Methods for determining the risk of acute graft versus host disease

Patent metadata

Field	Value
Publication number	US-10151745-B2
Application number	US-201314646486-A
Country	US
Kind code	B2
Filing date	Nov 21, 2013
Priority date	Nov 21, 2012
Publication date	Dec 11, 2018
Grant date	Dec 11, 2018

Abstract

The present invention relates to a method for determining whether a candidate human transplant donor is at risk of inducing acute graft versus host disease (aGVHD) in a human transplant recipient, which may in turn allow the selection of a donor exhibiting no risk for the recipient. The present invention also relates to a method for adjusting the immunosuppressive treatment administered to a human transplanted recipient following its graft transplantation after having performing the method for determining risk of the invention. The methods comprise expanding the candidate donor's iNKT cells (invariant NKT cells) and determining the presence or absence of expansion of the CD4(−) iNKT cell sub-population. In particular, CD3+CD4− TCRV[alpha]24V[beta]11 cells are determined. Kits are disclosed.

First claim

The invention claimed is:

1. A method for determining whether the population of invariant NKT (iNKT) cells from a biological sample of a candidate human transplant donor expands, comprising the following steps of:
i) measuring the CD4− iNKT cell subpopulation in the population of invariant NKT (iNKT) cells of a biological sample obtained from the candidate human transplant donor, wherein said candidate human transplant donor is a human leukocyte antigen (HLA)-identical donor, an HLA-matched donor, an HLA haploidentical donor, and/or a 4/6 to 6/6 HLA (A, B, DR) cord blood donor;
ii) expanding the population of iNKT cells from the biological sample obtained from the candidate human transplant donor; and
iii) detecting the presence or absence of expansion of the CD4− iNKT cell subpopulation in the population obtained at step ii).

2. The method according to claim 1 , wherein the step ii) of expanding the population of iNKT cells is carried out by (a) isolating peripheral blood mononuclear cells (PBMCs) from the biological sample obtained from said candidate human transplant donor, and (b) culturing said PBMCs in a medium comprising both an agent stimulating the proliferation of iNKT cells and an agent activating iNKT cells.

3. The method according to claim 1 or claim 2 , wherein the CD4− iNKT cell subpopulation is a population of CD3+CD4− TCR Vα24Vβ11 cells.

4. The method according to claim 2 , wherein the agent stimulating the proliferation of iNKT cells is interleukin 2 (IL-2).

5. The method according to claim 2 , wherein the agent activating iNKT cells is a glycolipid antigen selected from the group consisting of alpha-galactosylceramide (alpha-GalCer), alpha-glucuronosylceramide, phosphatidylinositoltetramannoside, isoglobotrihexosylceramide, ganglioside GD3, phosphatidylcholine, phosphatidylethanolamine, phosphatidylinositol, sulfatide, beta-galactosylceramide, lipophosphoglycan, glycoinositol phospholipid, beta-anomeric galactoceramide and alpha-anomeric galactosylceramide, and bacterial lipid antigens.

6. The method according to claim 5 , wherein the glycolipid antigen is alpha-GalCer.

7. The method according to claim 3 wherein the agent stimulating the proliferation of iNKT cells is interleukin 2 (IL-2).

8. The method according to claim 3 wherein the agent activating iNKT cells is a glycolipid antigen selected from the group consisting of alpha-galactosylceramide (alpha-GalCer), alpha-glucuronosylceramide, phosphatidylinositoltetramannoside, isoglobotrihexosylceramide, ganglioside GD3, phosphatidylcholine, phosphatidylethanolamine, phosphatidylinositol, sulfatide, beta-galactosylceramide, lipophosphoglycan, glycoinositol phospholipid, beta-anomeric galactoceramide and alpha-anomeric galactosylceramide, and bacterial lipid antigens.

9. The method according to claim 8 , wherein the glycolipid antigen is alpha-GalCer.

10. The method according to claim 4 wherein the agent activating iNKT cells is a glycolipid antigen selected from the group consisting of alpha-galactosylceramide (alpha-GalCer), alpha-glucuronosylceramide, phosphatidylinositoltetramannoside, isoglobotrihexosylceramide, ganglioside GD3, phosphatidylcholine, phosphatidylethanolamine, phosphatidylinositol, sulfatide, beta-galactosylceramide, lipophosphoglycan, glycoinositol phospholipid, beta-anomeric galactoceramide and alpha-anomeric galactosylceramide, and bacterial lipid antigens.

11. The method according to claim 10 , wherein the glycolipid antigen is alpha-GalCer.

12. A method of transplanting a tissue or organ comprising:
i) measuring the CD4− iNKT cell subpopulation in the population of invariant NKT (iNKT) cells of a biological sample obtained from a candidate human tissue or organ transplant donor, wherein said candidate human transplant donor is a human leukocyte antigen (HLA)-identical donor, an HLA-matched donor, an HLA haploidentical donor, and/or a 4/6 to 6/6 HLA (A, B, DR) cord blood donor;
ii) expanding the population of iNKT cells from the biological sample obtained from the candidate human transplant donor;
iii) detecting the presence or absence of expansion of the CD4− iNKT cell subpopulation in the population obtained at step ii); and
iv) transplanting a tissue or organ from the candidate donor into a transplant recipient if expansion of the CD4− iNKT cell subpopulation is detected in step iii).

Assignees

• Inst Nat Sante Rech Med
• Centre Nat Rech Scient
• Assist Publique—Hopitaux De Paris
• Univ Paris Descartes
• Imagine Inst Des Maladies Genetiques Necker Enfants Malades
• Inst Nat Sante Rech Med
• Univ Paris Descartes

Inventors

• Hermine Olivier
• Rubio Marie Thérèse
• Bouillie Marie
• Leite De Maraes Maria

Classifications

• A61P37/06

  Immunosuppressants, e.g. drugs for graft rejection · CPC title

• G01N33/505 Primary

  involving T-cells · CPC title

• G01N2800/52

  Predicting or monitoring the response to treatment, e.g. for selection of therapy based on assay results in personalised medicine; Prognosis · CPC title

• G01N2800/50

  Determining the risk of developing a disease · CPC title

• G01N2800/245

  Transplantation related diseases, e.g. graft versus host disease · CPC title