Abstract
Objectives: Endotoxin (ET) is a structural molecule of the Gram-negative bacilli extracellular membrance, which activates targeted cells including macrophages and neutrophils, and causes septic shock. But it is known that the conventional ET measurement method has many problems; for example, a discrepancy between plasma ET concentration and clinical manifestation in the septic patients has been reported. The purpose of this study was to evaluate the usefulness of a newly developed method (Endotoxin Activity Assay (EAA)) to measure the ET activity (EA) in patients under sepsis compared with the prior method of the limulus amebocyte lysate (LAL) assay and explore the association between EA levels and patients’ severity. Method: We measured the EA levels in 40 patients (aged 63.5 +/－ 17.7 years) admitted to the ICU. EA level was measured using a chemiluminometer (Autolumat LB953; EG & Berthold). Patients were divided into 5 groups: 1) control group; 2) systemic inflammatory response syndrome (SIRS) group; 3) sepsis (SIRS and infection) group; 4) severe sepsis group and 5) septic shock group. We then compared the EA level between each group and control group. We made the statistical evaluation by unpaired t test and significant difference was p Results: The EA levels were significantly increased as sepsis severity rises. The measured EA levels were (0.18 +/－ 0.09), (0.33 +/－ 0.19), (0.39 +/－ 0.16), (0.65 +/－ 0.25) and (0.78 +/－ 0.34) in control, SIRS, sepsis, severe sepsis and septic shock groups, respectively. In the EA level measured by EAA, severe patients had a tendency to exceed the cutoff value. Conclusion: The EA levels were significantly correlated as patients’ severity rise. Measuring EA levels on admission to ICU may provide a mechanism to identify and target severe septic patients.
Highlights
Endotoxin (ET), a complex lipopolysaccharide (LPS) that is present in the cell walls of Gram-negative bacteria, is a potent trigger of innate immunity [1]
The patients were divided into 5 groups in according to patients’ general condition: 1) control group: 12 patients, those who were preoperative status; 2) systemic inflammatory response syndrome (SIRS) group: 17 patients, Systemic Inflammatory Response Syndrome (SIRS) was considered to be present at least 2 of 3 criteria were met: temperature above 38 ̊C or below 36 ̊C, heart rate of more than 90 beats/min, respiratory rate of more than 20 breasts/min or partial pressure of carbon dioxide of less than 32 mmHg, or white blood cell count above 12,000 mm3 or below 4000 mm3 [16]; 3) sepsis group: 14 patients, those who were diagnosed with sepsis, episodes of infection were diagnosed by microbiologic, laboratory, radiologic and operative data; 4) severe sepsis group: 32 patients and 5) septic shock group: 16 patients
All patients were divided into 5 groups: 1) control group; 2) SIRS group; 3) sepsis group; 4) severe sepsis group and 5) septic shock group based on their clinical findings
Summary
Endotoxin (ET), a complex lipopolysaccharide (LPS) that is present in the cell walls of Gram-negative bacteria, is a potent trigger of innate immunity [1]. Isolated and purified from the wall of several gram-negative bacteria, it has been used to investigate many aspects of the immuno-inflammatory response of sepsis through inoculation in the laboratory animal or in humans [2]. The commonly used method, the chromogenic limulus amebocyte lysate (LAL) assay, is based on the ability of ET to induce coagulation of hemolymph in the horseshoe crab, limulus polyphemus. The utility of this assay has been limited because of circulating inhibitors of the coagulation reaction [12]. A novel assay for ET, based on the ability of antigen-antibody complex to prime neutrophils and augmented respiratory burst re-
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