Lipopolysaccharides (LPS) of Oral Black-Pigmented Bacteria Induce Tumor Necrosis Factor Production by LPS-Refractory C3H/HeJ Macrophages in a Way Different from That ofSalmonellaLPS

Abstract

Enterobacteriaceae and oral black-pigmented bacteria (Porphyromonas gingivalis and Prevotella intermedia) are known to activate LPS-refractory C3H/HeJ macrophages. When contaminating proteins are removed from R-form LPS of Enterobacteriaceae by repurification, however, this ability is lost. In the present study, we investigated the capacity of LPS from P. gingivalis, P. intermedia, Salmonella minnesota, and Salmonella abortusequi to induce production of tumor necrosis factor (TNF) in gamma interferon-primed C3H/HeJ macrophages before and after repurification. P. abortusequi S-LPS was fractionated by centrifugal partition chromatography into two LPS forms: SL-LPS, having homologous long O-polysaccharide chains, and SS-LPS having short oligosaccharide chains. Prior to repurification, all LPS forms except SL-LPS induced TNF production in both C3H/HeJ and C3H/HeN macrophages. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis showed that repurification removed contaminating protein from the preparations, and repurified SS-LPS and S. minnesota Ra-LPS no longer stimulated TNF production in C3H/HeJ macrophages, although C3H/HeN macrophages remained responsive. In contrast, repurified oral bacterial LPS retained the capacity to induce TNF production in C3H/HeJ macrophages. Oral bacterial LPS preparations also were not antagonized by excess inactive, repurified SL-LPS; Ra-LPS; Rhodobacter sphaeroides lipid A, a competitive LPS antagonist, or paclitaxel, an LPS agonist, and they were comparatively resistant to polymyxin B treatment. Nevertheless, oral bacterial LPS was less toxic to D-galactosamine-treated C3H/HeN mice than was LPS from Salmonella. These findings indicate that the active molecule(s) and mode of action of LPS from P. gingivalis and P. intermedia are quite different from those of LPS from Salmonella.

Immunobiological activities of a chemically synthesized lipid A of Porphyromonas gingivalis

“…2 and [24]). We and other research groups have previously demonstrated that LPS and its natural lipid A from P. gingivalis were mitogenic for splenic MNC and B-cells from both of these murine strains [23,24,33,34]. The total protein content in P. gingivalis lipid A specimen was scarcely detectable by amino acid analysis, as described previously [35].…”

Structural significance of the acyl group at the C‐10 position and the A ring of the taxane core of paclitaxel for inducing nitric oxide and tumor necrosis factor production by murine macrophages

“…1B and 2). We recently suggested that LPS from oral black-pigmented bacteria induces TNF production by C3H/HeJ MP as well as C3H/ HeN MP in a manner di¡erent from that of Salmonella LPS [21]. Oral black-pigmented bacterial LPS and taxoid 5 may use other functional receptor(s) di¡erent from TLR 4, such as other TLR families [35].…”

“…The Re-chemotype LPS from Salmonella minnesota R595 LPS was kindly provided by Dr. K. Hisatsune, Josai University, Sakado, Saitama, Japan. The Ra-chemotype LPS from S. minnesota R60 LPS was purchased from Sigma Chemical Co., and it was used only in the experiments using C3H/HeJ MP, because C3H/HeJ MP could hardly respond to Re-chemotype LPS [21]. Murine recombinant interferon Q (IFNQ) was donated from Shionogi Pharmaceutical Co., Osaka, Japan.…”

Toll-like receptor 4-dependent recognition of structurally different forms of chemically synthesized lipid As of Porphyromonas gingivalis

“…It was also shown that P. gingivalis LPS is heterogeneous, with a greater number of lipid A species, such as tri-, tetra-, and/or pentaacylated lipid As [4,6]. Furthermore, P. gingivalis LPS and its lipid A were reported to activate cells from LPS-hyporesponsive C3H/HeJ mice, as well as those from LPS-responsive C3H/HeN mice [7,8], which was thought to be due to its unique structure. To confirm this structure-activity relationship, we synthesized a counterpart of P. gingivalis strain 381-type tri-acylated lipid A, compound PG-381-3FA, and found that it induced interleukin (IL)-6 and tumour necrosis factor (TNF)-a production in peritoneal macrophages from C3H/HeN mice, but not C3H/HeJ mice [9].…”