Chemistry: molecular biology and microbiology – Enzyme – proenzyme; compositions thereof; process for...
Reexamination Certificate
1989-12-13
2001-06-19
Jones, W. Gary (Department: 1807)
Chemistry: molecular biology and microbiology
Enzyme , proenzyme; compositions thereof; process for...
C435S207000, C435S005000, C435S974000, C424S085100, C530S326000, C530S350000, C530S395000, C530S403000, C930S221000
Reexamination Certificate
active
06248574
ABSTRACT:
The present invention relates to the production and use, in diagnosis, treatment and prevention of HIV infection, of an antigenic protein that reacts specifically with antibodies against the Human Immunodeficiency Virus type 1 (HIV-1). The present invention also relates to a self-replicating cell that produces such a recombinant protein; to a vaccine comprising the recombinant protein; to an antigen-based screening test, based on the protein, for detecting antibodies to HIV; to monoclonal and polyclonal antibodies, produced using the protein, that protect against HIV-1 infection or disease; and to an antibody-based screening test.
HIV has been established as the primary etiologic agent in pathogenesis of acquired immunodeficiency syndrome (AIDS) and related disorders. See, e.g., Gruest, J., et al.,
Science
220: 863-71 (1983); Gallo, R. C., et al.,
Science
224: 500-03 (1984). Because AIDS can be transmitted by blood products, a highly accurate method of screening blood samples for presence of the virus is desirable. Infection of humans with HIV leads to production of antibodies directed against most of the viral structural antigens, forming the basis for screening via viral lysate based tests. However, these tests have several disadvantages, including false positives thought to arise from the presence of non-viral proteins in the viral lysate preparations used in the solid phase component of the current assays.
The antibodies produced upon infection include antibodies against both core and envelope proteins. The emergence of antibodies to envelope glycoproteins, e.g., gp160, and its subunits gp120 (the extracellular glycoprotein or EGP) and gp41 (the transmembrane protein or TMP), appears to precede the emergence of antibodies to core proteins, leading researchers to study these proteins as a possible basis for improved diagnostic assays. In addition, these antibodies to env proteins appear to be involved in induction of active immunity, suggesting their use in vaccine preparation.
Based on study of the envelope amino acid sequences, and in an attempt to reduce the rate of false positives, the art has proposed serological assays employing synthetic polypeptides that mimic naturally occurring antigenic determinants on viral proteins. Data generated in studies using synthetic peptides have indicated that some of the conserved domains in gp120 and gp41, respectively, contain immunodominant epitopes that may be appropriate for diagnosis. From analysis of conserved HIV domains from gp41 it appears that, within a linear sequence spanning about 40 amino acids (amino acid 570-612), numerous and highly immunodominant epitopes of HIV reside. Wang, J. J. G., et al.,
Proc. Nat'l Acad. Sci.
USA 83: 6159-63 (1986); Gnann, J. W., et al.,
J. Infect. Dis.
156: 261-67 (1987).
Conserved domains in gp120 and gp41 have also been identified that are involved in neutralization of different HIV isolates. Ho, D. D., et al., J.
Virol.
61: 2024-28 (1987);
Science
239: 1021-23 (1988). Neutralizing-specific antibodies to the major envelope glycoprotein are type-specific and have recently been mapped to a highly variable sequence of gp120. Putney, S. D., et al.,
J. Cell. Biochem.
12B: 5 (1988). Neutralization assays with immune sera from animals or humans, see Robert-Guroff, M., et al.,
Nature
316: 72-74 (1985); Weiss, R. A., et al.,
Nature
316: 69-71 (1985); Rasheed, S., et al.,
Virology
150: 1-6 (1986), have revealed that the envelope proteins contains epitopes that elicit antibodies capable of neutralizing HIV in vitro. But the presence of these antibodies in vivo has only a limited effect on progression of the disease, Robert-Guroff, M., et al., loc. cit.; Wendler, I., et al.,
AIDS Res. Human Retroviruses
3: 157-63 (1987), and no significant difference in titers which can be correlated with clinical status. In humans, there are no known patterns of antibody response indicative of immunity.
SUMMARY OF THE INVENTION
It is therefore an object of the present invention to provide a fusion protein, readily produced in commercially significant quantities, that is an effective vaccine for prevention of HIV infection.
It is a further object of the present invention to provide a fusion protein, readily produced in commercially significant quantities, that is an exceedingly sensitive and specific antigen for detection of HIV-1 antibodies.
It is yet another object of the present invention to provide a highly accurate diagnostic assay for detecting antibodies to HIV-1.
In accomplishing these and other objects, there has been provided, according to one aspect of the present invention, a fusion protein comprised of an amino acid sequence selected from the group consisting of NVTENFNMWKN, KAKRRVVQREKRAVG, ERYLKDQQLLGIWGCSGKLIC and EESQNQQEKNEQELLELDKWA; and a non-HIV polypeptide sequence, such that the amino-acid sequence and the polypeptide sequence comprise the backbone of the fusion protein, wherein the fusion protein reacts with an HIV-positive serum. In a preferred embodiment, the amino-acid sequence is joined via a peptide bond to an N-terminus of the non-HIV polypeptide sequence.
In accordance with another aspect of the present invention, a self-replicating cell is provided that expresses a polypeptide comprising an amino-acid sequence selected from the group consisting of NVTENFNMWKN, KAKRRVVQREKRAVG, ERYLKDQQLLGIWGCSGKLIC and EESQNQQEKNEQELLELDKWA.
Also provided, according to still another aspect of the present invention, is a diagnostic assay for detecting the presence of anti-HIV antibody in a sample, comprising the steps of (A) immobilizing on a solid matrix a fusion protein comprising an amino-acid sequence, ERYLKDQQLLGIWGCSGKLIC and a non-HIV polypeptide sequence, such that the amino-acid sequence is accessible to an antibody contacting a surface of the matrix; (B) bringing a sample into contact with the surface of the matrix; and (C) monitoring the surface for binding of HIV-specific antibody. In one preferred embodiment, step (C) comprises detecing the presence of anti-HIV antibody in a sample that tests HIV-negative when tested using a conventional whole-virus western blot assay.
Pursuant to another aspect of the present invention, a diagnostic assay is provided for detecting the presence of anti-HIV antibody in a sample, comprising the steps of (A) providing a fusion protein comprising an amino-acid sequence ERYLKDQQLLGIWGCSGKLIC and an amino acid sequence of an enzyme; (B) combining a sample with the fusion protein in a liquid; and (C) monitoring the combination for a modulation of activity of the enzyme.
A vaccine comprising a fusion protein, as described above, and a sterile, pharmacologically acceptable carrier therefor is also provided, in accordance with yet another aspect of the present invention.
In accordance with another aspect of the present invention, there has been provided an immunotherapy method that comprises the step of administering to a subject an immunostimulatory amount of a vaccine as described above. In a preferred embodiment, the subject is already infected with HIV-1 when the vaccine is administered.
Pursuant to another aspect of the present invention, an immunotherapy method is provided comprising the step of administering to a subject an immunostimulatory amount of a hyperimmune globulin prepared according to a method comprised of immunizing a plasma donor with a vaccine as described above, such that a hyperimmune globulin is produced which contains antibodies directed against HIV-1. According to another aspect of the present invention, an immunotherapy method is provided that comprises administering to a subject an immunostimulatory amount of a hyperimmune globulin prepared in the aforementioned manner.
Also provided is an immunotoxin conjugate comprising a fusion protein, as described above, conjugated to an immunotoxin. In addition, an immunotherapy method is provided, according to another aspect of the present invention, wherein a subject already infected with HIV-1 receives antibodies directed against a fusion protein of the invention, wherei
Arwine Elizabeth
Jones W. Gary
Moran John Francis
Sisson Bradley L.
LandOfFree
Polypeptides selectively reactive with antibodies against... does not yet have a rating. At this time, there are no reviews or comments for this patent.
If you have personal experience with Polypeptides selectively reactive with antibodies against..., we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Polypeptides selectively reactive with antibodies against... will most certainly appreciate the feedback.
Profile ID: LFUS-PAI-O-2516157