Biochemical and immunobiological properties of lipopolysaccharide (LPS) from Bacteroides gingivalis and comparison with LPS from Escherichia coli

Abstract

Lipopolysaccharides (LPSs) were isolated from Bacteroides gingivalis and Escherichia coli by the phenolwater and butanol-water procedures. The phenol-water-extracted LPS from B. gingivalis 381 was composed of 46% carbohydrate, 23% hexosamine, 18% fatty acid, and 5% protein. The major component sugars of this preparation were glucose, glucosamine, rhamnose, galactose, galactosamine, and mannose, and their molecular ratio was 1:0.9:0.7:0.6:0.6:0.4, respectively. Neither heptose nor 2-keto-3-deoxyoctonate was detected. the butanol-water-extracted LPS from this strain was composed of 76% glucose, 7% fatty acid, and 13% protein, and it was associated with a number of polypeptides (13 to 56 kilodaltons). The main fatty acid of both LPS preparations was palmitic acid. It was found that biological activities of LPS from B. gingivalis were comparable to those of LPS from E. coli in terms of activation of the clotting enzyme of Limulus amebocyte lysate, mitogenicity, polyclonal B cell activation, and stimulation of interleukin 1 production in BALB/c mice. Furthermore, LPS-nonresponsive C3H/HeJ spleen cells were found to yield good mitogenic responses to both phenol-water-extracted LPS and butanol-water-extracted LPS from B. gingivalis or butanol-water-extracted LPS from E. coli. On the other hand, spleen cells from LPS-responsive C3H/HeN mice responded well to all these LPS preparations. * Corresponding author. MATERIALS AND METHODS Bacterial strains and growth conditions. B. gingivalis 381 (originally isolated by S. Sockransky, Forsyth Dental Center, Boston, Mass.) and E. coli strain K235 (19) were obtained from Y.

Chemical composition and immunobiological properties of lipopolysaccharide and lipid‐associated proteoglycan from Actinobacillus actinomycetemcomitans

“…Similar preparations were obtained from whole cells of E. eoli K235 by the phenolwater and butanol-water extractions and were designated as E. eoli LPS and LPG, respectively. The chemical composition and immunobiological activities of the E. coli preparations were reported recently by Koga et al (1985). These preparations were used in this study for comparison with those o[ A. actinomycetemcomitans.…”

“…The crude extract was treated with nuclease, washed extensively with pyrogen-free water by ultracentrifugation, and lyophilized. Furthermore, LPG was prepared from whole cells by the butanolwater extraction method as described previously by Koga et al (1985). These extracts were not treated with proteolytic enzymes.…”

Role of Platelet‐Actlvatlng‐Factor (PAF) on Cellular Responses after Stimulation with Leptospire Lipopolysaccharide

“…(ii) They suggested that the classic LPS inhibitor polymyxin B prevented the adherence induced by the enterobacterial LPS but not the adherence by leptospires. However, the mitogenicity for spleen cells was unaffected by polymyxin B when Porphyromonas gingivalis LPS (18) or leptospire LPS (unpublished data) was used. (iii) They described that a total lipid extract gave negative results.…”

Inhibitory effects of rabbit antisera on mitogenic activity of Actinobacillus actinomycetemcomitans lipopolysaccharide

“…LPS was extracted from lyophilized cells of A. actinomyceiemcomilans by the hot phenol-water procedure of Westphal & Jann (19). The crude extract was treated with nuclease, washed extensively with pyrogen-free water by ultracentrifugation, and lyophilized (20). These materials from A. actinomyceiemcomilans strains ATCC 29523, Y4 and NCTC 9710 are referred to as ATCC 29523 LPS, Y4 LPS, NCTC 9710 LPS, respectively.…”

“…Murine spleen cells from C3H/HeN and C3H/HeJ mice were suspended in RPMI 1640 medium (GIB-CO Laboratories, Grand Island, NY) supplemented with penicillin (100 U/ml), streptomycin (100 g/ml), and A'-2-hydroxyethyl-piperazine-A''-2cthanesulfonic acid (HEPES) buffer (pH 7.2, 15 mM), and adjusted to 5 x lO'/ml. Spleen cells (5 x 10^) were culturd in triplicate in a 96-well microculture plate in a total volume of 0.2 ml ofthe medium containing various amounts of LPS, hpid A or concanavalin A (ConA; Sigma) at 37 °C in a humidified 5% COj atmosphere (20). Cultures were pulsed with 0.25 fiG of ['HJthymidine (['H]TdR; 2 Ci/mmol; ICN Radiochemicals, Irvine, CA) during the final 6 h of a 48-h incubation.…”