A new method for removing endotoxin from plasma using hemocompatible affinity chromatography technology, applicable for extracorporeal treatment of septic patients

Abstract

The pathogenesis of sepsis begins with the proliferation of micro-organisms at a site of infection, followed by invasion of the bloodstream and other organs. Gram-negative bacteria account for a large part of sepsis cases. The structural component of Gram-negative bacteria, endotoxin or lipopolysaccharide (LPS), induces the synthesis and release of endogenous mediators of sepsis. A growing number of investigations of the molecular mechanisms occurring in sepsis, point to endotoxin as a central mediator leading to multi-organ failure and death. In numerous clinical trials, attempts to target molecules downstream of endotoxin have been made, but have not been associated with improved survival. We describe an affinity-based system for the selective removal of endotoxin from plasma. The small-scale device, a 1.5 ml cartridge, contains beads that bind endotoxin with high specificity and efficiency. In addition, evidence is presented that this device does not affect plasma hemostasis, nor does it activate the complement system. Taken together, these results represent a proof of principle for endotoxin removal from plasma, which may be of clinical value to treat sepsis by extracorporeal circulation of the blood through a scaled-up version of this endotoxin-removing device.

Peptidoglycan and Bacterial DNA Induce Inflammation and Coagulation Markers in Synergy

“…Peripheral blood mononuclear cells were prepared by Ficoll separation of peripheral blood from healthy volunteers as previously described [ 13 ]. Mononuclear cells were plated at a density of 10 6 cells/mL in 1.5 mL RPMI 1640 (Irvine Scientific, Santa Ana, Calif), supplemented with 100 U/mL penicillin/streptomicin, 2 mM glutamine, and 3% fetal calf serum, and treated with PepG (Sigma, St Louis, Mich), bactDNA (KM Biomedicals, Aurora, Ohio; LPS < 0.2 EU/mL measured by the limulus amebocyte lysate assay [ 13 ]), or both. TNF- α was measured by ELISA from the culture supernatant after 5-hour incubation and elimination of nonadherent cells.…”

Endotoxin detection in full blood plasma in a theranostic approach to combat sepsis

“…Annually, an estimated 750 000 individuals are affected by sepsis in the United States (similar incidence is reported in Europe and around the world), 1 with approximately 200 000 casualties. [2][3][4] Sepsis indeed is a life-threatening condition, in reality, constituting one of the leading causes of death worldwide 4 and by extension responsible for tremendous medical expenditure. 3 The condition occurs when a local infection overcomes the host natural defence mechanism, 4 triggering a vigorous and dysregulated whole-body inammatory response that will cause afflicted persons to display the generic symptoms of the 'systemic inammatory response syndrome' (SIRS).…”

“…[2][3][4] Sepsis indeed is a life-threatening condition, in reality, constituting one of the leading causes of death worldwide 4 and by extension responsible for tremendous medical expenditure. 3 The condition occurs when a local infection overcomes the host natural defence mechanism, 4 triggering a vigorous and dysregulated whole-body inammatory response that will cause afflicted persons to display the generic symptoms of the 'systemic inammatory response syndrome' (SIRS). 1,[4][5][6] If not treated promptly and properly, this generalized, self-injurious complication 1,4,7 can worsen into stages of various severity, the last of which (i.e.…”

“…8,9 At acute states of the affliction, mortality is extremely high. 3,10,11 The pathogenesis of sepsis is relatively well known, one of the most potent immuno-activator being bacterial lipopolysaccharide (LPS) 1,7,10,12 also known as 'endotoxin'which is a major constituent of the outer membrane of Gram-negative bacteria (e.g. Escherichia coli).…”