Stevens R H, Sela M N, McArthur W P, Nowotny A, Hammond B F
Infect Immun. 1980 Jan;27(1):246-54. doi: 10.1128/iai.27.1.246-254.1980.
An endotoxin was isolated from Capnocytophaga sputigena strain 4 by a modification of the hot phenol-water method. The extraction procedure yielded a lipopolysaccharide which accounted for approximately 1.5% of the dry weight of the cells. The material was composed of 18.6% lipid (as C(15) fatty acid), 46.5% neutral sugar including 9.6% hexose, 18.3% 6-deoxy sugar, 1.0% 2-keto-3-deoxy sugar, and 4.8% heptose. Hexosamine, protein, and phosphorus were found in quantities amounting to 9.0, 2.9, and 2.0% of the dry weight, respectively. No pentose or nucleic acid was detected. Acid hydrolysis resulted in the release of the constituent sugars and the formation of an insoluble precipitate. The lipopolysaccharide was tested for numerous biological activities characteristic of endotoxins. The pyrogenicity was relatively low; the fever index 40 was 17 mug, and 10 mug was required to give the characteristic biphasic fever response. The toxicity of the extract was very low, with a 50% chicken embryo lethal dose of 15.6 mug and a 50% mouse embryo lethal dose of greater than 8 mg. Similarly, the C. sputigena endotoxin had modest effects on leukocytes when compared with endotoxin standards from other organisms. The extract exhibited little or no mitogenicity when tested on mouse spleen lymphocytes. It was not toxic to human peripheral polymorphonuclear leukocytes and caused the release of only a small (13%) portion of lysosomal enzymes. Although the C. sputigena lipopolysaccharide caused significant activation of mouse peritoneal macrophages, the dose required was twice that of an Escherichia coli endotoxic standard. However, the Limulus amoebocyte lysate clotting activity of the lipopolysaccharide was comparable to that of an Serratia marcescens lipopolysaccharide standard, and passive hemagglutination tests revealed that 1 mug of the lipopolysaccharide was capable of sensitizing 1 ml of a 2% sheep erythrocyte suspension for agglutination with an antiserum prepared against C. sputigena whole cells.
通过改良热酚 - 水法从生痰二氧化碳嗜纤维菌菌株4中分离出一种内毒素。提取过程得到一种脂多糖，其占细胞干重的约1.5%。该物质由18.6%的脂质（以C(15)脂肪酸计）、46.5%的中性糖组成，其中包括9.6%的己糖、18.3%的6 - 脱氧糖、1.0%的2 - 酮 - 3 - 脱氧糖和4.8%的庚糖。己糖胺、蛋白质和磷的含量分别占干重的9.0%、2.9%和2.0%。未检测到戊糖或核酸。酸水解导致组成糖的释放并形成不溶性沉淀。对该脂多糖进行了多种内毒素特有的生物活性测试。其致热原性相对较低；发热指数40时为17微克，产生典型双相发热反应需要10微克。提取物的毒性非常低，鸡胚半数致死剂量为15.6微克，小鼠胚半数致死剂量大于8毫克。同样，与生痰二氧化碳嗜纤维菌内毒素相比，来自其他生物体的内毒素标准品对白细胞的影响更大。在小鼠脾淋巴细胞上进行测试时，该提取物几乎没有或没有促有丝分裂活性。它对人外周多形核白细胞无毒，仅导致溶酶体酶释放一小部分（13%）。尽管生痰二氧化碳嗜纤维菌脂多糖可显著激活小鼠腹腔巨噬细胞，但所需剂量是大肠杆菌内毒素标准品的两倍。然而，该脂多糖的鲎试剂凝血活性与粘质沙雷氏菌脂多糖标准品相当，被动血凝试验表明，1微克脂多糖能够使1毫升2%的绵羊红细胞悬液对用生痰二氧化碳嗜纤维菌全细胞制备的抗血清发生凝集敏感。
