Warren H S, Glennon M L, Wainwright N, Amato S F, Black K M, Kirsch S J, Riveau G R, Whyte R I, Zapol W M, Novitsky T J
Department of Pediatrics, Massachusetts General Hospital, Boston.
Infect Immun. 1992 Jun;60(6):2506-13. doi: 10.1128/iai.60.6.2506-2513.1992.
In order to examine the ability of Limulus antilipopolysaccharide factor (LALF) to bind lipopolysaccharide (LPS), we purified LALF to homogeneity from Limulus amoebocyte lysate and coupled it covalently to agarose beads. LALF-coupled beads captured more tritiated LPS from rough and smooth strains of gram-negative bacteria than did control human serum albumin-coupled beads. Unlabeled homologous and heterologous LPS competed for the binding of 3H-LPS to LALF-coupled beads. LALF bound LPS in a dose-dependent manner as assessed by the precipitation of LPS-LALF complexes with 50% saturated ammonium sulfate. We also studied the ability of LALF to neutralize LPS. LPS preincubated with LALF was less mitogenic for murine splenocytes, was less pyrogenic in the rabbit fever assay, was less lethal in mice which had been sensitized to LPS with actinomycin D, and induced less fever, neutropenia, and pulmonary hypertension when infused into sheep. Our findings extend prior studies which suggested that LALF binds to and neutralizes LPS from multiple strains of gram-negative bacteria.
为了检测鲎抗脂多糖因子（LALF）结合脂多糖（LPS）的能力，我们从鲎血细胞裂解物中纯化出均一的LALF，并将其共价偶联到琼脂糖珠上。与对照人血清白蛋白偶联的珠子相比，LALF偶联的珠子从革兰氏阴性菌的粗糙型和平滑型菌株中捕获的氚标记LPS更多。未标记的同源和异源LPS竞争3H-LPS与LALF偶联珠子的结合。通过用50%饱和硫酸铵沉淀LPS-LALF复合物评估，LALF以剂量依赖方式结合LPS。我们还研究了LALF中和LPS的能力。与LALF预孵育的LPS对小鼠脾细胞的促有丝分裂作用较弱，在兔热原试验中致热作用较弱，在经放线菌素D致敏的小鼠中致死作用较弱，注入绵羊体内时引起的发热、中性粒细胞减少和肺动脉高压也较少。我们的研究结果扩展了先前的研究，这些研究表明LALF能结合并中和多种革兰氏阴性菌的LPS。
