Miltenyi Biotec, Gmbh

Marco Frentsch

Andreas Thiel

Anne Schönbrunn

Dan Robin Miller

Siegfried Kohler

Anne Schōnbrunn

Martin Rothe

The present invention relates to a method of identifying and separating non-regulatory T-cells (conventional T-cells) from a mixture comprising regulatory T-cells by using of the CD154 molecule (CD40 ligand) through depletion of CD154+ T-cells from the mixture or in combination with additional positive selection of Treg using markers that are specific for regulatory T-cells, such as for example, CD25, GITR, CTLA4 or markers which are specific for activated regulatory T-cells, such as, for example, CD137, 'latent TGF-beta (LAP)', GARP (LRRC32), CD121a/b, thereby generating a cell composition of activated Treg cells. The invention relates also to a kit comprising an antibody for detecting CD154 and at least one additional antibody for detecting markers for activated or non-activated regulatory T-cells. The antibodies can be coupled to a fluorescent dye or magnetic microparticles.

Method for the identification and separation of non-regulatory T-cells from a mixture of regulatory T-cells

The invention relates to the use of the 4-1BB receptor for identifying and/or separating specific regulatory Th cells after activation with an antigen, polyclonal regulatory Th cells after polyclonal activation and to a method for the identification and/or separation of regulatory Th cells, the regulatory T cells being detected on the basis of the expression of the 4-1BB receptor by an antibody, an antigen or ligands which are coupled to a fluorescent substance, haptenes or magnetic microparticles. The invention also relates to a kit comprising an antibody, an antigen or ligands for detecting the 4-1BB receptor for the identification and/or separation of a regulatory Th cell, the antibody being coupled to a fluorescent substance, haptenes or magnetic microparticles.

Use of the 4-1BB receptor for identifying and/or separating activated regulatory Th cells (Treg)

What is suggested herein is the use of CD154 for the detection and/or isolation of antigen-specific T lymphocytes and a method for the detection and/or isolation of antigen-specific T lymphocytes, wherein the suspension is contacted with a CD40/CD154 system inhibitor, intra- or extracellular determination of CD154 is effected, and the cells having CD154 are detected and/or isolated.

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Berlin, Germany

Marco Frentsch