Recombinant FIV glycoprotein 160 and P24 gag protein

Details Recombinant FIV glycoprotein 160 and P24 gag protein Recombinant FIV glycoprotein 160 and P24 gag protein

1991-02-28

2001-05-08

Achutamurthy, Ponnathapu (Department: 1652)

Chemistry: molecular biology and microbiology

Micro-organism, tissue cell culture or enzyme using process...

Recombinant dna technique included in method of making a...

C435S252300, C435S252330, C435S320100, C435S471000, C530S350000, C530S395000, C530S826000, C514S002600, C424S819000, C536S023720

Reexamination Certificate

active

06228608

ABSTRACT:

FIELD OF THE INVENTION
The present invention relates generally to the fields of hematology, immunology and recombinant genetics. The invention specifically relates to recombinant feline immunodeficiency virus glycoprotein 160 envelope protein and fragments thereof, and p24 gag protein and fragments thereof. In another aspect, it relates to the use of the FIV antigens and fragments to induce, in a feline, antibodies to FIV.
DESCRIPTION OF THE BACKGROUND ART
Feline immunodeficiency virus (FIV), formerly called feline T lymphotropic lentivirus (Pederson et al.,
Science
, 235:790 (1987)), has been identified in the United States, the United Kingdom (Harbour et al.,
Vet Rec
, 122:84 (1988)), Japan (Ishida et al.,
Jpn J Vet Sci
, 50:39 (1988)), Australia (Sabine et al.,
Aust Vet Practit
, 18:105 (1988)), and New Zealand (Swinney et al.,
NZ Vet J
, 37:41 (1989)). The virus appears to be spread by horizontal transmission, predominantly by bite wounds (Yamamoto et al.,
Am. J. Vet. Res
., 8:1246 (1988); Friend et al.,
Aust. Vet J
., 67:237 (1990). FIV has been classified as a member of the subfamily Lentivirinae in the family Retroviridae. This is the family that includes human and simian immunodeficiency viruses, equine infectious anaemia, maedi visna of sheep and caprine-arthritis encephalitis viruses (CAEV).
Cloning and sequencing of FIV has confirmed it to be a lentivirus by its genomic organization and antigenic similarity of its core proteins to those of visna virus and CAEV (Olmsted et al.,
Proc. Natl. Acad. Sci. USA
, 86:2448 (1989); Talbott et al.,
Proc. Natl. Acad. Sci. USA
, 86:5743 (1989); Dow et al.,
Journal Of Acquired Immune Deficiency Syndromes
, 3:658 (1990).
Until the present time, however, very little information has been available regarding the outer shell, or envelope, of the feline immunodeficiency virus. Inasmuch as an elucidation of envelope proteins would be of great value in understanding and modulating the immunological characteristics of FIV, a need has continued to exist for such data.
SUMMARY OF THE INVENTION
Accordingly, the present inventors have succeeded in cloning and expressing novel envelope proteins and protein fragments of FIV, which are useful in the diagnosis and treatment of this immune condition. Specifically, the inventors report the FIV glycoprotein 160 envelope protein, as well as the FIV 0.4 envelope protein, which is an amplified product of the FIV gp160 envelope protein. The inventors also report the novel FIV 1.2 envelope protein that is an amplified product obtained from the region of FIV envelope from position 6996 to 8129.
In addition to the novel envelope proteins and fragments, the inventors have successfully identified p24 gag protein from FIV which is also useful in the diagnosis and treatment of FIV. The inventors have further identified FIV 0.6 which is an amplified product obtained from FIV gag protein.
It is an object of the present invention, then, to provide for a method of diagnosing the infection of a feline by FIV. The present invention thus provides an important advance in the study and therapy of feline immune deficiency syndromes.
The work presented here demonstrates that recom-binant FIV 0.4 envelope protein may be expressed by a transformed cell. The production of recombinant FIV 0.4 envelope protein makes possible new assays and treatments for FIV. It is therefore an object of the present invention to use recombinant proteins from the gp160 envelope protein and the gag protein to develop vaccines in the prevention of FIV infection in cats.
Thus in one embodiment, there is provided according to the invention recombinant functionally active feline immunodeficiency virus 0.4 envelope protein, or a functional or chemical derivative thereof.
In yet another embodiment is provided the FIV gp160 derived 0.4 envelope fragment which is produced by eukaryotic cells. Yet another embodiment of the invention comprises the plasmid pLCBCOFIVO.4. There is also provided according to this invention, methods for producing FIV gp160 derived 0.4 envelope protein, comprising culturing the transformed cell under conditions allowing expression of the gp160 derived 0.4 envelope fragment, and recovering said gp160 derived 0.4 envelope protein.
In yet another embodiment, the present invention provides for an antibody against the FIV envelope protein of the invention.
Further a method of purifying FIV from a sample is provided according to the present invention, comprising contacting said sample with the antibody of the invention, so as to form a complex between said antibody and FIV in said sample, and removing the FIV from said antibody so as to obtain purified FIV.
Moreover, a method of detecting FIV in a sample comprising contacting said sample with the antibody of the invention, wherein said antibody is detectably labeled, so as to form a complex between FIV in said sample and said detectably labeled antibody, and detecting the complexed or uncomplexed detectably labeled antibody.
An additional embodiment of the current invention comprises a pharmaceutical preparation comprising the antibody of the invention. In another embodiment, there is provided a process for the preparation of a pharmaceutical composition useful for inducing the production in a cat of antibodies to FIV, the process comprising admixing an immunologically effective amount of the polypeptide comprising the amino acid sequence of the gp160, gp120 or FIV 0.4 envelope protein together with a pharmaceutically acceptable carrier.
These and other non-limiting embodiments of the present invention will be apparent to those of skill from the following detailed description of the invention.


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