Production of chimeric human papillomavirus
View Patent ↗This invention relates to infectious chimeric papillomaviruses, and especially those where the early genes are from human papillomavirus (HPV) 18, and the late genes are from another HPV. Also presented are methods of culturing the virus in raft cell cultures, and to assays utilizing these chimeric viruses.
1. An infectious chimeric papillomavirus comprising early genes and late genes, wherein the early genes are from HPV18 and the late genes are from a second papillomavirus source.
2. An infectious chimeric papillomavirus according to claim 1 wherein the late genes are from a source selected from the group consisting of: HPV 6a, HPV 6b, HPV 11, HPV 16, HPV 18, HPV 31, HPV 33, HPV 35, HPV 39, HPV 41, HPV 42, HPV 47, HPV 51, HPV 57, HPV 58, bovine papillomavirus, cottontail rabbit papillomavirus, and canine papillomavirus.
3. A vector encoding the chimeric infectious papillomavirus of claim 1 .
4. A host cell comprising the chimeric infectious papillomavirus of claim 1 .
5. A method of obtaining an infectious chimeric papillomavirus comprising:
(a) preparing a chimeric papillomavirus which comprises early genes and late genes, wherein the early genes are from HPV18 and the late genes are from a second papillomavirus source;
(b) preparing a raft cell culture of differentiating epithelial cells;
(c) infecting the cell culture with the chimeric papillomavirus;
(d) incubating the cell culture under conditions which support viral replication; and
(e) harvesting the resultant chimeric papillomavirus.
6. A method according to claim 5 wherein the late genes are from a source selected from the group consisting of: HP 6a, HPV 6b, HPV 11, HPV 16, HPV 18, bovine papilloma virus, cottontail rabbit papillomavirus, and canine papillomavirus.
7. An assay to detect the presence of a human papillomavirus neutralizing antibody in a serum comprising:
(a) contacting a cell culture comprising an infectious chimeric papillomavirus with the serum, wherein the infectious chimeric papillomavirus comprises early genes and late genes, wherein the early genes are from HPV18 and late genes are from a second papillomavirus source; and
(b) determining if chimeric papillomavirus replication occurs within the cell culture in the presence of the serum.
8. An assay according to claim 7 wherein step (b) comprises contacting nucleic acids from the chimeric virus with a reverse transcriptase, and amplifying the resultant cDNA.
9. An assay according to claim 8 further comprising comparing the amount of cDNA obtained to that from a control cell.