LPS interactions with immobilized and soluble antimicrobial peptides.
A promising approach in sepsis therapy is the use of peptides truncated from serum- and membrane-proteins with binding domains for LPS: antimicrobial peptides (AMPs). AMPs can be useful in combination with conventional antibiotics to increase killing and neutralize LPS. Although many AMPs show a high specificity towards bacterial membranes, they can also exhibit toxicity, i.e. non-specific membrane lysis, of mammalian cells such as erythrocytes and therefore, unsuitable as systemic drugs. A way to overcome this problem may be an extracorporeal therapy with immobilized peptides. This study will compare neutralization of LPS using different AMPs in solution and when immobilized on to solid phases. The peptides ability to neutralize LPS-induced cytokine release in whole blood will also be tested. The peptides are truncated derivates from the known AMPs LL-37, SC4, BPI, S3 Delta and CEME. Two different methods were used to immobilize peptides, biomolecular interaction analysis, and Pierce SulfoLink Coupling Gel. To investigate LPS binding in solution the LAL test was used. After whole blood incubation with LPS and AMPs ELISA was used to measure TNFalpha, IL-1 beta and IL-6 production. The results suggest that immobilization of antimicrobial peptides does not inhibit their capacity to neutralize LPS, although there are differences between the peptides tested. Thus, peptides derived from LL-37 and CEME were more efficient both in LPS binding and neutralizing LPS-induced cytokine production.
Department of Biomedical Laboratory Science, Malmö University, Malmö, Sweden. Anna.Gustafsson@mah.se
Anna
A
Gustafsson
Anders I
AI
Olin
Lennart
L
Ljunggren
D000327:Adsorption / Q000187:drug effects; D000595:Amino Acid Sequence; D023181:Antimicrobial Cationic Peptides / Q000737:chemistry / Q000378:metabolism* / Q000494:pharmacology; D016207:Cytokines / Q000037:antagonists & inhibitors; D006801:Humans; D055767:Immobilized Proteins / Q000737:chemistry / Q000378:metabolism*; D007526:Isoelectric Point; D008070:Lipopolysaccharides / Q000378:metabolism*; D008969:Molecular Sequence Data; D009500:Neutralization Tests; D012995:Solubility / Q000187:drug effects; D020349:Surface Plasmon Resonance
D023181:Antimicrobial Cationic Peptides; D016207:Cytokines; D055767:Immobilized Proteins; D008070:Lipopolysaccharides