Method of transducing non-dividing myeloid cells utilizing chimeric murine leukemia viruses containing Vpx
The invention is directed to a chimeric gammaretrovirus comprising an gammaretroviral virion which contains a lentiviral Vpx protein and methods of use thereof. In a particular aspect, the chimeric gammaretrovirus is a chimeric murine leukemia virus (MLV) comprising an MLV virion which contains a lentiviral Vpx protein. The invention is also directed to use of the chimeric gammaretrovirus to produce a MLV that can transduce a non-dividing cell (G1/S/G2), transduce a non-dividing cell; enhance the ability of a MLV to transduce a non-dividing cell; transduce a quiescent (G0) cell; and enhance the ability of a human immunodeficiency virus 1 (HIV-1) to transduce a quiescent (G0) cell.
1. A method of transducing a non-dividing myeloid cell comprising contacting the cell with a chimeric murine leukemia virus (MLV) comprising an MLV virion which contains a lentiviral Vpx protein and maintaining the cell under conditions in which nucleic acid of the chimeric MLV is transferred to the cell, thereby transducing the non-dividing myeloid cell.
2. The method of claim 1 wherein the chimeric MLV further comprises an exogenous sequence that is expressed upon transduction of the chimeric MLV into the non-dividing myeloid cell.
3. A method of enhancing the ability of a murine leukemia virus (MLV) to transduce a non-dividing myeloid cell comprising contacting the cell with a chimeric MLV comprising an MLV virion which contains a lentiviral Vpx protein and maintaining the cell under conditions in which nucleic acid of the chimeric MLV is transferred to the cell, thereby enhancing the ability of a MLV to transduce the non-dividing myeloid cell.
4. The method of claim 3 wherein the chimeric MLV further comprises an exogenous sequence that is expressed upon transduction of the chimeric MLV into the non-dividing myeloid cell.
5. The method of claim 1 wherein the lentiviral Vpx protein is a primate lentiviral Vpx protein.
6. The method of claim 5 wherein the primate lentiviral Vpx protein is a Simian Immunodeficiency Virus (SIV) Vpx protein.
7. The method of claim 6 wherein the SIV Vpx protein is fused to a C terminus of a MLV gag protein.
8. The method of claim 7 further comprising all or a portion of a SIV gag protein fused to the C terminus of the MLV gag protein.
9. The method of claim 8 wherein the portion of the SIV gag protein is a p6 domain.
10. The method of claim 6 wherein the SIV Vpx has an amino acid sequence comprising:
MSXPRERIPP GNSGEETXGE AFDWLHRTXE EINRAAVNHL PRELIFQVWR RXWEYWHDEM GMSVSYTKXR YLCLIQKALF MHCKKGCRCL GGEHGAGGWR PGPPPPPPPG LA (SEQ ID NO: 1);
or
MSDPRERIPP GNSGEETIGE AFDWLHRTVE EINRAAVNHL PRELIFQVWR RSWEYWHDEM GMSVSYTKYR YLCLIQKAMF MHCKKGCRCL GGEHGAGGWR PGPPPPPPPG LA (SEQ ID NO: 2).
11. The method of claim 7 wherein the MLV gag protein has an amino acid sequence comprising
(SEQ ID NO: 3)
MFIPLIFLPQLLGNVLVCVLAHHFGKEFMGQTVTTPLSLTLGHWKDVE
RIAHNQSVDVKKRRWVTFCSAEWPTFNVGWPRDGTFNRDLITQVKIKV
FSPGPHGHPDQVPYIVTWEALAFDPPPWVKPFVHPKPPPPLPPSAPSL
PLEPPRSTPPRSSLYPALTPSLGAKPKPQVLSDSGGPLIDLLTEDPPP
YRDPRPPPSDRDGNGGEATPAGEAPDPSPMASRLRGRREPPVADSTTS
QAFPLRAGGNGQLQYWPFSSSDLYNWKNNNPSFSEDPGKLTALIESVL
ITHQPTWDDCQQLLGTLLTGEEKQRVLLEARKAVRGDDGRPTQLPNEV
DAAFPLERPDWDYTTQAGRNHLVHYRQLLLAGLQNAGRSPTNLAKVKG
ITQGPNESPSAFLERLKEAYRRYTPYDPEDPGQETNVSMSFIWQSAPD
IGRKLERLEDLKNKTLGDLVREAEKIFNKRETPEEREERIRRETEEKE
ERRRTEDEQKEKERDRRRHREMSKLLATVVSGQKQDRQGGERRRSQLD
RDQCAYCKEKGHWAKDCPKKPRGPRGPRPQPVATMASSEDVIKEFMRF
KVRMEGSVNGHEFEIEGEGEGRPYEGTQTAKLKVTKGGPLPFAWDILS
PQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGVVTVTQD
SSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALK
GEIKMRLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNE
DYTIVEQYERAEGRHSTGALYK.
12. The method of claim 1 wherein the non-dividing myeloid cell is a terminally differentiated cell.
13. The method of claim 12 wherein the terminally differentiated cell is selected from the group consisting of: a macrophage, a dendritic cell.
14. The method of claim 3 wherein the lentiviral Vpx protein is a primate lentiviral Vpx protein.
15. The method of claim 14 wherein the primate lentiviral Vpx protein is a Simian Immunodeficiency Virus (SIV) Vpx protein.
16. The method of claim 15 wherein the SIV Vpx protein is fused to a C terminus of a MLV gag protein.
17. The method of claim 16 further comprising all or a portion of a SIV gag protein fused to the C terminus of the MLV gag protein.
18. The method of claim 17 wherein the portion of the SIV gag protein is a p6 domain.
19. The method of claim 15 wherein the SIV Vpx has an amino acid sequence comprising:
MSXPRERIPP GNSGEETXGE AFDWLHRTXE EINRAAVNHL PRELIFQVWR RXWEYWHDEM GMSVSYTKXR YLCLIQKALF MHCKKGCRCL GGEHGAGGWR PGPPPPPPPG LA (SEQ ID NO: 1);
or
MSDPRERIPP GNSGEETIGE AFDWLHRTVE EINRAAVNHL PRELIFQVWR RSWEYWHDEM GMSVSYTKYR YLCLIQKAMF MHCKKGCRCL GGEHGAGGWR PGPPPPPPPG LA (SEQ ID NO: 2).
20. The method of claim 16 wherein the MLV gag protein has an amino acid sequence comprising
(SEQ ID NO: 3)
MFIPLIFLPQLLGNVLVCVLAHHFGKEFMGQTVTTPLSLTLGHWKDVE
RIAHNQSVDVKKRRWVTFCSAEWPTFNVGWPRDGTFNRDLITQVKIKV
FSPGPHGHPDQVPYIVTWEALAFDPPPWVKPFVHPKPPPPLPPSAPSL
PLEPPRSTPPRSSLYPALTPSLGAKPKPQVLSDSGGPLIDLLTEDPPP
YRDPRPPPSDRDGNGGEATPAGEAPDPSPMASRLRGRREPPVADSTTS
QAFPLRAGGNGQLQYWPFSSSDLYNWKNNNPSFSEDPGKLTALIESVL
ITHQPTWDDCQQLLGTLLTGEEKQRVLLEARKAVRGDDGRPTQLPNEV
DAAFPLERPDWDYTTQAGRNHLVHYRQLLLAGLQNAGRSPTNLAKVKG
ITQGPNESPSAFLERLKEAYRRYTPYDPEDPGQETNVSMSFIWQSAPD
IGRKLERLEDLKNKTLGDLVREAEKIFNKRETPEEREERIRRETEEKE
ERRRTEDEQKEKERDRRRHREMSKLLATVVSGQKQDRQGGERRRSQLD
RDQCAYCKEKGHWAKDCPKKPRGPRGPRPQPVATMASSEDVIKEFMRF
KVRMEGSVNGHEFEIEGEGEGRPYEGTQTAKLKVTKGGPLPFAWDILS
PQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGVVTVTQD
SSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALK
GEIKMRLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNE
DYTIVEQYERAEGRHSTGALYK.
21. The method of claim 3 wherein the non-dividing myeloid cell is a terminally differentiated cell.
22. The method of claim 21 wherein the terminally differentiated cell selected from the group consisting of: a macrophage, a dendritic cell.