HPV vaccines
Provided herein are genetically modified arenaviruses suitable as vaccines against neoplastic diseases or cancer. The invention also relates to pharmaceutical compositions and methods for the prevention or treatment of certain infections causing neoplastic diseases or cancer, such as infections with oncogenic viruses. Specifically, provided herein are pharmaceutical compositions, vaccines, and methods of preventing or treating diseases and conditions caused by and associated with infections with Human Papillomavirus (HPV), such as cervical cancer, anogenital cancer, head and neck cancer and skin cancers. Also provided herein are immunotherapies for the treatment of a neoplastic disease, such as a neoplastic disease caused by infection with oncogenic viruses.
1. A method of generating a tri-segmented arenavirus viral vector comprising one L segment and two S segments and a nucleotide sequence encoding a polypeptide comprising an amino acid sequence that is at least 99% identical to SEQ ID NO: 10, wherein the method comprises:
a. transfecting into a host cell one or more complementary deoxyribonucleic acid (cDNA) of the one L segment and two S segments;
b. maintaining the host cell under conditions suitable for virus formation; and
c. harvesting the tri-segmented arenavirus viral vector,
wherein one of the two S segments is selected from the group consisting of:
(i) an S segment wherein the ORF encoding the NP is under control of an arenavirus 5′ UTR,
(ii) an S segment wherein the ORF encoding the Z protein is under control of an arenavirus 5′ UTR,
(iii) an S segment wherein the ORF encoding the L protein is under control of an arenavirus 5′ UTR,
(iv) an S segment wherein the ORF encoding the GP is under control of an arenavirus 3′ UTR,
(v) an S segment wherein the ORF encoding the L protein is under control of an arenavirus 3′ UTR, and
(vi) an S segment wherein the ORF encoding the Z protein is under control of an arenavirus 3′ UTR.
2. The method of claim 1 , wherein the method further comprises transcribing the one or more cDNAs of the one L segment and two S segments using a promoter.
3. The method of claim 2 , wherein the promoter is a bidirectional promoter.
4. The method of claim 1 , wherein the method further comprises transfecting into the host cell one or more nucleic acids encoding an arenavirus polymerase.
5. The method of claim 4 , wherein the arenavirus polymerase is the L protein.
6. The method of claim 1 , wherein the method further comprises transfecting into the host cell one or more nucleic acids encoding the NP protein.
7. The method of claim 2 , wherein the promoter is selected from the group consisting of:
an RNA polymerase I promoter;
(ii) an RNA polymerase II promoter; and
(iii) a T7 promoter.
8. The method of claim 1 , wherein the tri-segmented arenavirus viral vector is derived from a lymphocytic choriomeningitis virus (LCMV) or a Pichinde virus.
9. The method of claim 8 , wherein the LCMV is MP strain, WE strain, Armstrong strain, or Armstrong Clone 13 strain.
10. The method of claim 1 , wherein the tri-segmented arenavirus viral vector comprises all four arenavirus ORFs, and wherein the tri-segmented arenavirus viral vector is infectious and replication competent.
11. The method of claim 1 , wherein the tri-segmented arenavirus viral vector is attenuated.
12. The method of claim 1 , wherein one of the two S segments comprises a nucleotide sequence encoding a polypeptide comprising an amino acid sequence that is at least 99% identical to SEQ ID NO: 10.
13. The method of claim 1 , wherein one of the two S segments comprises a nucleotide sequence encoding a polypeptide comprising an amino acid sequence that is 100% identical to SEQ ID NO: 10.
14. The method of claim 1 , wherein the two S segments each comprise a nucleotide sequence encoding a polypeptide comprising an amino acid sequence that is at least 99% identical to SEQ ID NO: 10.
15. The method of claim 1 , wherein the two S segments each comprise a nucleotide sequence encoding a polypeptide comprising an amino acid sequence that is 100% identical to SEQ ID NO: 10.
16. The method of claim 1 , wherein inter-segmental recombination of the two S segments, uniting two arenavirus ORFs on only one instead of two separate S segments, abrogates viral promoter activity.
17. A method of generating a tri-segmented arenavirus viral vector comprising one L segment and two S segments, wherein the method comprises:
a. transfecting into a host cell one or more complementary deoxyribonucleic acid (cDNA) of the one L segment and two S segments;
b. maintaining the host cell under conditions suitable for virus formation; and
c. harvesting the tri-segmented arenavirus viral vector,
wherein a first S segment of the two S segments is engineered to carry an ORF encoding GP in a position under control of an arenavirus 3′ UTR and an ORF encoding a polypeptide comprising an amino acid sequence that is at least 99% identical to SEQ ID NO: 10 in a position under control of an arenavirus 5′ UTR and a second S segment of the two S segments is engineered to carry an ORF encoding NP in a position under control of an arenavirus 3′ UTR and an ORF encoding a polypeptide comprising an amino acid sequence that is at least 99% identical to SEQ ID NO: 10 in a position under control of an arenavirus 5′ UTR, wherein the arenavirus 3′ UTR is a 3′ UTR of an arenavirus S segment and the arenavirus 5′ UTR is a 5′ UTR of an arenavirus S segment.
18. The method of claim 17 , wherein the tri-segmented arenavirus viral vector is attenuated.
19. The method of claim 17 , wherein the tri-segmented arenavirus viral vector is derived from a LCMV or a Pichinde virus.
20. The method of claim 17 , wherein the polypeptide encoded on the first S segment and the polypeptide encoded on the second S segment each comprise an amino acid sequence that is 100% identical to SEQ ID NO: 10.