MODIFIED MATRIX PROTEINS OF VESICULAR STOMATITIS VIRUS
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IPC분류정보
국가/구분
United States(US) Patent
공개
국제특허분류(IPC7판)
A61K-039/39
C12N-007/00
C07K-014/005
출원번호
US-0901339
(2014-06-26)
공개번호
US-0144022
(2016-05-26)
국제출원번호
PCT/CA2014/050614
(2014-06-26)
발명자
/ 주소
KANG, Chil-Yong
KIM, Gyoung Nyoun
출원인 / 주소
KANG, Chil-Yong
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. A nucleotide sequence that encodes for 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 a nucleotide sequence that encodes for a modified matrix (M) protein, the nucleotide sequence comprising a sequence selected from SEQ ID NO:2, SEQ ID NO:6 and SEQ ID NO:7. 8. A recombinant vesicular stomatitis virus (rVSV), wherein said rVSV comprises a modified matrix (M) protein, the modified M protein comprising 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. 9. The rVSV of claim 7, 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 7, 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 7, 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 substitution: G21E/L111A/M51R. 12. The rVSV of claim 10, 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 a nucleotide sequence of SEQ ID NO: 2. 14. The rVSV according to any one of claims 10-12, wherein the rVSVInd is capable of producing VSVInd particles at permissible temperatures and incapable of producing the particles at non-permissible temperatures. 15. The rVSV of claim 7, 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 14, wherein the modified M protein is encoded by a gene comprising a nucleotide sequence of SEQ ID NO: 7 17. The rVSV of claim 14, wherein the modified M protein comprises the amino acid sequence SEQ ID NO: 10. 18. The rVSV of claims 7-17, wherein the rVSV is a chimeric rVSV that expresses a protein of a foreign pathogen. 19. The rVSV of claim 17, wherein said pathogen is a viral, fungal, bacterial or parasitic pathogen. 20. The rVSV of claims 7-19, wherein the rVSV is essentially non-cytolytic and avirulent. 21. A vaccine, the vaccine including a modified matrix (M) protein of a vesicular stomatitis virus (VSV) wherein the modified M protein is encoded by a nucleotide sequence comprising a sequence selected from: SEQ ID NO:2, SEQ ID NO:6 and SEQ ID NO:7. 22. A vaccine, wherein the vaccine comprises an effective amount of one or more attenuated recombinant vesicular stomatitis virus (rVSV), the one or more attenuated rVSVs including a modified matrix (M) protein, the modified M protein comprising 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. 23. The vaccine of claim 22, wherein the 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 substitution: G21E/L111A/M51R. 24. The vaccine of claim 23, wherein the modified M protein comprises the amino acid sequence of SEQ ID NO: 4. 25. The vaccine of claim 23, wherein the modified M protein is encoded by a gene comprising a nucleotide sequence of SEQ ID NO: 2. 26. The vaccine of any one of claims 23 to 25, wherein the rVSVInd is capable of producing VSVInd particles at permissible temperatures and incapable of producing the particles at non-permissible temperatures. 27. The vaccine of claims 23-26, wherein the rVSVInd is a full length rVSVInd. 28. The vaccine of claim 22, 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. 29. The vaccine of claim 28, wherein the modified M protein comprises the amino acid sequence SEQ ID NO: 10. 30. The vaccine of claim 28, wherein the modified M protein is encoded by a gene comprising a nucleotide sequence of SEQ ID NO: 7. 31. The vaccine of claims 28-30, wherein the rVSVNJ is a full length rVSVNJ. 32. The vaccine of any one of claims 21 to 31, wherein the rVSV is a chimeric rVSV that expresses a protein of a foreign pathogen, and wherein said chimeric rVSV is capable of inducing an immune response to said protein. 33. The vaccine of claim 32, wherein the vaccine comprises a mixture of attenuated chimeric rVSVs, wherein at least two chimeric rVSVs in the mixture express a different protein of the foreign pathogen. 34. The vaccine of claim 32, wherein the pathogen is a viral, a fungal, a bacterial or a parasitic pathogen. 35. The vaccine of claim 32, wherein the pathogen is a lentivirus. 36. The vaccine according to claim 35, wherein the lentivirus is a HIV and the protein of the foreign pathogen is a HIV protein. 37. The vaccine of claim 32, wherein the pathogen is HCV and the protein of the foreign pathogen is a HCV protein. 38. The vaccine of claim 22, 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. 39. The vaccine of claim 22, wherein the A at position 111 in (i) and the A at position 110 in (ii) are encoded by a gca codon. 40. The vaccine of any one of claims 21 to 39, wherein said vaccine is capable of inducing a humoral, cellular and mucosal immune response. 41. The vaccine of any one of claims 21 to 40, wherein said vaccine further includes an adjuvant. 42. 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. 43. The prime boost combination vaccine of claim 42, wherein SEQ ID NO:4 is encoded by a gene comprising SEQ ID NO:2. 44. The prime boost combination vaccine of claim 42, 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. 45. The prime boost combination vaccine of claim 42, wherein (a) is a priming vaccine and (b) is a booster vaccine. 46. The prime boost combination vaccine of claim 42, wherein (b) is a priming vaccine and (a) is a booster vaccine. 47. The prime boost combination vaccine of claim 42, 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. 48. The prime boost combination vaccine of claim 47, wherein the pathogen is a viral, a fungal, a bacterial or a parasitic pathogen. 49. The prime boost combination vaccine of claim 47, wherein the pathogen is a lentivirus. 50. The prime boost combination vaccine of claim 49, wherein the lentivirus is a HIV and the protein is a HIV protein. 51. The prime boost combination vaccine of claim 50, 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. 52. The prime boost combination vaccine of claim 51, wherein the HIV gene is selected from the group of HIV genes consisting of env, gag and pol. 53. The prime boost combination vaccine of claim 49, wherein the pathogen is HCV and the epitope is a HCV protein. 54. The prime boost combination vaccine of claim 53, 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. 55. The prime boost combination vaccine of claim 54, wherein the HCV protein is a structural or a non-structural HCV protein. 56. The prime boost combination vaccine of any one of claims 47 to 55, wherein each one of the two vaccines comprise 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. 57. The prime boost combination vaccine of any one of claims 42 to 54, wherein each one of the two vaccines is capable of inducing humoral, cellular and mucosal immune responses. 58. The prime boost combination vaccine of any one of claims 42 to 57, wherein the serotype of vaccine (a) is Indiana and the serotype of vaccine (b) is New Jersey. 59. The prime boost combination vaccine of any one of claims 42 to 58, wherein each one of vaccine (a) and vaccine (b) further comprises an adjuvant. 60. 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. 61. The kit of claim 60, wherein (a) and (b) are formulated in a pharmaceutically acceptable carrier. 62. The kit of claim 60, wherein SEQ ID NO:4 is encoded by a gene comprising SEQ ID NO:2. 63. The kit of claim 60, 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. 64. An isolated peptide comprising an amino acid sequence selected from the group of amino acid sequences listed as SEQ ID NOs: 4 and 10. 65. An isolated nucleotide sequences comprising a nucleotide sequence selected from the group SEQ ID NO:2, SEQ ID NO:6 and SEQ ID NO:7. 66. A use of the vaccine of claim 21 or 22 to induce an immune response in a subject. 67. A use of the prime boost combination vaccine of claim 42 to induce an immune response in a subject. 68. A method of inducing an immune response in a subject, wherein the method comprises administering to the subject: (a) an effective amount of a vaccine comprising an attenuated recombinant vesicular stomatitis virus (rVSV) of one serotype having a first modified M protein, the first modified M protein comprising the amino acid sequence of SEQ ID NO: 3 including at least the following substitutions: G21E/L111A/M51R; and (b) an effective amount of another vaccine comprising an attenuated rVSV of another serotype having a second modified M protein, the second modified M protein comprising the amino acid sequence of SEQ ID NO: 8 including at least the following substitutions: G22E/L110A/M48R/M51R. 69. The method of claim 68, wherein (a) is administered to the subject before (b) is administered to the subject. 70. The method of claim 69, wherein (b) is administered to the subject more than one time over the course of inducing. 71. The method of claim 68, wherein (a) is administered to the subject and (b) is administered to the subject at about weeks three, eight and sixteen post-administration of (a). 72. The method of claim 68, wherein (b) is administered to the subject before (a) is administered to the subject. 73. The method of claim 68, wherein (a) is administered to the subject more than one time over the course of inducing. 74. The method of claim 72, wherein (b) is administered to the subject and (a) is administered to the subject at about weeks three, eight and sixteen post-administration of (b). 75. The method of claim 68, wherein the two rVSVs are chimeric rVSVs that express a protein of a foreign pathogen, and wherein the two rVSVs are capable of inducing an immune response to the protein. 76. The method of claim 75, wherein the pathogen is a viral, a fungal, a bacterial or a parasitic pathogen. 77. The method of claim 75, wherein the pathogen is a lentivirus. 78. The method according to claim 77, wherein the lentivirus is a human immunodeficiency virus (HIV) and the protein is a HIV protein. 79. The method of claim 78, 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 a HIV gene inserted in between the G gene and the L gene. 80. The method of claim 79, wherein the HIV gene is selected from the group of HIV genes consisting of gag, env and pol. 81. The method of claim 75, wherein the pathogen is hepatitis C virus (HCV) and the protein is a HCV protein. 82. The method of claim 81, 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 of the rVSV. 83. The method of claim 75, wherein the two vaccines comprise a mixture of attenuated chimeric rVSVs, wherein at least two of the attenuated chimeric rVSVs in the mixture express a different protein of the pathogen. 84. The method of any one of claims 68 to 83, wherein each one of the two vaccines (a) and (b) induces humoral, cellular and mucosal immune responses. 85. The method of any one of claims 68 to 84, wherein the one serotype of vaccine (a) is Indiana and the other serotype of vaccine (b) is New Jersey. 86. The method of any one of claims 68 to 85, wherein each of vaccine (a) and vaccine (b) further comprises an adjuvant.
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