DbpA compositions and methods of use

• US 6,312,907 B1
• Filed: 01/21/2000
• Issued: 11/06/2001
• Est. Priority Date: 04/24/1995
• Status: Expired due to Fees

• Current Assignee: Texas A&M University System
• Original Assignee: Texas A&M University System
• Inventors: Guo, Betty P., H{umlaut over (oo)}k, Magnus
• Primary Examiner(s): Zitomer, Stephanie W.
• Application Number:
  US09/489,352
• Time in Patent Office: 655 Days
• Field of Search: 435/6, 435/320.1, 435/252.3, 435/810, 435/252.33, 435/255.2, 435/254.11, 435/325, 435/352, 435/849, 435/942, 435/101, 435/134, 435/147, 435/195, 435/197, 536/23.1, 536/24.32
• US Class Current: 435/6.16
• CPC Class Codes: A01K 2217/05 Animals comprising random inserted nucleic acids (transgenic)
• CPC Class Codes: A61K 38/00 Medicinal preparations containing peptides peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00
• CPC Class Codes: A61K 39/00 Medicinal preparations containing antigens or antibodies materials for immunoassay G01N33/53
• CPC Class Codes: A61P 31/04 Antibacterial agents
• CPC Class Codes: A61P 37/00 Drugs for immunological or allergic disorders
• CPC Class Codes: C07K 14/20 from Spirochaetales (O), e.g. Treponema, Leptospira
• CPC Class Codes: Y02A 50/30 Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
• CPC Class Codes: Y10S 530/806 Antigenic peptides or proteins
• CPC Class Codes: Y10S 530/825 Bacteria

Abstract

Disclosed are the dbp gene and dbp-derived nucleic acid segments from Borrelia burgdorferi, the etiological agent of Lyme disease, and DNA segments encoding dbp from related borrelias. Also disclosed are decorin binding protein compositions and methods of use. The DBP protein and antigenic epitopes derived therefrom are contemplated for use in the treatment of pathological Borrelia infections, and in particular, for use in the prevention of bacterial adhesion to decorin. DNA segments encoding these proteins and anti-(decorin binding protein) antibodies will also be of use in various screening, diagnostic and therapeutic applications including active and passive immunization and methods for the prevention of Borrelia colonization in an animal. These DNA segments and the peptides derived therefrom are contemplated for use in the preparation of vaccines and, also, for use as carrier proteins in vaccine formulations, and in the formulation of compositions for use in the prevention of Lyme disease.

Claims

1. An isolated nucleic acid segment comprising a dbpA gene that encodes a decorin binding protein or peptide that includes the contiguous amino acid sequence of SEQ ID NO:

34;

a contiguous amino acid sequence of at least about 100 amino acids from SEQ ID NO;

42 or SEQ ID NO;

46;

or a contiguous amino acid sequence of at least about 25 amino acids from SEQ ID NO;

50.

2. The isolated nucleic acid segment of claim 1, wherein said dbpA gene encodes a decorin binding protein that includes the contiguous amino acid sequence of SEQ ID NO:

34, SEQ ID NO;

42, SEQ ID NO;

46 or SEQ ID NO;

50.

3. The isolated nucleic acid segment of claim 2, wherein said dbpA gene comprises the contiguous nucleic acid sequence of SEQ ID NO:

33, SEQ ID NO;

41, SEQ ID NO;

45 or SEQ ID NO;

49.

4. The isolated nucleic acid segment of claim 1, comprising a dbpA gene that encodes a decorin binding protein that includes the contiguous amino acid sequence of SEQ ID NO:

34.

5. The isolated nucleic acid segment of claim 4, wherein said dbpA gene comprises the contiguous nucleic acid sequence of SEQ ID NO:

33.

6. The isolated nucleic acid segment of claim 1, comprising a dbpA gene that encodes a decorin binding protein that includes a contiguous amino acid sequence of at least about 100 amino acids from SEQ ID NO:

42.

7. The isolated nucleic acid segment of claim 6, comprising a dbpA gene that encodes a decorin binding protein that includes the contiguous amino acid sequence of SEQ ID NO:

42.

8. The isolated nucleic acid segment of claim 7, wherein said dbpA gene comprises the contiguous nucleic acid sequence of SEQ ID NO:

41.

9. The isolated nucleic acid segment of claim 1, comprising a dbpA gene that encodes a decorin binding protein that includes a contiguous amino acid sequence of at least about 100 amino acids from SEQ ID NO:

46.

10. The isolated nucleic acid segment of claim 9, comprising a dbpA gene that encodes a decorin binding protein that includes the contiguous amino acid sequence of SEQ ID NO:

46.

11. The isolated nucleic acid segment of claim 10, wherein said dbpA gene comprises the contiguous nucleic acid sequence of SEQ ID NO:

45.

12. The isolated nucleic acid segment of claim 1, comprising a dbpA gene that encodes a decorin binding protein that includes a contiguous amino acid sequence of at least about 25 amino acids from SEQ ID NO:

50.

13. The isolated nucleic acid segment of claim 12, comprising a dbpA gene that encodes a decorin binding protein that includes a contiguous amino acid sequence of at least about 50 amino acids from SEQ ID NO:

50.

14. The isolated nucleic acid segment of claim 13, comprising a dbpA gene that encodes a decorin binding protein that includes a contiguous amino acid sequence of at least about 100 amino acids from SEQ ID NO:

50.

15. The isolated nucleic acid segment of claim 14, comprising a dbpA gene that encodes a decorin binding protein that includes the contiguous amino acid sequence of SEQ ID NO:

50.

16. The isolated nucleic acid segment of claim 15, wherein said dbpA gene comprises the contiguous nucleic acid sequence of SEQ ID NO:

49.

17. The isolated nucleic acid segment of claim 1 operatively linked to a promoter that directs the expression of said nucleic acid segment in a host cell.

18. The isolated nucleic acid segment of claim 17, further defined as a recombinant vector.

19. The isolated nucleic acid segment of claim 1, further defined as an RNA segment.

20. The isolated nucleic acid segment of claim 1, comprised within a recombinant host cell.

35. The isolated nucleic acid segment of claim 1, dispersed in a pharmaceutically acceptable carrier.

21. A recombinant host cell comprising an isolated nucleic acid segment comprising a dbpA gene that encodes a decorin binding protein or peptide that includes the contiguous amino acid sequence of SEQ ID NO:

34;

a contiguous amino acid sequence of at least about 100 amino acids from SEQ ID NO;

42 or SEQ ID NO;

46;

or a contiguous amino acid sequence of at least about 25 amino acids from SEQ ID NO;

50.

22. The recombinant host cell of claim 21, further defined as a prokaryotic host cell.

23. The recombinant host cell of claim 21, further defined as a eukaryotic host cell.

24. The recombinant host cell of claim 21, wherein said isolated nucleic acid segment is comprised within a recombinant vector.

25. The recombinant host cell of claim 24, wherein said recombinant vector is integrated into the host cell chromosome.

26. The recombinant host cell of claim 21, wherein said dbpA gene encodes a decorin binding protein that includes the contiguous amino acid sequence of SEQ ID NO:

34, SEQ ID NO;

42, SEQ ID NO;

46 or SEQ ID NO;

50.

27. The recombinant host cell of claim 26, wherein said dbpA gene comprises the contiguous nucleic acid sequence of SEQ ID NO:

33, SEQ ID NO;

41, SEQ ID NO;

45 or SEQ ID NO;

49.

28. A nucleic acid detection kit comprising, in a suitable container, a detection reagent and an isolated nucleic acid segment comprising a dbpA gene that encodes a decorin binding protein or peptide that includes the contiguous amino acid sequence of SEQ ID NO:

34;

a contiguous amino acid sequence of at least about 100 amino acids from SEQ ID NO;

42 or SEQ ID NO;

46;

or a contiguous amino acid sequence of at least about 25 amino acids from SEQ ID NO;

50.

29. The nucleic acid detection kit of claim 28, wherein said dbpA gene encodes a decorin binding protein that includes the contiguous amino acid sequence of SEQ ID NO:

34, SEQ ID NO;

42, SEQ ID NO;

46 or SEQ ID NO;

50.

30. The nucleic acid detection kit of claim 29, wherein said dbpA gene comprises the contiguous nucleic acid sequence of SEQ ID NO:

33, SEQ ID NO;

41, SEQ ID NO;

45 or SEQ ID NO;

49.

31. A method for detecting a nucleic acid sequence encoding a DbpA protein or peptide, comprising the steps of:

(a) contacting sample nucleic acids suspected of encoding a DbpA protein or peptide with an isolated nucleic acid segment encoding a DbpA protein or peptide that includes the contiguous amino acid sequence of SEQ ID NO;

34;

a contiguous amino acid sequence of at least about 100 amino acids from SEQ ID NO;

42 or SEQ ID NO;

46;

or a contiguous amino acid sequence of at least about 25 amino acids from SEQ ID NO;

50, under conditions effective to allow hybridization of substantially complementary nucleic acids; and

(b) detecting the hybridized complementary nucleic acids thus formed, thereby detecting a nucleic acid sequence encoding a DbpA protein or peptide.

32. The method of claim 31, wherein the isolated nucleic acid segment encoding a DbpA protein comprises a detectable label, and the hybridized complementary nucleic acids are detected by detecting said label.

33. The method of claim 31, wherein said isolated nucleic acid segment encodes a DbpA protein that includes the contiguous amino acid sequence of SEQ ID NO:

34, SEQ ID NO;

42, SEQ ID NO;

46 or SEQ ID NO;

50.

34. The method of claim 33, wherein said isolated nucleic acid segment comprises the contiguous nucleic acid sequence of SEQ ID NO:

33, SEQ ID NO;

41, SEQ ID NO;

45 or SEQ ID NO;

49.