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WO1989001628A1 - Anticorps monoclonaux reagissant avec la glycoproteine a du cytomegalovirus - Google Patents

Anticorps monoclonaux reagissant avec la glycoproteine a du cytomegalovirus Download PDF

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Publication number
WO1989001628A1
WO1989001628A1 PCT/US1988/002644 US8802644W WO8901628A1 WO 1989001628 A1 WO1989001628 A1 WO 1989001628A1 US 8802644 W US8802644 W US 8802644W WO 8901628 A1 WO8901628 A1 WO 8901628A1
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Prior art keywords
hcmv
monoclonal antibody
gpa
binding
epitope
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PCT/US1988/002644
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English (en)
Inventor
Nancy O. Lussenhop
Bruce E. Kari
Richard C. Gehrz
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Childrens Hospital Inc
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Childrens Hospital Inc
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Priority claimed from US07/083,502 external-priority patent/US5126130A/en
Application filed by Childrens Hospital Inc filed Critical Childrens Hospital Inc
Priority to KR1019890700600A priority Critical patent/KR890702036A/ko
Publication of WO1989001628A1 publication Critical patent/WO1989001628A1/fr
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

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    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/569—Immunoassay; Biospecific binding assay; Materials therefor for microorganisms, e.g. protozoa, bacteria, viruses
    • G01N33/56983—Viruses
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/08—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from viruses
    • C07K16/081—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from viruses from DNA viruses
    • C07K16/085—Herpetoviridae, e.g. pseudorabies virus, Epstein-Barr virus
    • C07K16/089—Cytomegalovirus
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P21/00—Preparation of peptides or proteins
    • C12P21/02—Preparation of peptides or proteins having a known sequence of two or more amino acids, e.g. glutathione
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides

Definitions

  • HCMV human cytomegalovirus
  • HCMV is a member of the family herpesvirdae, and is composed of a nuclear complex of nucleic acid and proteins surrounded by an external membrane envelope containing glycopeptides and glycolipids.
  • envelope glycoproteins characterized by our laboratory and others, several are associated by disulfide bonds in high molecular weight complexes.
  • Commonly assigned U.S. patent application Serial No. 933,789, filed November 24, 1986, entitled, "Immunogenic Glycopeptides of Human Cytomegalovirus” describes the isolation and characterization of anti- genically related glycopeptides of molecular weights 9X,000 daltons and 50,000-52,000 altons.
  • mcAb monoclonal antibodies
  • 41C2 and 9B7 immunopreci- pcit ' ate these individual glycoproteins, as well as the antrgenically-related glycoprotein complexes.
  • McAb 9B7 also exhibits neutralizing activity against HCMV ⁇ n_ vitro in the presence of complement. Convalescent sera from patients who have recovered from HCMV infection also contain antibodies that immunoprecipitate these HCMV glycoproteins, suggesting that antibodies reactive with these glycoproteins may participate in neutraliza- tion of virus iji vivo.
  • HCMV-specific cAbs that have practical clinical utility, in the diagnosis and treatment of HCMV infections.
  • mcAbs that are useful both in direct detection of HCMV in clinical specimens and rapid identification of the virus in tissue culture.
  • Such antibodies may be used either to capture and concen ⁇ trate a viral antigen, or alternatively, for immuno- logical detection of the virus using a variety of known immunoassay techniques.
  • hyperimmune HCMV globulin to immunosuppressed patients prior to organ or bone marrow transplantation may possibly attenuate or prevent life-threatening HCMV infections in the post- transplant period.
  • the present invention provides 13 related murine mcAbs which can be generated by the immunization of mice with whole Towne HCMV virions and/or detergent extracted envelope glycoproteins of Towne strain HCMV.
  • the spleen cells of the immunized mice are then fused with a murine myeloma cell line to generate antibody- producing murine B-cell hybridomas.
  • the antibodies produced by these murine hybridomas immunoprecipitate glycoproteins of molecular weights 130,000, 93,000, and 50,000-52,000 daltons.
  • glycoproteins are asso ⁇ ciated with one or more high molecular weight glycopro ⁇ tein complexes in the viral envelope.
  • glycopro ⁇ tein complexes have been referred to as the gCI family by D. R. Gretch et al., 3. Virol., _62, 875 (1988).
  • the 130kD glycoprotein complex disclosed in patent application Serial No. 933,789 is referred to ? herein as human cytomegalovirus glycoprotein A (HCMV GCA).
  • HCMV GCA human cytomegalovirus glycoprotein A
  • ELISA ELISA assays were performed to ' study the relation ⁇ ships among the antigenic sites recognized by the various mcAbs. Surprisingly, either inhibition or augmentation in the binding of the antibodies to the virus was observed when a second antibody was present simultaneously.
  • the mcAbs were assigned to at least 7 groups which recognize distinct antigenic sites within 3 domains of HCMV GCA.
  • the present invention is directed to generation of an HCMV-specific murine mcAb produced by a process comprising: a. immunizing a mouse with a composition co pris- ing HCMV virions or purified HCMV envelope glycopeptides derived from HCMV GCA having molecular weights of about 130,000, 93,000, or 50,000-52,000 daltonsj
  • the mcAbs of the present invention were characterized with respect to their specificity, binding affinity, and their ability to neutralize HCMV in the presence of complement, either alone or in combination with other murine mcAbs -of the invention which are also obtained from the hybridomas of step (c) above.
  • the present invention also provides an effec ⁇ tive method for detecting HCMV in a sample of physio ⁇ logical material infected with HCMV. This method com ⁇ prises: a. reacting said physiological material with a first monoclonal antibody which binds to a first epitope of HCMV GPA to form a complex between said HCMV GPA and said first mono ⁇ clonal antibody; and
  • detecting the presence of said label or alternatively, reacting said binding site with a detectable label prior to said detection.
  • the concentration of said label provides a measure of the concentration of HCMV GPA pre- sent in said physiological material.
  • the detectable label is a radio ⁇ active isotope or an enzyme which has been chemically linked to the second antibody.
  • the binding site for said label is provided by a molecule capable of binding a * free enzyme or a radioisotope, e.g., biotin or deferoxamine. These molecules can be chemi ⁇ cally linked to mcAbs by methods known to the art. For example, see U.S. Patent No. 4,680,338.
  • the enzyme preferably is linked to a substance which binds strongly to the binding site, for example, an avidin- enzyme conjugate will bind to biotin.
  • Steps (a) and (b) can be performed simultaneously or the order of addition of the first and second mcAb can be inverted. Therefore, the pr.esent invention also provides a " method for detecting HCMV in a sample of physiologi ⁇ cal material infected with HCMV comprising: a. reacting said physiological material with a first monoclonal antibody to a first epitope of HCMV GPA, said first monoclonal antibody
  • ⁇ comprises a detectable label or a binding site for said label, to yield a complex between HCMV GPA and said first monoclonal antibody;
  • detecting the presence of said label or alternatively, reacting said binding site with a detectable label prior to said detection.
  • the concentration of said label provides a measure of the concentration of HCMV GPA pre ⁇ sent in said physiological material.
  • the present invention is also directed to a method for raising plasma immunogenicity to HCMV GPA by administering a pharmaceutical unit dosage form com ⁇ prising one or more of the present antibodies to a patient who has been exposed to HCMV. It is believed that the antibodies of the present invention will have therapeutic potential, either as a prophylactic agent to prolong the life expectancy of infected and/or diseased patients by retarding the clinical progression of the disease, or possibly, as a curative agent which can act to eliminate the virulence of the virus. The present antibodies might also represent a valuable adjunct to chemical antiviral agents.
  • the present invention is also directed to a composition consisting essentially of a mixture of:
  • a first monoclonal antibody e.g., 9B7, which binds to a first epitope on HCMV GPA, wherein said first monoclonal anti ⁇ body neutralizes HCMV infectivity in vitro in the presence of complement;
  • a second monoclonal antibody e.g., 41C2 which binds to a second epitope on HCMV
  • HCMV GCA-specific monoclonal antibodies were also isolated and characterized which possessed the unexpected ability to substantially inhibit or com ⁇ pletely block the ability of these therapeutically- useful first or second classes of HCMV GCA-specific mcAbs. Therefore, it is highly preferred that the HCMV GCA-neutralizing composition of the invention be pre ⁇ pared to be free of these interfering mcAbs, i.e., the composition should be free of a monoclonal antibody which binds to a third epitope on HCMV GPA, does not neutralize HCMV infectivity in the presence or absence of complement, and which inhibits the binding of the first antibody and the second antibody. Specific examples of such mcAbs are exemplified hereinbelow.
  • the isolated mcAbs preferably are diluted with a pharmaceutically-acceptable liquid carrier, such as an aqueous IV fluid, prior to being assayed for bioac- tivity or administered as a unit dosage form _i vivo.
  • a pharmaceutically-acceptable liquid carrier such as an aqueous IV fluid
  • the resultant solution is steril ⁇ ized, e.g., by filtration.
  • Preservatives commonly employed with IgG preparations, such as maltose, gly- cine or thimerosal, may be added in pharmaceutically- acceptable amounts.
  • the resulting solutions are preferably admin ⁇ istered parenterally, e.g., by intravenous infusion or injection.
  • the amount of mcAb composition administered will vary widely, and will depend on the physique and physical condition of the im unosuppressed or HCMV- infected patient. Such factors are necessarily empiri ⁇ cal, and can ' be determined by the clinician, employing known HCMV staging criteria. In some clinical situa ⁇ tions, it may be necessary to administer a plurality of doses of the mcAb composition, in order to neutralize the infectivity of viral particles as they are released from infected target cells.
  • Figure 1 depicts immunoprecipitates formed by reacting unreduced (UR) or reduced (R) HCMV GCA pro ⁇ teins with GCA-specific monoclonal antibodies (mcAbs) and convalescent sera.
  • UR unreduced
  • R reduced
  • mcAbs GCA-specific monoclonal antibodies
  • Figure 2 depicts a Western Blot of the HCMV glycoproteins recognized by GCA-specific mcAbs.
  • Figure 3 is a diagrammatic represen ⁇ tation of the epitopes recognized by the GCA-specific mcAbs. Inhibition is represented by overlapping solid lines, while augmentation and the direction of augmen ⁇ tation are indicated by arrows. Domains I, II and III are shown to the right and left. Panel B schematically depicts a possible arrangement of the epitopes on the HCMV GCA peptide backbone.
  • Figure 4 is a graphic depiction of plaque reduction assays using Towne strain HCMV and combina ⁇ tions of antibodies.
  • HCMV Towne strain and AD169 strain were grown with or without 3 H-glucosamine on human skin fibroblast cultures, harvested and purified on sucrose gradients as described previously (Kari et al *» J- Virol., 60, 345-352 (1986)). The purified virus was resuspended in Tris-NaCl buffer (50 M Tris hydrochloride, 150 mM NaCl pH 7.4), and extracted with 1% Nonidet P-40
  • NP-40 Sigma Chem. Co., St. Louis, MO
  • Tris-NaCl buffer 50 M Tris hydrochloride, 10 M NaCl, 2 mM phenylmethyl sulfonylfluoride pH 7.5
  • Uninfected skin fibroblasts were extracted in a similar fashion for use as negative controls. Reduc ⁇ tion and alkylation of HCMV Towne NP-40 crude extracted material was performed as described by Kari et al., J. Virol., 6Q_, 345-352 (1986). All detergent extracted viral or control fibroblast materials were passed over an Extracti-Gel D column and eluted with water to remove the detergent.
  • mice hybridomas secreting mcAbs to HCMV proteins was performed as previously described (Kari et al., 3 ⁇ _
  • mice 15 Virol., £0, 345-352 (1986)).
  • the antibodies described in this patent application involved three separate fusion experiments using either AD169 or Towne strain purified HCMV virions.
  • Balb/C mice were immunized for 2, 5 or 10
  • Spleen cells from immunized mice were fused with SP2-2-Agl4 myeloma cells (American Tissue Culture Collection) using polyethylene glycol as the fusing agent.
  • Resulting hybrid cells were screened for specific antibody production to HCMV using an enzyme-linked immunosorbent assay (ELISA). (See Section G below.) Antigens used in the ELISA assay were either purified HCMV Towne
  • the clonality of the mcAbs was established by twice subcloning of the hybridomas producing each antibody. Furthermore, each antibody had a unique isotype (i.e., 41C2-IgGl; 9B7 IgG-2b), and each appears as an individual antibody by isoelectric focusing.
  • the mcAbs in ascites fluid were diluted 1/500 in 1% gelatin in TBS, and allowed to bind to the paper overnight at room temperature.
  • the paper was washed with PBS-0.05% Tween 20 (polysorbate 20), and alkaline phosphatase- labelled goat anti-mouse IgG (KPL) diluted 1/2000 with 1% gelatin in TBS was added and allowed to incubate for one hour at room tem ⁇ perature.
  • the paper was washed once again and the substrate 5-bromo-4-chloro-3-indolyl- phosphate/nitroblue tetrazolium in 0.1 M Tris buffer solution (KPL) was added. Reaction of the antigen-antibody complex with the sub ⁇ strate resulted in the formation of an insol- uble purple product. The reaction was stopped by immersing the paper in water.
  • HCMV antigens were coated onto 96 well microtiter Immulon II plates by incubating 200 ng of total protein in 50 ⁇ l of 0.05 M carbonate buffer (pH 9.5) per well for 18 hours at 4°C. The wells were then washed three times with pho"sphafee-buffered saline with 0.05% Tween 20 (PB$T), once with distilled water, and then were air dried. The plates were stored at 4°C with dessicant until use.
  • the antigens used included purified HCMV Towne strain whole virion, unreduced and reduced proteins extracted from skin fibroblasts infected with - 13-
  • HCMV Towne strain for 8-12 days See Section A, preparation of viruses
  • NP-40 extracted proteins derived from uninfected skin fibro ⁇ blasts See Section A, preparation of 5 viruses.
  • the mcAbs in ascites fluids were purified by HPLC and biotinylated using biotin N- hydroxysuccinimide ester. Twenty-five ⁇ l of a
  • the biotinylated antibody was titered in 10-fold dilutions beforehand to determine the amount that would give an O.D. of >0.5 and ⁇ 1.5 in the absence of a second antibody.
  • the "unlabelled second antibody was shown to be active by titering against HCMV whole virus antigen. The amount of binding was quanti- tated by adding peroxidase-labelled goat anti-
  • tissue culture media fluid is removed from the fibro- blast monolayer by vacuum suction.
  • PBS phosphate buffered saline
  • Microtiter wells in 96- well microtiter plates are pre-coated with HCMV GCA-specific mcAb 41C2 (IVI-10119) as the capture antibody. Included in each plate are
  • Sample solutions are solu- bilized in PBS-1% NP-40 in each tissue culture well, and are then dispensed into individual microtiter wells. Blank, negative skin fibro- blast and HCMV positive control samples are
  • biotinylated GCA-specific mcAb 9B7 (IVI-10117), which recognizes a different epi ⁇ tope on GCA than that recognized by 41C2, are then added to each microtiter well as a detec- 5 tion antibody.
  • the microtiter plate is then placed in a plastic bag and incubated at 37°C for 1 hour. Wells are then washed 4 times with wash buffer (PBS-0.05% Tween), 100 ⁇ l of streptavidin-peroxidase is added and the 10 plate is incubated for 30 minutes at 37°C in a plastic bag. Wells are then washed 4 times with wash buffer and then one time with dis ⁇ tilled water.
  • Ortho-pheylenediamine (OPD) substrate 100 ⁇ l is added and reacted for 15 15 minutes at room temperature. The reaction is then stopped with 25 ⁇ l of 5 N H2SO . After " 10 minutes, the microtiter plate is read with a Dynatech plate reader at 490 nm or by visual colorometric reading. This method has been 20. shown to detect HCMV at a total protein con ⁇ centration of 10 nanograms.
  • HCMV GCA opes of HCMV virions by anion-exchange HPLC
  • a gene has been identified in the unique long (U [ _) segment of the HCMV genome,
  • glycoproteins derived from a precursor glycoprotein of molecular weight 138,000 daltons found in infected cells but not in mature virions.
  • the 138 kD glycopro- - teio in turn is derived from the 95,000
  • glycoprotein A intermediate of 138,000 daltons as well as the three mature A-type glycoproteins of 130,000, 93,000 and 50,000-52,000 daltons. 5
  • Monoclonal antibodies 41C2, 9B7 and 11B4, and additional mcAbs have been shown to immunoprecipitate the same three glyco ⁇ proteins from detergent extracts of HCMV 10 virions.
  • the presence of the precursor glycoprotein reflects contamination of virion preparations with cell-associated viral glycoprotein intermediates that result from the method used to obtain 15 large quantities of HCMV for these stu ⁇ dies.
  • HCMV contains additional glycoproteins of similar molecular weights to those comprising GCA
  • the pattern of 3 antige- 20 nically related glycoproteins recognized by the mcAbs described herein is unique to GCA and thus it can be concluded that all 13 monoclonals recognize the same glycoprotein complex, GCA. 25
  • the envelope glycoproteins were presented as antigen on whole virions 5 adsorbed onto 96 well microtiter polystyrene plates.
  • topographical analysis of the anti ⁇ genic sites recognized by 7 of these mcAbs was carried out in greater detail by adding vary ⁇ ing amounts of unlabelled mcAb to the same or different biotinylated mcAbs to determine the
  • Group I consisted of anti ⁇ bodies 41C2, 26B11 and 39E11, and was consti ⁇ tuted o ⁇ _the basis of the ability of each mcAb within this group to mutually inhibit binding
  • group III consisted
  • Antibody 18F9 (group II) enhanced the binding of 39E11 and 26B11 to GCA on the whole virus, whereas 39E11 and 26B11 reciprocally inhibited the binding of 18F9.
  • 18F9 was noted to inhibit the binding of 39E11 and 26B11, while
  • neutralizing activity may reflect non-specific effect of high IgG concentration.
  • Group I mcAb 41C2 did not exhibit neutralizing activity at any protein concentration, whereas 26B11 and 39E11 neutralized HCMV at protein concentrations of 6.5 ⁇ g/well and 0.5 ⁇ g/well, 5 respectively.
  • group II mcAbs 34G7 was non-neutralizing; 9B7 and 18F9 both neutra ⁇ lized HCMV in the presence of complement at concentrations of 0.55 ⁇ g/well and 5 ⁇ g/well, respectively.
  • the group III mcAb 11B4 did not 10 neutralize Towne HCMV.
  • 11B4 domain III
  • 11B4 was " capable of inhibiting the binding of all other GCA-specific anti ⁇ bodies and, in addition, was non-neutralizing (Fig. 4C).
  • the effect of 11B4 on the neutral- izing activity of 9B7 was then examined. In the presence of complement, 9B7 showed signif ⁇ icant neutralizing activity at 1 ⁇ g/well or greater (Fig. 4B).
  • epitope 39E11 of domain I and 9B7 and 18F9 of domain II mutually inhibit binding as a result of their close proximity on the linear structure of the peptide backbone, which is not altered by reduction.
  • This hypothetical model suggests that the epitope(s) on GCA responsible for neutralization is located at the base of the crypt, since monoclonals recognizing these epitopes, while not recognizing other epitopes within the same domain, neutralize HCMV in the presence of complement. It also appears that augmen ⁇ tation of binding of a neutralizing antibody to its epitope may increase its biological activity. More ⁇ over, changes in conformation of so-called "non-neutralizing" epitopes brought about by simultane- ous binding of two non-neutralizing antibodies may result in significant viral neutralization associated with these same epitopes.
  • Antibody (11B4) was placed in domain III. This antibody had the ability to inhibit the binding of all other antibodies tested and visa versa. The ability of 11B4 to inhibit the binding of other antibodies was retained when GCA was extracted with a non-ionic deter ⁇ gent, but this ability appeared diminished after reduc ⁇ tion of disulfide bonds. The sensitivity of the epi- tope recognized by 11B4 to denaturation was also reflected in the lack of reactivity of 11B4 in the Western Blot system. Monoclonal antibody 11B4 was an IgG2b subclass. This subclass is capable of binding complement. However, 11B4 failed to neutralize HCMV Towne strain ' even in the presence of complement.
  • 11B4 is a non-neutralizing antibody which inhibited the binding of the neutraliz ⁇ ing antibodies 39E11, 18F9 and 9B7.
  • Monoclonal anti ⁇ body 11B4 also inhibited the neutralizing activity of 9B7 in a plaque reduction assay.
  • HCMV-specific murine mcAbs are provided in which particular combinations bind synergistically to the virus and enhance the biological activity over that of either mcAb used alone. This is of particular importance in determining the clinical utility of HCMV-specific murine mcAbs for diagnostic and therapeutic purposes, given that amounts required for any individual mcAb alone may either lack sen- sitivity or be required in greater quantity than that practical for commercial production.
  • mcAbs may play an important role in the development of viral diagnostic tests * because different mcAtis can be used in a non-competing format. For example, one mcAb recognizing a unique epitope of a particular HCMV protein can be used to capture and con ⁇ centrate the virus, whereas a second mcAb which recog ⁇ nizes a different, non-competing epitope of the same protein can be used for immunologic detection of the viral antigen. Moreover, two mcAbs which mutually augment the binding of the other antibody to its par ⁇ ticular antigenic site may be used to increase the sen ⁇ sitivity of immunologic detection of the viral antigen.
  • HCMV-specific mcAbs described in this application, these prototypes can detect HCMV in nanogra amounts.
  • Monoclonal antibodies to HCMV may also be use ⁇ ful in the treatment of potentially life-threatening opportunistic HCMV infections in immunosuppressed patients. Among these patients are organ transplant patients, bone marrow transplant patients, patients with congenital or acquired immune deficiency diseases, patients on immunosuppressive drugs, and patients with congenital HCMV infection.
  • HCMV-specific mcAbs can be generated in large quantity at low cost, and thus represent a significant advantage over hyperimmune glo ⁇ bulin.
  • Therapeutic administration of HCMV hyperimmune globulin to patients with active infection may also be useful, although success in this regard has been limited to date.
  • Monoclonal antibodies developed in our labora ⁇ tory have been shown to have high specific neutralizing activity against Towne strain HCMV in vitro.
  • synergistic neutralizing activity using two or more monoclonal antibodies directed against different epitopes of the same glycoprotein may allow for high therapeutic activity with relatively low quantities of mixtures of mcAbs compared to that required using indi- vidual mcAbs.
  • synergistic neutralizing activity is observed over a wide range of relative con ⁇ centrations of augmenting mcAbs. This should greatly simplify the selection of a pharmaceutical unit dose that will exhibit the desired therapeutic effect in patients.
  • HCMV hyperimmune globulin may contain certain HCMV-specific antibodies which com ⁇ pete with the neutralizing antibodies and " alter their biological activity, thereby potentially promoting HCMV infection.
  • murine or human HCMV-specific mcAbs to be used for immunotherapy should be selected on the basis of their ability to augment the binding and neutralizing activity of other HCMV antibodies. Equally important, it is critical to exclude those mcAbs that may inhibit the binding and/or neutralizing activity of other HCMV-specific antibod- ies.
  • HCMV monoclonal antibodies described herein are murine monoclonals. They potentially can elicit an allergic response in humans. However, it is likely that most patients with serious HCMV infections will require only one or two treatments with HCMV- specific mcAbs. Therefore, the chance of significant allergic response to the mcAb is lessened. In clinical trials using other murine mcAbs, hypersensitivity reac ⁇ tions have not presented a major problem. Furthermore, the FDA has already approved the use of murine 0KT3 mcAb (ORTHOCLONE, Ortho Pharmaceutical Co.) for cancer treatment, indicating that murine monoclonals will be an acceptable form of therapy. The following are rationales for use of murine versus human mcAbs for human therapeutics:
  • the murine mcAbs of the present invention are well-characterized and exhibit the appropriate immunogenic responses.
  • the present invention provides compositions which may be used in clinical trials.
  • the present invention provides mixtures of murine anti-HCMV GPC mcAbs for prophylactic and thera ⁇ Treatment of patients with HCMV. It would not be practical or advisable to use human monoclonals because of the lack of stability of human hybridomas and lack of the necessary production levels for obtaining a therapeutic quantity of human mcAbs. It is becoming routine for transplant patients to receive immunoprophylaxis, both pre- and post-transplant.

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Abstract

Composition pouvant neutraliser l'infectivité du cytomégalovirus humain (HCMV), comprenant un mélange de (a) un premier anticorps monoclonal qui se lie à un premier épitope sur le GPA du HCMV, ledit premier anticorps monoclonal neutralisant l'infectivité du HCMV in vitro en présence d'un complément; et (b) un deuxième anticorps monoclonal qui se lie à un deuxième épitope sur le GPA du HCMV, ledit deuxième anticorps monoclonal ne neutralisant pas l'infectivité du HCMV in vitro en présence d'un complément. L'activité de neutralisation du mélange est plus intense que l'activité de neutralisation du premier anticorps monoclonal et la composition est éventuellement exempte de quantités significatives d'un anticorps monoclonal qui se lie à un troisième épitope sur le GPA du HCMV, et qui interfère avec la liaison des premier et deuxième anticorps monoclonaux.
PCT/US1988/002644 1987-08-07 1988-08-03 Anticorps monoclonaux reagissant avec la glycoproteine a du cytomegalovirus Ceased WO1989001628A1 (fr)

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Application Number Priority Date Filing Date Title
KR1019890700600A KR890702036A (ko) 1987-08-07 1988-08-03 사이토메갈로 바이러스 당단백질 a와 반응성인 모노클로날 항체

Applications Claiming Priority (6)

Application Number Priority Date Filing Date Title
US07/083,502 US5126130A (en) 1986-11-24 1987-08-07 Monoclonal antibodies reactive with specific antigenic sites on human cytomegalovirus glycoprotein a
US083,502 1987-08-07
US8381887A 1987-08-10 1987-08-10
US083,818 1987-08-10
US14476088A 1988-01-19 1988-01-19
US144,760 1988-01-19

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EP (1) EP0376973A1 (fr)
JP (1) JPH02504551A (fr)
AU (1) AU2324988A (fr)
WO (1) WO1989001628A1 (fr)

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1991004277A1 (fr) * 1989-09-14 1991-04-04 Children's Biomedical Research Institute Anticorps monoclonaux specifiques contre une glycoproteine de cytomegalovirus
EP0534102A1 (fr) * 1991-08-29 1993-03-31 BEHRINGWERKE Aktiengesellschaft Peptides spécifiques pour le cytomégalovirus humain et leurs utilisations
US5744298A (en) * 1991-08-29 1998-04-28 Behringwerke Aktiengesellschaft DNA sequences which encode immunoreactive HCMV-specific peptides

Citations (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0122841A1 (fr) * 1983-03-31 1984-10-24 Institut Pasteur Anticorps monoclonaux anti-cytomégalovirus humains, hybridomes secréteurs de ces anticorps et polypeptides porteurs d'un déterminant antigénique séquentiel de cytomégalovirus humains
EP0162533A2 (fr) * 1984-02-20 1985-11-27 Cogent Limited Détection d'IgM spécifique pour le cytomégalovirus humain
EP0180288A2 (fr) * 1984-11-02 1986-05-07 Institut Merieux Vaccin contre le cytomégalovirus
EP0236145A1 (fr) * 1986-03-07 1987-09-09 Cogent Limited Procédés de production de glycoprotéines du HCMV, anticorps contre celle-ci et vaccins HCMV, ainsi que des vecteurs recombinants à cet effet
EP0252302A1 (fr) * 1986-06-12 1988-01-13 BEHRINGWERKE Aktiengesellschaft Glycoprotéine principale du cytomégalovirus humain, sa préparation et son utilisation
US4743562A (en) * 1984-08-21 1988-05-10 The Board Of Trustees Of The Leland Stanford Junior University Purified human cytomegalovirus protein
WO1988003952A2 (fr) * 1986-11-24 1988-06-02 The Children's Hospital, Incorporated Glycopeptides immunogeniques du cytomegalovirus humain

Patent Citations (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0122841A1 (fr) * 1983-03-31 1984-10-24 Institut Pasteur Anticorps monoclonaux anti-cytomégalovirus humains, hybridomes secréteurs de ces anticorps et polypeptides porteurs d'un déterminant antigénique séquentiel de cytomégalovirus humains
EP0162533A2 (fr) * 1984-02-20 1985-11-27 Cogent Limited Détection d'IgM spécifique pour le cytomégalovirus humain
US4743562A (en) * 1984-08-21 1988-05-10 The Board Of Trustees Of The Leland Stanford Junior University Purified human cytomegalovirus protein
EP0180288A2 (fr) * 1984-11-02 1986-05-07 Institut Merieux Vaccin contre le cytomégalovirus
EP0236145A1 (fr) * 1986-03-07 1987-09-09 Cogent Limited Procédés de production de glycoprotéines du HCMV, anticorps contre celle-ci et vaccins HCMV, ainsi que des vecteurs recombinants à cet effet
EP0252302A1 (fr) * 1986-06-12 1988-01-13 BEHRINGWERKE Aktiengesellschaft Glycoprotéine principale du cytomégalovirus humain, sa préparation et son utilisation
WO1988003952A2 (fr) * 1986-11-24 1988-06-02 The Children's Hospital, Incorporated Glycopeptides immunogeniques du cytomegalovirus humain

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
Biological Abstracts, vol. 80, 1985 (Philadelphia, PA, US) L. Rasmussen et al.: "Viral polypeptides detected by a complement-dependent neutralizing murine monoclonal anti-body to human cytomegalovirus" *
Biological Abstracts, vol. 83, 1987 (Philadelphia, PA, US) B. Kari et al.: "Characterization of monoclonal antibodies reactive to several biochemically distinct human cytomegalovirus glycoprotein complexes" *
Biological Abstracts, vol. 84, 1987 (Philadelphia , PA, US) Y. Masuho et al.: "Human monoclonal antibodies neutralizing human cytomegalovirus" *

Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1991004277A1 (fr) * 1989-09-14 1991-04-04 Children's Biomedical Research Institute Anticorps monoclonaux specifiques contre une glycoproteine de cytomegalovirus
EP0534102A1 (fr) * 1991-08-29 1993-03-31 BEHRINGWERKE Aktiengesellschaft Peptides spécifiques pour le cytomégalovirus humain et leurs utilisations
US5744298A (en) * 1991-08-29 1998-04-28 Behringwerke Aktiengesellschaft DNA sequences which encode immunoreactive HCMV-specific peptides
US5859185A (en) * 1991-08-29 1999-01-12 Behringwerke Aktiengesellschaft HCMV-specific peptides, agents therefor and the use thereof
US6143493A (en) * 1991-08-29 2000-11-07 Dade Behring Marburg Gmbh HCMV-specific peptides, agents therefor and the use thereof

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AU2324988A (en) 1989-03-09
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