Proteins expressed by mycobacterium tuberculosis and not by BCG and their use as diagnostic reagents and vaccines

Patent metadata

Field	Value
Publication number	US-9482671-B2
Application number	US-201614989135-A
Country	US
Kind code	B2
Filing date	Jan 6, 2016
Priority date	May 4, 1999
Publication date	Nov 1, 2016
Grant date	Nov 1, 2016

Abstract

The present invention is directed to reagents useful for generating immune responses to Mycobacterium tuberculosis and for diagnosing infection and disease in a subject that has been exposed to M. tuberculosis.

First claim

What is claimed:

1. A method of in vitro diagnosis that discriminates between infection by Mycobacterium tuberculosis -complex and vaccination by Bacille Calmette Guerin (BCG) strain of Mycobacterium bovis comprising:
providing a population of cells comprising CD4 T lymphocytes from a subject;
contacting cells of the population with at least two different antigens, wherein the antigens are isolated polypeptides of the Mycobacterium tuberculosis -complex that are not encoded by BCG, including at least one isolated polypeptide selected from the group consisting of (i) a first amino acid sequence comprising the sequence of MTBN4 (SEQ ID NO: 4), (ii) a second amino acid sequence that is an antigenic segment of MTBN4 and (iii) a third amino acid sequence that is identical to said first or second amino acid sequence but that has conservative substitutions and that retains Mycobacterium tuberculosis -complex specific antigenic properties; and
determining whether or not there has been an immune response to said at least two different antigens, wherein CD4 T lymphocytes from a subject that has been infected by Mycobacterium tuberculosis -complex respond to said at least two different antigens, and CD4 T lymphocytes from a subject vaccinated with the BCG strain of Mycobacterium bovis but not infected by Mycobacterium tuberculosis -complex do not respond to said at least two different antigens.

2. The method of claim 1 , wherein said at least one isolated polypeptide comprises said second or third amino acid sequence.

3. The method of claim 1 , wherein the step of contacting is contacting said cells with a composition containing said at least two different antigens.

4. The method of claim 3 , wherein the determining step comprises testing for production of at least one cytokine.

5. The method of claim 4 , wherein the at least one cytokine includes interferon-γ (IFN-γ).

6. The method of claim 1 , wherein the determining step comprises testing for production of at least one cytokine.

7. The method of claim 6 , wherein the at least one cytokine includes interferon-γ (IFN-γ).

8. The method of claim 1 , wherein the isolated polypeptides of the Mycobacterium tuberculosis -complex are encoded within the RD1, RD2, and RD3 regions.

Assignees

• Univ Rutgers

Inventors

• Gennaro Maria L

Classifications

• G01N2800/26

  Infectious diseases, e.g. generalised sepsis · CPC title

• A61K39/04

  Mycobacterium, e.g. Mycobacterium tuberculosis · CPC title

• A61K38/00

  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 title

• G01N2333/35

  from Mycobacteriaceae (F) · CPC title

• A61P31/06

  for tuberculosis · CPC title