Biophysical mechanisms of endotoxin neutralization by cationic amphiphilic peptides.

Bacterial endotoxins (lipopolysaccharides (LPS)) are strong elicitors of the human immune system by interacting with serum and membrane proteins such as lipopolysaccharide-binding protein (LBP) and CD14 with high specificity. At LPS concentrations as low as 0.3 ng/ml, such interactions may lead to severe pathophysiological effects, including sepsis and septic shock. One approach to inhibit an uncontrolled inflammatory reaction is the use of appropriate polycationic and amphiphilic antimicrobial peptides, here called synthetic anti-LPS peptides (SALPs). We designed various SALP structures and investigated their ability to inhibit LPS-induced cytokine secretion in vitro, their protective effect in a mouse model of sepsis, and their cytotoxicity in physiological human cells. Using a variety of biophysical techniques, we investigated selected SALPs with considerable differences in their biological responses to characterize and understand the mechanism of LPS inactivation by SALPs. Our investigations show that neutralization of LPS by peptides is associated with a fluidization of the LPS acyl chains, a strong exothermic Coulomb interaction between the two compounds, and a drastic change of the LPS aggregate type from cubic into multilamellar, with an increase in the aggregate sizes, inhibiting the binding of LBP and other mammalian proteins to the endotoxin. At the same time, peptide binding to phospholipids of human origin (e.g., phosphatidylcholine) does not cause essential structural changes, such as changes in membrane fluidity and bilayer structure. The absence of cytotoxicity is explained by the high specificity of the interaction of the peptides with LPS.

[Resurgence of mumps in the vaccine era. Factors involved in an outbreak in Navarre, Spain, 2006-2007]. / Rebrote de parotiditis en la era vacunal. Factores implicados en un brote en Navarra, 2006-2007.

BACKGROUND AND OBJECTIVE: We analysed a mumps outbreak that occurred in Navarre between August 2006 and December 2007, in which vaccinated persons were widely affected. PATIENTS AND METHODS: Reports of mumps cases were completed by searching primary, emergency and hospital records and laboratory reports. Factors that could affect the occurrence of cases were analysed by birth cohort. RESULTS: A total of 2866 mumps cases were detected (attack rate 4.7/1000), with 61% of cases in men and a peak incidence at age 19 (inter-quartile range 16-25 years). 14% of cases were confirmed by laboratory: 59 by virus isolation, 14 by PCR and 333 by IgM. The G1 genotype was identified in 7 cases. 21% of cases had been born before 1980 (pre-vaccine cohorts), and 0.2% had not yet reached the vaccination age (15 months). In the cohorts born between 1980 and 2000 (with the opportunity for vaccination), 94.5% of cases had received at least one dose and 88.3%, two doses. 31% of cases occurred in cohorts vaccinated with a first (1995-1997) or second (1986-1988) dose of the Rubini strain. There was also a record of 772 cases who had received two doses of the Jeryl Lynn strain. CONCLUSIONS: This widespread outbreak is explained by the concurrence of various factors. The current vaccine has substantially reduced the incidence of mumps, but appears unable to totally eliminate virus circulation.

Free thiol group of MD-2 as the target for inhibition of the lipopolysaccharide-induced cell activation.

MD-2 is a part of the Toll-like 4 signaling complex with an indispensable role in activation of the lipopolysaccharide (LPS) signaling pathway and thus a suitable target for the therapeutic inhibition of TLR4 signaling. Elucidation of MD-2 structure provides a foundation for rational design of inhibitors that bind to MD-2 and inhibit LPS signaling. Since the hydrophobic binding pocket of MD-2 provides little specificity for inhibitors, we have investigated targeting the solvent-accessible cysteine residue within the hydrophobic binding pocket of MD-2. Compounds with affinity for the hydrophobic pocket that contain a thiol-reactive group, which mediates covalent bond formation with the free cysteine residue of MD-2, were tested. Fluorescent compounds 2-(4'-(iodoacetamido)anilino)naphthalene-6-sulfonic acid and N-pyrene maleimide formed a covalent bond with MD-2 through Cys(133) and inhibited LPS signaling. Cell activation was also inhibited by thiol-reactive compounds JTT-705 originally targeted against cholesterol ester transfer protein and antirheumatic compound auranofin. Oral intake of JTT-705 significantly inhibited endotoxin-triggered tumor necrosis factor alpha production in mice. The thiol group of MD-2 also represents the target of environmental or endogenous thiol-reactive compounds that are produced in inflammation.