Methods for stimulating antigen-specific T cell responses

Patent metadata

Field	Value
Publication number	US-9708583-B2
Application number	US-201414785924-A
Country	US
Kind code	B2
Filing date	Apr 22, 2014
Priority date	Apr 23, 2013
Publication date	Jul 18, 2017
Grant date	Jul 18, 2017

Abstract

The present invention relates to methods for stimulating antigen-specific T cell responses. In particular, the invention relates to a method for stimulating antigen (Ag)-specific T cell responses in a blood sample or PBMC sample isolated from a subject comprising the step consisting in culturing said blood or PBMC sample in a appropriate culture medium which comprises an amount of IL-1beta and an amount of a least one antigen.

First claim

The invention claimed is:

1. A method for stimulating antigen (Ag)-specific T cell responses in a blood sample or an unfractionated peripheral blood mononuclear cell (PBMC) sample isolated from a subject comprising the steps of
culturing said blood or unfractionated PBMC sample in an appropriate culture medium which comprises an amount of interleukin-1 beta (IL-1β) and an amount of a least one antigen,

wherein the culture medium does not contain a differentiating agent selected from the group consisting of Granulocyte/Macrophage Colony-Stimulating Factor (GM-CSF), interleukin-4 (IL-4) and FMS-like tyrosine kinase 3 (Flt-3) ligand, and
detecting amplification of a T cell response to said antigen in said blood sample or said unfractionated PBMC sample.

2. The method of claim 1 wherein said amount of IL-10 is from about 0.1 to about 1,000 ng/ml.

3. The method of claim 2 , wherein said amount of IL-1β is from about 1 to about 100 ng/ml.

4. The method of claim 2 , wherein said amount of IL-1β is about 10 ng/ml.

5. The method of claim 1 wherein the antigen is selected from the group consisting of proteins, peptides, nucleic acids, tissue preparations and cell preparations.

6. The method according to claim 1 wherein the antigen is a disease associated antigen.

7. The method according to claim 1 wherein the antigen is a therapeutic protein.

8. The method of claim 1 , wherein said step of detecting is performed by detecting at least one of a CD4+ T cell response and a CD8+ T cell response.

9. An in vitro method for determining the immunogenic potential of a therapeutic protein in a subject in need thereof, comprising,
stimulating antigen (Ag)-specific T cell responses in a blood sample or an unfractionated PBMC sample isolated from said subject by culturing said blood sample or said unfractionated PBMC sample in an appropriate culture medium which comprises an amount of IL-1β and an amount of said therapeutic protein, wherein the culture medium does not contain a differentiating agent selected from the group consisting of Granulocyte/Macrophage Colony-Stimulating Factor (GM-CSF), IL-4 and FMS-like tyrosine kinase 3 (Flt-3) ligand, and
detecting amplification of a T cell response to said therapeutic protein in said blood sample or said unfractionated PBMC sample, wherein if an amplified T cell response is detected, then said therapeutic protein will be immunogenic in said subject, and if a T cell response is not detected, then said therapeutic protein will not be immunogenic in said subject.

10. An in vitro method for determining the immunogenic potential of a vaccine adjuvant in a subject, comprising,
stimulating antigen (Ag)-specific T cell responses in a blood sample or an unfractionated PBMC sample isolated from said subject by culturing said blood sample or said unfractionated PBMC sample in an appropriate culture medium which comprises an amount of IL-1β and an amount of said vaccine adjuvant, wherein the culture medium does not contain a differentiating agent selected from the group consisting of Granulocyte/Macrophage Colony-Stimulating Factor (GM-CSF), IL-4 and FMS-like tyrosine kinase 3 (Flt-3) ligand, and
detecting amplification of a T cell response to said vaccine adjuvant in said blood sample or said unfractionated PBMC sample, wherein if an amplified T cell response is detected, then said vaccine adjuvant will be immunogenic in said subject, and if a T cell response is not detected, then said vaccine adjuvant will not be immunogenic in said subject.

11. An in vitro method for evaluating potential of an agent to induce a tolerogenic effect in a subject in need thereof, comprising,
stimulating antigen (Ag)-specific T cell responses in a blood sample or an unfractionated PBMC sample isolated from said subject by culturing said blood sample or said unfractionated PBMC sample in an appropriate culture medium which comprises an amount of IL-1β and an amount of said agent, wherein the culture medium does not contain a differentiating agent selected from the group consisting of Granulocyte/Macrophage Colony-Stimulating Factor (GM-CSF), IL-4 and FMS-like tyrosine kinase 3 (Flt-3) ligand, and
detecting amplification of a T cell response to said agent in said blood sample or said unfractionated PBMC sample, wherein if an amplified T cell response is detected, then said agent will not induce a tolerogenic effect in said subject, and if a T cell response is not detected, then said agent will induce a tolerogenic effect in said subject.

12. An in vitro method for screening Ags and epitopes in order to select those eliciting an Ag-specific T cell response, comprising
stimulating antigen (Ag)-specific T cell responses in a blood sample or an unfractionated PBMC sample isolated from a subject by culturing an amount of IL-1β and an amount of Ag or epitopes in culture medium that does not contain a differentiating agent selected from the group consisting of Granulocyte/Macrophage Colony-Stimulating Factor (GM-CSF), IL-4 and FMS-like tyrosine kinase 3 (Flt-3) ligand,
detecting amplification of a T cell response to said Ag or epitope in said culture medium, and
selecting Ags and epitopes which elicit said T cell response.

13. An in vitro method for producing polyclonal T cells and T cell lines or clones recognizing a Ag or combination of Ags, comprising
stimulating antigen (Ag)-specific T cell responses in a blood sample or an unfractionated PBMC sample of isolated from said subject by culturing said sample in an appropriate culture medium which comprises an amount of IL-1β and an amount of said Ag or combination of Ags, wherein the culture medium does not contain a differentiating agent selected from the group consisting of Granulocyte/Macrophage Colony-Stimulating Factor (GM-CSF), IL-4 and FMS-like tyrosine kinase 3 (Flt-3) ligand, and
detecting amplification of a T cell response to said Ag or combination of Ags in said T cells, and, if an amplified T cell response is detected in a T cell, then selecting said T cell for producing polyclonal T cells and T cell lines or clones which recognize said Ag or combination of Ags.

Assignees

• Inserm (Institut Nat De La Sante Et De La Rech Medicale)
• Univ Paris Descartes

Inventors

• Mallone Roberto
• Afonso Georgia

Classifications

• G01N33/575

  for cancer · CPC title

• G01N33/564 Primary

  for pre-existing immune complex or autoimmune disease {, i.e. systemic lupus erythematosus, rheumatoid arthritis, multiple sclerosis, rheumatoid factors or complement components C1-C9} · CPC title

• G01N33/569

  for microorganisms, e.g. protozoa, bacteria, viruses · CPC title

• C12N5/0636 Primary

  T lymphocytes · CPC title

• C12N2501/115

  Basic fibroblast growth factor (bFGF, FGF-2) · CPC title