CMV glycoproteins and recombinant vectors
This invention also relates to recombinant vectors expressing one or more of the human CMV (HCMV) glycoproteins US2, US3, US6 and US11 or corresponding functional rhesus CMV (RhCMV) homologues Rh182, Rh184, Rh185 or Rh189, methods of making them, uses for them, expression products from them, and uses for the expression products. This invention also relates to recombinant cytomegalovirus vectors vectors lacking one or more of the glycoproteins, methods of making them, uses for them, expression products from them, and uses for the expression products.
1. A method of determining efficacy of a cytomegalovirus (CMV) immunogenic composition comprising or expressing at least one CMV immunogen, wherein the method comprises the steps of:
(a) detecting the presence of a pre-challenge CD8+ T cell response of a test subject to the at least one CMV immunogen;
(b) challenging the test subject with a recombinant CMV challenge virus expressing the at least one CMV immunogen, wherein the glycoproteins US2, US3, US6 and US11 or orthologs thereof are deleted from the challenge virus, and wherein the challenge virus is capable of eliciting a CD8+ T cell response to the at least one CMV immunogen within a CMV seronegative host; and
(c) detecting the presence or absence of a post-challenge CD8+ T cell response of the test subject to the at least one CMV immunogen;
wherein the CMV immunogenic composition is efficacious in eliciting a protective immune response against CMV if a post-challenge boost to the CD8+ T cell response of the test subject to the at least one CMV immunogen is absent.
2. The method of claim 1 , wherein:
(i) the CMV seronegative test subject is a human CMV (HCMV) seronegative test subject,
(ii) the at least one CMV immunogen is a HCMV immunogen, and
(iii) the recombinant CMV challenge virus is a recombinant HCMV challenge virus.
3. The method of claim 1 , wherein:
(i) the CMV seronegative test subject is a rhesus CMV (RhCMV) seronegative test subject,
(ii) the at least one CMV immunogen is a RhCMV immunogen, and
(iii) the recombinant CMV challenge virus is a recombinant RhCMV challenge virus.
4. The method of claim 1 , wherein the CMV immunogenic composition comprises an attenuated CMV strain.
5. The method of claim 4 , wherein the CMV immunogenic composition lacks the transactivator pp71.
6. The method of claim 1 , wherein the recombinant CMV challenge virus is a recombinant HCMV challenge virus, and wherein all glycoproteins within the US2 to US11 region are deleted from the recombinant HCMV challenge virus.
7. The method of claim 1 , wherein the recombinant CMV challenge virus is a recombinant RhCMV challenge virus, and wherein all glycoproteins within the Rh182-189 region are deleted from the recombinant RhCMV challenge virus.
8. A method of determining efficacy of a CMV immunogenic composition comprising or expressing at least one CMV immunogen, wherein the method comprises the steps of:
(a) detecting the presence of a pre-challenge CD8+ T cell response of a test subject to the at least one CMV immunogen and the absence of a pre-challenge CD8+ T cell response of the test subject to at least one additional immunogen expressed by a recombinant CMV challenge virus, wherein the CMV immunogenic composition does not comprise or express the at least one additional immunogen;
(b) challenging the test subject with the recombinant CMV challenge virus expressing the at least one CMV immunogen and the at least one additional immunogen, wherein the glycoproteins US2, US3, US6 and US11 or orthologs thereof are deleted from the challenge virus, and wherein the recombinant CMV challenge virus is capable of eliciting a CD8+ T cell response to the at least one CMV immunogen and the at least one additional immunogen within a CMV seronegative host; and
(c) detecting the presence or absence of a post-challenge CD8+ T cell response of the test subject to the at least one additional immunogen;
wherein the CMV immunogenic composition is efficacious in eliciting a protective immune response against CMV if a de novo post-challenge CD8+ T cell response of the test subject to the at least one additional immunogen is absent.
9. The method of claim 8 , wherein:
(i) the CMV seronegative test subject is a HCMV seronegative test subject,
(ii) the at least one CMV immunogen is a HCMV immunogen, and
(iii) the recombinant CMV challenge virus is a recombinant HCMV challenge virus.
10. The method of claim 8 , wherein:
(i) the CMV seronegative test subject is a RhCMV seronegative test subject,
(ii) the at least one CMV immunogen is a RhCMV immunogen, and
(iii) the recombinant CMV challenge virus is a recombinant RhCMV challenge virus.
11. The method of claim 8 , wherein the CMV immunogenic composition comprises an attenuated CMV strain.
12. The method of claim 11 , wherein the CMV immunogenic composition lacks the transactivator pp71.
13. The method of claim 8 , wherein the recombinant CMV challenge virus is a recombinant HCMV challenge virus, and wherein all glycoproteins within the US2 to US11 region are deleted from the recombinant HCMV challenge virus.
14. The method of claim 8 , wherein the recombinant CMV challenge virus is a recombinant RhCMV challenge virus, and wherein all glycoproteins within the Rh182-189 region are deleted from the recombinant RhCMV challenge virus.
15. The method of claim 8 , wherein the at least one additional immunogen is a bacterial-derived immunogen, a parasite-derived immunogen, viral-derived immunogen, or a cancer-derived immunogen.
16. A method of determining efficacy of a CMV immunogenic composition comprising or expressing at least one CMV immunogen and at least one additional immunogen, wherein the method comprises the steps of:
(a) detecting the presence of a pre-challenge CD8+ T cell response of a test subject to the at least one additional immunogen;
(b) challenging the test subject with a recombinant CMV challenge virus expressing the at least one CMV immunogen and the at least one additional immunogen, wherein the glycoproteins US2, US3, US6 and US11 or orthologs thereof are deleted from the challenge virus, and wherein the recombinant CMV challenge virus is capable of eliciting a CD8+ T cell response to the at least one CMV immunogen and the at least one additional immunogen within a CMV seronegative host; and
(c) detecting the presence or absence of a post-challenge CD8+ T cell response of the test subject to the at least one additional immunogen;
wherein the CMV immunogenic composition is efficacious in eliciting a protective immune response against CMV if a boost to the CD8+ T cell response of the test subject to the at least one additional immunogen is absent.
17. The method of claim 16 , wherein:
(i) the CMV seronegative test subject is a HCMV seronegative test subject,
(ii) the at least one CMV immunogen is a HCMV immunogen, and
(iii) the recombinant CMV challenge virus is a recombinant HCMV challenge virus.
18. The method of claim 16 , wherein:
(i) the CMV seronegative test subject is a RhCMV seronegative test subject,
(ii) the at least one CMV immunogen is a RhCMV immunogen, and
(iii) the recombinant CMV challenge virus is a recombinant RhCMV challenge virus.
19. The method of claim 16 , wherein the CMV immunogenic composition comprises an attenuated CMV strain.
20. The method of claim 19 , wherein the CMV immunogenic composition lacks the transactivator pp71.
21. The method of claim 16 , wherein the recombinant CMV challenge virus is a recombinant HCMV challenge virus, and wherein all glycoproteins within the US2 to US11 region are deleted from the recombinant HCMV challenge virus.
22. The method of claim 16 , wherein the recombinant CMV challenge virus is a recombinant RhCMV challenge virus, and wherein all glycoproteins within the Rh182-189 region are deleted from the recombinant RhCMV challenge virus.
23. The method of claim 16 , wherein the at least one additional immunogen is a bacterial-derived immunogen, a parasite-derived immunogen, viral-derived immunogen, or a cancer-derived immunogen.
24. A method of determining efficacy of a CMV immunogenic composition comprising or expressing at least one CMV immunogen, wherein the method comprises the steps of:
(a) administering the immunogenic composition to a CMV seronegative test subject; and
(b) challenging the test subject with a recombinant CMV challenge virus if a pre-challenge CD8+ T cell response to the at least one CMV immunogen is detected within the test subject, wherein the glycoproteins US2, US3, US6 and US11 or orthologs thereof are deleted from the challenge virus, and wherein the challenge virus expresses the at least one CMV immunogen and is capable of eliciting a CD8+ T cell response to the at least one CMV immunogen within a CMV seronegative host;
wherein the CMV immunogenic composition is efficacious in eliciting a protective immune response against CMV if a post-challenge boost to the CD8+ T cell response of the test subject to the at least one CMV immunogen is absent.
25. The method of claim 24 , wherein:
(i) the CMV seronegative test subject is a HCMV seronegative test subject,
(ii) the at least one CMV immunogen is a HCMV immunogen, and
(iii) the recombinant CMV challenge virus is a recombinant HCMV challenge virus.
26. The method of claim 24 , wherein:
(i) the CMV seronegative test subject is a RhCMV seronegative test subject,
(ii) the at least one CMV immunogen is a RhCMV immunogen, and
(iii) the recombinant CMV challenge virus is a recombinant RhCMV challenge virus.
27. The method of claim 24 , wherein the CMV immunogenic composition comprises an attenuated CMV strain.
28. The method of claim 27 , wherein the CMV immunogenic composition lacks the transactivator pp71.
29. The method of claim 24 , wherein the recombinant CMV challenge virus is a recombinant HCMV challenge virus, and wherein all glycoproteins within the US2 to US11 region are deleted from the recombinant HCMV challenge virus.
30. The method of claim 24 , wherein the recombinant CMV challenge virus is a recombinant RhCMV challenge virus, and wherein all glycoproteins within the Rh182-189 region are deleted from the recombinant RhCMV challenge virus.
31. A method of determining efficacy of a CMV immunogenic composition comprising or expressing at least one CMV immunogen, wherein the method comprises the steps of:
(a) administering the CMV immunogenic composition to a HCMV seronegative test subject, wherein the CMV immunogenic composition does not comprise or express at least one additional immunogen expressed by a recombinant CMV challenge virus; and
(b) challenging the test subject with the recombinant CMV challenge virus if a pre-challenge CD8+ T cell response to the at least one CMV immunogen is detected within the test subject, wherein the glycoproteins US2, US3, US6 and US11 or orthologs thereof are deleted from the challenge virus, and wherein the recombinant CMV challenge virus is capable of eliciting a CD8+ T cell response to the at least one CMV immunogen and the at least one additional immunogen within a CMV seronegative host;
wherein the CMV immunogenic composition is efficacious in eliciting a protective immune response against CMV if a de novo post-challenge CD8+ T cell response of the test subject to the at least one additional immunogen is absent.
32. The method of claim 31 , wherein:
(i) the CMV seronegative test subject is a HCMV seronegative test subject,
(ii) the at least one CMV immunogen is a HCMV immunogen, and
(iii) the recombinant CMV challenge virus is a recombinant HCMV challenge virus.
33. The method of claim 31 , wherein:
(i) the CMV seronegative test subject is a RhCMV seronegative test subject,
(ii) the at least one CMV immunogen is a RhCMV immunogen, and
(iii) the recombinant CMV challenge virus is a recombinant RhCMV challenge virus.
34. The method of claim 31 , wherein the CMV immunogenic composition comprises an attenuated CMV strain.
35. The method of claim 34 , wherein the CMV immunogenic composition lacks the transactivator pp71.
36. The method of claim 31 , wherein the recombinant CMV challenge virus is a recombinant HCMV challenge virus, and wherein all glycoproteins within the US2 to US11 region are deleted from the recombinant HCMV challenge virus.
37. The method of claim 31 , wherein the recombinant CMV challenge virus is a recombinant RhCMV challenge virus, and wherein all glycoproteins within the Rh182-189 region are deleted from the recombinant RhCMV challenge virus.
38. The method of claim 31 , wherein the at least one additional immunogen is a bacterial-derived immunogen, a parasite-derived immunogen, viral-derived immunogen, or a cancer-derived immunogen.
39. A method of determining efficacy of a CMV immunogenic composition comprising or expressing at least one CMV immunogen and at least one additional immunogen, wherein the method comprises the steps of:
(a) administering the CMV immunogenic composition to a CMV seropositive test subject; and
(b) challenging the test subject with a recombinant CMV challenge virus if pre-challenge CD8+ T cell responses to the at least one CMV immunogen and the at least one additional immunogen are detected within the test subject, wherein the glycoproteins US2, US3, US6 and US11 or orthologs thereof are deleted from the challenge virus, and wherein the recombinant CMV challenge virus expresses the at least one CMV immunogen and the at least one additional immunogen and is capable of eliciting CD8+ T cell responses to the at least one CMV immunogen and the at least one additional immunogen within a CMV seronegative host;
wherein the CMV immunogenic composition is efficacious eliciting a protective immune response against CMV if a boost to the CD8+ T cell response of the test subject to the at least one additional immunogen is absent.
40. The method of claim 39 , wherein:
(i) the CMV seronegative test subject is a HCMV seronegative test subject,
(ii) the at least one CMV immunogen is a HCMV immunogen, and
(iii) the recombinant CMV challenge virus is a recombinant HCMV challenge virus.
41. The method of claim 39 , wherein:
(i) the CMV seronegative test subject is a RhCMV seronegative test subject,
(ii) the at least one CMV immunogen is a RhCMV immunogen, and
(iii) the recombinant CMV challenge virus is a recombinant RhCMV challenge virus.
42. The method of claim 39 , wherein the CMV immunogenic composition comprises an attenuated CMV strain.
43. The method of claim 42 , wherein the CMV immunogenic composition lacks the transactivator pp71.
44. The method of claim 39 , wherein the recombinant CMV challenge virus is a recombinant HCMV challenge virus, and wherein all glycoproteins within the US2 to US11 region are deleted from the recombinant HCMV challenge virus.
45. The method of claim 39 , wherein the recombinant CMV challenge virus is a recombinant RhCMV challenge virus, and wherein all glycoproteins within the Rh182-189 region are deleted from the recombinant RhCMV challenge virus.
46. The method of claim 39 , wherein the at least one additional immunogen is a bacterial-derived immunogen, a parasite-derived immunogen, viral-derived immunogen, or a cancer-derived immunogen.