WO1996039424A1 - Facteur c stimulant les cellules tueuses naturelles - Google Patents

Facteur c stimulant les cellules tueuses naturelles Download PDF

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Publication number
WO1996039424A1
WO1996039424A1 PCT/US1995/007200 US9507200W WO9639424A1 WO 1996039424 A1 WO1996039424 A1 WO 1996039424A1 US 9507200 W US9507200 W US 9507200W WO 9639424 A1 WO9639424 A1 WO 9639424A1
Authority
WO
WIPO (PCT)
Prior art keywords
polypeptide
nkef
polynucleotide
cells
dna
Prior art date
Application number
PCT/US1995/007200
Other languages
English (en)
Inventor
Jian Ni
Guo-Liang Yu
Reiner Gentz
Craig A. Rosen
Original Assignee
Human Genome Sciences, Inc.
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Human Genome Sciences, Inc. filed Critical Human Genome Sciences, Inc.
Priority to PCT/US1995/007200 priority Critical patent/WO1996039424A1/fr
Priority to AU28186/95A priority patent/AU2818695A/en
Publication of WO1996039424A1 publication Critical patent/WO1996039424A1/fr

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Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/52—Cytokines; Lymphokines; Interferons
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides

Definitions

  • NK cells are also important controlling viral infection and the regulation of hematopoiesis (Trinchieri).
  • Figure 4 illustrates the growth inhibitory activity of NKEF C against Jurkat human T-cell leukemia cells.
  • Figure 5 illustrates the effect of NKEF C on VSV lytic infection.
  • the polynucleotide of the present invention may be in the form of RNA or in the form of DNA, which DNA includes cDNA, genomic DNA, and synthetic DNA.
  • the DNA may be double- stranded or single-stranded, and if single stranded may be the coding strand or non-coding (anti-sense) strand.
  • the coding sequence which encodes the mature polypeptide may be identical to the coding sequence shown in Figure 1 (SEQ ID NO:l) or that of the deposited clone or may be a different coding sequence which coding sequence, as a result of the redundancy or degeneracy of the genetic code, encodes the same mature polypeptide as the DNA of Figure 1 (SEQ ID NO:l) or the deposited cDNA.
  • Fragments of the full length NKEF C gene may be used as a hybridization probe for a cDNA library to isolate the full length gene and to isolate other genes which have a high sequence similarity to the NKEF C gene or similar biological activity.
  • Probes of this type preferably have at least 30 bases and may contain, for example, 50 or more bases.
  • the probe may also be used to identify a cDNA clone corresponding to a full length transcript and a genomic clone or clones that contain the complete NKEF C gene including regulatory and promotor regions, exons, and introns.
  • An example of a screen comprises isolating the coding region of the NKEF C gene by using the known DNA sequence to synthesize an oligonucleotide probe. Labeled oligonucleotides having a sequence complementary to that of the gene of the present invention are used to screen a library of human cDNA, genomic DNA or mRNA to determine which members of the library the probe hybridizes to.
  • Microbial cells employed in expression of proteins can be disrupted by any convenient method, including freeze-thaw cycling, sonication, mechanical disruption, or use of cell lysing agents, such methods are well known to those skilled in the art.
  • the present invention also relates to a diagnostic assay for detecting altered levels of NKEF C protein in various tissues since over-expression compared to normal control tissue samples can detect the presence of a tumor or viral infection.
  • Assays used to detect levels of NKEF C protein in a sample derived from a host are well-known to those of skill in the art and include radioimmunoassays, competitive-binding assays, Western Blot analysis and preferably an ELISA assay.
  • An ELISA assay initially comprises preparing an antibody specific to the NKEF C antigen, preferably a monoclonal antibody. In addition a reporter antibody is prepared against the monoclonal antibody.
  • Transformants are identified by their ability to grow on LB plates and ampicillin/kanamycin resistant colonies were selected. Plasmid DNA was isolated and confirmed by restriction analysis. Clones containing the desired constructs were grown overnight (0/N) in liquid culture in LB media supplemented with both Amp (100 ug/ml) and Kan (25 ug/ml) . The O/N culture is used to inoculate a large culture at a ratio of 1:100 to 1:250. The cells were grown to an optical density 600 (O.D. 600 ) of between 0.4 and 0.6. IPTG ( Isopropyl-B-D-thiogalacto pyranoside”) was then added to a final concentration of 1 mM.
  • O.D. 600 optical density 600
  • Fibroblasts are obtained from a subject by skin biopsy.
  • the resulting tissue is placed in tissue-culture medium and separated into small pieces. Small chunks of the tissue are placed on a wet surface of a tissue culture flask, approximately ten pieces are placed in each flask.
  • the flask is turned upside down, closed tight and left at room temperature over night. After 24 hours at room temperature, the flask is inverted and the chunks of tissue remain fixed to the bottom of the flask and fresh media (e.g., Ham's F12 media, with 10% FBS, penicillin and streptomycin, is added. This is then incubated at 37°C for approximately one week. At this time, fresh media is added and subsequently changed every several days.
  • fresh media e.g., Ham's F12 media, with 10% FBS, penicillin and streptomycin

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  • Chemical & Material Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Organic Chemistry (AREA)
  • Biochemistry (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Zoology (AREA)
  • Biophysics (AREA)
  • General Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Medicinal Chemistry (AREA)
  • Molecular Biology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Toxicology (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)

Abstract

La présente invention concerne un facteur C stimulant les cellules tueuses naturelles et l'ADN (ARN) codant ce polypeptide, ainsi qu'une procédure de production dudit polypeptide par des techniques de recombinaison. Elle concerne également des procédés d'utilisation de ce polypeptide pour la prévention et/ou le traitement d'infections virales, d'une inflammation, de la néoplasie et de lésions dues à des radicaux de superoxydes. L'invention a en outre pour objet des dosages de diagnostic permettant d'identifier des mutations dans la séquence d'acide nucléique codant un polypeptide du type de celui qu'elle décrit, et de détecter des modifications du niveau dudit polypeptide pour déceler la présence de maladies telles que, par exemple, le cancer.
PCT/US1995/007200 1995-06-06 1995-06-06 Facteur c stimulant les cellules tueuses naturelles WO1996039424A1 (fr)

Priority Applications (2)

Application Number Priority Date Filing Date Title
PCT/US1995/007200 WO1996039424A1 (fr) 1995-06-06 1995-06-06 Facteur c stimulant les cellules tueuses naturelles
AU28186/95A AU2818695A (en) 1995-06-06 1995-06-06 Natural killer cell enhancing factor c

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
PCT/US1995/007200 WO1996039424A1 (fr) 1995-06-06 1995-06-06 Facteur c stimulant les cellules tueuses naturelles

Publications (1)

Publication Number Publication Date
WO1996039424A1 true WO1996039424A1 (fr) 1996-12-12

Family

ID=22249265

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US1995/007200 WO1996039424A1 (fr) 1995-06-06 1995-06-06 Facteur c stimulant les cellules tueuses naturelles

Country Status (2)

Country Link
AU (1) AU2818695A (fr)
WO (1) WO1996039424A1 (fr)

Cited By (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
FR2798672A1 (fr) * 1999-09-17 2001-03-23 Commissariat Energie Atomique Formes acides des peroxyredoxines et leur utilisation comme moyen de diagnostic
US6294164B1 (en) 1995-06-06 2001-09-25 Human Genome Sciences, Inc. Natural killer cell enhancing factor C
WO2001090177A1 (fr) * 2000-05-24 2001-11-29 Shanghai Biowindow Gene Development Inc. Nouveau polypeptide, activateur humain de la mort naturelle des cellules b13.64, et polynucleotide codant ce polypeptide
US6605278B1 (en) * 1998-08-18 2003-08-12 Research Development Foundation Uses of trank, a novel secretory cytokine
EP1370697A1 (fr) * 2001-03-23 2003-12-17 The General Hospital Corporation Medicaments a base de peroxyredoxine pour le traitement d'infections par le vih-1, et leurs methodes d'utilisation
US7510828B2 (en) 2001-01-26 2009-03-31 Ralf Geiben Lynn Method of decreasing the infectivity of HIV in a biological sample through the administration of S-antithrombin
US8563693B2 (en) 2001-01-26 2013-10-22 Acceleration Biopharmaceuticals, Inc. Method of treating human immunodeficiency virus infection in a mammal comprising administering heparin-activated antithrombin III

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5185431A (en) * 1988-08-31 1993-02-09 Eisai Co., Ltd. Recombinant natural killer cell activator
WO1993008827A1 (fr) * 1991-11-04 1993-05-13 The Regents Of The University Of California Facteur de renfort des cellules tueuses naturelles

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5185431A (en) * 1988-08-31 1993-02-09 Eisai Co., Ltd. Recombinant natural killer cell activator
US5316933A (en) * 1988-08-31 1994-05-31 Eisai Co., Ltd. Recombinant natural killer cell activator
WO1993008827A1 (fr) * 1991-11-04 1993-05-13 The Regents Of The University Of California Facteur de renfort des cellules tueuses naturelles

Cited By (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6294164B1 (en) 1995-06-06 2001-09-25 Human Genome Sciences, Inc. Natural killer cell enhancing factor C
US6605278B1 (en) * 1998-08-18 2003-08-12 Research Development Foundation Uses of trank, a novel secretory cytokine
FR2798672A1 (fr) * 1999-09-17 2001-03-23 Commissariat Energie Atomique Formes acides des peroxyredoxines et leur utilisation comme moyen de diagnostic
WO2001090177A1 (fr) * 2000-05-24 2001-11-29 Shanghai Biowindow Gene Development Inc. Nouveau polypeptide, activateur humain de la mort naturelle des cellules b13.64, et polynucleotide codant ce polypeptide
US7510828B2 (en) 2001-01-26 2009-03-31 Ralf Geiben Lynn Method of decreasing the infectivity of HIV in a biological sample through the administration of S-antithrombin
US8563693B2 (en) 2001-01-26 2013-10-22 Acceleration Biopharmaceuticals, Inc. Method of treating human immunodeficiency virus infection in a mammal comprising administering heparin-activated antithrombin III
EP1370697A1 (fr) * 2001-03-23 2003-12-17 The General Hospital Corporation Medicaments a base de peroxyredoxine pour le traitement d'infections par le vih-1, et leurs methodes d'utilisation
EP1370697A4 (fr) * 2001-03-23 2004-04-14 Gen Hospital Corp Medicaments a base de peroxyredoxine pour le traitement d'infections par le vih-1, et leurs methodes d'utilisation

Also Published As

Publication number Publication date
AU2818695A (en) 1996-12-24

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