Influenza antigens, vaccine compositions, and related methods

• US 20070275014A1
• Filed: 02/13/2007
• Published: 11/29/2007
• Est. Priority Date: 02/13/2006
• Status: Active Grant

• Current Assignee: iBio, Inc.
• Original Assignee: Fraunhofer USA Inc. (Fraunhofer-Institut für Nachrichtentechnik HHI)
• Inventors: Yusibov, Vidadi, Mett, Vadim, Musiychuck, Konstantin
• Granted Patent: US 8,124,103 B2
• Time in Patent Office: Days
• US Class Current: 424/209.100
• CPC Class Codes: A61K 2039/517 Plant cells
• CPC Class Codes: A61K 2039/542 oral/gastrointestinal
• CPC Class Codes: A61K 2039/543 intranasal
• CPC Class Codes: A61K 2039/55505 Inorganic adjuvants
• CPC Class Codes: A61K 2039/55566 Emulsions, e.g. Freund's adjuvant, MF59
• CPC Class Codes: A61K 2039/6031 Proteins
• CPC Class Codes: A61K 2039/70 Multivalent vaccine
• CPC Class Codes: A61K 39/00 Medicinal preparations containing antigens or antibodies materials for immunoassay G01N33/53
• CPC Class Codes: A61K 39/12 Viral antigens
• CPC Class Codes: A61K 39/145 Orthomyxoviridae, e.g. influenza virus
• CPC Class Codes: A61K 39/385 Haptens or antigens, bound to carriers
• CPC Class Codes: A61P 31/16 for influenza or rhinoviruses
• CPC Class Codes: C07K 14/005 from viruses
• CPC Class Codes: C07K 2319/00 Fusion polypeptide
• CPC Class Codes: C12N 15/62 DNA sequences coding for fusion proteins
• CPC Class Codes: C12N 15/8257 for the production of primary gene products, e.g. pharmaceutical products, interferon
• CPC Class Codes: C12N 15/8258 for the production of oral vaccines (antigens) or immunoglobulins
• CPC Class Codes: C12N 2760/16122 New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
• CPC Class Codes: C12N 2760/16134 Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein
• CPC Class Codes: C12N 9/2402 hydrolysing O- and S- glycosyl compounds (3.2.1)
• CPC Class Codes: C12N 9/2448 : Licheninase (3.2.1.73)
• CPC Class Codes: C12Y 302/01018 : Exo-alpha-sialidase (3.2.1.18), i.e. trans-sialidase
• CPC Class Codes: C12Y 302/01073 : Licheninase (3.2.1.73)

Abstract

The present invention relates to the intersection of the fields of immunology and protein engineering, and particularly to antigens and vaccines useful in prevention of infection by influenza virus. Provided are recombinant protein antigens, compositions, and methods for the production and use of such antigens and vaccine compositions.

Claims

1. An isolated antigen comprising a component of an influenza A integral membrane protein fused to a thermostable protein;

wherein the integral membrane protein component comprises at least one domain selected from the group consisting of hemagglutinin (HA) a domain of neuraminidase (NA) and a domain of M2.

2-11. -11. (canceled)

12. A vaccine composition comprising an antigen comprising a component of an influenza A integral membrane protein fused to a thermostable protein and a pharmaceutically acceptable carrier;

wherein the integral membrane protein component comprises at least one domain selected from the group consisting of a domain of hemagglutinin (HA) a domain of neuraminidase (NA) and a domain of M2; and

wherein the composition is capable of eliciting an immune response upon administration to a subject.

13-31. -31. (canceled)

32. A vaccine composition comprising at least two antigens, each of which comprises a component of an influenza A integral membrane protein, wherein the integral membrane protein component comprises at least one domain selected from the group consisting of a domain of hemagglutinin (HA) a domain of neuraminidase (NA) and a domain of M2;

wherein at least one antigen is fused to a thermostable protein and a pharmaceutically acceptable carrier;

wherein the composition is capable of eliciting an immune response upon administration to a subject.

33. The vaccine composition of claim 32, wherein at least one integral membrane protein component comprises of at least one domain hemagglutinin (HA), wherein the domain is selected from the group consisting of SEQ ID NO.:

1, SEQ ID NO.;

3, SEQ ID NO.;

7, SEQ ID NO.;

8, SEQ ID NO.;

33, SEQ ID NO.;

11, SEQ ID NO.;

12, SEQ ID NO.;

13, SEQ ID NO.;

20 and SEQ ID NO.;

24.

34. The vaccine composition of claim 32, wherein at least one integral membrane protein component comprises at least one domain of neuraminidase (NA) selected from the group consisting of SEQ ID NO.:

2, SEQ ID NO.;

4, SEQ ID NO.;

16, and SEQ ID NO.;

18.

35. The vaccine composition of claim 32, wherein at least one integral membrane protein component consists of full length hemagglutinin (HA) selected from SEQ ID NO.:

1 or SEQ ID NO.;

3.

36. The vaccine composition of claim 32, wherein at least one integral membrane protein component consists of full length neuraminidase (NA) selected from SEQ ID NO.:

2 or SEQ ID NO.;

4.

37. The vaccine composition of claim 32, wherein the thermostable protein comprises a modified lichenase protein sequence.

38. The vaccine composition of claim 32, wherein the thermostable protein comprises a modified lichenase protein sequence from Clostridium thermocellum.

39. The vaccine composition of claim 37, wherein the coding sequence for lichenase has been optimized for protein expression in plants.

40. The vaccine composition of claim 37, wherein the lichenase protein sequence comprises the N-terminal domain, the C-terminal domain, and the surface loop domain of lichenase.

41. The vaccine composition of claim 40, wherein the integral membrane protein component fused to lichenase is any one of an N-terminal fusion, a C-terminal fusion, or a surface loop insertion fusion protein.

42. The vaccine composition of claim 32, wherein the integral membrane protein component comprises at least two domains selected from the group consisting of SEQ ID NO.:

1, SEQ ID NO.;

3, SEQ ID NO.;

7, SEQ ID NO.;

8, SEQ ID NO.;

33, SEQ ID NO.;

11, SEQ ID NO.;

12, SEQ ID NO.;

13, SEQ ID NO.;

20, SEQ ID NO.;

24, SEQ ID NO.;

2, SEQ ID NO.;

4, SEQ ID NO.;

16, and SEQ ID NO.;

18 and wherein the composition is capable of eliciting an immune response upon administration to a subject.

43. The vaccine composition of claim 42, wherein the integral membrane protein component comprises a domain of hemagglutinin (HA) and a domain of neuraminidase (NA).

44. The vaccine composition of claim 42, wherein the integral membrane protein component comprises full length hemagglutinin (HA) and full length neuraminidase (NA).

45. The vaccine composition of claim 32, further comprising a third antigen, wherein the composition is capable of eliciting an immune response upon administration to an animal.

46. The vaccine composition of claim 45, wherein the third antigen comprises a component of influenza A integral membrane protein fused to a thermostable protein and a pharmaceutically acceptable carrier;

wherein the integral membrane protein component of the first antigen comprises at least one domain selected from the group consisting of a domain of hemagglutinin (HA) a domain of neuraminidase (NA) and a domain of M2, and the integral membrane protein component of the second antigen comprises at least one domain distinct from the first antigen selected from the group consisting of a domain of hemagglutinin (HA) a domain of neuraminidase (NA) and a domain of M2; and

wherein the composition is capable of eliciting an immune response upon administration to a subject.

47. The vaccine composition of claim 32 wherein the antigen is produced in a plant selected from a transgenic plant and a plant transiently expressing the antigen.

48. The vaccine composition of claim 32 wherein the composition comprises antigen which is purified, partially purified, or unpurified from plant cells, a plant, seeds, fruit, or an extract thereof.

49. The vaccine composition of claim 32, further comprising at least one vaccine adjuvant.

50. The vaccine composition of claim 49 wherein the adjuvant is selected from the group consisting of alum, MF59, saponin, and MALP2.

51. A method for inducing a protective immune response against influenza A infection in a subject comprising administering to a subject an effective amount of an anti-influenza A vaccine composition, wherein the administration is sufficient to stimulate production of antigen specific antibodies or stimulate a cellular immune response by the subject;

thereby inducing a protective immune response;

wherein the vaccine composition comprises an antigen comprising a component of an influenza A integral membrane protein fused to a thermostable protein; and

wherein the integral membrane protein component comprises at least one domain selected from the group consisting of a domain of hemagglutinin (HA) a domain of neuraminidase (NA) and a domain of M2.

52-55. -55. (canceled)

56. A method for producing an antigen protein comprising a component of an influenza A integral membrane protein fused to a thermostable protein, comprising:

a. preparing a nucleic acid construct encoding an antigen comprising a component of an influenza A integral membrane protein fused to a thermostable protein;

b. introducing the nucleic acid of step a into a cell; and

c. incubating the cell under conditions favorable for expression of the antigen protein;

thereby producing the antigen protein;

wherein the integral membrane protein component comprises at least one domain selected from the group consisting of a domain of hemagglutinin (HA) and a domain of neuraminidase (NA).

57-75. -75. (canceled)

76. An isolated nucleic acid construct comprising nucleic acid sequence encoding a component of an influenza A integral membrane protein fused to a thermostable protein;

and wherein the integral membrane protein component comprises at least one domain selected from the group consisting of a domain of hemagglutinin (HA) and a domain of neuraminidase (NA).

91. A host cell comprising the nucleic acid construct of claim 76.

77-90. -90. (canceled)

92-94. -94. (canceled)