Abstract
Mycobacterium bovis bacillus Calmette-Guérin (BCG) efficacy as an immunotherapy tool can be influenced by the genetic background or immune status of the treated population and by the BCG substrain used. BCG comprises several substrains with genetic differences that elicit diverse phenotypic characteristics. Moreover, modifications of phenotypic characteristics can be influenced by culture conditions. However, several culture media formulations are used worldwide to produce BCG. To elucidate the influence of growth conditions on BCG characteristics, five different substrains were grown on two culture media, and the lipidic profile and physico-chemical properties were evaluated. Our results show that each BCG substrain displays a variety of lipidic profiles on the outermost surface depending on the growth conditions. These modifications lead to a breadth of hydrophobicity patterns and a different ability to reduce neutral red dye within the same BCG substrain, suggesting the influence of BCG growth conditions on the interaction between BCG cells and host cells.
Highlights
One hundred years after being developed, Mycobacterium bovis bacillus CalmetteGuerin (BCG) is still the preferred prophylactic option for preventing tuberculosis (TB)worldwide
We evaluated the presence of lipids such as phthiocerol dimycocerosates (PDIMs) or phenol glycolipid (PGL), which are distinctively expressed among BCG substrains [7,8], physicochemical properties such as hydrophobicity, and the neutral red reducing power of five different BCG substrains grown on two Sauton formulations currently used for BCG production
As shown by thin-layer chromatography (TLC) (Figure 1A,B and Table 1), BCG-Pasteur and BCG-Phipps, which synthesize PDIM and PGL when cultured in 7H10 [8], produced these lipids when grown on A60
Summary
One hundred years after being developed, Mycobacterium bovis bacillus CalmetteGuerin (BCG) is still the preferred prophylactic option for preventing tuberculosis (TB)worldwide. BCG vaccine effectiveness appears to be influenced by several factors including the genetic background and/or immune status of the vaccinated population, BCG substrain used as a vaccine, interference with environmental nontuberculous mycobacteria, etc. To understand the variability of efficacy, a comparison of BCG vaccines needs to be performed using exactly the same parameters through in vitro, ex vivo, and in vivo experiments and in clinical trials to obtain reliable and comparable results. BCG vaccines comprise several substrains that present genetic differences [1] and, different phenotypic characteristics. Modifications to phenotypic characteristics can be influenced by culture conditions [2]. Mycobacteria are prone to lose the ability to synthesize cell wall lipids, which are costly and unnecessary in vitro
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