To optimize the protocol for the detection of intracellular cytokines within activated epitope-specific CD4+ T cells, we tested different procedures in splenocytes from mice parenterally immunized with two different antigens, the chimeric TB vaccine antigen H56 or OVA, combined with the liposome adjuvant CAF01, 5 days after the booster immunization. The gene discussed is CD4; the disease is tuberculosis.