Chemistry: natural resins or derivatives; peptides or proteins; – Proteins – i.e. – more than 100 amino acid residues – Blood proteins or globulins – e.g. – proteoglycans – platelet...
Reexamination Certificate
1998-07-30
2001-07-31
Park, Hankyel (Department: 1648)
Chemistry: natural resins or derivatives; peptides or proteins;
Proteins, i.e., more than 100 amino acid residues
Blood proteins or globulins, e.g., proteoglycans, platelet...
C424S192100, C424S208100, C435S005000, C435S007100, C435S339100
Reexamination Certificate
active
06268484
ABSTRACT:
BACKGROUND OF THE INVENTION
1. Technical field
The present invention is in the field of immunology, especially detection, prevention and treatment of HIV-1 infection and AIDS therapy. More particularly, it concerns monoclonal antibodies, drugs and vaccines made from these antibodies and methods based on the use of these antibodies, drugs and vaccines for analytical and/or clinical applications.
2. Description of Related Art
In the sera of human immunodeficiency virus type 1 (HIV-1) infected patients, anti-virus antibodies can be detected over a certain period after infection without any clinical manifestations of the acquired immunodeficiency syndrome (AIDS). At this state of active immune response, high numbers of antigen-specific B-cells are expected in the circulation. These B-cells are used as fusion partners for the generation of human monoclonal anti-HIV antibodies.
Monoclonal antibodies can be produced by known procedures, e.g., as described by R. Kennet et al. in “Monoclonal Antibodies and Functional Cell Lines; Progress and Applications”. Plenum Press (New York), 1984.
Further materials and methods applied are based on known procedures, e.g., such as described in J. Virol. 67:6642-6647, 1993.
Monoclonal antibodies and in particular human monoclonal antibodies have been widely used in the last few years in order to improve the understanding of HIV-1 neutralization by antibodies released upon immunization with HIV-1 derived immunogens or upon infection in afflicted patients (J. Virol. 62:2107-2114, 1988; Immunology 76;515-534, 1992; J. Virol. 67:6642-6647, 1993; U.S. Pat. No. 5,087,557). Many efforts have been made to overcome the detrimental capability of the HIV-1 virus to rapidly charge its morphology under immunological pressure and thereby to escape the capture by antibodies released from a patient's immune systems or developed and applied by researchers, As a result thereof, there is presently no reliable antibody-leased (nor any other) vaccine for active or passive immunization on the market. One significant step forward has been made when an antigens determinant on the smaller subunit gp41 of the HIV-1 envelope glycoprotein gp160 was found (EP 570 357 A2), which corresponds to the amino acid sequence “ELDKWA” (SEQ ID NO:11) located at amino acid position number 662 to 667 of gp41 of HIV-1 isloate BH10. The authors report therein an HIV-1 neutralizing human monoconal antibody specifically binding to said antigenic determinant. The antibody proved to be a powerful tool for biochemical analysis of the binding epitome and its variability, The discovery of the highly conserved state of said gp41-epitope gave rise to the hope of possibly finding a vaccine composition suitable for more reliable prevention of human individuals from HIV-1 infection and/or for more successful therapeutic treatment of infected patients.
The results reported in EP 570 357 A2 motivated the present inventors to intensify their research activities which finally led them to the novel and inventive findings herein disclosed.
However, in spite of promising results of the art relating to the use of HIV-1 neutralizing monoclonal antibodies, there is at least one major drawback to this sort of approach. It lies in the wide-spread use of laboratory strains of HIV-1 isolates, which have become adapted to lab-conditions and are more or less attenuated and hence only poorly—if at all—representative of the properties and behaviour of primary HIV-1 isolates. Consequently, promising vaccine compositions drawn against laboratory HIV-1 strains frequently proved non-efficacious when applied against primary HIV-1 isolates, e.g., of blood samples of infected persons (see J. Cohen, Science 262:980-981, 1993).
The second major drawback was and still is the ability of the HIV-1 virus to escape antibody capture by morphological variation, which very often renders the remarkable efforts of the researchers almost useless. Such escape mutants may be characterized by a change of only one or several of the amino acids within one of the targeted antigenic determinants and may occur, e.g., as a result of spontaneous or induced mutation.
SUMMARY OF THE INVENTION
The present invention therefore provides antibodies which have been found to overcome the disadvantages of the prior art and which can be used for the manufacture of vaccines for active and/or passive immunization of persons in need of such treatment. Such beneficial antibodies are, for instance, produced by any one of the cell lines CL1 through CL6 listed below. The invention also provides for human monoclonal antibodies that are functionally equivalent to the antibodies of CL1 through CL6. These functionally equivalent antibodies substantially share at least one major functional property with an antibody of CL1 to CL6 as herein described, comprising: binding specificity to gp160; bindinig dependence on glycosylation; reactivity in the presence of tunicamycin; inhibition of infections of human lymphocytes by primary HIV-1 isolates; reactivity towards antiidiotypes; competition for same binding site; reduction of the HIV-1 level in blood serum after intravenous administration to an infected patient; and/or specific binding to HIV-1 neutralizing antibodies.
It is also an object of the present invention to provide for the hybridoma and/or CHO cell lines producing any one of the antibodies disclosed and claimed herein.
The invention is further directed to mixtures of antibodies according to the present invention, as well as to methods of using individual antibodies or mixtures thereof for the prevention and/or therapeutical treatment of HIV-1 infections in vitro and in vivo, and/or for improved detection of HIV-1 infections.
The cell lines CL1 to CL4 produce monoclonal antibodies recognizing HIV-envelope glycoproteins, and in particular specific antigenic determinants of gp160. The antibodies of CL1 and CL4 recognize and bind to an amino acid sequence of gp41/gp160 corresponding to the epitope located at amino acid position number 662 to 667 (“ELDKWA”) of gp41 of HIV-1 isloate BH10 (GenBank accession M15654; (SEQ ID NOS:1-10) numbering as described in the Swissprot database entry ENV$HIV10). The monoclonal antibodies of CL2 and CL3 bind to two different antigenic determinants, more particularly to fragments of gp120/gp160 corresponding to the epitope sequences located at amino acid positions 79 to 184 and 326 to 400 respectively, of processed gp120 of HIV-1 isolate BH10 (GenBank accession M15654; numbering as described in the Swissprot database entry ENV$HIV10).
While the idiotypic antibodies produced by CL1 to CL4 are directed to the capture and neutralization of HIV-1 viruses in vitro and in vivo, the antiidiotypic antibodies released from CL5 and CL6 take an opposite role, i.e., they mimic the viruses, more particularly they mimic the corresponding antigenic determinant(s) of the HIV-1 viruses. The anti-idiotypic antibodies of CL5 and CL6 are of a nature such that they bind to the idiotypic antibody of CL2 at essentially the same location(s) (antigenic determinants) on gp160 as does the virus itself.
REFERENCES:
patent: 4761470 (1988-08-01), Emini et al.
patent: 5087557 (1992-02-01), McClure
patent: 5245015 (1993-09-01), Fung et al.
patent: 5516657 (1996-05-01), Murphy et al.
Fahey et al., Status of immune-based therapies in HIV infection and AIDS Clin. Exp. Immunol. (1992) 88, 1-5 01/92.
Luckow et al., Trends in the development of Baculovirus expression vectors, Bio/Tech vol. 6, pp. 47-55, see Abstract and p. 47, col. 1, sentence 4. (01/88).
Ratner et al., Complete nucleotide sequence of the AIDS virus, HTLV-III, Nature 313:277-284 (01/85).
Ballaun Claudia
Buchacher Andrea
Ernst Wolfgang
Katinger Hermann
Klima Annelies
Birch Stewart Kolasch & Birch, LLP.
Park Hankyel
Polymun Scientific Immunbiologische Forschung GmbH
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