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Pore formation, polymerization, hemolytic and leukotoxic effects of a new Enterobacter cloacae toxin neutralized by antiserum.
Microbiological Research ( IF 6.1 ) Pub Date : 2005-05-11 , DOI: 10.1016/j.micres.2005.01.005
María Gabriela Paraje 1 , Alberto Jorge Eraso , Inés Albesa
Affiliation  

A new toxin of Enterobacter cloacae was purified and studied by SDS-PAGE electrophoresis with the purpose of investigating its ability to generate polymers and their molecular mass. Monomer of 13.3 kDa and structures of multimeric mass were detected. The toxin of 66 kDa was the most abundant form of toxin. This polymer and the monomer were selected to examine blood cells damage. Membrane pores caused by both toxin forms seemed to be of similar dimension (estimated in 3.6 nm by experiments with osmotic protectors) and were able to lyse erythrocytes and leukocytes. The results obtained indicate that polymerization and pore formation are involved in the molecular events that participate in the cytotoxic effects of E. cloacae toxin. Immunization of rabbits with 13.3kDa toxin generated antibody response capable of inhibiting oxidative stress as well as hemolytic and leukotoxic effects. Immunoblotting indicated that monomer and polymer reacted with antihemolysin serum. The importance of E. cloacae toxin "in vivo" was studied in animals by means of assays performed in peritoneum of rats, inoculated with the hemolytic strain (C1) and a non-hemolytic variant (C4). Both strains stimulated infiltration of leukocytes in peritoneum, but C1 caused cell death and lysis wheras assays with C4 maintained the viability of leukocytes even within 5 h after extraction of samples.

中文翻译:

抗血清中和的新型阴沟肠杆菌毒素的毛孔形成,聚合,溶血和白细胞毒性作用。

阴沟肠杆菌的一种新毒素被纯化并通过SDS-PAGE电泳进行研究,目的是研究其产生聚合物的能力及其分子量。检测到13.3 kDa的单体和多聚体质量的结构。66 kDa的毒素是最丰富的毒素形式。选择该聚合物和单体以检查血细胞损伤。由两种毒素形式引起的膜孔似乎具有相似的尺寸(通过渗透性保护剂的实验在3.6 nm处估计),并且能够裂解红细胞和白细胞。获得的结果表明,聚合反应和孔形成与参与阴沟肠杆菌毒素的细胞毒性作用的分子事件有关。免疫兔子13。3kDa毒素产生的抗体反应能够抑制氧化应激以及溶血和白细胞毒性作用。免疫印迹表明单体和聚合物与抗溶血素血清反应。通过在大鼠腹膜中进行的试验(溶血菌株(C1)和非溶血性变体(C4)接种),在动物中研究了泄殖大肠杆菌毒素“体内”的重要性。两种菌株均刺激白细胞渗入腹膜,但C1引起细胞死亡,C4裂解裂解试验甚至在提取样品后5小时内仍保持了白细胞的活力。通过在大鼠腹膜中进行的试验对动物进行了研究,该试验接种了溶血菌株(C1)和非溶血性变体(C4)。两种菌株均刺激白细胞渗入腹膜,但C1引起细胞死亡,C4裂解裂解试验甚至在提取样品后5 h内仍保持了白细胞的活力。通过在大鼠腹膜中进行的试验对动物进行了研究,该试验接种了溶血菌株(C1)和非溶血性变体(C4)。两种菌株均刺激白细胞渗入腹膜,但C1引起细胞死亡,C4裂解裂解试验甚至在提取样品后5 h内仍保持了白细胞的活力。
更新日期:2019-11-01
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