Fig. 2

Detection of murine LN DCs Ag-presenting activity ex vivo. Suspensions of inguinal LN cells from C57BL/6 mice were stained with anti-CD11c MicroBeads and separated by autoMACS, resulting in a population of 94.7% CD11c⁺ cells (a). To investigate the dynamics of LN DC Ag-presenting activity, we harvested and sorted LN DCs from the inguinal LN at various time points groups after s.c. injection of mice (n = 6) with BCG or heat-killed BCG. Then, these cells were serially diluted and used to directly stimulate DE10 T-cell hybridomas. In vivo formation of Ag85A peptide-MHC complexes on LN DCs from mice injected with BCG (b) or heat-killed BCG (c) were estimated by measuring IL-2 production in DE10 T-cell hybridoma culture supernatants ex vivo. The experiment was repeated at least three times