity in a tumor necrosis factor (TNF) inducing assay with phytohemagglutinin (PHA)-activated T lymphocytes; [0019] (4) co-mitogenic activity with peripheral blood T lymphocytes; and [0020] (5) synergizes with IL-2 in inducing .gamma. IFN production in PBLs and maintaining PBL proliferation. [0021] Another aspect of the invention includes DNA sequences comprising cDNA sequences encoding the expression of a human NKSF polypeptide, a human NKSF larger subunit polypeptide, and a human NKSF smaller subunit polypeptide. Such sequences include a sequence of nucleotides encoding one or more of the subunits and peptide sequences described above. [0022] Also provided by the present invention is a vector containing a DNA sequence encoding NKSF or a subunit of NKSF in operative association with an expression control sequence. Host cells transformed with such vectors for use in producing recombinant NKSF or its recombinant subunits are also provided by the present invention. Continue reading about Antibodies to natural killer stimulatory factor... Full patent description for Antibodies to natural killer stimulatory factor Brief Patent Description - Full Patent Description - Patent Application Claims Click on the above for other options relating to this Antibodies to natural killer stimulatory factor patent application. ###  How KEYWORD MONITOR works... a FREE service from FreshPatents1. Sign up (takes 30 seconds). 2. Fill in the keywords to be monitored. 3. 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