WO1989010966A1 - Vectors encoding hcmv glycoprotein and expression products - Google Patents
Vectors encoding hcmv glycoprotein and expression products Download PDFInfo
- Publication number
- WO1989010966A1 WO1989010966A1 PCT/US1989/001850 US8901850W WO8910966A1 WO 1989010966 A1 WO1989010966 A1 WO 1989010966A1 US 8901850 W US8901850 W US 8901850W WO 8910966 A1 WO8910966 A1 WO 8910966A1
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- Prior art keywords
- hcmv
- expression vector
- polypeptide
- hxlf1
- hxlf2
- Prior art date
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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
- C07K14/005—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from viruses
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
-
- 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
- C12N2710/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA dsDNA viruses
- C12N2710/00011—Details
- C12N2710/16011—Herpesviridae
- C12N2710/16111—Cytomegalovirus, e.g. human herpesvirus 5
- C12N2710/16122—New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
Definitions
- This invention relates to a family of antigenically related glycoprotein complexes contained within the envelope of human cytomegalovirus (HCMV), and in particular to a multi-gene family encoding a principal glycoprotein component of these complexes, gp47-52.
- This glycoprotein has been shown to elicit humoral and cell-mediated immune responses in humans, and is useful for the diagnosis and treatment of HCMV infection.
- cytomegalovirus is the most common cause of congenital infection leading to mental retardation, deafness and other neurological handicaps in the United States. Moreover, this virus is frequently associated with life-threatening opportunistic infection in organ and bone marrow transplant recipients, patients with congenital and acquired immunodeficiency diseases, and patients receiving immunosuppressive therapy for cancer, autoimmune diseases and the like. Because HCMV is ubiquitous in nature, measures to limit exposure of individuals at risk of serious infection have met with limited success.
- antiviral chemotherapy is of little value to infants who have already suffered the devastating consequences of intrauterine HCMV infection, and thus far has shown little promise in ameliorating the progression of systemic HCMV infection and HCMV-associated interstitial pneumonia in the immunocompromised host.
- a safe, effective HCMV vaccine is urgently needed to protect against congenital and acquired HCMV infection.
- improved diagnostic tests can play an important role in detection of subclinical HCMV infection, thereby facilitating early therapeutic intervention to decrease the incidence or severity of clinical disease.
- HCMV is a member of the family herpesviridae.
- HCMV virions consist of a linear double stranded DNA molecule enclosed within an icosahedral nucleocapsid, surrounded by a tegument and a lipid-containing envelope containing a number of viral glycoproteins and glycoprotein complexes.
- Three major antigenically distinct sets of HCMV envelope glycoprotein complexes have been described thus far, each made up of two or more glycoproteins associated by disulfide bonds.
- the first set of complexes, designated gel contains glycoproteins with molecular weights of 55,000 (gp55], 93,000 (gp93) and 130,000 (gp130).
- HSV herpes simplex virus
- a second set of antigenically related complexes appears to represent multimeric forms of one or more gene product(s).
- the isolation and characterization of the principal glycoprotein contained within this family of complexes, gp47-52, has been described in detail in U.S. patent application Serial No. 933,789, wherein it is referred to as GLP-B, and in U.S. patent application Serial No. 158,389, filed February 22, 1988, wherein it is referred to as gp52(II). The disclosure of these applications are incorporated by reference herein.
- gp86 and 145,000 (gp145) have been isolated from a third complex, designated gcIII.
- the gene coding for gp86 has been identified and exhibits homology with gH of HSV-1.
- a single monoclonal antibody (mcAb) reactive with the gcII family of complexes, 9E10 (IVI-10118), has been extensively characterized, and is described in detail in U.S. patent application Serial No. 933,789.
- This monoclonal antibody exhibits cross-reactivity among several strains of HCMV, adenovirus type 2, and herpes simplex virus types 1 and 2.
- this monoclonal neutralizes Towne strain HCMV without complement, suggesting the possibility that gp47-52 plays an integral role in the biological events required for replication of the virus.
- the present invention provides a recombinant expression vector, such as a plasmid, comprising DNA derived from the HCMV genome.
- This HCMV DNA can code a polypeptide which reacts with gcII-specific monoclonal antibody 9E10 (IVI-10118), and corresponds to a portion of gp47-52, an immunogenic glycoprotein present in the major HCMV envelope glycoprotein complex gcll. Weston and Barrell, J . Mol. Biol., 192, 177
- HXLF Hind III X left reading frame
- the present invention is based upon our discovery that the gcll glycoprotein, gp47-52, is encoded by ohe or more of the genes encompassed by the HXLF gene family. Therefore, the present invention is also directed to the isolated gene family, e.g., to a substantially pure composition of matter consisting essentially of at least one of the members of the HXLF gene family of HCMV, e.g., HXLF1, HXLF2, HXLF3, HXLF4 and HXLF5.
- the term "DNA derived from the HCMV genome” refers to both DNA which is isolated from HCMV virions, such as Towne strain virions, and to DNA- which is chemically synthesized.
- gp47-52 refers to either fully-, partially-, or non-glycosylated polypeptide segments of gp47-52 which comprise epitopes which are reactive with 9E10 or with patient sera comprising antibodies reactive with gp47-52.
- Non-glycosylated segments of gp47-52 are also referred to as parts of the "polypeptide backbone" of gp47-52.
- the present recombinant expression vectors can be employed to produce immunogenic polypeptides which formally correspond to a portion of gp47-52.
- a plasmid containing all or a portion of the HXLF gene family can be linearized, mRNA synthesized in vitro, and then translated under conditions whereby the corresponding polypeptides are either glycosylated, or nonglycosylated.
- the translation products can be either analyzed by SDS-PAGE and extracted therefrom, or solubilized for immunoprecipitation as described in the examples hereinbelow.
- the present invention also provides a number of new gp47-52-related polypeptides (p20, p21-25) and glycoproteins (gp30, gp25) which comprise epitopes which react with 9E10, as well as with antibodies in the HCMV-positive sera of human patients.
- HCMV gp47-52 The role of HCMV gp47-52 in virus infection is unknown, as is the role of its parent glycoprotein complex, gcll.
- gcII is a major component of the virion envelope and is a target for antibody-mediated neutralization.
- these data strongly support the inclusion of gp47-52 and/or the present "subunit proteins" of gp47-52 into HCMV vaccines, which may also include other classes of HCMV envelope glycoproteins, such as gel.
- the gp47-52-related polypeptides and glycoproteins can be readily prepared in quantity by transformation or transfection of suitable host cells with the present vectors; or by known chemical methods of polypeptide synthesis. After isolation and purification of the polypeptide, an immunologically-effective amount thereof is combined with a pharmaceutically-acceptable vehicle, such as a sterile liquid carrier, and parenterally administered to a mammal, in order to raise the titer of anti-HCMV antibodies in the blood of the recipient.
- a pharmaceutically-acceptable vehicle such as a sterile liquid carrier
- the present compositions are useful in the production of monoclonal antibodies, which in turn can be used either directly or indirectly to diagnose HCMV, or to treat HCMV infections.
- the present immunogenic compositions can also be used as diagnostic reagents, e.g., to detect antigen antibodies in physiological fluids, e.g., by RIA or ELISA procedures.
- the present compositions are also useful to produce clonal populations of antigenspecific T-helper lymphocytes, which in turn can be used for HCMV therapy.
- the present invention also provides RNA probes, a number of which are exemplified hereinbelow, which are complementary to sequences in the HXLF gene family. These probes can be used to detect HCMV in clinical specimens using hybridization methods known in the art.
- the five HXLF genes were cloned into an SP6 expression vector in both the sense and antisense orientations (Fig. 1C).
- Monocistronic, bicistronic, and polycistronic RNAs synthesized in vitro using SP6 polymerase were translated in rabbit reticulocyte lysates with or without canine pancreatic microsomal membranes.
- the HXLF1 or the HXLF1 plus HXLF2 translation products were detected when using the above mRNAs.
- the HXLF3, 4 and 5 gene products were not detected by in vitro translation of the SP6 derived polycistronic mRNA.
- Nonglycosylated or glycosylated HXLF1 and HXLF2 gene products were immunoprecipitated by monoclonal antibody 9E10 which is specific for a virion envelope glycoprotein complex (gc) designated gcII.
- the monoclonal antibody 9E10 immunoprecipitated a diffuse glycoprotein (gp) band designated gp47-52 from HCMV-infected cell lysates.
- the amino acid composition of gp47-52 purified from virion envelopes has the highest similarity to the predicted amino acid composition of the product of the HXLF1 plus HXLF2 open reading frames, but it is more similar to that of HXLF2 than to that of HXLF1.
- the Northern blot results imply that gp47-52 is synthesized predominantly from the abundant 1.62 kb bicistronic mRNA encoded by the HXLF1 and HXLF2 genes.
- the glycoprotein could also be synthesized by the monocistronic 0.8 kb mRNA encoded by the HXLF2 gene as well as the mRNAs predicted from the other HXLF genes.
- Figure 1 schematically depicts the location of the HXLF homologous gene family of human cytomegalovirus and its cloning into an SP6 expression vector.
- Panel (A) depicts the prototype arrangement of the human cytomegalovirus (HCMV) genome showing large unique (U L ), short unique (U S ), terminal repetitive (TR) and internal repetitive (IR) regions.
- Panel (B) depicts the HindIII-XbaI fragment derived from the U s region and designates the map units on the viral genome.
- the HXLF family of homologous ORFs are depicted by closed bars labelled HXLF1 through HXLF5.
- Dashed arrows indicate predicted mRNA transcripts with approximate sizes in kb based on the locations of TATA boxes and polyadenylation signals (stars) relative to the ORFs.
- Beneath each bar representing an ORF is the molecular weight (X10 3 ) of the protein product predicted by the respective ORF. Restriction endonuclease sites are indicated by arrows.
- Panel (C) depicts the HXLF gene family cloned into an SP6 expression vector in both the sense (SP65IT) and antisense (SP64IT) orientations.
- Bs BstE II
- HIII HindIII
- Xb XbaI
- Xh XhoI
- SP SP6 promoter
- Amp ampicillin resistance gene
- Plasmid pSP65IT has been deposited at In Vitro International, 611(P) Hammond's Ferry Road, Linthicum, MD 21090, under the access code IVI-10168.
- Virus and cells The culturing of human foreskin fibroblasts and the propagation of human cytomegalovirus Towne strain have been described previously by M. F. Stinski in J. Virol., 26, 686 (1978).
- the viral DNA polymerase inhibitor phosphonoacetic acid was used as described by M. W. Wathen et al., 3. Virol., 41, 462 (1982).
- the pSP64 and pSP65 cloning vectors contain the Salmonella typhimurium phage SP6 promoter upstream of a polylinker sequence containing unique restriction sites for insertion of DNA sequences.
- the bacteriophage SP6 RNA polymerase initiates transcription exclusively at an SP6 promoter, and thus allows for synthesis of pure, full-length single stranded RNA from a DNA template cloned downstream of the SP6 promoter.
- the polylinker is cloned in opposite directions relative to the SP6 promoter in pSP64 and pSP65 to allow cloning of the DNA sequences in sense and anti-sense orientations, respectively.
- the SP6 expression system used in these studies was adapted from an original cloning vector derived by insertion of the SP6 promoter into pBR322.
- a particular disadvantage of the original cloning vector was the presence of 250 base pairs upstream of the restriction site used for cloning, resulting in transcription of extraneous sequences between the transcription start site and the restriction sites of the polylinker.
- pSP64 and pSP65 have only six bases and nine bases, respectively, between the transcription start site and the restriction enzyme insertion site of the polylinkers.
- a particular advantage of these cloning vectors is that the major RNA transcripts are derived from transcription of the DNA template strand to the last and penultimate nucleotide.nicks in the DNA template do not serve as transcription initiation sites, nor does SP6 RNA polymerase terminate at nicks in the template and thus premature termination resulting in RNA transcripts shorter than full-length, is relatively rare.
- RNA in rabbit reticulocyte lysates was accomplished as described by M. Stinski et al., 3. Virol., 46, 1 (1983). Translation products were radiolabelled with 35 S-methionine (>1000 Ci/mmol; Amersham). For some experiments, translation was supplemented with canine pancreatic microsomal membranes as recommended by the manufacturer (Amersham).
- Antigens were solubilized in RIPA buffer (PBS, pH 7.4, containing 0.1% SDS, 1% NP-40, 0.5% sodium deoxycholate, 100 ⁇ g/ml PMSF and 0.02% sodium azide) and immunoprecipitated with monoclonal or polyclonal antibodies plus protein A sepharose (Pharmacia; Uppsala, Sweden) as described by D. R. Gretch et al., Anal. Biochem., 163, 270 (1987).
- SDS-PAGE SDS-polyacrylamide gel electrophoresis
- autoradiography has been described by M. F. Stinski et al., 3. Virol., 12, 594 (1976).
- RNA samples were purchased from Promega (Madison, WI).
- Antisense RNA probes were synthesized from linearized pSP64IT templates as directed by the supplier (Amersham) except that 150 ⁇ Ci of ⁇ 32 P-UTP (800 Ci/mmol; Amersham) was supplemented with cold UTP to a final concentration of 50 ⁇ M to ensure full length probe synthesis.
- gp47-52 was purified from virion and dense body envelopes using ion exchange and gel exclusion high performance liquid chromatography (HPLC) as described previously by B. Kari et al., cited above. After strong acid hydrolysis, amino acid compositions were determined by reverse phase HPLC on a C-18 column (E. M. Science, Elmsford, NY) as described by H. Scholze, 3. Chromat., 350, 453 (1985).
- Example I Northern blot analysis of the HXLF steady state mRNA in infected cells.
- the HXLF gene family which consists of five homologous ORFs in tandem, was cloned into the SP6 expression vectors in sense and antisense orientations as outlined in Figure 1.
- Four different mRNA transcripts were predicted from the HXLF gene family based on the locations of TATA boxes and polyadenylation signals relative to the ORFs (see Figure IB).
- a 1.6 kb bicistronic RNA was predicted from the HXLFl and HXLF2 genes while a 2.7 kb tricistronic RNA was predieted from the HXLF3 through HXLF5 genes.
- monocistronic transcripts of 0.8 and 1.0 kb genes were predicted from HXLF2 and HXLF5 genes, respectively.
- PAA treatment increases the steady state level of viral RNA.
- a 4.5 kb antisense RNA probe which covered all five HXLF genes was synthesized from pSP64IT by linearizing the antisense template with restriction endonuclease Xbal (see Figure 1C).
- the 1.62 kb transcript is presumably the bicistronic mRNA predicted from the HXLF1 and HXLF2 genes. Both probes would be expected to hybridize to the 1.62 kb mRNA given the relative homology between the various HXLF genes.
- the full length probe detected minor mRNAs of 0.8 and 1.0 kb which correspond to the monocistronic RNAs predicted from the HXLF2 and HXLF5 genes, respectively.
- Neither probe detected the 2.7 kb polycistronic transcript predicted from the HXLF3, HXLF4 and HXLF5 genes.
- the predominant mRNA observed was a 1.62 kb mRNA predicted to be encoded by the HXLF1 and HXLF2 genes which accumulated in the cytoplasm of virusinfected cells at early times after infection.
- the level of this transcript relative to total cellular and viral RNA is decreased at late times after infection.
- HXLF1 In vitro synthesis of the HXLF gene products.
- HXLF1 the first ORF
- HXLF2 the first two ORFs together
- HXLF1 through HXLF5 the sense plasmid pSP65IT was completely linearized with either restriction endonuclease Xhol, BstEII or HindIII, respectively (see Figure 1C).
- the approximate sizes of the in vitro synthesized mRNAs were determined by agarose-formaldehyde gel electrophoresis (data not shown) and the RNAs were translated in rabbit reticulocyte lysates in the presence or absence of dog pancreas membranes (DPM). DPM allow for core glycosylation with N-acetylglucosamine and mannose sugars during glycoprotein translation in vitro. [J. E. Rothman et al., Nature, 269, 775 (1977)] The translation products were either analyzed by SDS-PAGE or solubilized for immunoprecipitation as described hereinbelow.
- the diffuse nature of p21-25 is characteristic of the HXLF1 gene product only and may be due to internal initiation or premature termination during translation of the HXLF1 mRNA. Alternatively, the diffuse nature of p21-25 may be due to the higher proline content predicted to be present in HXLF1 (see Table I). Proline rich regions can disrupt part of the protein's alpha helical structure leading to anomolous mobilities in denaturing polyacrylamide gels.
- HXLF5 ORFs were predicted to make proteins of 28, 27.5 and 26.2 kDa, respectively (see Figure IB). Thus, the products of the downstream HXLF ORFs were apparently not expressed in vitro even though full-length RNA was detected by agarose-formaldehyde gel electrophoresis. It is unlikely that the protein products migrated in gels the same as the HXLF1 and HXLF2 products.
- the HXLF ORFs were predicted by Weston et al., cited above, to have sites for N- as well as O- glycosylation. Therefore, the in vitro synthesized mRNAs were also translated in the presence of dog pancrease membranes (DPM) to allow for core glycosylation of the nascent polypeptides [J. E. Rothman et al., Nature, 269, 775 (1977)]. After translation of mRNA from the HXLF1 gene in the presence of DPM, a 30 kDa glycoprotein (gp30) was detected. When mRNA from the HXLF1 plus HXLF2 genes was translated in the presence of DPM, a major 30 kDa glycoprotein band was again detected.
- DPM dog pancrease membranes
- HXLF1 and HXLF2 proteins were glycosylated to approximately the same apparent molecular weight, the DPM suppressed translation of the HXLF2 mRNA, or the HXLF2 glycosylated protein is present in low amounts.
- DPM deoxyribonucleic acid
- Example III Immunoprecipitation of the HXLF gene products with a HCMV-specific monoclonal antibody or with human convalescent serum.
- Monoclonal antibodies that react with envelope glycoprotein complexes gcI (gp55, gp93-130), II (gp47-52) and III (gp86, gp145) were used for immunoprecipitation.
- Monoclonal antibody 9E10 which recognizes a major glycoprotein component of the gcll family, gp47-52, reacted strongly with the diffuse P21-25 translated from the HXLF1 mRNA.
- monoclonal antibody 9E10 immunoprecipitated both the diffuse p21-25 and non-diffuse p20 specified by mRNA from HXLF1 and HXLF2.
- monoclonal antibody 9E10 When mRNA from HXLFl through HXLF5 was translated, monoclonal antibody 9E10 again immunoprecipitated p21-25 and p20. However, neither monoclonal 41C2, which reacts with the gcI family, nor monoclonal antibody 1G6, which reacts with the gcIII family, immunoprecipitated the HXLF mRNA translation products.
- p21-25 from the HXLF1 ORF is gcII specifie, since immunoprecipitation with 9E10 detects the entire diffuse band p21-25.
- the p20 product of the HXLF2 ORF is apparently also recognized by monoclonal antibody 9E10 even though there is only a low degree of homology between these two ORFs.
- the possibility that p20 was immunoprecipitated via disulfide-linkage with p21-25 is unlikely because immunoprecipitated p20 was also detected by non-reducing SDS-PAGE.
- Example IV DPM-mediated translation.
- mRNA from all five ORFs was translated in the presence of DPM and the products were immunoprecipitated with either monoclonal antibody 9E10 or human convalescent serum 218. Both reacted with the 30 kDa product of the HXLF1 ORF.
- both immunoprecipitated a second 25 kDa species which may represent either unglycosylated HXLF1 ORF product or glycosylated product from the HXLF2 ORF.
- Monoclonal antibodies to gel or gcIII, as well as HCMV negative human serum reacted weakly or failed to react with these translation products. The weak reactivity with monoclonal antibody 41C2 could be prevented by adding 1 mg/ml of ovalbumin (data not shown).
- gp47-52 The synthesis of gp47-52 was investigated using infected human fibroblast cells in the presence or absence of the ionophore monensin, which inhibits golgi transport and allows the addition of high mannose N-linked but not O-linked oligosaccharides [D. C. Johnson et al., 3. Virol., 43, 1102 (1982) and Cell, 32, 987 (1983)].
- Infected cells were radiolabelled with 3 H-glucosamine at various times after infection and in the presence of monensin, and the resultant antigenic products were solubilized and immunoprecipitated with 9E10 as described hereinabove.
- a characteristically diffuse glycoprotein band (gp47-52) was detected at 48 and 72 hr p.i. but not at 24 hr p.i. or in uninfected cells.
- glycoproteins designated gp32 and gp25 were immunoprecipitated by 9E10. These glycoproteins are probably simple N-linked precursors of gp47-52 since monensin blocks the processing of simple N-linked sugars as well as the addition of 0-linked oligosaccharide chains. It is unlikely that gp32 represents golgi-mediated processing of gp25 or vice versa since the concentration of monensin used completely blocked processing of the HCMV gB homolog precursor glycoprotein (data not shown). These results are consistent with the immunoprecipitation of 30 and 25 kDa HXLF gene products from in vitro translations supplemented with DPM since DPM allows simple Nglycosylation in vitro. In addition, the identification of multiple precursors in the presence of monensin implies that gp47-52 is encoded by multiple HXLF genes.
- Example VI Comparison of the amino acid composition of gp47-52 with the predicted amino acid compositions of the HXLF gene products.
- gp47-52 is a product of the HXLF genes, this glycoprotein was purified from virion envelopes by a combination of ion exchange and gel exclusion high performance liquid chromatography (HPLC) followed by immunoprecipitation as described by B. Kari et al., J. Virology, 60, 345 (1986), the disclosure of which is incorporated by reference herein.
- the reduced and alkylated gp47-52 was subjected to strong acid hydrolysis and its amino acid composition was determined by reverse phase HPLC as described by Scholze, in 3. Chromatography, 350, 453 (1985).
- the observed amino acid composition of gp47-52 was then compared to the predicted amino acid compositions of the products coded by the HXLF genes as shown in Table I, below.
- gp47-52 is composed of multiple HXLF gene products.
- the heterogeneous nature of gp47-52 and the different mRNA size classes detected by Northern blot analysis support this possibility. Therefore, gp47-52, which is a major component of the virion complex gcII, is specified by the HXLF gene family between map units of 0.857 and 0.879 in the short unique component of the viral genome.
- the virion envelope glycoprotein gp47-52 is a product of the HXLF gene family of human cytomegalovirus.
- the evidence for this is as follows: (i) monoclonal antibody 9E10, which is specific for gp47-52, immunoprecipitated the in vitro synthesized HXLF1 and HXLF2 gene products; (ii) the mean amino acid composition of gp47-52 is most similar to the predicted composition of the HXLF1 plus HXLF2 genes; (iii) carbohydrate analysis of gp47-52 has revealed a high level of 0-linked sugars and low amounts of N-linked sugars.
Abstract
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Cited By (3)
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EP0436537A1 (en) * | 1988-01-29 | 1991-07-17 | Chiron Corporation | Recombinant cmv neutralizing proteins |
WO1996004383A1 (en) * | 1994-07-29 | 1996-02-15 | American Cyanamid Company | Identification of a human cytomegalovirus gene region involved in down-regulation of mhc class i heavy chain expression |
US9346874B2 (en) | 2009-12-23 | 2016-05-24 | 4-Antibody Ag | Binding members for human cytomegalovirus |
-
1989
- 1989-05-01 AU AU35504/89A patent/AU3550489A/en not_active Abandoned
- 1989-05-01 WO PCT/US1989/001850 patent/WO1989010966A1/en not_active Application Discontinuation
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- 1989-05-01 EP EP89905553A patent/EP0413738A1/en not_active Withdrawn
Non-Patent Citations (7)
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ARCHIVES OF VIROLOGY, Vol. 98, 1988, SPRINGER VERLAG, B. KARI et al., "Isolation and Characterization of a Human Cytomegalovirus Glycoprotein Containing a High Content of O-linked Oligosaccharides", pages 171-188. * |
BIOLOGICAL ABSTRACTS/RRM, No. BR35:92649, D.R. GRETCH et al., "In-vitro Translation of a Bicistronic Messenger RNA Encoding a Human Cytomegalovirus Envelope Glycoprotein Complex GCII"; & SYMPOSIUM ON THE MOLECULAR BIOLOGY OF RNA HELD AT THE 17TH ANNUAL UCLA (UNIVERSITY OF CALIFORNIA-LOS ANGELES) SYMPOSIA ON MOLECULAR AND CELLULAR BIOLOGY, KEYSTONE, COLORADO, USA, 4-10 April 1988; & J. CELL. BIOCHEM., Suppl. 0 (12 Part D), 1988. * |
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JOURNAL OF VIROLOGY, Vol. 60, No. 2, November 1986, AMERICAN SOCIETY FOR MICROBIOLOGY, B. KARI et al., "Characterization of Monoclonal Antibodies Reactive to Several Biochemically Distinct Human Cytomegalovirus Glycoprotein Complexes", pages 345-352. * |
JOURNAL OF VIROLOGY, Vol. 62, No. 3, March 1988, AMERICAN SOCIETY FOR MICROBIOLOGY, D.R. GRETCH et al., "Identification and Characterization of Three Distinct Families of Glycoprotein Complexes in the Envelopes of Human Cytomegalovirus", pages 875-881. * |
JOURNAL OF VIROLOGY, Vol. 62, No. 6, June 1988, AMERICAN SOCIETY FOR MICROBIOLOGY, D.R. GRETCH et al., "A Multigene Family Encodes the Human Cytomegalovirus Glycoprotein Complex gcII (gp47-52 Complex)", pages 1956-1962. * |
Cited By (11)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
EP0436537A1 (en) * | 1988-01-29 | 1991-07-17 | Chiron Corporation | Recombinant cmv neutralizing proteins |
EP0436537A4 (en) * | 1988-01-29 | 1992-04-08 | Chiron Corporation | Recombinant cmv neutralizing proteins |
WO1996004383A1 (en) * | 1994-07-29 | 1996-02-15 | American Cyanamid Company | Identification of a human cytomegalovirus gene region involved in down-regulation of mhc class i heavy chain expression |
WO1996004384A1 (en) * | 1994-07-29 | 1996-02-15 | American Cyanamid Company | Identification of a human cytomegalovirus gene involved in down-regulation of mhc class i heavy chain expression |
US5720957A (en) * | 1994-07-29 | 1998-02-24 | American Cyanamid Company | Recombinant human cytomegalovirus vaccine |
US5753476A (en) * | 1994-07-29 | 1998-05-19 | American Cyanamid Company | Identification of a human cytomegalovirus gene region involved in down regulation of MHC class I heavy chain expression |
US5843458A (en) * | 1994-07-29 | 1998-12-01 | American Cyanamid Company | Recombinant human cytomegalovirus having a US2 deletion |
US5846806A (en) * | 1994-07-29 | 1998-12-08 | American Cyanamid Company | Identification of a human cytomegalovirus gene region involved in down-regulation of MHC class I heavy chain expression |
US5906935A (en) * | 1994-07-29 | 1999-05-25 | American Cyanamid Company | Cells transformed or transfected with HCMV US2-US5, US10-US11 genes |
US5908780A (en) * | 1994-07-29 | 1999-06-01 | American Cyanamid Company | Cells transformed or transfected with HCMV US2 gene |
US9346874B2 (en) | 2009-12-23 | 2016-05-24 | 4-Antibody Ag | Binding members for human cytomegalovirus |
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EP0413738A1 (en) | 1991-02-27 |
AU3550489A (en) | 1989-11-29 |
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