IMMUNOTHERAPY AND DIAGNOSIS OF MUCORMYCOSIS USING CotH

• US 20130108642A1
• Filed: 09/14/2012
• Published: 05/02/2013
• Est. Priority Date: 09/15/2011
• Status: Active Grant

• Current Assignee: Los Angeles Biomedical Research Institute At Harbor-Ucla Medical Center (University of California)
• Original Assignee: Mingfu Liu
• Inventors: Ibrahim, Ashraf S., Fu, Yue, Edwards, John E., Filler, Scott, Liu, Mingfu, Ghebremariam, Teklegiorgis
• Granted Patent: US 9,279,002 B2
• Time in Patent Office: Days
• US Class Current: 424/139.1
• CPC Class Codes: A61K 2039/505 comprising antibodies
• CPC Class Codes: A61P 31/10 Antimycotics
• CPC Class Codes: C07K 14/37 from fungi
• CPC Class Codes: C07K 16/14 against material from fungi, algea or lichens
• CPC Class Codes: C07K 2317/24 containing regions, domains or residues from different species, e.g. chimeric, humanized or veneered
• CPC Class Codes: C07K 2317/34 Identification of a linear epitope shorter than 20 amino acid residues or of a conformational epitope defined by amino acid residues
• CPC Class Codes: C07K 2317/622 Single chain antibody (scFv)
• CPC Class Codes: C07K 2317/76 Antagonist effect on antigen, e.g. neutralization or inhibition of binding
• CPC Class Codes: G01N 2333/37 from fungi
• CPC Class Codes: G01N 2469/10 Detection of antigens from microorganism in sample from host
• CPC Class Codes: G01N 33/56961 Plant cells or fungi

Abstract

The invention provided Mucorales CotH polypeptides and encoding nucleic acid molecules. The Mucorales CotH polypeptides and encoding nucleic acids can be advantageously used to diagnose, treat or prevent fungal conditions, in particular mucormycosis.

Claims

1. An isolated nucleic acid encoding a Mucorales CotH polypeptide, an immunogenic fragment thereof, or a functional fragment thereof.

2. The isolated nucleic acid of claim 1, wherein said nucleic acid comprises a nucleic acid selected from:

(a) nucleic acid encoding an amino acid sequence set forth in SEQ ID NOS;

1, 3, 5, 11, 15, 16, 18, 20, 22, 24, 26, 28, 30 or 32, or

(b) nucleic acid that hybridizes to the nucleic acid of (a) under moderately stringent conditions, wherein said nucleic acid contiguously encodes biologically active Mucorales CotH polypeptide, or

(c) nucleic acid degenerate with respect to either (a) or (b) above, wherein said nucleic acid encodes biologically active Mucorales CotH polypeptide.

4. The nucleic acid of claim 2, wherein said amino acid sequence comprises least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98% or 99% sequence identity to the amino acids sequence set forth in SEQ ID NOS:

1, 3, 5, 11, 15, 16, 18, 20, 22, 24, 26, 28, 30 or 32.

7. A vector containing the nucleic acid of claim 2.

8. A recombinant cell containing the nucleic acid of claim 2.

11. An antisense-nucleic acid capable of specifically binding to mRNA encoded by said nucleic acid according to claim 2.

24. A method for expression of a Mucorales CotH polypeptide, said method comprising culturing cells of claim 8 under conditions suitable for expression of said Mucorales CotH.

30. A composition comprising an amount of the antisense-nucleic acid according to claim 11 effective to inhibit expression of a Mucorales CotH polypeptide and an acceptable carrier capable of delivering said antisense-nucleic acid to a cell.

40. A method of diagnosing mucormycosis infection in a subject, comprising the steps of:

(a) providing a test sample from the subject;

(b) contacting said sample with an agent that can binds said nucleic acid of claim 1 under suitable conditions, wherein said conditions allow specific binding of said agent to said nucleic acid or polypeptide; and

(c) comparing the amount of said specific binding in said test sample with the amount of specific binding in a control sample, wherein an increased or decreased amount of said specific binding in said test sample as compared to said control sample is diagnostic of mucormycosis infection.

3. (canceled)

5-6. -6. (canceled)

9. A CotH oligonucleotide, comprising between 15 and 300 contiguous nucleotides of SEQ ID NOS:

2, 4, 6, 9, 10, 12-14, 17, 19, 21, 23, 25, 27, 29 or 31 or the anti-sense strand thereof.

12. A kit for detecting the presence of a Mucorales CotH nucleic acid sequence comprising at least one oligonucleotide according to claim 9.

31. A method for identifying nucleic acids encoding a Mucorales CotH polypeptide, comprising contacting a sample containing nucleic acids with one or more oligonucleotides according to claim 9, wherein said contacting is effected under highly stringent hybridization conditions, and identifying a nucleic acid that hybridizes to said oligonucleotide.

32. A method of detecting a Mucorales CotH nucleic acid molecule in a sample, comprising contacting said sample with two or more oligonucleotides of claim 9, amplifying a nucleic acid molecule, and detecting said amplification.

10. (canceled)

13. (canceled)

14. An isolated Mucorales CotH polypeptide, an immunogenic fragment thereof, or a functional fragment thereof.

15. The Mucorales CotH polypeptide of claim 14, wherein said polypeptide comprises least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98% or 99% sequence identity to the amino acids sequence set forth in SEQ ID NOS:

1, 3, 5, 11, 15, 16, 18, 20, 22, 24, 26, 28, 30 or 32.

25. An isolated anti-Mucorales CotH antibody having specific reactivity with a Mucorales CotH polypeptide according to claim 14.

35. A method for detecting the presence of a Mucorales organism in a sample, comprising contacting a sample with an antibody according to claim 25, and detecting the presence of specific binding of said antibody to said sample, thereby detecting the presence of Mucorales organism in said sample.

36. A pharmaceutical composition comprising a pharmaceutically acceptable carrier and the anti-Mucorales CotH antibody of claim 25.

37. A vaccine composition for immunization of a subject against a fungal condition, comprising an immunogenic amount of the Mucorales CotH polypeptide of claim 14, or an immunogenic fragment of said polypeptide, and a pharmaceutically acceptable carrier.

39. A method of treating or preventing mucormycosis in a subject in need thereof comprising administering an therapeutically effective amount of a pharmaceutical composition of claim 36.

16-23. -23. (canceled)

26-29. -29. (canceled)

33-34. -34. (canceled)

38. (canceled)

41. (canceled)