No currently licensed vaccine reliably prevents pulmonary tuberculosis, a leading cause of infectious disease mortality with a growing rate of extensive drug resistance. Developing effective new vaccines will likely require identifying Mycobacterium tuberculosis (Mtb)-derived antigens presented on MHC class II (MHC-II) by infected phagocytes that CD4+ T cells can recognize to mediate protective immunity. To prime T cells capable of recognizing infected cells, vaccines will also need to deliver antigens to appropriate processing pathways. Here, we use mass spectrometry (MS) to identify 27 Mtb-derived MHC-II peptides presented by infected human dendritic cells, derived from 13 source proteins. Using targeted quantitative MS, we assess and optimize the ability of vaccines to drive presentation of MHC-II epitopes that match those presented by Mtb-infected cells, including the existing Bacillus Calmette Guerin (BCG) vaccine and new mRNA-encoded immunogens. Our results identify potential antigenic targets and immunogen design principles for tuberculosis vaccine development.