Modified matrix proteins of vesicular stomatitis virus
원문보기
IPC분류정보
국가/구분
United States(US) Patent
등록
국제특허분류(IPC7판)
A61K-039/39
A61K-039/12
C07K-014/005
C12N-015/86
C12N-007/00
A61K-039/00
출원번호
US-0901339
(2014-06-26)
등록번호
US-9943593
(2018-04-17)
국제출원번호
PCT/CA2014/050614
(2014-06-26)
국제공개번호
WO2014/205579
(2014-12-31)
발명자
/ 주소
Kang, Chil-Yong
Kim, Gyoung Nyoun
출원인 / 주소
Kang, Chil-Yong
대리인 / 주소
Krupnik, Eduardo
인용정보
피인용 횟수 :
0인용 특허 :
0
초록▼
The present invention relates to vesicular stomatitis virus (VSV) matrix (M) protein mutants. One mutant M protein includes a glycine changed to a glutamic acid at position (21), a leucine changed to alanine at position (111) and a methionine changed to an arginine at position (51). Another M protei
The present invention relates to vesicular stomatitis virus (VSV) matrix (M) protein mutants. One mutant M protein includes a glycine changed to a glutamic acid at position (21), a leucine changed to alanine at position (111) and a methionine changed to an arginine at position (51). Another M protein mutant includes a glycine changed to a glutamic acid at position (22) and a methionine changed to an arginine at positions (48) and (51). These new rVSVs having the mutant M are significantly attenuated and lost virulence, including neurovirulence, and are capable of inducing an immune responses against an antigen of interest. In addition, a rVSV serotype Indiana having the first described M mutant is capable of efficient replication at 31° C., and of poor replication or incapable of replication at about 37° C. or higher.
대표청구항▼
1. A modified matrix (M) protein of a vesicular stomatitis virus (VSV), wherein the modified M protein comprises an amino acid sequence selected from: (i) SEQ ID NO: 3 including at least the following substitutions: G21E/L111A/M51R; and (ii) SEQ ID NO: 8 including at least the following substitution
1. A modified matrix (M) protein of a vesicular stomatitis virus (VSV), wherein the modified M protein comprises an amino acid sequence selected from: (i) SEQ ID NO: 3 including at least the following substitutions: G21E/L111A/M51R; and (ii) SEQ ID NO: 8 including at least the following substitutions: G22E/L110A/M48R/M51R. 2. The modified M protein of claim 1, wherein the modified M protein of (i) comprises an amino acid sequence SEQ ID NO:4 and the modified M protein of (ii) comprises the amino acid sequence SEQ ID NO:10. 3. The modified M protein of claim 1, wherein the E at position 21 in (i) and the E at position 22 in (ii) are encoded by a gaa codon, and wherein the R at position 51 in (i) and (ii) and the R at position 48 in (ii) is encoded by a cga codon. 4. The modified M protein of claim 1, wherein the A at position 111 in (i) and the A at position 110 in (ii) is encoded by a gca codon. 5. The modified M protein of claim 1, wherein the amino acid sequence of (i) is encoded by a gene comprising SEQ ID NO:2, and the amino acid sequence of (ii) is encoded by a gene comprising SEQ ID NO:7. 6. An isolated nucleotide sequence that encodes a modified matrix protein of a vesicular stomatitis virus, wherein the nucleotide sequence comprises a sequence selected from: SEQ ID NO:2, SEQ ID NO:6 and SEQ ID NO:7. 7. A recombinant vesicular stomatitis virus (rVSV), wherein said rVSV includes the nucleotide sequence of claim 6. 8. A recombinant vesicular stomatitis virus (rVSV), wherein said rVSV comprises the modified matrix (M) protein of claim 1. 9. The rVSV of claim 8, wherein the E at position 21 in (i) and the E at position 22 in (ii) are encoded by a gaa codon, and wherein the R at position 51 in (i) and (ii) and the R at position 48 in (ii) is encoded by a cga codon. 10. The rVSV of claim 8, wherein the A at position 111 in (i) and the A at position 110 in (ii) are encoded by a gca codon. 11. The rVSV of claim 8, wherein said rVSV is a recombinant vesicular stomatitis virus Indiana serotype (rVSVInd), and wherein the modified M protein comprises the amino acid sequence of SEQ ID NO: 3 including at least the following substitutions: G21E/L111A/M51R. 12. The rVSV of claim 11, wherein the modified M protein comprises the amino acid sequence of SEQ ID NO: 4. 13. The rVSV of claim 10, wherein the modified M protein is encoded by a gene comprising the nucleotide sequence of SEQ ID NO: 2. 14. The rVSV according to claim 11, wherein the rVSVInd is capable of producing VSVInd particles at permissive temperatures and incapable of producing the particles at non-permissive temperatures. 15. The rVSV of claim 8, wherein said rVSV is a recombinant vesicular stomatitis virus New Jersey serotype (rVSVNJ), and wherein the modified M protein comprises the amino acid sequence of SEQ ID NO: 8 including at least the following substitutions: G22E/L110A/M48R/M51R. 16. The rVSV of claim 15, wherein the modified M protein is encoded by a gene comprising the nucleotide sequence of SEQ ID NO: 7. 17. The rVSV of claim 15, wherein the modified M protein comprises the amino acid sequence of SEQ ID NO: 10. 18. The rVSV of claim 8, wherein the rVSV is a chimeric rVSV that expresses a protein of a foreign pathogen. 19. The rVSV of claim 18, wherein said pathogen is a viral, fungal, bacterial or parasitic pathogen. 20. The rVSV of claim 8, wherein the rVSV is non-cytolytic and avirulent. 21. A vaccine, the vaccine including the modified matrix (M) protein of claim 5. 22. A vaccine, wherein the vaccine comprises an effective amount of an attenuated form of the recombinant vesicular stomatitis virus (rVSV) of claim 8. 23. The vaccine of claim 22, wherein said vaccine is capable of inducing a humoral, cellular and mucosal immune response. 24. The vaccine of claim 22, wherein said vaccine further includes an adjuvant. 25. A prime boost combination vaccine, wherein the prime boost combination vaccine comprises: (a) an effective amount of a vaccine comprising an attenuated recombinant vesicular stomatitis virus (rVSV) of one serotype having a first modified matrix (M) protein comprising the amino acid sequence of SEQ ID NO:4; and (b) an effective amount of a vaccine comprising a rVSV of another serotype having a second modified M protein comprising the amino acid sequence of SEQ ID NO:9 or SEQ ID NO:10. 26. The prime boost combination vaccine of claim 25, wherein SEQ ID NO:4 is encoded by a gene comprising SEQ ID NO:2. 27. The prime boost combination vaccine of claim 25, wherein SEQ ID NO:9 is encoded by a gene comprising SEQ ID NO:6, and SEQ ID NO:10 is encoded by a gene comprising SEQ ID NO:7. 28. The prime boost combination vaccine of claim 25, wherein (a) is a priming vaccine and (b) is a booster vaccine. 29. The prime boost combination vaccine of claim 25, wherein (b) is a priming vaccine and (a) is a booster vaccine. 30. The prime boost combination vaccine of claim 25, wherein the two attenuated rVSVs are chimeric rVSVs that express a protein of a foreign pathogen, and wherein the two chimeric rVSVs are capable of inducing an immune response to the protein. 31. The prime boost combination vaccine of claim 30, wherein the pathogen is a viral, a fungal, a bacterial or a parasitic pathogen. 32. The prime boost combination vaccine of claim 30, wherein the pathogen is a lentivirus. 33. The prime boost combination vaccine of claim 32, wherein the lentivirus is a HIV and the protein is a HIV protein. 34. The prime boost combination vaccine of claim 33, wherein the rVSV of one serotype and the rVSV of the other serotype include a surface glycoprotein (G) gene and a RNA dependent RNA polymerase (L) gene, and wherein a gene for expressing the HIV protein is inserted in between the G gene and the L gene. 35. The prime boost combination vaccine of claim 34, wherein the HIV gene is selected from the group of HIV genes consisting of env, gag and pol. 36. The prime boost combination vaccine of claim 30, wherein the pathogen is HCV and the epitope is a HCV protein. 37. The prime boost combination vaccine of claim 36, wherein the rVSV of one serotype and the rVSV of the other serotype include a surface glycoprotein (G) gene and a RNA dependent RNA polymerase (L) gene, and wherein a gene for expressing the HCV protein is inserted in between the G gene and the L gene. 38. The prime boost combination vaccine of claim 37, wherein the HCV protein is a structural or a non-structural HCV protein. 39. The prime boost combination vaccine of claim 30, wherein each one of the two vaccines comprises a mixture of the attenuated chimeric rVSVs, and wherein at least two of the attenuated chimeric rVSVs in the mixture have a different protein of the pathogen. 40. The prime boost combination vaccine of claim 25, wherein each one of the two vaccines is capable of inducing humoral, cellular and mucosal immune responses. 41. The prime boost combination vaccine of claim 25, wherein the serotype of vaccine (a) is Indiana and the serotype of vaccine (b) is New Jersey. 42. The prime boost combination vaccine of claim 25, wherein each one of vaccine (a) and vaccine (b) further comprises an adjuvant. 43. A kit comprising: (a) at least one dose of an effective amount of a vaccine comprising a recombinant vesicular stomatitis virus Indiana serotype (rVSVInd) having a modified matrix (M) protein comprising the amino acid sequence of SEQ ID NO:4, and (b) at least one dose of an effective amount of a vaccine comprising a recombinant vesicular stomatitis virus New Jersey serotype (rVSVNJ) having a modified M protein comprising the amino acid sequence of SEQ ID NO:9 or the amino acid sequence of SEQ ID NO:10. 44. The kit of claim 43, wherein (a) and (b) are formulated in a pharmaceutically acceptable carrier. 45. The kit of claim 43, wherein SEQ ID NO:4 is encoded by a gene comprising SEQ ID NO:2. 46. The kit of claim 43, wherein SEQ ID NO:9 is encoded by a gene comprising SEQ ID NO:6 and SEQ ID NO:10 is encoded by a gene comprising SEQ ID NO:7. 47. A method of inducing an immune response in a subject comprising administering to the subject an effective amount of the vaccine of claim 22. 48. A method of inducing an immune response in a subject comprising administering to the subject the prime boost combination vaccine of claim 25. 49. A method of inducing an immune response in a subject comprising administering to the subject an effective amount of the vaccine of claim 21. 50. The vaccine of claim 21, wherein said vaccine is capable of inducing a humoral, cellular and mucosal immune response, and wherein said vaccine further includes an adjuvant.
※ AI-Helper는 부적절한 답변을 할 수 있습니다.