Miltenyi, 130-094-299, CD4+ Recent Thymic Emigrant Isolation Kit, human

Background information
RTEs are a subpopulation of naive T cells characterized by their content of T cell receptor excision circles (TRECs).
CD31 (platelet endothelial cell adhesion molecule-1, PECAM-1) has been described as a marker to identify the CD4+ RTEs among the naive CD4+ T cell population, as its expression correlates with the TREC content of CD4+ T cells1. CD4+ T cells lacking CD31 expression harbor little or no TRECs, suggesting that they already proliferated in the periphery.
Therefore, the magnetic isolation of CD4+ RTEs is possible by direct isolation of CD31+ cells from pre-enriched, untouched naive CD4+ T cells.

Detailed separation procedure
The isolation is performed in a two-step procedure. First, non-CD4+ and memory CD4+ T cells are indirectly magnetically labeled with a cocktail of biotinylated antibodies and Anti-Biotin MicroBeads. The labeled cells are subsequently depleted over a MACS® Column. In the second step, the flow-through fraction of pre-enriched naive CD4+ T cells is labeled with CD31 MicroBeads for subsequent positive selection of CD4+CD45RA+CD31+ RTEs.

Columns
For the first magnetic separation (depletion): LS or autoMACS® Columns. For the second magnetic separation (positive selection): MS or autoMACS Columns.

Overview
The CD4+ Recent Thymic Emigrant Isolation Kit was developed for the isolation of CD4+CD45RA+CD31+ recent thymic emigrants (RTE) cells from human PBMCs in a two-step procedure.