T denticola

Bacterial strains and plasmids used in this study are shown in Table 1. A. actinomycetemcomitans (ATCC 43718), F. nucleatum subsp. nucleatum (ATCC 25586), P. gingivalis (ATCC 33277), T. forsythia (ATCC 43037), and T. denticola (ATCC 33521) were obtained from American Type Culture Collection (Manassas, VA, USA). Bacterial cultures were grown under anaerobic growth conditions at 37 °C using an anaerobic growth chamber AS‐580 (Anaerobe Systems, Morgan Hill, CA, USA) supplied with anaerobic gas (90% N₂, 5% H₂, and 5% CO₂). F. nucleatum and P. gingivalis were cultured in brain heart infusion, T. forsythia was cultured using brain heart infusion supplemented with 2.5 mM N‐acetylmuramic acid (Sigma, St. Louis, MO, USA). A. actinomycetemcomitans and T. denticola were cultured in MTGE‐anaerobic enrichment broth (Anaerobe Systems).

Anaerobic bacteria T. denticola (ATCC 35405), P. gingivalis (ATCC 33277) and F. nucleatum (ATCC 25586) were purchased from ATCC. T. denticola, P. gingivalis and F. nucleatum were grown as described previously at 37°C under oxygen deprived anaerobic conditions in New Oral Spirochete Medium or in Brain-heart Infusion (BHI) broth supplemented with hemin (5 μg ml^-1) and vitamin K (1 μg ml^-1) [143 , 144 (link)]. Purity of spirochete cultures was confirmed by dark field microscopy prior to use in experiments. Oral commensal bacteria S. gordonii (ATCC 10558), S. salivarius (ATCC 13419) and V. parvula (ATCC 10790) were grown overnight at 37°C under anaerobic conditions in BHI broth then harvested for subsequent assays. Gram staining followed by microscopic evaluation and colony morphology on Mitis Salivarius plates were used to confirm purity of cultures. T.denticola lipoologosaccharide (Td-LOS) was a gift from Daniel Grenier, Universite Laval, Quebec, Canada, P.gingivalis lipopolysaccharide (Pg-LPS) and F.nucleatum lipopolysaccharide (Fn-LPS) were provided by Richard P. Darveau, University of Washington, Seattle and V. parvula lipopolysaccharide (Vp-LPS) was provided by Christopher Fenno, University of Michigan, Ann Arbor, MI.