NLF20: an antimicrobial peptide with therapeutic potential against invasive<i>Pseudomonas aeruginosa</i>infection

Papareddy, Praveen; Kasetty, Gopinath; Kalle, Martina; Bhongir, Ravi K. V.; Mörgelin, Matthias; Schmidtchen, Artur; Malmsten, Martin

doi:10.1093/jac/dkv322

Cited by 15 publications

(15 citation statements)

References 50 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…HCII belongs to the serpin family that has increasingly been shown to have non-inhibitory roles in inflammation and angiogenesis. 10,15,46 Therefore it is also possible that HCII-N also may have a non-inhibitory role.…”

Section: Discussionmentioning

confidence: 99%

Identification and characterization of a novel isoform of heparin cofactorIIin human liver

et al. 2020

View full text Add to dashboard Cite

Heparin cofactor II (HCII) is predominantly expressed in the liver and inhibits thrombin in blood plasma to influence the blood coagulation cascade. Its deficiency is associated with arterial thrombosis. Its cleavage by neutrophil elastase produces fragment that helps in neutrophil chemotaxis in the acute inflammatory response in human. In the present study, we have identified a novel alternatively spliced transcript of the HCII gene in human liver. This novel transcript includes an additional novel region in continuation with exon 3 called exon 3b. Exon 3b acts like an alternate last exon, and hence its inclusion in the transcript due to alternative splicing removes exon 4 and encodes for a different C‐terminal region to give a novel protein, HCII‐N. MD simulations of HCII‐N and three‐dimensional structure showed a unique 51 amino acid sequence at the C‐terminal having unique RCL‐like structure. The HCII‐N protein purified from bacterial culture showed a protein migrating at lower molecular weight (MW 55 kDa) as compared to native HCII (MW 66 kDa). A fluorescence‐based analysis revealed a more compact structure of HCII‐N that was in a more hydrophilic environment. The HCII‐N protein, however, showed no inhibitory activity against thrombin. Due to large conformational variation observed in comparison with native HCII, HCII‐N may have alternate protease specificity or a non‐inhibitory role. Western blot of HCII purified from large plasma volume showed the presence of a low MW 59 kDa band with no thrombin activity. This study provides the first evidence of alternatively spliced novel isoform of the HCII gene.

show abstract

Section: Discussionmentioning

confidence: 99%

Identification and characterization of a novel isoform of heparin cofactorIIin human liver

et al. 2020

View full text Add to dashboard Cite

show abstract

“…Upon its interaction with cell surface glycosaminoglycans, the inert HCII undergoes conformational changes that activate it and induce novel biological functions, including endotoxin binding and antimicrobial properties (45,46). Peptide epitope mapping localized the antimicrobial properties of HCII to helices A and D (45,47). In vivo analysis revealed that HCII-deficient mice were more susceptible to P. aeruginosa infection than their wild-type counterparts, while challenge of wild-type mice with P. aeruginosa reduced the level of HCII within their bodies (45).…”

Section: Discussionmentioning

confidence: 99%

“…In vivo analysis revealed that HCII-deficient mice were more susceptible to P. aeruginosa infection than their wild-type counterparts, while challenge of wild-type mice with P. aeruginosa reduced the level of HCII within their bodies (45). Using a murine model of P. aeruginosa sepsis, Papareddy et al (47) demonstrated that intraperitoneal injection of mice with synthetic NFL20, a peptide that corresponds to the D helix of HCII, significantly enhanced their survival. Additionally, the number of P. aeruginosa bacteria recovered from the liver, kidney, and spleen of NLF20-treated mice was significantly reduced compared with the number recovered from those organs of untreated infected mice (47).…”

Section: Discussionmentioning

confidence: 99%

“…Using a murine model of P. aeruginosa sepsis, Papareddy et al (47) demonstrated that intraperitoneal injection of mice with synthetic NFL20, a peptide that corresponds to the D helix of HCII, significantly enhanced their survival. Additionally, the number of P. aeruginosa bacteria recovered from the liver, kidney, and spleen of NLF20-treated mice was significantly reduced compared with the number recovered from those organs of untreated infected mice (47). Thus, it is possible that in our model of P. aeruginosa sepsis, the HepP heparinase produced by PA14 during bacteremia cleaves HSPGs and prevents HCII activation.…”

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

Heparinase Is Essential for Pseudomonas aeruginosa Virulence during Thermal Injury and Infection

et al. 2018

View full text Add to dashboard Cite

The opportunistic pathogen is a major cause of sepsis in severely burned patients. If it is not eradicated from the wound, it translocates to the bloodstream, causing sepsis, multiorgan failure, and death. We recently described the heparinase-encoding gene, , whose expression was significantly enhanced when strain UCBPP_PA14 (PA14) was grown in whole blood from severely burned patients. Further analysis demonstrated that contributed to the virulence of PA14 in the model. In this study, we utilized the murine model of thermal injury to examine the contribution of to the pathogenesis of during burn wound infection. Mutation of reduced the rate of mortality from 100% for mice infected with PA14 to 7% for mice infected with PA14:: While comparable numbers of PA14 and PA14:: bacteria were recovered from infected skin, only PA14 was recovered from the livers and spleens of infected mice. Despite its inability to spread systemically, PA14:: formed perivascular cuffs around the blood vessels within the skin of the thermally injured/infected mice. Intraperitoneal inoculation of the thermally injured mice, bypassing the need for translocation, produced similar results. The rate of mortality for mice infected with PA14:: was 0%, whereas it was 66% for mice infected with PA14. As before, only PA14 was recovered from the livers and spleens of infected mice. These results suggest that plays a crucial role in the pathogenesis of PA14 during burn wound infection, most likely by contributing to PA14 survival in the bloodstream of the thermally injured mouse during sepsis.

show abstract

“…This example of additive cooperation of AMPs provides valuable information for considering future therapeutic strategies regarding the combination of AMPs. Notably, the therapeutic efficacy of AMPs was evaluated in the clinical context not only for their direct antibacterial activity, but also the indirect immunomodulatory activity to promote the clearance of bacterial infection [3,12]. The aims of this study were to compare the antibacterial and antibiofilm activities of the four cathelicidins BMAP-27, BMAP-34, mCRAMP, and LL-37, to determine their immunomodulatory activity on neutrophils, and to evaluate the therapeutic efficacy of the combination of BMAP-27 and LL-37 against P. aeruginosa infection.…”

Section: Introductionmentioning

confidence: 99%

Differential Abilities of Mammalian Cathelicidins to Inhibit Bacterial Biofilm Formation and Promote Multifaceted Immune Functions of Neutrophils

Xie

Zan

Zhang

et al. 2020

IJMS

View full text Add to dashboard Cite

Mammalian cathelicidins act as the potent microbicidal molecules for controlling bacterial infection, and are considered promising alternatives to traditional antibiotics. Their ability to modulate host immune responses, as well as their bactericidal activities, is essential for therapeutic interventions. In this study, we compared the bactericidal activities, antibiofilm activities and immune-modulatory properties of cathelicidins BMAP-27, BMAP-34, mCRAMP, and LL-37, and evaluated the therapeutic efficacy of the combination of BMAP-27 and LL-37 using a mouse pulmonary infection model. Our results showed that all of the four cathelicidins effectively killed bacteria via rapid induction of membrane permeabilization, and BMAP-27 exhibited the most excellent bactericidal activity against diverse bacterial pathogens. BMAP-27, mCRAMP, and LL-37 effectively inhibited biofilm formation, while BMAP-34, mCRAMP and LL-37 exerted immunomodulatory functions with varying degrees of efficacy by stimulating the chemotaxis of neutrophils, inducing the production of reactive oxygen species, and facilitating the formation of neutrophil extracellular traps. Of note, the combination of BMAP-27 and LL-37 effectively enhanced the clearance of Pseudomonas aeruginosa and reduced the organ injury in vivo. Together, these findings highlight that identifying the appropriate synergistic combination of mammalian cathelicidins with different beneficial properties may be an effective strategy against bacterial infection.

show abstract

NLF20: an antimicrobial peptide with therapeutic potential against invasivePseudomonas aeruginosainfection

Abstract: Together, these results indicate that functional epitopes of HCII may have therapeutic potential against bacterial infection.

Cited by 15 publications

References 50 publications

Identification and characterization of a novel isoform of heparin cofactorIIin human liver

Identification and characterization of a novel isoform of heparin cofactorIIin human liver

Heparinase Is Essential for Pseudomonas aeruginosa Virulence during Thermal Injury and Infection

Differential Abilities of Mammalian Cathelicidins to Inhibit Bacterial Biofilm Formation and Promote Multifaceted Immune Functions of Neutrophils

Contact Info

Product

Resources

About