Analysis of production levels of InlA and InlB invasion factors in Listeria monocytogenes isolates collected in the Russian Federation

Abstract

Background. Listeria monocytogenes is characterized by the presence of epidemic hypervirulent clones. A key feature of L. monocytogenes is its capacity to invade non-professional phagocytic cells. Hypervirulent clones are strongly associated with the increased production and/or the presence of certain isoforms of invasion factors InlA and InlB. The purpose of the study is to create a test system for InlA and InlB detection and to measure the InlA and InlB production levels in L. monocytogenes isolates belonging to clonal groups with different virulence potential. Materials and methods. The study was performed using 32 L. monocytogenes strains belonging to epidemic clones ECII, ECIV, ECVII (clonal complexes CC1, CC2, CC7) and hypovirulent clonal complex CC9. Sequencing of inlA and inlB genes was performed. The indirect enzyme-linked immunosorbent assay was used to analyze the production levels of InlA and InlB proteins. Results. The variability of InlA was revealed among strains belonging to the same clonal complex: 3 InlA isoforms were identified among strains belonging to CC7; out of 8 strains belonging to CC9, one strain had a stop codon in the inlA gene, leading to the loss of function of the InlA protein. The differences between inlB alleles correlated with the specificity of strains belonging to a certain clonal complex. Differences in production levels of invasion factors were measured. In strains belonging to CC9, the InlA production level was 2.5 times as low compared to strains belonging to CC1, CC2, and CC7. In strains belonging to phylogenetically related CC1 and CC2, the InlB production level was on average 4 times as high compared to strains belonging to CC7 and CC9. Conclusion. The obtained results confirm the variability of major invasion factors both among clonal complexes and strains of the same complex. The increased production of invasion factors InlA and InlB correlates with the potential virulence of strains.