Peptides derived from the lymphadenopathy-associated virus...

Drug – bio-affecting and body treating compositions – Antigen – epitope – or other immunospecific immunoeffector – Amino acid sequence disclosed in whole or in part; or...

Reexamination Certificate

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C424S208100, C530S300000, C530S324000

Reexamination Certificate

active

06555112

ABSTRACT:

This invention relates to cloned DNA sequences indistinguishable from genomic RNA and DNA of lymphadenopathy-associated virus (LAV), a process for their preparation and their uses. It relates more particularly to stable probes including a DNA sequence which can be used for the detection of the LAV virus or related viruses or DNA proviruses in any medium, particularly biological samples containing any of them. The invention also relates to polypeptides, whether glycosylated or not, encoded by said DNA sequences.
Lymphadenopathy-associated virus (LAV) is a human retrovirus first isolated from the lymph node of a homosexual patient with lymphadenopathy syndrome, frequently a prodrome or a benign form of acquired immune deficiency syndrome (AIDS). Subsequently, other LAV isolates were recovered from patients with AIDS or pre-AIDS. All available data are consistent with the virus being the causative agent of AIDS.
A method for cloning such DNA sequences has already been disclosed in British Patent Application Nr. 84 23659, filed on Sep. 19, 1984. Reference is hereafter made to that application as concerns subject matter in common with the further improvements to the invention disclosed herein.
SUMMARY OF THE INVENTION
The present invention aims at providing additional new means which are not only useful for the detection of LAV or related viruses, (hereafter more generally referred to as “LAV viruses” or “Human Immunodeficiency Virus Type 1” or simply “HIV-1” but also new means that have more versatility, particularly in detecting specific parts of the genomic RNA of said viruses whose expression products are not always directly detectable by immunological methods.
The present invention further aims at providing polypeptides containing sequences in common with polyptides encoded by the LAV genomic RNA. It relates even more particularly to polypeptides comprising antigenic determinants included in the proteins encoded and expressed by the LAV genome occuring in nature. An additional object of the invention is to further provide means for the detection of proteins related to LAV virus, particularly for the diagnosis of AIDS or pre-AIDS or, to the contrary, for the detection of antibodies against the LAV virus or proteins related therewith, particularly in patients afflicted with AIDS or pre-AIDS or more generally in asymtomatic carriers and in blood-related products. Finally, the invention also aims at providing immunogenic polypeptides, and more particularly protective is polypeptides for use in the preparation of vaccine compositions against AIDS or related syndromes.
The present invention relates to additional DNA fragments, hybridizable with the genomic RNA of LAV as they will be disclosed hereafter as well as with additional cDNA variants corresponding to the whole genomes of LAV viruses. It further relates to DNA recombinants containing said DNAs or cDNA fragments.
The invention relates more particularly to a cDNA variant corresponding to the whole of LAV retroviral genomes, which is characterized by a series of restriction sites in the order hereafter (from the 5′ end to the 3′ end).
The coordinates of the successive sites of the whole LAV genome (restriction map) are indicated hereafter too, with respect to the Hind III site (selected as of coordinate 1) which is located in the R region. The coordinates are estimated with an accuracy of ±200 bp:
Hind
III
0
Sac
I
50
Hind
III
520
Pst
I
800
Hind
III
1 100
Bgl
II
1 500
Kpn
I
3 500
Kpn
I
3 900
Eco
RI
4 100
Eco
RI
5 300
Sal
I
5 500
Kpn
I
6 100
Bgl
II
6 500
Bgl
II
7 600
Hind
III
7 850
Bam
HI
8 150
Xho
I
8 600
Kpn
I
8 700
Bgl
II
8 750
Bgl
II
9 150
Sac
I
9 200
Hind
III
9 250
Another DNA variant according to this invention optionally contains an additional Hind III approximately at the 5 550 coordinate.
Reference is further made to
FIG. 1
which shows a more detailed restriction map of said whole DNA (&lgr;J19).
An even more detailed nucleotide sequence of a preferred DNA according to the invention is shown in
FIGS. 4-12
hereafter.
The invention further relates to other preferred DNA fragments which will be referred to hereafter.


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