Title:  Cloned DNA Sequences Related To The Entire Genomic RNA Of Human Immunodeficiency Virus II (HIV-2), Polypeptides Encoded By These DNA Sequences And Use Of These DNA Clones And Polypeptides In Diagnostic Kits

United States Patent:  6,355,789

Assignee:  Institut Pasteur (Paris, FR)

Filed:  June 6, 1995

Foreign Application Priority Data:  Jan 22, 1986[FR] (86 00911); Feb 06, 1986[FR] (86 01635); Feb 13, 1986[FR] (86 01985); Mar 18, 1986[FR] (86 03881); Mar 24, 1986[FR] (86 04215).

The present invention is directed toward nucleic acids containing the full-length human immunodeficiency virus type 2 ROD (HIV-2_ROD) pol gene. HIV-2, which was originally designated lymphadenopathy-associated virus type II (LAV-II), was isolated from AIDS patients in West Africa. The virus is genotypically and phenotypically distinct from HIV-1 and bears a closer genetic relationship to the simian immunodeficiency virus (SIV). The present invention describes the preparation of HIV-2_ROD proviral molecular clones from a genomic lambda phage library of CD4⁺ -infected cells. The complete nucleotide sequence of the full-length genome was determined and the putative gag, pol, env, vif (Q), vpr (R), vpx (X), nef (F), tat, and rev (art) genes identified. The claimed invention is directed toward nucleic acids containing the full-length HIV-2_ROD pol gene (nt 1829-4936). These nucleic acids should prove useful as diagnostic reagents for the detection of HIV-2 and facilitate expression of the pol gene product.

SUMMARY OF THE INVENTION

As noted previously, the present invention relates to the cloned nucleotide sequences homologous or identical to at least a portion of the genomic RNA of HIV-2 viruses and to polypeptides encoded by the same. The present invention also relates to kits capable of diagnosing an HIV-2 infection.

Thus, a main object of the present invention is to provide a kit capable of diagnosing an infection caused by the HIV-2 virus. This kit may operate by detecting at least a portion of the RNA genome of the HIV-2 virus or the provirus present in the infected cells through hybridization with a DNA probe or it may operate through the immunodiagnostic detection of polypeptides unique to the HIV-2 virus.

Additional objects and advantages of the present invention will be set forth in part in the description which follows, or may be learned from practice of the invention. The objects and advantages may be realized and attained by means of the instrumentalities and combinations particularly pointed out in the appended claims.

To achieve these objects and in accordance with the purposes of the present invention, cloned DNA sequences related to the entire genomic RNA of the LAV-II virus are set forth. These sequences are analogous specifically to the entire genome of the LAV-II ROD strain.

To further achieve the objects and in accordance with the purposes of the present invention, a kit capable of diagnosing an HIV-2 infection is described. This kit, in one embodiment, contains the cloned DNA sequences of this invention which are capable of hybridizing to viral RNA or analogous DNA sequences to indicate the presence of an HIV-2 infection. Different diagnostic techniques can be used which include, but are not limited to: (1) Southern blot procedures to identify viral DNA which may or may not be digested with restriction enzymes; (2) Northern blot techniques to identify viral RNA extracted from cells; and (3) dot blot techniques, i.e., direct filtration of the sample through an ad hoc membrane such as nitrocellulose or nylon without previous separation on agarose gel. Suitable material for dot blot technique could be obtained from body fluids including, but not limited to, serum and plasma, supernatants from culture cells, or cytoplasmic extracts obtained after cell lysis and removal of membranes and nuclei of the cells by ultra-centrifugation as accomplished in the "CYTODOT" procedure as described in a booklet published by Schleicher and Schull.

In an alternate embodiment, the kit contains the polypeptides created using these cloned DNA sequences. These polypeptides are capable of reacting with antibodies to the HIV-2 virus present in sera of infected individuals, thus yielding an immunodiagnostic complex.

To further achieve the objects of the invention, a vaccinating agent is provided which comprises at least one peptide selected from the polypeptide expression products of the viral DNA in admixture with suitable carriers, adjuvents stabilizers.

Claim 1 of 1 Claim

What is claimed is:

1. A nucleic acid of HIV-2 having the nucleotide sequence of a full length pol gene as set forth in FIG. 6.

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