Resting human CD4+T cell isolation from TOM. (Top left) Flow cytometric analysis of cells pooled from the different tissues of a TOM prior to magnetic negative selection showed the presence of both CD4+ and CD4neg cells. (Bottom left) Prior to negative selection CD4+ T cells expressed various levels of CD25 and HLA-DR. (Top right) After isolation 99% of the cells obtained were CD4+. (Bottom right) Consistent with a resting phenotype, isolated cells were CD3+CD4+ and did not express CD25 or HLA-DR. Gating strategy: (Top) live→hCD45+→hCD3+. (Bottom) live→hCD45+→hCD3+→CD4+.