Retrovirus isolated from mantle histiocytes in mantle cell lymphoma

Abstract

The present invention features an isolated, intact virus associated with human lymphoma, and originally isolated from a mantle cell lymphoma, referred to herein as a mantle histiocyte retrovirus (MHRV). The invention also features compositions and methods for detecting MHRV, as well as methods and compositions for propagating MHRV in vitro, screening for anti-MHRV agents, and generation of attenuated MHRV strains.

1 .- 13 . (canceled)  
     
     
         14 . An isolated mammalian cell comprising a polynucleotide comprising a sequence encoding a polypeptide comprising an amino acid sequence of at least 4 contiguous amino acid residues of amino acid residues 1-22 of SEQ ID NO:8 or a complement thereof.  
     
     
         15 . The isolated mammalian cell of  claim 14 , wherein the polynucleotide comprises a sequence encoding a polypeptide comprising an amino acid sequence of at least 10 contiguous amino acid residues of amino acid residues 1-22 of SEQ ID NO:8 or a complement thereof.  
     
     
         16 . An isolated mammalian cell comprising a polynucleotide comprising a sequence of at least 12 contiguous residues of nucleic acid residues 1-66 of SEQ ID NO:7 or a complement thereof.  
     
     
         17 . The isolated mammalian cell of  claim 16 , wherein the polynucleotide comprises a sequence of at least 33 contiguous residues of nucleic acid residues 1-66 of SEQ ID NO:7 or a complement thereof.  
     
     
         18 . The isolated mammalian cell of  claim 17 , wherein the polynucleotide comprises a sequence of at least 50 contiguous residues of nucleic acid residues 1-66 of SEQ ID NO:7 or a complement thereof.  
     
     
         19 . (canceled)  
     
     
         20 . The isolated mammalian cell of  claim 14 , wherein the polynucleotide is operably linked to a heterologous promoter element.  
     
     
         21 . An isolated mammalian cell comprising a polynucleotide comprising a sequence that hybridizes under conditions of high stringency to at least a portion of the polynucleotide sequence of nucleotides 1-66 of SEQ ID NO:7 or a complement thereof.  
     
     
         22 . An isolated mammalian cell comprising a nucleic acid sequence having at least 65% identity to at least 12 contiguous nucleotides of nucleic acid residues 1-66 of SEQ ID NO:7 or a complement thereof.  
     
     
         23 .- 24 . (canceled)  
     
     
         25 . An isolated recombinant MHRV GAG polypeptide.  
     
     
         26 . An isolated recombinant MHRV GAG polypeptide encoded by the polynucleotide of any of claims  14 ,  18 ,  20 ,  21  and  22 .  
     
     
         27 . An isolated antibody that specifically binds the isolated MHRV GAG polypeptide of  claim 25 .  
     
     
         28 . (canceled)  
     
     
         29 . A method for detecting mantle histiocyte retrovirus (MHRV) in a mammalian cell, the method comprising: 
 contacting a biological sample comprising a mammalian cell suspected of containing an MHRV polynucleotide with an MHRV-specific nucleic acid probe, said contacting being for a time sufficient for binding of the MHRV-specific nucleic acid probe to the sample to form complexes between the probe and a nucleic acid probe target; and    detecting the presence or absence of complexes of the MHRV-specific probe and the probe target in the sample;    wherein detection of complexes in the sample indicates the mammalian cell containing an MHRV polynucleotide is present in the sample.    
     
     
         30 . The method of  claim 29 , wherein the MHRV-specific nucleic acid probe comprises at least 8 contiguous nucleotide residues of SEQ ID NO:1 or a complement thereof.  
     
     
         31 . The method of  claim 29 , wherein the MHRV-specific probe and the probe target are nucleic acid, and wherein the MHRV-specific probe comprises at least 8 contiguous nucleotide residues of residues 1-66 of SEQ ID NO:7 or a complement thereof.  
     
     
         32 .- 33 . (canceled)  
     
     
         34 . The method of  claim 29 , wherein the biological sample comprising the mammalian cell is obtained from blood or a blood-derived product.  
     
     
         35 . The method of  claim 29 , wherein the biological sample comprises a mammalian tissue containing a macrophage or a macrophage-derived tumor cell.  
     
     
         36 . The method of  claim 29 , wherein the method comprises: 
 contacting the biological sample comprising a mammalian cell suspected of containing an MHRV polynucleotide with a first MHRV-specific nucleic acid probe and with a second MHRV-specific nucleic acid probe, wherein the first probe and the second probe each comprise at least 15 contiguous nucleotides of a nucleic acid sequence of SEQ ID NO:7 or a complement thereof, said contacting being under conditions effective to produce an amplified DNA product; and    detecting the presence or absence amplified DNA product;    wherein detection of amplified DNA product corresponding to an amplified DNA product expected from a nucleic acid sequence comprising SEQ ID NO:7 indicates an MHRV polynucleotide is present in the sample.    
     
     
         37 . The method of  claim 36 , wherein the first probe comprises a sequence HERV-9 (SEQ ID NO:3) or HERV-10 (SEQ ID NO:4).  
     
     
         38 . The method of  claim 36 , wherein the second probe comprises a sequence of HERV-8 (SEQ ID NO:2), HERV-11 (SEQ ID NO:5), or HERV-12 (SEQ ID NO:6).  
     
     
         39 . The method of  claim 36 , wherein the first probe is HERV-8 (SEQ ID NO:2), the second probe is HERV-9 (SEQ ID NO:3), wherein detection of an amplified product of about 304 bp indicates an MHRV polynucleotide is in the sample.  
     
     
         40 . The method of  claim 36 , wherein the first probe is HERV-9 (SEQ ID NO:3), the second probe is HERV-12 (SEQ ID NO:6), wherein detection of an amplified product of about 1321 bp indicates an MHRV polynucleotide is in the sample.  
     
     
         41 . The method of  claim 36 , wherein the first probe is HERV-10 (SEQ ID NO:4) the second probe is HERV-11 (SEQ ID NO:5), wherein detection of an amplified product of about 1966 bp indicates an MHRV polynucleotide is in the sample.  
     
     
         42 . A kit for detection of mantle histiocyte retrovirus (MHRV), the kit comprising an MHRV-specific probe in a labeled vial, wherein the probe is: 
 an MHRV-specific nucleic acid probe that specifically hybridizes to a sequence encoding an MHRV GAG polypeptide;    an MHRV-specific GAG antibody; and    an MHRV polypeptide that specifically binds an anti-MHRV GAG polypeptide.    
     
     
         43 . (canceled)  
     
     
         44 . A method for detecting an MHRV-associated disease, the method comprising: 
 contacting a biological sample comprising a mammalian cell with an MHRV-specific probe, wherein the biological sample was obtained from a subject suspected of having an MHRV-associated disease, said contacting being for a time sufficient for binding of the MHRV-specific probe to the sample to form complexes between the probe and a probe target; and    detecting complexes of the MHRV-specific probe and the probe target in the sample;    wherein detection of complexes in the sample indicates MHRV is present in the sample, which indicates the subject may have an MHRV-associated disease.    
     
     
         45 . The method of  claim 44 , wherein the MHRV-associated disease is an MHRV-associated lymphoma.  
     
     
         46 . The method of  claim 44 , wherein the MHRV-associated disease is teratocarcinoma, multiple sclerosis, autoimmune rheumatic diseases, and schizophrenia.  
     
     
         47 . A method for producing an MHRV GAG polypeptide of  claim 25 , the method comprising the steps of: 
 recovering a polypeptide from a culture of recombinant host cells containing a recombinant MHRV GAG polypeptide-encoding polynucleotide, said culture being under conditions suitable for the expression of the polypeptide.    
     
     
         48 .- 55 . (canceled)  
     
     
         56 . The isolated mammalian cell of  claim 15 , wherein the polynucleotide is operably linked to a heterologous promoter element.  
     
     
         57 . The isolated mammalian cell of  claim 16 , wherein the polynucleotide is operably linked to a heterologous promoter element.  
     
     
         58 . The isolated mammalian cell of  claim 17 , wherein the polynucleotide is operably linked to a heterologous promoter element.  
     
     
         59 . The isolated mammalian cell of  claim 18 , wherein the polynucleotide is operably linked to a heterologous promoter element.