Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving virus or bacteriophage
Patent
1995-11-22
2000-10-24
Stucker, Jeffrey
Chemistry: molecular biology and microbiology
Measuring or testing process involving enzymes or...
Involving virus or bacteriophage
435 6, 435 71, 4353201, 536 231, 536 2372, 536 243, 530326, 530327, C12Q 170, C12Q 168, C12N 1509, C12N 1563
Patent
active
061365289
DESCRIPTION:
BRIEF SUMMARY
The present invention relates to multiple sclerosis and in particular to a virus associated with this disease.
Multiple sclerosis (MS) is a debilitating wasting disease which generally strikes its victims at some time after adolescence. The disease is associated with degeneration of the myelin sheaths surrounding nerve cells which leads to a loss of motor and sensory function.
There are no cures for MS and even attempts to ameliorate the symptoms and slow the progress of the disease have up to now met with only limited success.
Many different theories over the years have been expounded to explain the cause of MS, such as environmental and/or genetic factors. A viral origin has also been postulated although no conclusive data which would support this theory over any others have been demonstrated.
We have now demonstrated that MS is associated with a retrovirus. We have used material from patients with MS, made cDNA and amplified this material by PCR with degenerate oligonucleotide primers based upon a highly condensed region of retroviral polymerase (pol) genes. By this means we have obtained retroviral sequences from a previously unknown retrovirus which we have called human multiple sclerosis virus (HMSV). Our isolate of HMSV is described below as HMSV-1.
The present invention relates to nucleic acids such as DNA or RNA encoding all or part of HMSV genome. The invention also provides polypeptides and fragments thereof of HMSV. The invention further provides antibodies (including polyclonal, monoclonal, single domain and humanised antibodies) and fragments thereof (especially fragments containing an antigen binding site such as Fab' or F(ab).sub.2 fragments) against such peptides.
The peptides, antibodies and fragments thereof of the invention may be incorporated into kits for immunodiagnosis of HMSV.
Portions of the nucleic acid sequences derived from HMSV are useful as probes to diagnose the presence of virus in samples, and to isolate naturally occurring variants of the virus. These sequences also make available polypeptide sequences of HMSV antigens encoded within the HMSV genome(s) and permits the production of polypeptides which are useful as standards or reagents in diagnostic tests and/or as components of vaccines. Antibodies, both polyclonal and monoclonal, directed against HMSV epitopes contained within these polypeptide sequences are also useful for diagnostic tests, as therapeutic agents, for screening of antiviral agents, and for the isolation of the HMSV agent from which these sequences derive. In addition, by utilizing probes derived from these sequence, it is possible to isolate and sequence other portions of the HMSV genome, thus giving rise to additional probes and polypeptides which are useful in the diagnosis and/or treatment, both prophylactic and therapeutic, of MS.
Thus the invention provides a polynucleotide in substantially isolated form comprising a sequence of nucleotides which is capable of selectively hybridizing to the genome of the human multiple sclerosis virus (HMSV) or the complement thereof, wherein HMSV is characterized by: protein(s) or polyprotein(s); selectively hybridizable with any one of the nucleotide sequences illustrated in SEQ ID NOS: 1 to 6.
Nucleotide sequences homologous to the sequences of any one of SEQ ID NOS: 1 to 6 will be those in which there is at least 60%, for example 70, 75, 80, 85, 90, or 95% homology over the length of the sequence exemplified. Those of skill in the art will of course appreciate that because the figures illustrate cDNA sequences and the virus is an RNA virus, the bases represented by the letters T and U of the genetic code must be considered equivalent for the purposes of homology measurement.
Similarly, a sequence which is selectively hybridizable to any one of the sequences of SEQ ID NOS: 1-6 will have a degree of homology to the complementary strand of the sequence to which they can hybridize similar to the homology mentioned above.
Accordingly, the features (i) to (iv) provide a "fingerprint" which can be read by those of ski
REFERENCES:
Poiesz, et al. : Polymerase chain reaction and the detection . . . : J. Ceel. Biochem. :Suppl. 13E: abstracts WH 224, Apr. 1989.
Maniatis, et al. : Molecular Cloning a laboratory manual: Cold Spring Harbor: pp. 431-433, 1982.
Garson Jeremy
Tuke Philip
Biomerieux S.A.
Nelson Brett
Stucker Jeffrey
University College London
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