ABSTRACT
This project has 3 aims: 1) Determine if different BCG produce variable amounts of AM and DG. 2) Determine if variations in AM and DGcorrelate with differential BCG activation of host innate and adaptive immunity. 3) Determine if modifications to culture media affects BCGAM and DG levels. In this reporting period, CP analysis of additional BCG strains (including the six strains initially proposed) led to theidentification of strains with significant defects in AM and DG production. The variations are more pronounced when BCG are cultured inSautons medium. Culturing in 7H9 medium results in increased AM production consistently for all BCG strains, while DG production isvariably increased among different BCG strains. Although variations in CP among BCG exist when cultured in 7H9, infection of macrophagecells with BCG cultured under the same condition induces pro-inflammatory responses that do not correlate with CP composition. As such,the relationship between CP composition and induction of innate immune responses may be more complex than initially perceived. We havealso discovered that culturing under low glycerol conditions uniformly increases AM production but variably increases DG production amongthe twelve BCG strains. Inclusion of additional exogenous cues to low glycerol, specifically lowered phosphate and SHX enhances AM andDG production in only some BCG strains. Collectively, the results of our investigation indicates that CP production is far more variable inBCG than previously appreciated and could certainly impact their effectiveness as vaccines. We have also discovered that CP production isnegatively regulated by glycerol as well as by low phosphate and SHX in some but not all BCG strains.