Ex vivo generation of MHCII restricted CD4

1. A method for generating ex vivo MHCII restricted CD4 + Foxp3 + regulatory T cells having the following phenotype: CD3 + CD4 + Foxp3 + comprising

culturing in a first culture medium CD3 + CD4 + CD25 − T cells in the presence of a TCRαβ cell activator and adding regulatory T cell differentiation agents to the first culture medium, wherein said regulatory T cell differentiation agents consist of:

i) PGE2,

ii) TGFβ (Transforming growth factor beta),

iii) rapamycin and

iv) IL-2,

for at least 5 days, and

expanding in a second culture medium the MHCII restricted CD4 + Foxp3 + regulatory T cells in the presence of the TCRαβ cell activator and adding regulatory T cell differentiation agents to the second culture medium, wherein said regulatory T cell differentiation agents consist of:

i) PGE2,

ii) TGFβ (Transforming growth factor beta),

iii), rapamycin and

iv) IL-2,

for at least 5 days;

wherein

the TCRαβ cell activator is a polyclonal TCR αβ cell activator or an antigen-specific TCRαβ cell activator, with the caveat that the TCRαβ cell activator is not tolerogenic dendritic cells (DCs) pulsed with at least one self-peptide antigen;

and wherein said expanding step produces MHCII restricted CD4 + Foxp3 + regulatory T cells that remain stable when placed in inflammatory conditions.

2. The method according to claim 1 , wherein the polyclonal TCRαβ cell activator is an anti-CD3 antibody or an anti-TCRαβ antibody.

3. The method of claim 1 , wherein the MHCII restricted CD4 + Foxp3 + regulatory T cells produced in said expanding step lack IL-1R1 expression.