Methods and materials for the generation of regulatory T cells
View Patent ↗Methods are disclosed for the generation of immunosuppressive regulatory T cells. The methods can include contacting a population of CD4+CD25− T cells with a T cell receptor (TCR)/CD3 activator, a TCR co-stimulator activator, and rapamycin. Kits for the generation of immunosuppressive regulatory T cells, methods of use, and cell populations are also disclosed.
1. A method of generating immunosuppressive regulatory T cells from a sample containing a mixed population of CD4+CD25+ and CD4+CD25− T cells, the method comprising:
(a) preparing a mixture comprising:
(i) the sample containing the mixed population of CD4+CD25+ and CD4+CD25− T cells,
(ii) an anti-CD3 antibody,
(iii) an anti-CD28 antibody,
(iv) interleukin 2, and
(v) rapamycin; and
(b) incubating the mixture prepared in (a) under conditions suitable for the selective expansion of CD4+CD25+FOXP3− T cells over other T cell subtypes,
wherein both the anti-CD3 antibody and the anti-CD28 antibody are immobilized on a solid phase, and
wherein interleukin 4 is not added to the mixture of (a).
2. The method of claim 1 , wherein the anti-CD3 antibody and the anti-CD28 antibody are immobilized on the same solid phase.
3. The method of claim 1 , wherein the anti-CD3 antibody and the anti-CD28 antibody are immobilized on different solid phases.
4. The method of claim 1 , wherein the solid phase is a bead.
5. The method of claim 1 , wherein the solid phase comprises glass, silica, latex, polymeric materials, plastic, tissue culture plastic, dextran, cellulose, polyethylene glycol, iron, or combinations thereof.
6. The method of claim 1 , wherein the solid phase comprises magnetic beads, superparamagnetic beads, or magnetizable beads.