EP0981606A2 - Procede et construction pouvant inhiber une migration cellulaire - Google Patents

Procede et construction pouvant inhiber une migration cellulaire

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Publication number
EP0981606A2
EP0981606A2 EP98923197A EP98923197A EP0981606A2 EP 0981606 A2 EP0981606 A2 EP 0981606A2 EP 98923197 A EP98923197 A EP 98923197A EP 98923197 A EP98923197 A EP 98923197A EP 0981606 A2 EP0981606 A2 EP 0981606A2
Authority
EP
European Patent Office
Prior art keywords
nucleic acid
acid molecule
domain
recombinant nucleic
protein
Prior art date
Legal status (The legal status 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 status listed.)
Withdrawn
Application number
EP98923197A
Other languages
German (de)
English (en)
Inventor
Paulus Hubertus Andreas Quax
Johan Hendrikus Verheijen
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Nederlandse Organisatie voor Toegepast Natuurwetenschappelijk Onderzoek TNO
Original Assignee
Nederlandse Organisatie voor Toegepast Natuurwetenschappelijk Onderzoek TNO
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.)
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Publication date
Application filed by Nederlandse Organisatie voor Toegepast Natuurwetenschappelijk Onderzoek TNO filed Critical Nederlandse Organisatie voor Toegepast Natuurwetenschappelijk Onderzoek TNO
Priority to EP98923197A priority Critical patent/EP0981606A2/fr
Publication of EP0981606A2 publication Critical patent/EP0981606A2/fr
Withdrawn legal-status Critical Current

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Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/14—Hydrolases (3)
    • C12N9/48—Hydrolases (3) acting on peptide bonds (3.4)
    • C12N9/50—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25)
    • C12N9/64—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from animal tissue
    • C12N9/6421—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from animal tissue from mammals
    • C12N9/6424—Serine endopeptidases (3.4.21)
    • C12N9/6456—Plasminogen activators
    • C12N9/6459—Plasminogen activators t-plasminogen activator (3.4.21.68), i.e. tPA
    • 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
    • C07K14/475—Growth factors; Growth regulators
    • C07K14/485—Epidermal growth factor [EGF], i.e. urogastrone
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/81—Protease inhibitors
    • C07K14/8107—Endopeptidase (E.C. 3.4.21-99) inhibitors
    • C07K14/811—Serine protease (E.C. 3.4.21) inhibitors
    • C07K14/8114—Kunitz type inhibitors
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/81—Protease inhibitors
    • C07K14/8107—Endopeptidase (E.C. 3.4.21-99) inhibitors
    • C07K14/811—Serine protease (E.C. 3.4.21) inhibitors
    • C07K14/8114—Kunitz type inhibitors
    • C07K14/8117—Bovine/basic pancreatic trypsin inhibitor (BPTI, aprotinin)
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/81—Protease inhibitors
    • C07K14/8107—Endopeptidase (E.C. 3.4.21-99) inhibitors
    • C07K14/8146—Metalloprotease (E.C. 3.4.24) inhibitors, e.g. tissue inhibitor of metallo proteinase, TIMP
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Y—ENZYMES
    • C12Y304/00—Hydrolases acting on peptide bonds, i.e. peptidases (3.4)
    • C12Y304/21—Serine endopeptidases (3.4.21)
    • C12Y304/21069—Protein C activated (3.4.21.69)
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2319/00—Fusion polypeptide

Definitions

  • the domain with an effector function has protease inhibitor activity and comprises a protease inhibitor or active part thereof, said protease inhibitor being selected from the group consisting of (bovine) pancreatic trypsin inhibitor, (bovine) splenic trypsin inhibitor, urinary trypsin inhibitor, tissue inhibitor of matrix metalloproteinase 1, tissue inhibitor of matrix metalloproteinase 2, tissue inhibitor of matrix metallo- proteinase 3, and elastase inhibitor.
  • the domain with an effector function may comprise (an active part of) two or more different protease inhibitors, or two or more copies of (an active part of) a protease inhibitor, or both.
  • the present invention relates to the use of hybrid proteins in which a receptor binding domain is linked to a functional protein in order to induce a local action of this protein and to prevent systemic effects and/or diffusion.
  • this invention relates to such hybrid proteins that might be produced by a subset of cells as target cells after transfection or transduction with expression vectors. More specifically the invention relates to the use of such expression vectors, coding for hybrid proteins consisting of a receptor binding domain and a protease inhibitor domain, for the prevention of cell migration and tissue remodeling by inhibition of proteases at the surface of migrating or invading cells.
  • the method and construct described in the present invention can be applied as therapy in diseases in which cell migration and/or tissue remodeling occurs.
  • Diffusion of the inhibitor and systemic side effects are prevented by binding the hybrid protein (by its receptor binding domain) to the cell surface of the target cell.
  • Local expression of this hybrid protein also contributes to the reduction of systemic side effects, while the negative effect of diffusion of the protein is reduced by the production at the site where action is required.
  • the local expression of the hybrid protein in specific sub- populations of cells e.g. endothelial cells prone to migrate during angiogenesis, can be enhanced using cell type-specific or tissue-specific expression vectors, in which the expression of the protein is under control of a promoter with cell type-specific or tissue-specific regulatory elements.
  • - Binding of a protease inhibitor to a cell surface receptor can locate the inhibitor close to its molecular target, the cell surface bound proteolytic enzyme.
  • a protease inhibitor to a cell surface receptor for a proteolytic enzyme, such as the urokinase receptor, may have an additional inhibitory effect. It prevents the binding of the proteolytic enzyme to its receptor, and thus strongly reduces the action of this enzyme as has been shown for blocking the binding of u-PA to its receptor which can strongly inhibit cell migration.
  • Hybrid proteins for which the expression vectors (e.g. adenoviral or retroviral expression vectors) contain the encoding DNA sequences, might contain a region that binds to a cell surface receptor and that is not subsequently internalized.
  • Receptor binding domains that can be used for this purpose are e.g. the u-PAR binding domain of urokinase plasminogen activator, the receptor binding domain of epidermal growth factor, the receptor associated protein (RAP) that binds to the LDL-R related protein (LRP) , also called ⁇ 2 -macroglobulin receptor, and the VLDL-receptor .
  • the inhibitor part of the encoded hybrid protein might consist of various protease inhibitors such as bovine pancreatic trypsin inhibitor, also called aprotinin or TrasyloA, other trypsin inhibitors such as urinary trypsin inhibitor, inhibitors for matrix-degrading metalloproteinases such the tissue inhibitors of metalloproteinases TIMP-1, TIMP-2 and TIMP-3, or variants thereof. Also inhibitors for other proteases like elastase are very suitable for being incorporated into the expression vector containing the DNA sequences encoding the hybrid proteins. Multiple copies of the DNA sequences encoding the functional protein part of the hybrid protein e.g.
  • the inhibitor part, or combinations of different inhibitors or derivatives thereof might be incorporated into the DNA construct in the expression vector.
  • Another very attractive possibility would be to use such an expression vector encoding hybrid receptor binding protein to apply any functional protein that should exert its action in the local environment of the target cell, e.g. a protease involved in the activation of a growth factor or an other e.g. vasoregulatory component.
  • the action of the functional protein or protein domains of the hybrid protein is localized to the direct microenvironment of the target cells by binding of the receptor binding domain to a receptor at the surface of the target cells.
  • Production of the hybrid protein in the direct environment of the target cells or even by the target cells themselves can be obtained by transfection or transduction of these cells by the use of expression vectors that might be based on a non-viral or an adeno- or retroviral vector system. Expression in specific cell or tissue types might be achieved by the use of specific promoter elements in the expression vectors.
  • EXAMPLE 1 An expression plasmid encoding the aminoterminal fragment of urokinase plasminogen activator (u-PA) , amino acids 1-138, hereafter referred to as ATF, can be constructed by deleting the DNA sequences encoding amino acids 139 till 401 in an expression plasmid for the full length u-PA using a polymerase chain reaction (PCR) with the following oligo- nucleotides: 5 ' -cccgggctttttttccatctgcgcagtc-3 ' and 5 ' -agggtcaccaaggaagagaatggc-3 ' .
  • PCR polymerase chain reaction
  • the newly formed DNA fragment can be circularized by ligation to restore the circular character of the expression plasmid.
  • an expression plasmid encoding the ATF and the C terminal last 11 amino acid residues including the stop codon can be constructed.
  • sequence of the thus formed DNA construct encoding the u-PA ATF fragment then is determined and compared with the predicted sequence as a control for possible mutations introduced during the construction procedure.
  • FIG. 1 The construction pCRII-ATF from pCRII-uPA using PCR is shown in Figure 1.
  • FIG 1 the area indicated between the lines was removed during the PCR amplification, resulting in the ATF plasmid.
  • the plasmid pCRII-uPA is shown to the left, plasmid pCRII-ATF to the right.
  • DNA fragments encoding amino acid residues 36-93 of bovine pancreatic trypsin inhibitor (BPTI) and the homologous amino acid residues of bovine spleen trypsin inhibitor (BSTI) can be isolated by performing a PCR reaction on genomic DNA isolated for bovine aortic endothelial cells using the following oligonucleotides : 5 ' - tcgcqacctgacttctqcctaqaqc-3 ' covering nucleotides 2509 to 2533 (with modifications, indicated in i talics, in the 5 1 region of the oligonucleotide to introduce a Nrul site (underlined) for cloning purposes) of the BPTI gene according to the published sequence (GENBANK, BTBPTIG) , and nucleotide 2442 to 2462 of the BSTI gene according to the published sequence (GENBANK, BTBSTIG) and 5 ' -crqrtcacccaqqqccca
  • amplified DNA fragments then were cloned into an appropriate plasmid vector, pCRII or pUC13, and then the exact sequence of the amplified DNA fragments in the isolated clones was analyzed to differentiate between BPTI and BSTI which have a very high degree of homology.
  • the DNA fragment encoding amino acids 1 to 207 of the human tissue inhibitor of metalloproteinase type 1 is isolated by performing a reverse transcriptase polymerase chain reaction on total RNA isolated from human foreskin fibroblasts by using the following oligonucleotides
  • EXAMPLE 4 For construction of a recombinant adenovirus containing sequences encoding the ATF. BPTI hybrid protein, this sequence is cloned in the adenoviral vector construction adapter and expression plasmid pMAD5.
  • This plasmid contains part of the wildtype adenovirus type 5 DNA sequences, a Major Late Promoter (MLP) , and a poly-adenylation (polyA) signal and can be used as either an expression vector or a shuttle vector to construct a recombinant adenovirus.
  • MLP Major Late Promoter
  • polyA poly-adenylation
  • This plasmid was derived from plasmid pMLPIO as follows.
  • First pMLPlO-lin was constructed by insertion of a synthetic DNA fragment with unique sites for the restriction endonucleases Mlul , Spll, SnaBl, Spel, AsuII and Muni into the Hindlll site of pMLPIO. Subsequently, the adenovirus Bglll fragment spanning nt 3328 to 8914 of the Ad5 genome was inserted into the Muni site of pMLPlO-lin. Finally, the Sall-BamHI fragment was deleted to inactivate the tetracycline resistance gene, resulting in plasmid pMAD5. To clone the ATF. BPTI sequence into the pMAD5 plasmid between the MLP promoter and the polyA signal the following strategy has been followed.
  • this pCRII-ATF plasmid was digested with the restriction enzymes Smal and Bstell.
  • the pCRII- BPTI plasmid was digested with the restriction enzymes Nrul and Bstell and the BPTI containing fragment was cloned into the pCRII-ATF plasmid (see figure 2) .
  • the construction pCRII- ATF-BPTI is shown in Fig. 2.
  • the ATF-BPTI sequence was cloned into pMAD5. This was done by digestion of the pCRI I -ATF-BPTI plasmid with the restriction enzymes EcoRV and Spel, isolation of the ATF-BPTI encoding DNA fragment and cloning of this fragment into the SnaBI and Spel digested pMAD5 plasmid. The cloning was tested by restriction analysis and sequence analysis.
  • the pMAD5 -ATF-BPTI shuttle vector for the construction of ATF-BPTI adenoviral vector is shown in Figure 3.
  • this sequence is cloned in the pMAD5 expression plasmid.
  • This plasmid contains part of the wildtype adenovirus type 5 DNA sequences, a Major Late Promoter (MLP) , and a polyadenylation (polyA) signal and can be used as either an expression vector or a shuttle vector to construct a recombinant adenovirus.
  • MLP Major Late Promoter
  • polyA polyadenylation
  • oligonucleotides contain recognition sites for the restriction enzymes Nrul (first oligonucleotide, underlined) and BstEII and Sspl respectively (second oligonucleotide, underlined) ; these sites are needed for the cloning procedure.
  • the amplified DNA fragment was cloned into a pCRII vector and called pCRII-TIMPl.
  • This vector was subsequently digested with the restriction enzymes Nrul and Bstell and the TIMP1 containing DNA fragment was cloned into the pCRII-ATF plasmid (see figure 1) .
  • the ATF-TIMP sequence was cloned into pMAD5. This was done by digestion of the pCRII -ATF-TIMP plasmid with the restriction enzymes EcoRV and Spel, isolation of the ATF-TIMP encoding DNA fragment and cloning of this fragment into the Snabl and Spel digested pMAD5 plasmid. The cloning was tested by restriction analysis and sequence analysis.
  • this sequence is cloned in the pMAD5 expression plasmid.
  • This plasmid contains part of the wildtype adenovirus type 5 DNA sequences, a Major Late Promoter (MLP) , and a poly- adenylation (polyA) signal and can be used as either an expression vector or a shuttle vector to construct a recombinant adenovirus.
  • MLP Major Late Promoter
  • polyA poly- adenylation
  • a PCR reaction with the oligonucleotides 5 ' -cccgggctttttccatctgcgcagtc-3 ' and 5 ' -agggtcaccaaggaagagaatggc-3 ' was performed as described in example 1 to make a pCRII-ATF plasmid (see figure 1) .
  • pCRIIATF* 5 ' -gactctagagcaaaaatgacaaccag-3 ' and the resulting DNA fragment was cloned into the pCRII cloning vector.
  • the signal peptide of u-PA is removed and a Sspl restriction enzyme recognition site is introduced (underlined) .
  • the resulting plasmid DNA is designated pCRIIATF*.
  • This vector was subsequently digested with the restriction enzymes Sspl and EcoRV and the TIMP1 containing DNA fragment was cloned into a EcoRV-Sspl digested pCRII-ATF* plasmid.
  • the resulting plasmid containing the TIMP-ATF DNA fragment was called pCRII -TIMP-ATF.
  • the TIMP- ATF sequence was cloned into pMAD5.
  • EXAMPLE 7 Vectors encoding hybrid proteins containing multiple copies of the BPTI unit coupled to the ATF domain have been constructed. To construct these multiple BPTI vectors, the following strategy is followed.
  • the pMAD5-ATF-BPTI described in EXAMPLE 4 is digested with the restriction enzymes Sspl and BstEII. In this way the vector is opened exactly in the open reading frame at the end of the BPTI sequence.
  • the pCRII-BPTI plasmid described in EXAMPLE 2 is digested with Nrul and BstEII resulting in a BPTI encoding DNA fragment with one blunt end (Nrul) . The fragment was then monodirectionally cloned into the Sspl BstEII pMAD5-ATF-BPTI vector.
  • the thus constructed plasmid named pMAD5 -ATF-BPTI -BPTI was used as a shuttle vector for the construction of recombinant adenoviruses .
  • This approach can be repeated multiple times to construct vectors containing multiple BPTI -domains .
  • a vector encoding a hybrid protein containing both a BPTI unit and a TIMPl unit coupled to the ATF domain has been constructed.
  • This BPTI-TIMP vector the following strategy is followed.
  • ATF-BPTI as an inhibitor for plasmin bound to the cell surface via the interaction of the ATF domain with the u-PA receptor (uPAR) was tested using mouse cell lines that are either or not transfected with the human uPA receptor gene. These cells were incubated for 6 hrs with diluted medium of the ATF-BPTI virus-infected CHO cells. Cell extracts were made of the uPAR-transfected cells and the parental mouse cells lacking the human uPAR. Parallel cultures underwent a short acid treatment (pH 3 , 3 min) before the cell extracts were made. This treatment will remove any u-PA or ATF bound to the u-PA receptor.
  • the cell extracts were incubated with InM plasmin and the plasmin activity was determined. Plasmin activity could only be inhibited by the cell extract of the u-PAR containing cell line. No inhibition of plasmin activity was observed in the cell extracts of parental cell line, lacking the u-PA receptor, and in the acid-treated u-PAR containing cell line. This clearly indicates that ATF-BPTI can function as a u-PAR bound plasmin inhibitor.
  • ATF-BPTI in endothelial cells e.g. to specifically inhibit the migration of endothelial cells during angiogenesis
  • cloning sequences of the promoter of the human pre-pro-endothelin 1 gene (nucleotide 2180-3680 of HUMEDN1B (GENBANK)) in front of the ATF-BPTI encoding DNA in an adenoviral vector.
  • highly endothelial cell-specific expression of the ATF- BPTI hybrid protein can be obtained.
  • Proteolytic degradation of the extracellular matrix is a key event in many cell migration and tissue remodeling processes. This proteolytic matrix degradation is often found to be mediated by urokinase-type plasminogen activation.
  • an experiment was performed using human synoviocytes . These cells were infected with an ATF-BPTI adenovirus while they were seeded on an 3 H- labeled extracellular matrix existing of bovine cartilage material. Profound inhibition of matrix degradation could be observed in the virus treated cells (figure 4) indicating that matrix degradation can be inhibited by infecting cells with the ATF- BPTI encoding virus.
  • Figure 4 shows the degradation of cartilage matrix by human synoviocytes in the presence of plasminogen.
  • Matrix is incubated with control medium (lane 1) , synoviocytes (lane 2) , synoviocytes infected with ATF-BPTI adenovirus (lane 3) , and synoviocytes incubated with Trasylol (lOOKIU/ml) (lane 4) .
  • EXAMPLE 12 In the process of restenosis smooth muscle cell migration and vessel wall remodeling are key events in which plasmin mediated proteolysis of extracellular matrix components is involved. In vivo application of general plasmin inhibitors to interfere in this process may have systemic side effects. Application of a plasmin inhibitor to the surface of the migrating cells might prevent these side effects. Infection of the blood vessel wall with an ATF-BPTI adenovirus at a site where neointima formation can be expected, e.g.
  • TPL tripartite leader sequence

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  • Life Sciences & Earth Sciences (AREA)
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  • General Health & Medical Sciences (AREA)
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  • Medicinal Chemistry (AREA)
  • Gastroenterology & Hepatology (AREA)
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  • Engineering & Computer Science (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Biophysics (AREA)
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Abstract

L'invention concerne une molécule d'acide nucléique recombinante qui comprend un vecteur convenant pour une transfection ou une transduction de cellules mammaliennes. Lesdits vecteurs contiennent une insertion d'acide nucléique qui code un polypeptide (ou une protéine) hybride pouvant être exprimé et comprend deux domaines ayant, l'un une fonction de fixation, et l'autre une fonction effectrice. Le domaine ayant une fonction de fixation peut comprendre un domaine de fixation d'un récepteur, et le domaine ayant une fonction effectrice peut avoir une activité enzymatique, en particulier une activité d'inhibition de protéase. Le vecteur peut être un vecteur viral (adénovirus ou rétrovirus) ou non viral convenant pour une transfection ou une transduction de cellules mammaliennes. L'insertion d'acide nucléique qui code un polypeptide (ou une protéine) hybride pouvant être exprimé peut être assujettie à un promoteur spécifique d'une cellule ou d'un tissu. On décrit en outre un processus destiné à prévenir une activité protéolytique locale, une dégradation matricielle extracellulaire, une migration cellulaire, une invasion cellulaire ou un remodelage tissulaire, l'opération consistant à transfecter ou transduire les cellules en cause ou des cellules dans leur environnement avec la molécule d'acide nucléique recombinante pour obtenir localement l'expression du polypeptide (ou de la protéine) hybride codé par ladite molécule. On décrit enfin un processus de production du polypeptide ou de la protéine hybride par transfection ou transduction de cellules mammaliennes avec la molécule d'acide nucléique recombinante pour obtenir l'expression du polypeptide (ou de la protéine) hybride ainsi produit, et éventuellement le récupérer.
EP98923197A 1997-05-12 1998-05-11 Procede et construction pouvant inhiber une migration cellulaire Withdrawn EP0981606A2 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
EP98923197A EP0981606A2 (fr) 1997-05-12 1998-05-11 Procede et construction pouvant inhiber une migration cellulaire

Applications Claiming Priority (4)

Application Number Priority Date Filing Date Title
EP97201423 1997-05-12
EP97201423 1997-05-12
EP98923197A EP0981606A2 (fr) 1997-05-12 1998-05-11 Procede et construction pouvant inhiber une migration cellulaire
PCT/NL1998/000259 WO1998051788A2 (fr) 1997-05-12 1998-05-11 Procede et construction pouvant inhiber une migration cellulaire

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EP0981606A2 true EP0981606A2 (fr) 2000-03-01

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EP (1) EP0981606A2 (fr)
JP (1) JP2001525669A (fr)
AU (1) AU7553698A (fr)
CA (1) CA2289117A1 (fr)
NO (1) NO995564L (fr)
NZ (1) NZ500656A (fr)
WO (1) WO1998051788A2 (fr)

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NO995564L (no) 2000-01-11
WO1998051788A2 (fr) 1998-11-19
JP2001525669A (ja) 2001-12-11
NO995564D0 (no) 1999-11-12
AU7553698A (en) 1998-12-08
NZ500656A (en) 2001-11-30
WO1998051788A3 (fr) 1999-05-20
CA2289117A1 (fr) 1998-11-19

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