Aggregatibacter actinomycetemcomitans Outer Membrane Proteins 29 and 29 Paralogue Induce Evasion of Immune Response

Aggregatibacter actinomycetemcomitans (Aa) is abundant within the microbial dysbiotic community of some patients with periodontitis. Aa outer membrane protein 29 (OMP29), a member of the OMPA family, mediates the invasion of Aa to gingival epithelial cells (GECs). This study evaluated the effect of OMP29 and its paralogue OMP29(par) on the response of GECs to Aa. The omp29 or/and omp29(par) deletion mutants Aa Delta 29, Aa Delta 29P, and Aa Delta 29 Delta 29P were constructed, and recombinant Aa OMP29(His) was obtained. Microarray analysis and the evaluation of cxcl-8 gene expression were performed to examine the response of GECs line OBA-09 to Aa and its mutants. The expression of cxcl-8 and its product CXCL-8 was examined in LPS-stimulated OBA-09 cells with Aa OMP29(His). Proteomics analysis showed that the deletion of omp29 led to overexpression of both OMP29(par) and another membrane protein OMP39, the expression of which was further increased in Aa Delta 29 Delta 29P. OBA-09 cells challenged with Aa Delta 29 Delta 29P exhibited a higher expression of cxcl-8 in comparison to wildtype Aa strain AaD7S or single-deletion mutants Aa Delta 29 or Aa Delta 29P. LPS-stimulated OBA-09 cells challenged with Aa OMP29(His) showed reduced expressions of cxcl-8 and its product CXCL-8. OBA-09 cells challenged with Aa Delta 29 Delta 29P in comparison to Aa strain AaD7S resulted in higher expressions of genes involved in apoptosis and inflammatory response such as bcl2, birc3, casp3, c3, ep300, fas, fosb, grb2, il-1 alpha, il-1 beta, il-6, cxcl-8, nr3c1, prkcq, socs3, and tnfrsf1 beta and reduced expressions of cd74, crp, faslg, tlr1, and vcam1. The results suggested a novel strategy of Aa, mediated by OMP29 and OMP29(par), to evade host immune response by inhibiting CXCL-8 expression and modulating the genes involved in apoptosis and inflammatory response in GECs. Pending further confirmation, the strategy might interfere with the recruitment of neutrophils and dampen the host inflammatory response, leading to a more permissive subgingival niche for bacterial growth.