Yersinia pestis Δ ail Mutants Are Not Susceptible to Human Complement Bactericidal Activity in the Flea

• Published in: Applied and environmental microbiology Vol. 89; no. 2; p. e0124422
• Main Authors: Kolodziejek, Anna M, Bearden, Scott W, Maes, Sarah, Montenieri, John M, Gage, Kenneth L, Hovde, Carolyn J, Minnich, Scott A
• Format: Journal Article
• Language: English
• Published: United States American Society for Microbiology 28.02.2023
• Subjects: Animals; Anti-Bacterial Agents - metabolism; Bacterial Outer Membrane Proteins - metabolism; Bactericidal activity; Colonization; Complement; Complement C3b - metabolism; Complement C3b - pharmacology; Complement component C3b; Environmental Microbiology; Host-Microbial Interactions; Humans; Lipooligosaccharides; Mammals; Mice; Mutants; Plague; Plague - microbiology; Rats; Siphonaptera - metabolism; Siphonaptera - microbiology; Spotlight Selection; Virulence factors; Virulence Factors - genetics; Virulence Factors - metabolism; Yersinia pestis - genetics; Yersinia pestis - metabolism; transmission; Ail; complement; plague; flea; LPS; Yersinia pestis; serum resistance
• ISSN: 0099-2240; 1098-5336
• DOI: 10.1128/aem.01244-22

Ail confers serum resistance in humans and is a critical virulence factor of Y. pestis, the causative agent of plague. Here, the contribution of Ail for Y. pestis survival in the flea vector was examined. Rat or human but not mouse sera were bactericidal against a Y. pestis Δ mutant at 28°C . Complement components deposited rapidly on the Y. pestis surface as measured by immunofluorescent microscopy. Ail reduced the amount of active C3b on the Y. pestis surface. Human sera retained bactericidal activity against a Y. pestis Δ mutant in the presence of mouse sera. However, in the flea vector, the serum protective properties of Ail were not required. Flea colonization studies using murine sera and Y. pestis KIM6 wild type, a Δ mutant, and the Δ control showed no differences in bacterial prevalence or numbers during the early stage of flea colonization. Similarly, flea studies with human blood showed Ail was not required for serum resistance. Finally, a variant of Ail (Ail ) from a human serum-sensitive Y. pestis subsp. microtus bv. Caucasica 1146 conferred resistance to human complement when expressed in the Y. pestis KIM6 Δ mutant. This indicated that Ail activity was somehow blocked, most likely by lipooligosaccharide, in this serum sensitive strain. This work contributes to our understanding of how highly virulent Y. pestis evolved from its innocuous enteric predecessor. Among identified virulence factors is the attachment invasion locus protein, Ail, that is required to protect Y. pestis from serum complement in all mammals tested except mice. Murine sera is not bactericidal. In this study, we asked, is bactericidal sera from humans active in Y. pestis colonized fleas? We found it was not. The importance of this observation is that it identifies a protective niche for the growth of serum sensitive and nonsensitive Y. pestis strains.