Method for monitoring HIV specific T cell responses

Patent metadata

Field	Value
Publication number	US-9709577-B2
Application number	US-201314386940-A
Country	US
Kind code	B2
Filing date	Mar 22, 2013
Priority date	Mar 23, 2012
Publication date	Jul 18, 2017
Grant date	Jul 18, 2017

Abstract

The invention relates to a method and a diagnostic kit for monitoring HIV specific T cell responses and identifying subjects capable of controlling HIV progression or preventing HIV infection altogether. The method is based on the combined use of boosted flow cytometry and toggle peptides and can cover a large set of effector functions. The method is also suitable to detect T cell responses of any desirable cytokine or combination of cytokines to any pathogen.

First claim

The invention claimed is:

1. A method for assaying the T cell response specific against a pathogen in a subject which comprises: i) contacting a sample comprising T cells from the subject with a composition comprising an antigen from the pathogen and ii) determining the levels of a plurality of cytokines produced by the T cells in the sample,
wherein the levels of the plurality of cytokines are determined by boosted flow cytometry, said boosted flow cytometry comprising detecting several cytokines in the same channel of fluorescence.

2. The method according to claim 1 wherein the composition comprising an antigen is an antigen or a library of antigens.

3. The method according to claim 2 wherein the antigen or the library of antigens from the pathogen is a peptide variant or a library of peptides variants derived from a polypeptide of the pathogen.

4. The method according to claim 3 wherein the library of peptides variants is a toggled peptide.

5. The method according to claim 4 wherein the pathogen is HIV or EBV.

6. The method according to claim 5 wherein the peptide variants derive from the Gag or the Nef polyprotein.

7. The method according to claim 6 wherein the peptide variants derive from p24 or p17.

8. The method according to claim 1 wherein the plurality of cytokines are selected from the group consisting of cytokines characteristic of a Th1 response, cytokines characteristic of a Th2 response and both.

9. The method according to claim 8 wherein the cytokines characteristic of a Th1 response are selected from the group consisting of IFN-γ, TNF-α, MIP1-β, IL-2, IL-12 and IL-1B.

10. The method according to claim 8 wherein the cytokines characteristic of a Th2 response are selected from the group consisting of IL-4, IL-5, IL-6, IL-10, and IL-13.

11. The method according to claim 8 wherein the cytokines characteristic of a Th1 response, the cytokines characteristic of a Th2 response or both are determined in CD4+ cells and/or in CD8+ cells.

12. The method according to claim 8 further comprising determining the levels of cytokines characteristic of a Th9, Th17 or Th22 response.

13. The method according to claim 1 wherein the sample containing T cells is a PBMC preparation.

Assignees

• Esteve Labor Dr
• Fundació Privada Inst De Recerca De La Sida—Caixa
• Institució Catalana De Recerca I Estudis Avançats
• Fund Privada Inst De Recerca De La Sida—Caixa
• Institució Catalana De Recerca I Estudios Avançats

Inventors

• Ruiz Riol Marta
• Brander Christian
• Ibarrondo Javier

Classifications

• G01N2800/56

  Staging of a disease; Further complications associated with the disease · CPC title

• G01N2800/60

  Complex ways of combining multiple protein biomarkers for diagnosis · CPC title

• G01N33/6866 Primary

  Interferon · CPC title

• G01N2333/525

  Tumor necrosis factor [TNF] · CPC title

• G01N2333/54

  Interleukins [IL] · CPC title