Activity of the de novo engineered antimicrobial peptide WLBU2 against Pseudomonas aeruginosa in human serum and whole blood:: Implications for systemic applications
Activity of the de novo engineered antimicrobial peptide WLBU2 against Pseudomonas aeruginosa in human serum and whole blood:: Implications for systemic applications
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DOI：
10.1128/aac.49.8.3208-3216.2005
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发表时间：
2005-08-01
影响因子：
4.9
通讯作者：
Mietzner, TA
中科院分区：
文献类型：
--
作者：
Deslouches, B;Islam, K;Mietzner, TA
阳离子两亲性多肽作为一种新的抗菌剂的潜在来源已被广泛研究，这些抗菌剂可以补充现有的抗生素方案，以应对新出现的耐药细菌。然而，在某些生物相关的条件下(如血清和生理盐浓度)，抗菌活性的抑制阻碍了开发安全有效的抗菌肽用于临床的努力。我们分析了人源肽LL37和从头工程抗菌肽WLBU2在几种生物相关条件下的活性和选择性。宿主来源的合成肽LL37对铜绿假单胞菌表现出很高的活性，但对50-300 mM的氯化钠浓度表现出葡萄球菌特异性的敏感性。此外，在1~6 mM的镁和钙离子存在下，LL37的活性受到不同程度的抑制。相反，WLBU2在氯化钠中保持其活性，并维持生理血清中的镁和钙浓度。WLBU2对人血清中的铜绿假单胞菌(10(6)cfu/ml)有杀灭作用，最低杀菌浓度为9mU。血清杀菌动力学测定显示，WLBU2在20min内即可达到完全杀菌。与这些结果一致的是，WLBU2(15至20亩M)能够从体外全血样本中清除细菌。WLBU2的选择性进一步证明，它能够在与人单核细胞或皮肤成纤维细胞共同培养时特异性地消除铜绿假单胞菌，而不会对宿主细胞产生明显的不良影响。最后，在使用雌性瑞士韦氏小鼠的腹膜感染模型中，WLBU2对铜绿假单胞菌显示出强大的疗效。这些结果为WLBU2在治疗细菌性脓毒症中的潜在应用奠定了基础。
Cationic amphipathic peptides have been extensively investigated as a potential source of new antimicrobials that can complement current antibiotic regimens in the face of emerging drug-resistant bacteria. However, the suppression of antimicrobial activity under certain biologically relevant conditions (e.g., serum and physiological salt concentrations) has hampered efforts to develop safe and effective antimicrobial peptides for clinical use. We have analyzed the activity and selectivity of the human peptide LL37 and the de novo engineered antimicrobial peptide WLBU2 in several biologically relevant conditions. The host-derived synthetic peptide LL37 displayed high activity against Pseudomonas aeruginosa but demonstrated staphylococcus-specific sensitivity to NaCl concentrations varying from 50 to 300 mM. Moreover, LL37 potency was variably suppressed in the presence of 1 to 6 mM Mg2+ and Ca2+ ions. In contrast, WLBU2 maintained its activity in NaCl and physiologic serum concentrations of Mg2+ and Ca2+. WLBU2 is able to kill P. aeruginosa (10(6) CFU/ml) in human serum, with a minimum bactericidal concentration of < 9 mu M. Conversely, LL37 is inactive in the presence of human serum. Bacterial killing kinetic assays in serum revealed that WLBU2 achieved complete bacterial killing in 20 min. Consistent with these results was the ability of WLBU2 (15 to 20 mu M) to eradicate bacteria from ex vivo samples of whole blood. The selectivity of WLBU2 was further demonstrated by its ability to specifically eliminate P. aeruginosa in coculture with human monocytes or skin fibroblasts without detectable adverse effects to the host cells. Finally, WLBU2 displayed potent efficacy against P. aeruginosa in an intraperitoneal infection model using female Swiss Webster mice. These results establish a potential application of WLBU2 in the treatment of bacterial sepsis.