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Background  In vivo kinetics and frequencies of epitope-specific CD4 T cells in lymphoid compartments during  M. tuberculosis  infection and their resting memory pool after BCG vaccination remain unknown.    Methodology/Findings  Macaque DR*W201 tetramer loaded with Ag85B peptide 65 was developed to directly measure epitope-specific CD4 T cells in blood and tissues form macaques after  M. tuberculosis  infection or BCG vaccination via direct staining and tetramer-enriched approach. The tetramer-based enrichment approach showed that P65 epitope-specific CD4 T cells emerged at mean frequencies of ∼500 and ∼4500 per 10 7  PBL at days 28 and 42, respectively, and at day 63 increased further to ∼22,000/10 7  PBL after  M. tuberculosis  infection. Direct tetramer staining showed that the tetramer-bound P65-specific T cells constituted about 0.2–0.3% of CD4 T cells in PBL, lymph nodes, spleens, and lungs at day 63 post-infection. 10-fold expansion of these tetramer-bound epitope-specific CD4 T cells was seen after the P65 peptide stimulation of PBL and tissue lymphocytes. The tetramer-based enrichment approach detected BCG-elicited resting memory P65-specific CD4 T cells at a mean frequency of 2,700 per 10 7  PBL.    Significance  Our work represents the first elucidation of in vivo kinetics and frequencies for tetramer-bound epitope-specific CD4 T cells in the blood, lymphoid tissues and lungs over times after  M. tuberculosis  infection, and BCG immunization.

DR*W201/P65 Tetramer Visualization of Epitope-Specific CD4 T-Cell during M. tuberculosis Infection and Its Resting Memory Pool after BCG Vaccination