inlF Enhances Listeria monocytogenes Early-Stage Infection by Inhibiting the Inflammatory Response.

The internalin family proteins, which carry the leucine repeat region structural motif, play diverse roles in Listeria monocytogenes (Lm) infection and pathogenesis. Although Internalin F, encoded by inlF, was identified more than 20 years ago, its role in the Lm anti-inflammatory response remains unknown. Lm serotype 4b isolates are associated with the majority of listeriosis outbreaks, but the function of InlF in these strains is not fully understood. In this study, we aimed to elucidate the role of inlF in modulating the inflammatory response and pathogenesis of the 4b strain Lm NTSN. Strikingly, although inlF was highly expressed at the transcriptional level during infection of five non-phagocytic cell types, it was not involved in adherence or invasion. Conversely, inlF did contributed to Lm adhesion and invasion of macrophages, and dramatically suppressed the expression of pro-inflammatory cytokines interleukin (IL)-1ß and tumor necrosis factor (TNF-α). Consistent with the in vitro results, during Lm infection mice, inlF significantly inhibited the expression of IL-1ß and IL-6 in the spleen, as well as IL-1ß, IL-6, and TNF-α in the liver. More importantly, inlF contributed to Lm colonization in the spleen, liver, and ileum during the early stage of mouse infection via intragastric administration, inducing severe inflammatory injury and histopathologic changes in the late stage. To our knowledge, this is the first report to demonstrate that inlF mediates the inhibition of the pro-inflammatory response and contributes to the colonization and survival of Lm during the early stage of infection in mice. Our research partly explains the high pathogenicity of serovar 4b strains and will lead to new insights into the pathogenesis and immune evasion of Lm.

A hybrid sub-lineage of Listeria monocytogenes comprising hypervirulent isolates.

The foodborne pathogen Listeria monocytogenes (Lm) is a highly heterogeneous species and currently comprises of 4 evolutionarily distinct lineages. Here, we characterize isolates from severe ovine listeriosis outbreaks that represent a hybrid sub-lineage of the major lineage II (HSL-II) and serotype 4h. HSL-II isolates are highly virulent and exhibit higher organ colonization capacities than well-characterized hypervirulent strains of Lm in an orogastric mouse infection model. The isolates harbour both the Lm Pathogenicity Island (LIPI)-1 and a truncated LIPI-2 locus, encoding sphingomyelinase (SmcL), a virulence factor required for invasion and bacterial translocation from the gut, and other non-contiguous chromosomal segments from another pathogenic species, L. ivanovii. HSL-II isolates exhibit a unique wall teichoic acid (WTA) structure essential for resistance to antimicrobial peptides, bacterial invasion and virulence. The discovery of isolates harbouring pan-species virulence genes of the genus Listeria warrants global efforts to identify further hypervirulent lineages of Lm.

An essential role for hfq involved in biofilm formation and virulence in serotype 4b Listeria monocytogenes.

Regulator factor Hfq has been widely detected among both Gram-positive and Gram-negative bacteria; however, its role in Gram-positive bacteria is less well established and varies among species. In Listeria monocytogenes (Lm), an organism able to adapt to a range of environments and live both saprobiotic and parasitic lifestyles, the role of Hfq is not fully understood. Serotype 4b Listeria monocytogenes strains associated with the majority of listeriosis outbreak, while the function of hfq in serotype 4b strains still not referenced. Here, we constructed hfq deletion and reversion mutants of serotype 4b Lm NTSN and analysed the biological characteristics both in vitro and in vivo. The deletion of hfq resulted in a growth deficiency in medium containing 4.5% ethanol or 1% Triton X-100, and the growth of the mutant was significantly reduced at 4 °C. Furthermore, the hfq deletion dramatically decreased biofilm formation in BHI medium and gastric fluid medium, and reduced the invasion and replication rate into the Caco-2BBe cells and RAW264.7 cells. However, complementation restored the wild-type phenotype. Importantly, mouse infection experiments demonstrated that hfq played a more important role in the colonisation and virulence in serotype 4b strain Lm NTSN than in the serotype 1/2a strain Lm EGDe. Taken together, these results demonstrated that hfq is a novel factor associated with biofilm formation, and plays an essential role in the stress response and pathogenisis in serotype 4b strain Lm NTSN. Our data provide the basis for further research into the function of Hfq in serotype 4b Listeria monocytogenes.

Geographical and longitudinal analysis of Listeria monocytogenes genetic diversity reveals its correlation with virulence and unique evolution.

Listeria monocytogenes is one of the most important foodborne pathogens causing severe diseases with a mortality rate of 24%. However, the genetic diversity and evolution of L. monocytogenes, particularly at the worldwide level, are poorly defined. In this study, we performed multilocus sequence typing (MLST) and multi virulence locus sequence typing (MVLST) for 86 L. monocytogenes strains derived from 8 countries from 1926 to 2012 in order to better understand the molecular evolution and genetic characteristics of this pathogen. A total of 13 clonal complexes (CCs) were detected, of which CC1, CC2, CC3, CC7, CC9, CC4 are the most prevalent. Notably, polymorphism of housekeeping genes of isolates belong to CC1 (STs = 47) increased more rapidly over the time. MLST-based phylogenetic analysis showed that serotype 1/2b and 4b strains had an "interval-type" evolution pattern, while serotype 1/2a and 1/2c strains had a "progressive-type" evolution pattern. Furthermore, strains from temporally and geographically unrelated outbreaks in different countries were clustered in the same subgroup of phylogenetic tree, indicating that that L. monocytogenes developed highly similar virulence genes and genetic characteristics to adaptation in a special ecological niche. Interestingly, there was a high correlation between the population structure of MVLST and MLST among the isolates of cluster IA corresponding to CC1, CC2, CC4 and CC6 that had the highest potential to cause listeriosis outbreaks, strengthening that surveillance of these CCs is important for prevention of listeriosis. The present study offers insights into the internal relationships between the population structure, distribution and pathogenicity of L. monocytogenes.