Host-Specific Adaptation of Legionella pneumophila to Single and Multiple Hosts

Legionella pneumophila is an endosymbiotic bacterial species able to infect and reproduce in various protist and human hosts. Upon entry into human lungs, they may infect lung macrophages, causing Legionnaires' disease (LD), an atypical pneumonia, using similar mechanisms as in their protozoan hosts, despite the 2 hosts being separated by a billion years of evolution. In this study, we used experimental evolution to identify genes conferring host specificity to L. pneumophila. To this end, we passaged L. pneumophila in 2 different hosts-Acanthamoeba castellanii and the human macrophage-like cells U937-separately and by switching between the hosts twice a week for a year. In total, we identified 1,518 mutations present in at least 5% of the population at the time of sampling. Forty-nine mutations were fixed in the 18 populations at the end of the experiment. Two interesting groups of mutations included (i) mutations in 4 different strain-specific genes involved in lipopolysaccharide (LPS) synthesis, found only in the lineages passaged with A. castellanii and (ii) mutations in the gene coding for LerC, a key regulator of protein effector expression, which was independently mutated in 6 lineages grown in presence of the macrophage cells. We propose that the mutations degrading the function of the regulator LerC improve the fitness of L. pneumophila in human-derived cells and that modifications in the LPS are beneficial for growth in A. castellanii. This study is a first step in further investigating determinants of host specificity in L. pneumophila.