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Last Updated: March 28, 2024

Claims for Patent: 6,083,925


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Summary for Patent: 6,083,925
Title: Nucleic acid respiratory syncytial virus vaccines
Abstract:Non-replicating vectors containing a nucleotide sequence coding for an F protein of respiratory syncytial virus (RSV) and a promoter for such sequence, preferably a cytomegalovirus promoter, are described for in vivo immunization. The nucleotide sequence encloding the RSV F protein may lack a sequence encoding the homologous signal peptide but possessing a heterologous signal peptide enhancing RSV F protein expression. Such non-replicating vectors, including plasmids, also may contain a further nucleotide sequence located adjacent to the RSV F protein encoding sequence to enhance the immunoprotective ability of the RSV F protein when expressed in vivo. Such non-replicating vectors may be used to immunize a host against disease caused by infection with RSV, including a human host, by administration thereto, and may be formulated as immunogenic compositions with pharmaceutically-acceptable carriers for such purpose. Such vectors also may be used to produce antibodies for detection of RSV infection in a sample.
Inventor(s): Li; Xiaomao (Toronto, CA), Ewasyshyn; Mary E. (Thornhill, CA), Sambhara; Suryaprakash (Markham, CA), Klein; Michel H. (Willowdale, CA)
Assignee: Connaught Laboratories Limited (North York, CA)
Application Number:09/262,927
Patent Claims:1. A method of immunizing a host against disease caused by infection with respiratory syncytial virus (RSV), which comprises administering to said host an effective amount of a plasmid vector comprising a nucleotide sequence encoding an RSV F protein fragment that generates antibodies and/or cytotoxic T-lymphocytes (CTLs) that specifically react with RSV F protein, said RSV F protein fragment lacking an autologous RSV F signal peptide sequence and including a sequence encoding a heterologous signal peptide which enhances the level of expression of RSV F protein, and a promoter sequence operatively coupled to said nucleotide sequence for expression of said RSV F protein fragment in said host.

2. The method of claim 1 wherein said nucleotide sequence encodes a RSV F protein fragment lacking a transmembrane region.

3. The method of claim 1 wherein the nucleotide sequence encoding a signal peptide encodes a Herpes Simplex Virus I gD signal peptide.

4. The method of claim 1 wherein said host is a human.

5. The method of claim 1 wherein said plasmid vector further comprises a further nucleotide sequence located adjacent said nucleotide sequence to enhance the immunoprotective ability of said RSV F protein fragment when expressed in vivo from said vector in said host, wherein said further nucleotide sequence comprises a pair of splice sites to prevent aberrant mRNA splicing.

6. The method of claim 4 wherein said promoter sequence is an immediate early cytomegalovirus promoter.

7. The method of claim 5 wherein said promoter sequence is an immediate early cytomegalovirus promoter.

8. The method of claim 5 wherein said vector is p82M35B as shown in FIG. 10.

9. The method of claim 7 wherein said further nucleotide sequence is located between said nucleotide sequence and said promoter sequence.

10. The method of claim 8 wherein said further nucleotide sequence is that of rabbit .beta.-globulin intron II.

11. A method of using a gene encoding an RSV F protein fragment that generates antibodies and/or cytotoxic T-lymphocytes (CTLs) that specifically react with RSV F protein and which lacks an autologous RSV F signal peptide and includes a sequence encoding a heterologous signal peptide which enhances the level of expression of RSV F protein to produce an immune response in a host, which comprises:

isolating said gene;

operatively linking said gene to at least one promoter sequence to produce a plasmid vector, said promoter sequence directing expression of said RSV F protein fragment when said vector is introduced into a host to produce an immune response to said RSV F protein fragment; and

introducing said vector into the host.

12. The method of claim 11, wherein said gene encoding an RSV F protein fragment encodes an RSV F protein fragment lacking the transmembrane region.

13. The method of claim 11 wherein said sequence encoding a heterologous signal peptide encodes a Herpes Simplex Virus I gD signal peptide.

14. The method of claim 11 wherein said at least one promoter sequence comprises the immediately early cytomegalovirus promoter.

15. The method of claim 11 wherein said gene is contained within plasmid vector p82M35B (ATCC 203790).

16. The method of claim 12 wherein said at least one promoter sequence comprises the immediate early cytomegalovirus promoter.

17. The method of claim 14 including the step of:

operatively linking said gene to an immunoprotective enhancing sequence comprising a pair of splice sites to prevent aberrant mRNA splicing to produce an enhanced immunoprotection to said RSV F protein in said host.

18. The method of claim 17 wherein said immunoprotective enhancing sequence is introduced into said vector between said control sequence and said gene.

19. The method of claim 17 wherein said immunoprotective enhancing sequence is that of rabbit .beta.-globin intron II.

20. A method of producing a vaccine for protection of a host against disease caused by infection with respiratory syncytial virus (RSV), which comprises:

isolating a first nucleotide sequence encoding an RSV F protein fragment that generates antibodies and/or cytotoxic T-lymphocytes (CTLs) that specifically react with RSV F protein and which lacks an autologous RSV F signal peptide and includes a sequence encoding a heterologous signal peptide which enhances the level of expression of RSV F protein;

operatively linking said first nucleotide sequence to at least one promoter sequence to produce a plasmid vector, the promoter sequence directing expression of said RSV F protein when introduced into a host to produce an immune response to said RSV F protein;

operatively linking said first nucleotide sequence to a second nucleotide sequence to enhance the immunoprotective ability of said RSV F protein when expressed in vivo from the vector in a host, said second nucleotide sequence being introduced into said vector between said promoter sequence and said gene and comprising a pair of splice sites to prevent aberrant mRNA splicing, and

formulating said vector as a vaccine for in vivo administration.

21. The method of claim 20 wherein said plasmid vector is p82M35B ATCC 203790).

22. A vaccine produced by the method of claim 20.

23. A method of producing a vaccine for protection of a host against disease caused by infection with respiratory syncytial virus (RSV), which comprises:

isolating a nucleotide sequence encoding an RSV F protein fragment from which the transmembrane region is absent and which lacks an autologous RSV F signal peptide and includes a sequence encoding a heterologous signal peptide which enhances the level of expression of RSV F protein;

operatively linking said nucleotide sequence to at least one promoter sequence to produce a plasmid vector, the promoter sequence directing expression of said RSV F protein when introduced into a host to produce an immune response to said RSV F protein; and

formulating said vector as a vaccine for in vivo administration.

24. The method of claim 23 wherein said plasmid vector is p82M35B.

25. A vaccine produced by the method of claim 23.

Details for Patent 6,083,925

Applicant Tradename Biologic Ingredient Dosage Form BLA Approval Date Patent No. Expiredate
Merck Sharp & Dohme Corp. INTRON A interferon alfa-2b For Injection 103132 06/04/1986 ⤷  Try a Trial 2015-06-07
Merck Sharp & Dohme Corp. INTRON A interferon alfa-2b For Injection 103132 ⤷  Try a Trial 2015-06-07
Merck Sharp & Dohme Corp. INTRON A interferon alfa-2b Injection 103132 ⤷  Try a Trial 2015-06-07
>Applicant >Tradename >Biologic Ingredient >Dosage Form >BLA >Approval Date >Patent No. >Expiredate

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