Methods for detecting a mycobacterium tuberculosis infection

Patent metadata

Field	Value
Publication number	US-2016103127-A1
Application number	US-201514856352-A
Country	US
Kind code	A1
Filing date	Sep 16, 2015
Priority date	Nov 20, 2009
Publication date	Apr 14, 2016
Grant date	—

Abstract

Methods for detecting an infection with Mtb in a subject are disclosed. The methods include detecting the presence of CD8 + T cells that specifically recognize an Mtb polypeptide. The methods include in vitro assays for detecting the presence of CD8 + T cells in a biological sample, and in vivo assays that detect a delayed type hypersensitivity reaction. The methods also include detecting Mtb polypeptides and polynucleotides.

First claim

1 . A method for detecting Mycobacterium tuberculosis in a subject, comprising
contacting a biological sample from the subject comprising T cells ex vivo with one or more Mycobacterium polypeptides, and an antigen presenting cell presenting the one or more Mycobacterium polypeptides wherein the one or more Mycobacterium polypeptides (a) the amino acid sequence set forth as SEQ ID NO: 6; or (b) at least nine to twenty consecutive amino acids of at least one of the amino acid sequence set forth as SEQ ID NO: 6, wherein the nine to twenty consecutive amino acids specifically bind major histocompatibility complex (MHC) class I; and determining if the T cells specifically recognize the Mycobacterium polypeptide, wherein the presence of T cells that specifically recognize the Mycobacterium polypeptide detects Mycobacterium tuberculosis in the subject.

2 . (canceled)

3 . The method of claim 1 , wherein the T cells are CD8 + T cells.

4 . The method of claim 3 , in which determining if the CD8 + T cells specifically recognize the Mycobacterium polypeptide is determined by measuring secretion of a cytokine from the CD8 + T cells.

5 . The method according to claim 4 , wherein the cytokine is interferon (IFN)-γ.

6 . The method of claim 5 , wherein measuring secretion of IFN-γ is determined using an antibody that specifically binds IFN-γ.

7 . The method of claim 1 , wherein the biological sample is blood, isolated peripheral blood mononuclear cells, or isolated mononuclear cells

8 . The method of claim 1 , wherein the T cells are cultured in vitro with the Mycobacterium polypeptide.

9 . The method of claim 1 , wherein the polypeptide is administered to the subject.

10 . A method of detecting a Mycobacterium tuberculosis infection in a subject, comprising
detecting the presence of a Mycobacterium polypeptide or a polynucleotide encoding the polypeptide in a sample from the subject, wherein the Mycobacterium polypeptide comprises the amino acid sequence set forth SEQ ID NO: 6.

11 . (canceled)

12 . The method of claim 10 , comprising determining the presence of the Mycobacterium polypeptide.

13 . The method of claim 12 , wherein determining the presence of the Mycobacterium polypeptide comprises the use of an antibody that specifically binds the Mycobacterium polypeptide.

14 . The method of claim 10 , comprising determining the presence of the Mycobacterium polynucleotide.

15 . The method of claim 14 , wherein determining the presence of the Mycobacterium polypeptide comprises the use of polymerase chain reaction.

16 . The method of claim 10 , wherein the biological sample is blood, peripheral blood mononuclear cells, sputum, a lung biopsy, a lymph node biopsy, saliva, cerebral spinal fluid or isolated I cells.

17 . A method of detecting Mycobacterium tuberculosis in a subject, comprising;
administering to the subject an effective amount of a Mycobacterium polypeptide into the skin of the subject, wherein the Mycobacterium polypeptide comprises an amino acid sequence set forth as (a) the amino acid sequence set forth as SEQ ID NO: 6; or (b) at least nine to twenty consecutive amino acids of the amino acid sequence set forth as SEQ ID NO: 6, wherein the nine to twenty consecutive amino acids specifically bind major histocompatibility complex (MHC) class I; and detecting the presence of CD8 + T cells that specifically recognize the Mycobacterium polypeptide in the subject.

18 . (canceled)

19 . The method of claim 17 , wherein detecting the presence of CD8 + T cells comprises detecting the presence of CD8 + T cells in vivo.

20 . The method of claim 19 , wherein the detecting the presence of CD8 + T cells comprises detecting a delayed type hypersensitivity reaction.

21 . The method of claim 20 , wherein the Mycobacterium polypeptide is administered intradermally to the subject, and wherein the delayed type hypersensitivity reaction is measure by measuring redness, swelling or induration of the skin.

22 . The method of claim 1 , wherein the one or more Mycobacterium polypeptides comprises the amino acid sequence set forth as SEQ ID NO: 6.

23 . The method of claim 17 , wherein the one or more Mycobacterium polypeptides comprises the amino acid sequence set forth as SEQ ID NO: 6.

Assignees

• Univ Oregon Health & Science
• Us Dept Veterans Affairs

Inventors

• Lewinsohn David M
• Lewinsohn Deborah A

Classifications

• A61P37/04

  Immunostimulants · CPC title

• A61P31/06

  for tuberculosis · CPC title

• A61P31/16

  for influenza or rhinoviruses · CPC title

• A61K49/00

  Preparations for testing in vivo · CPC title

• A61K39/385

  Haptens or antigens, bound to carriers · CPC title