title: Isolation, purification, and partial characterization of a lipopolysaccharide from Haemophilus pleuropneumoniae.
abstract: Lipopolysaccharide (LPS) from Haemophilus pleuropneumoniae 1536, serotype 2, was isolated and purified by a procedure designed to be equally satisfactory for both smooth- and rough-type LPS. The LPS yield was 53%. Analysis of the preparations revealed that protein, nucleic acid, and cellular phospholipid contamination was negligible (less than 0.1%). Analysis of the sugar content of the LPS by gas-liquid chromatography and colorimetric analysis revealed the presence of rhamnose, mannose, galactose, glucose, heptose, glucosamine, galactosamine, and 3-deoxy-D-manno-2-octulosonic acid. The heptose and glucose contents appeared to be unusually high. The fatty acids of the LPS consisted of a mixture of C14:0 and C16:0 in a ratio of about 4.5:1 (50% of the total) and 3-hydroxy C14:0. When used as a preparatory dose for the dermal Shwartzman reaction, as little as 10 micrograms of the LPS injected intradermally in rabbits produced reddening and swelling. After intravenous injection of a 100-micrograms LPS provoking dose, necrosis was observed at all intradermal injection sites. Limulus amebocyte lysate gelation was observed with an LPS concentration as low as 0.5 ng/ml. A typical biphasic fever response was noted in rabbits injected with as little as 0.25 ng of LPS per kg of body weight.
mesh_terms: D000818:Animals; D005227:Fatty Acids / Q000032:analysis; D006190:Haemophilus / Q000032:analysis*; D008070:Lipopolysaccharides / Q000032:analysis / Q000302:isolation & purification* / Q000633:toxicity; D010743:Phospholipids / Q000032:analysis; D011817:Rabbits; D013400:Sugar Acids / Q000032:analysis
chemical_list: D005227:Fatty Acids; D008070:Lipopolysaccharides; D010743:Phospholipids; D013400:Sugar Acids; C002532:2-keto-3-deoxyoctonate