WO1994023582A9 - Vecteurs adenoviraux renfermant l'adn codant une proteine surfactant des poumons - Google Patents

Vecteurs adenoviraux renfermant l'adn codant une proteine surfactant des poumons

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Publication number
WO1994023582A9
WO1994023582A9 PCT/US1994/003831 US9403831W WO9423582A9 WO 1994023582 A9 WO1994023582 A9 WO 1994023582A9 US 9403831 W US9403831 W US 9403831W WO 9423582 A9 WO9423582 A9 WO 9423582A9
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WIPO (PCT)
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vector
adenoviral
surfactant protein
free
lung surfactant
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PCT/US1994/003831
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English (en)
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WO1994023582A1 (fr
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Priority to JP6523310A priority Critical patent/JPH09500782A/ja
Priority to EP94914075A priority patent/EP0701401A4/fr
Publication of WO1994023582A1 publication Critical patent/WO1994023582A1/fr
Publication of WO1994023582A9 publication Critical patent/WO1994023582A9/fr

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  • This invention relates to adenoviral vectors. More particularly, this invention relates to adenoviral vectors which include DNA encoding a lung surfactant protein and to the use of such vectors in treating disease states associated with lung surfactant protein deficiency, such as infant respiratory distress syndrome, and adult respiratory distress syndrome.
  • Surfactant proteins are natural endogenous proteins produced primarily within the alveolar and airway epithelial cells of the normal lung and interact with phospholipids to maintain the patency of the alveolar structures.
  • pulmonary surfactant protein concentration on the alveolar surface falls below critical levels, surface tension of the liquid-gas interface increases, thereby leading to alveolar collapse, pulmonary ventilation-perfusion mismatch, and hypoxia. In severe cases, this can lead to death.
  • Intermittent administration of exogenous bovine lung surfactant protein has shown partial, but not complete remission of the pathophysiology of the surfactant deficiency state. It is an object of the present invention to provide a recombinant expression vehicle for expressing pulmonary surfactant protein.
  • an adenoviral vector including a DNA sequence encoding a lung surfactant protein.
  • the adenoviral vector is a replication deficient adenoviral vector, i.e., such vector is free of a DNA sequence(s) which is (are) required for viral replication, such as, for example, the El DNA sequence or a portion thereof.
  • the adenoviral vector is free of at least a portion of the adenoviral El DNA sequence and is free of at least a portion of the adenoviral E3 DNA sequence.
  • the E3 region encodes several polypeptides which help the adenovirus to evade the immune surveillance of the host.
  • the adenoviral vector comprises an adenoviral 5' inverted terminal repeat, or ITR; an adenoviral 3' ITR; an adenoviral encapsidation signal; the DNA sequence encoding a lung surfactant protein; and a promoter controlling the expression of the DNA sequence encoding the lung surfactant protein.
  • the vector is free of at least the majority of the adenoviral El and E3 DNA sequences, but is not free of all of the E2 and E4 DNA sequences, and is not free of DNA sequences encoding adenoviral proteins expressed by the adenoviral major late promoter. In one embodiment, the vector is also free of at
  • the vector is free of the adenoviral El and E3 DNA sequences, and is free of one of the E2 and E4 DNA sequences, and is free of a portion of the other of the E2 and E4 DNA sequences.
  • the vector is free of at least the majority of the El and E3 DNA sequences, is free of at least a portion of at least one DNA sequence selected from the group consisting of the E2 and E4 DNA sequences, and is free of DNA sequences encoding adenoviral proteins expressed under control of the adenoviral major late promoter.
  • Suitable promoters which may be employed include, but are not limited to, adenoviral promoters, such as the adenoviral major late promoter; or heterologous promoters, such as the cytomegalovirus (CMV) promoter; the Rous Sarcoma Virus promoter; the respiratory syncytial virus promoter; inducible promoters, such as the mouse mammary tumor virus, or MMTV, promoter; the metallothionein promoter; and heat shock promoters.
  • tissue-specific promoters such as, but not limited to, lung surfactant protein promoters, may also be employed. It is to be understood, however, that the scope of the present invention is not to be limited to any specific promoter.
  • Lung surfactant proteins which may be encoded by the DNA sequence encoding a lung surfactant protein include surfactant protein A (SPA), surfactant protein B (SPB), and surfactant protein C (SPC).
  • SPA surfactant protein A
  • SPB surfactant protein B
  • SPC surfactant protein C
  • Surfactant protein A is described in Kuroki, et al., J. Biol. Chem., Vol. 263, No. 7, pgs. 3388-3394 (March 5, 1988).
  • Surfactant protein B and DNA encoding therefor are described in Pilot-Matias, et al., DNA. Vol. 8, No. 2, pgs. 75-86 (1989), Glasser, et al., Proc. Nat. Acad. Sci., Vol 84, pgs. 4007-4011 (June 1987); Revak, et al. , J. Clin. Invest.. Vol. 81, pgs.
  • the DNA sequence encoding a lung surfactant protein encodes lung surfactant protein B.
  • SPB is the most clinically important lung surfactant protein of those herein above described.
  • Such a vector in a preferred embodiment, is assembled first by constructing, according to standard techniques, a shuttle plasmid which contains, beginning at the leftward adenoviral genomic elements, the "critical left end elements", which include an adenoviral 5' ITR, an adenoviral encapsidation signal, and the Ela enhancer sequence; a promoter (which may be an adenoviral promoter or a foreign promoter); a tripartite leader sequence, a multiple cloning site; a poly A signal; and a DNA segment which corresponds to a segment of the adenoviral genome.
  • the critical left end elements which include an adenoviral 5' ITR, an adenoviral encapsidation signal, and the Ela enhancer sequence
  • a promoter which may be an adenoviral promoter or a foreign promoter
  • a tripartite leader sequence a multiple cloning site
  • poly A signal a DNA segment which corresponds to a
  • Such DNA segment serves as a substrate for homologous recombination with a modified or mutated adenovirus, and such sequence may encompass, for example, a segment of the adenoviral genome from base 3328 to base 6241 of the adenovirus 5 genome.
  • the plasmid may also include a selectable marker and an origin of replication.
  • the origin of replication may be, for example, a bacterial origin of replication.
  • a representative example of such a shuttle plasmid is pAVS6, shown in Figure 4.
  • An intron may be included within the transcribed portion to enhance the cytoplasmic mRNA accumulation levels.
  • the multiple cloning site facilitates the insertion of the DNA sequence encoding a lung surfactant protein into the plasmid.
  • restriction enzyme sites separating the above- mentioned components of the shuttle plasmid include "rare" restriction enzyme sites; i.e., sites which are found to occur randomly in eukaryotic genes at a frequency from about one in every 10,000 to about one in every 100,000 base pairs. This increases the flexibility and ease of rearranging components of the vectors in assembled shuttle plasmids.
  • Homologous recombination is then effected with a modified or mutated adenovirus in which at least the majority of the El and E3 adenoviral DNA sequences have been deleted, as shown, for example, in Figure 8.
  • a modified or mutated adenovirus in which at least the majority of the El and E3 adenoviral DNA sequences have been deleted, as shown, for example, in Figure 8.
  • Such homologous recombination may be effected through co- transfection of the shuttle plasmid and the modified adenovirus into a helper cell line, such as 293 (embryonic kidney epithelial) cells, by CaP0 4 precipitation.
  • a cloning vector is formed in which the modified adenovirus DNA which was 5' to the DNA segment in the shuttle plasmid corresponding to a similar segment of the modified adenoviral genome is replaced with the components in the shuttle plasmid which are 5' to such DNA segment.
  • This homologous recombination, or "crossing over" event can occur anywhere along the segment of the genome of the modified adenovirus which corresponds to the segment which is also contained within the shuttle plasmid (such as, for example, bases 3328 to 6241 of adenovirus 5 in Example 1 shown below) .
  • a vector which includes an adenoviral 5' ITR; an adenoviral encapsidation signal; an Ela enhancer sequence; a promoter; a tripartite leader sequence; a DNA sequence encoding a
  • SU8STITUTE SHEET (RULE *,) lung surfactant protein; a poly A signal; adenoviral DNA free of at least the majority of the El and E3 adenoviral DNA sequences; and an adenoviral 3' ITR.
  • This vector may then be introduced into a cell line such as the 293 cell line for production of large amounts of infectious recombinant adenoviral particles.
  • the 293 cell line is a human fetal kidney epithelial cell line into which has been permanently introduced 11% of the left end of the adenovirus 5 genome. This directs the synthesis of the adenoviral Ela and Elb proteins and allows trans- complementation of El-deleted vectors.
  • infectious viral particles may then be administered to a host in vivo as part of a gene therapy procedure.
  • infectious viral particles may be administered syste ically, such as by intravenous or intraperitoneal or intrasmuscular or subcutaneous administration, or may be administered topically, such as by intratracheal or intrabronchial administration, or, alternatively, the infectious viral particles may be administered in an aerosol formulation.
  • the infectious viral particles may be administered in an amount of up to about 10 13 pfu, preferably from about 10 7 pfu to about 10 12 pfu.
  • the infectious viral particles may be employed in the transduction of the epithelium of the respiratory tract or alveoli; whereby the lung epithelial cells will express lung surfactant protein in amounts sufficient to achieve clinical correction of lung surfactant protein deficiency.
  • infectious viral particles may be used to transduce eukaryotic cells in vitro.
  • Eukaryotic cells which may be transduced include, but are not limited to, macrop ages, lymphocytes, fibroblasts, liver cells, bronchial cells, and other epithelial or endothelial cells. Such eukaryotic cells then may be administered to a host as
  • SUBSTT ⁇ TE SHEET (RULE 26) part of a gene therapy procedure, or may be cultured in vitro whereby such cells produce lung surfactant protein.
  • the infectious viral particles may be used to transduce eukaryotic cells in vitro for the in vitro production of lung surfactant protein.
  • eukaryotic cells which may be transduced in vitro for the in vitro production of lung surfactant protein include, but are not limited to, those eukaryotic cells hereinabove described, as well as Chinese Hamster Ovary (CHO) cells, COS-7 cells, NIH 3T3 cells, vero cells, HeLa cells, MRC-5 cells, CN1 cells, W138 cells, and chicken ly phoma cells.
  • the lung surfactant protein produced by such cells may then be administered to a host in conjunction with an acceptable pharmaceutical carrier in order to treat lung surfactant protein deficiency states.
  • the vector comprises an adenoviral 5' ITR; an adenoviral 3' ITR; an adenoviral encapsidation signal; a DNA sequence encoding a lung surfactant protein; and a promoter controlling the DNA sequence encoding a lung surfactant protein.
  • the vector is free of the adenoviral El, E2, E3, and E4 DNA sequences, and the vector is free of DNA sequences encoding adenoviral proteins promoted by the adenoviral major late promoter; i.e., the vector is free of DNA encoding adenoviral structual proteins.
  • Such vector is sometimes hereinafter referred to as a "gutless adenoviral vector," or "GLAd" vector.
  • Promoters which are contained in the vector may be those hereinabove described.
  • Such vectors may be constructed by removing the adenoviral 5' ITR, the adenoviral 3' ITR, and the adenoviral encapsidation signal, from an adenoviral genome by standard techniques.
  • a promoter which may be an adenoviral promoter or a non- adenoviral promoter
  • tripartite leader sequence poly A
  • SUBSTIT ⁇ TESHEET(RULE 26) signal may, by standard techniques, be ligated into a base plasmid or "starter" plasmid such as, for example, pKSII" (Strategene) , to form an appropriate cloning vector.
  • the cloning vector may include a multiple cloning site, as hereinabove described, to facilitate the insertion of the foreign DNA sequence into the cloning vector.
  • An appropriate vector in accordance with the present invention is thus formed by cutting the cloning vector by standard techniques at appropriate restriction sites in the multiple cloning site, and then ligating the DNA sequence encoding a lung surfactant protein into the cloning vector.
  • the GLAd vector may then be packaged into infectious viral particles using a helper adenovirus or cell line which provides the necessary packaging materials.
  • a helper virus is used, in one embodiment, preferably it has a defective encapsidation signal in order that the helper virus will not package itself. Examples of such encapsidation-defective helper viruses which may be employed are described in Grable, et al., J. Virol.. Vol. 66, pgs. 723-731 (1992), and in Grable, et al., J. Virol. , Vol. 64, pgs. 2047-2056 (1990).
  • the helper virus has a normal packaging signal.
  • DNA for the vector and the encapsidation-defective helper virus are transfected into an appropriate cell line for the generation of infectious viral particles. Transfection may take place by electroporation, calcium phosphate precipitation, microinjection, or through proteoliposomes. Examples of appropriate cell lines include, but are not limited to, HeLa cells, A549 cells, or 293 (embryonic kidney epithelial) cells.
  • the infectious viral particles may then be purified away from helper virus by CsCl isopycnic density centrifugation and transduced into lung epithelial cells lining the respiratory tract or alveoli, as hereinabove described, whereby such cells express lung surfactant protein.
  • the vector is transfected into the cells, followed by infection of the cells with the encapsidation-defective helper virus.
  • the adenoviral construction shuttle plasmid pAvS6 was constructed in several steps using standard cloning techniques including polymerase chain reaction based cloning techniques.
  • Ad-dl327 (Thimmappaya, et al., Cell, Vol. 31, pg.
  • adenovirus 543 (1983) is identical to adenovirus 5 except that an Xbal fragment including bases 28593 to 30470 (or map units 78.5 to 84.7) of the adenovirus 5 genome, and which is located in the E3 region, has been deleted.
  • Genbank accession #M73260 The comlete Adenovirus 5 genome is registered as Genbank accession #M73260, incorporated herein by reference, and the virus is available from the American Type Culture Collection, Rockville, Maryland, U.S.A. under accession number VR-5.
  • Ad-dl327 was constructed by routine methods from Adenovirus 5 (Ad5). The method is outlined briefly as follows and previously described by Jones and Shenk, Cell 13:181-188 (1978).
  • Ad5 DNA is isolated by proteolytic digestion of the virion and partially cleaved with Xba I restriction endonuclease. The Xba I fragments are then reassembled by ligation as a mixture of fragments. This results in some ligated genomes with a sequence similar to Ad5, except excluding sequences 28593 bp to 30470 bp.
  • This DNA is then transfected into suitable cells (e.g. KB cells, HeLa cells, 293 cells) and overlaid with soft agar to allow plaque formation. Individual plaques are then isolated.
  • SUBST ⁇ TU7E SHEET (RULE 26) amplified, and screened for the absence of the 1878 bp E3 region Xba 1 fragment.
  • the orientation of this fragment was such that the Bglll site was nearest the T7 RNA polymerase site of pKSII " and the HindiII site was nearest the T3 RNA polymerase site of pKSII " .
  • This plasmid was designated pHR. ( Figure 1).
  • the ITR, encapsidation signal, Rous Sarcoma Virus promoter, the adenoviral tripartite leader (TPL) sequence and linking sequences were assembled as a block using PCR amplification ( Figure 2).
  • the ITR and encapsidation signal (sequences 1-392 of Ad-dl327 [identical to sequences from Ad5, Genbank accession #M73260]) were amplified (amplification 1) together from Ad-dl327 using primers containing NotI or Ascl restriction sites.
  • the Rous Sarcoma Virus LTR promoter was amplified (amplification 2) from the plasmid pRC/RSV (sequences 209 to 605; Invitrogen, San Diego, CA) using primers containing an Ascl site and an Sfil site. DNA products from amplifications 1 and 2 were joined using the "overlap" PCR method (amplification 3) with only the NotI primer and the Sfil primer. Complementarity between the Ascl containing end of each initial DNA amplification product from reactions 1 and 2 allowed joining of these two pieces during amplification.
  • the TPL was amplified (amplification 4) (sequences 6049 to 9730 of Ad-dl327 [identical to similar sequences from Ad5, Genbank accession #M73260]) from cDNA made from mRNA isolated from 293 cells infected for 16 hours with Ad-dl327 using primers containing Sfil and Xbal sites respectively. DNA fragments from amplification reactions 3 and 4 were then joined using PCR (amplification 5) with the NotI- and Xbal-site- containing primers, thus creating the complete gene block.
  • amplification 4 sequences 6049 to 9730 of Ad-dl327 [identical to similar sequences from Ad5, Genbank accession #M73260]
  • DNA fragments from amplification reactions 3 and 4 were then joined using PCR (amplification 5) with the NotI- and Xbal-site- containing primers, thus creating the complete gene block.
  • the ITR-encapsidation signal-TPL fragment was then purified, cleaved with NotI and Xbal and inserted into the NotI, Xbal cleaved pHR plasmid. This plasmid was
  • SUBSTITIITE SHEET (RULE 26) designated pAvS6A and the orientation was such that the NotI site of the fragment was next to the T7 RNA polymerase site ( Figure 3) .
  • the SV40 early polyA signal was removed from SV40 DNA as an Hpal-BamHI fragment, treated with T4 DNA polymerase and inserted into the Sail site of the plasmid pAvS6A-( Figure 3) to create pAvS6 ( Figures 3 and 4).
  • pAVS6SPB#7 Such clones are named pAVS6SPB#7, pAVS6SPB#12, and pAVS6SPB#13.
  • pAVS6 SPB#7 is shown in Figure 7. The orientation of the SPB DNA within the shuttle plasmid was obtained by evaluating the DNA sequences of the two termini of the SPB cDNA insert in the plasmid with primers derived from pAVS6.
  • the recombinant adenoviral vector AV1SPB1 ( Figure 8), containing SPB cDNA was constructed through homologous recombination between the Ad5 deletion mutant Ad-dl327 ( Figure 8), and pAVS6SPB#7. Homologous recombination, or "crossing over,” occurs between Ad-dl327 and pAVS6.SPB#7, along the segment common to both Ad-dl327 and pAVS6.SPB#7 which corresponds to bases 3328 to 6241 (or map units 9.24 to 17.34) of the adenovirus 5 genome.
  • Ad-dl327 has a deleted E3 region in which base pairs 28593 to 30470 are absent (Thimmappaya, et al, Cell, Vol. 31, pgs. 543-551 (1982)).
  • pAVS6SPB#7 contains an adenoviral 5' ITR, an origin of replication contained completely within the 5' ITR, an Ela enhancer and encapsidation signal, a Rous Sarcoma Virus promoter, an adenovirus 5 tripartite leader
  • SUBSJ1TUTE SHEET (RULE 26) sequence and the 2kb human SPB cDNA including the entire protein coding sequence (nucleotides 1 to 1172), and the SV40 poly A signal.
  • 293 cells (ATCC No. CRL 1573) were infected with AV1SPB1 at a multiplicity of infection (MOI) of 50 MOI units. At 12 hours post-infection, the cells were radiolabeled with 35 S-methionine (50 ⁇ Ci/ml) overnight. Identical amounts of labeled protein were used for immunoprecipitation with antisera against SPB. Immunopreciptates were analyzed by SDS-polyacrylamide gel electrophoresis on 16% gel. The gels were fluorographed. C 14 -labeled molecular weight markers and BioRad broad range molecular weight markers were used as size markers.
  • MOI multiplicity of infection
  • lane 1 shows an uninfected control
  • lane 2 shows AVISPBl-infected 293 cells in which electrophoresis of immunoprecipitates occurred under reducing conditions
  • lane 3 shows AVISPBl-infected 293 cells in which electrophoresis of immunoprecipitates occurred under non- reducing conditions.
  • the precursor protein also was detected in both reduced and unreduced conditions.
  • the filter was crosslinked (UV Crosslink, Stratagene), and hybridized with a 32 P-labeled 2.0 kb human SP-B cDNA probe prepared by random priming (Loftstrand) and evaluated by autoradiography. Lungs from uninfected control rats and from rats infected with AvlLacZ4 also were subjected to the above hybridization procedure.
  • the animals were sacrificed 48 hours after infection, and the lungs were prepared for in situ hybridization analysis according to the method of Wert, et al.. Development Biology. Vol. 156, pgs. 426-443 (1993), using human SP-BcRNA. An uninfected rat was used as a control.
  • ADDRESSEE Carella, Byrne, Bain, Gilfillan,
  • GCTGCTGCCC ACGCTCTGTG GCCCAGGCAC TGCTGCCTGG ACCACCTCAT CCTTGGCCTG 120 CGACGACGGG TGCGAGACAC CGGGTCCGTG ACGACGGACC TGGTGGAGTA GGAACCGGAC
  • CTACTCCGTC ATCCTGCTCG ACACGCTGCT GGGCCGCATG CTGCCCCAGC TGGTCTGCCG 840 GATGAGGCAG TAGGACGAGC TGTGCGACGA CCCGGCGTAC GACGGGGTCG ACCAGACGGC
  • GGAAAAGTGC AAGCAATTTG TGGAGCAGCA CACGCCCCAG CTGCTGACCC TGGTGCCCAG 1080 CCTTTTCACG TTCGTTAAAC ACCTCGTCGT GTGCGGGGTC GACGACTGGG ACCACGGGTC
  • TCTCCAGTGT ATCCACAGCC CCGACCTTTG ATGAGAACTC AGCTGTCCAG GTGCAAAGGA 1200 AGAGGTCACA TAGGTGTCGG GGCTGGAAAC TACTCTTGAG TCGACAGGTC CACGTTTCCT
  • TGTCTCAGCT CAACCACAGT CTGACACCAG AGCCCACTTC CATCCTCTCT GGTGTGAGGC 1440 ACAGAGTCGA GTTGGTGTCA GACTGTGGTC TCGGGTGAAG GTAGGAGAGA CCACACTCCG
  • CCTCACACCC ACCCCCATGC ACTCAAAGAT TGGATTTTAC AGCTACTTGC AATTCAAAAT 1620 GGAGTGTGGG TGGGGGTACG TGAGTTTCTA ACCTAAAATG TCGATGAACG TTAAGTTTTA
  • GAGCTATTGC TTTGTTAAGA TATAAAAAGG GGTTTCTTTT TGTCTTTCTG TAAGGTGGAC 1860 CTCGATAACG AAACAATTCT ATATTTTTCC CCAAAGAAAA ACAGAAAGAC ATTCCACCTG

Abstract

L'invention se rapporte à un vecteur adénoviral renfermant une séquence d'ADN codant une protéine surfactant des poumons. Le vecteur adénoviral peut être un vecteur adénoviral déficient de réplication qui est exempt au moins de la majorité des séquences d'ADN E1 et E3. Ces vecteurs peuvent être utilisés dans la génération de particules virales infectieuses capables de transduire des cellules épithéliales des poumons in vivo afin de faciliter l'expression de la protéine surfactant des poumons par ces cellules.
PCT/US1994/003831 1993-04-08 1994-04-07 Vecteurs adenoviraux renfermant l'adn codant une proteine surfactant des poumons WO1994023582A1 (fr)

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JP6523310A JPH09500782A (ja) 1993-04-08 1994-04-07 肺界面活性タンパク質をコード化するｄｎａを含むアデノウイルスベクター
EP94914075A EP0701401A4 (fr) 1993-04-08 1994-04-07 Vecteurs adenoviraux renfermant l'adn codant une proteine surfactant des poumons

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US08/044,406 1993-04-08

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Publication number Priority date Publication date Assignee Title
US5585362A (en) * 1989-08-22 1996-12-17 The Regents Of The University Of Michigan Adenovirus vectors for gene therapy
FR2705686B1 (fr) * 1993-05-28 1995-08-18 Transgene Sa Nouveaux adénovirus défectifs et lignées de complémentation correspondantes.
AU4743196A (en) * 1994-12-30 1996-07-24 Chiron Corporation Non-traumatic administration of gene delivery vehicles
DE69638058D1 (de) 1995-06-15 2009-11-26 Crucell Holland Bv Verpackungssysteme für humane rekombinante Adenoviren zur Gentherapie
US6783980B2 (en) 1995-06-15 2004-08-31 Crucell Holland B.V. Packaging systems for human recombinant adenovirus to be used in gene therapy
US6265212B1 (en) 1995-06-15 2001-07-24 Introgene B.V. Packaging systems for human recombinant adenovirus to be used in gene therapy
EP0927263A1 (fr) * 1996-01-05 1999-07-07 Genetic Therapy, Inc. Generation de vecteurs adenoviraux a mediation de recombinase
WO1998032860A1 (fr) * 1997-01-28 1998-07-30 Baxter International Inc. Procede de production a haut rendement de vecteurs adenoviraux
US6403370B1 (en) 1997-02-10 2002-06-11 Genstar Therapeutics Corporation Oncolytic/immunogenic complementary-adenoviral vector system
JP2001509375A (ja) * 1997-07-10 2001-07-24 ヘパヴェック アーゲー フュール ゲンテルアピエ 最小アデノウイルスベクターを作製するためのクローニングベクター
WO1999027101A1 (fr) * 1997-11-25 1999-06-03 Princeton University Procede de preparation de vecteurs d'adenovirus, vecteurs ainsi prepares et leurs utilisations
US6670188B1 (en) 1998-04-24 2003-12-30 Crucell Holland B.V. Packaging systems for human recombinant adenovirus to be used in gene therapy
ES2311307T3 (es) * 1998-10-20 2009-02-01 Children's Hospital Medical Center Proteina d surfactante para la prevencion y el diagnostico del enfisema pulmonar.
US8933032B2 (en) 1998-10-20 2015-01-13 Children's Hospital Medical Center Surfactant protein D for the treatment of disorders associated with lung injury
US6838428B2 (en) 1998-10-20 2005-01-04 Children's Hospital Medical Center Surfactant protein D for the prevention and diagnosis of pulmonary emphysema
US20120220531A1 (en) 2011-02-04 2012-08-30 Cincinnati Children's Hospital Medical Center Surfactant protein d for the treatment of disorders associated with lung injury

Family Cites Families (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5585362A (en) * 1989-08-22 1996-12-17 The Regents Of The University Of Michigan Adenovirus vectors for gene therapy
AU680459B2 (en) * 1992-12-03 1997-07-31 Genzyme Corporation Gene therapy for cystic fibrosis
FR2705686B1 (fr) * 1993-05-28 1995-08-18 Transgene Sa Nouveaux adénovirus défectifs et lignées de complémentation correspondantes.
FR2707664B1 (fr) * 1993-07-13 1995-09-29 Centre Nat Rech Scient Vecteurs viraux et utilisation en thérapie génique.

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