ABSTRACT
Abstract The present study investigated the effects of valerian methanolic extract and valerenic acid on the expression of LL-37 gene and protein in A549 and MRC5 line cells. After preparing Valerian seeds, sowing them in March 2020, and harvesting the rhizome in October 2020, the extract was prepared from the valerian rhizome by maceration method. Valerian acid content was determined using high performance liquid chromatography (HPLC). Two cell lines (A549 and MRC-5) were used to study the effects of valerian extract, and the MTT test was used to evaluate cell viability. The expression of LL-37 mRNA and protein was assessed by Real-Time PCR and western blot, respectively. In vivo safety assessments and histopathological analysis were also conducted. Data was analyzed by Graphpad Prism 8 software. Valerian methanolic extract and valerenic acid upregulated the LL-37 mRNA and protein expression in both treated cell lines. Valerenic acid showed a greater effect on upregulating LL-37 expression than valerian methanolic extract. A549 cells were more sensitive to valerian methanolic extract compared to MRC5 cells, and its cell viability was reduced. Furthermore, liver and kidney-related safety assessments showed that valerian methanolic extract had no toxic effects. In general, it was concluded that the methanolic extract of valerian as well as the resulting valerenic acid as the most important component of the extract has the ability to upregulate LL-37expression. Therefore, methanolic extract of valerian and valerenic acid can be considered for improving the immune system.
Subject(s)
Valerian/adverse effects , Plant Extracts/adverse effects , Cathelicidins/adverse effects , Blotting, Western/instrumentation , Chromatography, High Pressure Liquid/methods , Antimicrobial Cationic Peptides/agonists , A549 Cells/classification , Genes/genetics , Liver/abnormalitiesABSTRACT
Objective: To fabricate TiO2 nanotube material functionalized by antimicrobial peptide LL-37, and to explore its effects on biological behaviors such as adhesion and migration of human keratinocytes (HaCaT) and its antibacterial properties. Methods: The TiO2 nanotube array (NT) was constructed on the surface of polished titanium (PT) by anodization, and the antimicrobial peptide LL-37 was loaded on the surface of TiO2 nanotube (LL-37/NT) by physical adsorption. Three samples were selected by simple random sampling in each group. Surface morphology, roughness, hydrophilicity and release characteristics of LL-37 of the samples were analyzed with a field emission scanning electron microscope, an atomic force microscope, a contact angle measuring device and a microplate absorbance reader. HaCaT cells were respectively cultured on the surface of three groups of titanium samples. Each group had 3 replicates. The morphology of cell was observed by field emission scanning electron microscope. The number of cell adhesion was observed by cellular immunofluorescence staining. Cell counting kit-8 (CCK-8) assay was used to detect cell proliferation. Wound scratch assay was used to observe the migration of HaCaT. The above experiments were used to evaluate the effect of each group on the biological behavior of HaCaT cells. To evaluate their antibacterial effects, Porphyromonas gingivalis (Pg) was respectively inoculated on the surface of three groups of titanium samples. Each group had 3 replicates. The morphology of bacteria was observed by field emission scanning electron microscope. Bacterial viability was determined by live/dead bacterial staining. Results: A uniform array of nanotubes could be seen on the surface of titanium samples in LL-37/NT group, and the top of the tube was covered with granular LL-37. Compared with PT group [the roughness was (2.30±0.18) nm, the contact angle was 71.8°±1.7°], the roughness [(20.40±3.10) and (19.10±4.11) nm] and hydrophilicity (the contact angles were 22.4°±3.1° and 25.3°±2.2°, respectively) of titanium samples increased in NT and LL-37/NT group (P<0.001). The results of in vitro release test showed that the release of antimicrobial peptide LL-37 was characterized by early sudden release (1-4 h) and long-term (1-7 d) slow release. With the immunofluorescence, more cell attachment was found on NT and LL-37/NT than that on PT at the first 0.5 and 2.0 h of culture (P<0.05). The results of CCK-8 showed that there was no significant difference in the proliferation of cells among groups at 1, 3 and 5 days after culture. Wound scratch assay showed that compared with PT and NT group, the cell moved fastest on the surface of titanium samples in LL-37/NT group at 24 h of culture [(96.4±4.9)%] (F=35.55, P<0.001). A monolayer cells could be formed and filled with the scratch in 24 h at LL-37/NT group. The results of bacterial test in vitro showed that compared with the PT group, the bacterial morphology in the NT and LL-37/NT groups was significantly wrinkled, and obvious bacterial rupture could be seen on the surface of titanium samples in LL-37/NT group. The results of bacteria staining showed that the green fluorescence intensity of titanium samples in LL-37/NT group was the lowest in all groups (F=66.54,P<0.001). Conclusions: LL-37/NT is beneficial to the adhesion and migration of HaCaT cells and has excellent antibacterial properties, this provides a new strategy for the optimal design of implant neck materials.
Subject(s)
Humans , Titanium/chemistry , Antimicrobial Peptides , Cathelicidins , Sincalide , Anti-Bacterial Agents/pharmacology , Nanotubes/chemistry , Dental Materials , Bacteria , Keratinocytes , Surface PropertiesABSTRACT
Abstract Background Antimicrobial peptides are components of the innate immune system. Cathelicidin LL-37 plays an important role in antimicrobial defense, exerts proinflammatory effect and strongly affects the immune system functioning. Our recent study revealed that serum concentration of LL-37 is increased in patients with bipolar disorder. Objectives The aim of this study is to re-evaluate serum LL-37 levels in patients with euthymic bipolar disorder and in healthy controls, matched for anthropometric and body composition parameters. Methods 36 adult patients with euthymic bipolar disorder and 68 non-depressed adults were included into the study. Concentration of LL-37 in serum was assessed using ELISA method. Detailed anthropometric measurements, body composition and biochemical analyses were performed. Results There was a statistically significant difference (p = 0.01) in serum LL-37 level between patients with bipolar disorder (4.97 ± 7.98 ng/mL) and control subjects (1.78 ± 2.69 ng/mL). Discussion Results of this study indicate that LL-37 serum level is increased in euthymic bipolar disorder patients. We found that this increase could not be attributed to analyzed anthropometric or body composition parameters.
Subject(s)
Humans , Male , Female , Adult , Bipolar Disorder , Body Composition/drug effects , Cathelicidins/blood , Tobacco Use Disorder , Bipolar Disorder/metabolism , Body Weights and Measures , Linear Models , Laboratory TestABSTRACT
Campylobacter is a worldwide foodborne pathogen, associated with human gastroenteritis. The efficient translocation of Campylobacter and its ability to secrete toxins into host cells are the 2 key features of Campylobacter pathophysiology which trigger inflammation in intestinal cells and contribute to the development of gastrointestinal symptoms, particularly diarrhoea, in humans. The purpose of conducting this literature review is to summarise the current understanding of: i) the human immune responses involved in the elimination of Campylobacter infection and ii) the resistance potential in Campylobacter against these immune responses. This review has highlighted that the intestinal epithelial cells are the preliminary cells which sense Campylobacter cells by means of their cell-surface and cytosolic receptors, activate various receptors-dependent signalling pathways, and recruit the innate immune cells to the site of inflammation. The innate immune system, adaptive immune system, and networking between these systems play a crucial role in bacterial clearance. Different cellular constituents of Campylobacter, mainly cell membrane lipooligosaccharides, capsule, and toxins, provide protection to Campylobacter against the human immune system mediated killing. This review has also identified gaps in knowledge, which are related to the activation of following during Campylobacter infection: i) cathelicidins, bactericidal permeability-increasing proteins, chemokines, and inflammasomes in intestinal epithelial cells; ii) siglec-7 receptors in dendritic cell; iii) acute phase proteins in serum; and iv) T-cell subsets in lymphoid nodules. This review evaluates the existing literature to improve the understanding of human immunity against Campylobacter infection and identify some of the knowledge gaps for future research.
Subject(s)
Humans , Acute-Phase Proteins , Antigen-Presenting Cells , Campylobacter Infections , Campylobacter , Cathelicidins , Cell Membrane , Chemokines , Cytosol , Dendritic Cells , Epithelial Cells , Gastroenteritis , Guillain-Barre Syndrome , Homicide , Immune System , Inflammasomes , Inflammation , T-Lymphocyte Subsets , Toll-Like ReceptorsABSTRACT
Abstract Introduction Methicillin-resistant staphylococcus aureus is an emerging problem for the treatment of chronic suppurative otitis media, and also for pediatric tympanostomy tube otorrhea. To date, there are no effective topical antibiotic drugs to treat methicillin-resistant staphylococcus aureus otorrhea. Objective In this study, we evaluated the ototoxicity of topical KR-12-a2 solution on the cochlea when it is applied topically in the middle ear of guinea pigs. Methods The antimicrobial activity of KR-12-a2 against methicillin-resistant staphylococcus aureus strains was examined by using the inhibition zone test. Topical application of KR-12-a2 solution, gentamicin and phosphate buffered saline were applied in the middle ear of the guinea pigs after inserting ventilation tubes. Ototoxicity was assessed by auditory brainstem evoked response and scanning electron microscope examination. Results KR-12-a2 produced an inhibition zone against methicillin-resistant staphylococcus aureus from 6.25 µg. Hearing threshold in the KR-12-a2 and PBS groups were similar to that before ventilation tube insertion. However, the gentamicin group showed elevation of the hearing threshold and there were statistically significant differences compared to the phosphate buffered saline or the KR-12-a2 group. In the scanning electron microscope findings, the KR-12-a2 group showed intact outer hair cells. However, the gentamicin group showed total loss of outer hair cells. In our experiment, topically applied KR-12-a2 solution did not cause hearing loss or cochlear damage in guinea pigs. Conclusion In our experiment, topically applied KR-12-a2 solution did not cause hearing loss or cochlear damage in guinea pigs. The KR-12-a2 solution can be used as ototopical drops for treating methicillin-resistant staphylococcus aureus otorrhea; however, further evaluations, such as the definition of optimal concentration and combination, are necessary.
Resumo Introdução O staphylococcus aureus resistente à meticilina é um problema emergente não só para a otite média supurativa crônica, mas também para casos de otorreia crônica em crianças com tubo de ventilação. Até o momento, não há antibióticos tópicos efetivos para a otorreia causada por staphylococcus aureus resistente à meticilina. Objetivo Nesse estudo, avaliamos a ototoxicidade da solução tópica de KR-12-a2 na cóclea quando aplicada topicamente na orelha média de cobaias. Método A atividade antimicrobiana de KR-12-a2 contra cepas de staphylococcus aureus resistente à meticilina foi avaliada utilizando-se o teste de zona de inibição de crescimento. Foram aplicados na orelhas médias de 3 grupos de cobaias, ou solução tópica de KR-12-a2, ou gentamicina ou solução salina tamponada com fosfato após timpanostomia. A ototoxicidade foi avaliada através do exame auditivo de potencial evocado auditivo de tronco encefálico e por microscopia eletrônica de varredura. Resultados O KR-12-a2 produziu uma zona de inibição contra o staphylococcus aureus resistente à meticilina a partir de 6,25 µg. Alterações do limiar de audição no grupo KR-12-a2 e no grupo com solução salina foram semelhantes aos observados antes da inserção do tubo de ventilação. No entanto, o grupo gentamicina apresentou um limiar auditivo mais elevado, estatisticamente significativo em comparação ao grupo solução salina ou ao grupo KR-12-a2. Nos achados da microscopia eletrônica, o grupo KR-12-a2 apresentou células ciliadas externas intactas. No entanto, o grupo gentamicina apresentou perda total das células ciliadas externas. Em nosso experimento, a solução de KR-12-a2 aplicada topicamente não causou perda auditiva ou dano coclear em cobaias. Conclusão Em nosso experimento, a solução de KR-12-a2 aplicada topicamente não causou perda auditiva ou dano coclear em cobaias. A solução de KR-12-a2 pode ser utilizada como gotas otológicas para o tratamento da otorreia causada por staphylococcus aureus resistente à meticilina; no entanto, são necessárias outras avaliações, para a definição da concentração e das associações ideais.
Subject(s)
Animals , Male , Peptide Fragments/toxicity , Cochlea/drug effects , Cathelicidins/toxicity , Methicillin-Resistant Staphylococcus aureus/drug effects , Anti-Bacterial Agents/toxicity , Otitis Media, Suppurative/microbiology , Peptide Fragments/administration & dosage , Auditory Threshold , Staphylococcal Infections/drug therapy , Microscopy, Electron, Scanning , Microbial Sensitivity Tests , Reproducibility of Results , Administration, Topical , Evoked Potentials, Auditory, Brain Stem , Treatment Outcome , Cochlea/physiopathology , Disease Models, Animal , Cathelicidins/administration & dosage , Guinea Pigs , Hair Cells, Auditory/drug effects , Anti-Bacterial Agents/administration & dosageABSTRACT
The platelet-rich plasma (PRP) and antimicrobial peptides neutrophil extract (AMP extract) were prepared from rabbit neutrophils as two autologous blood-derived preparations, which could be applied locally to enhance healing process of tissues. Both preparations were analyzed using the MALDI TOF method for accurate qualitative assay. Growth factors (PDGF and VEGF) and microbicidal protein were reported in PRP. In AMP extract a-defensins, namely; NP-1, -2, -3a, -3b, -4, and -5 and cathelicidins represented among other by 15-kDa antibacterial protein (p15s) were detected. In the second part of our study the influence of antimicrobial extract on macrophages in vitro was tested. Then, degranulation of neutrophils in vitro and generation of reactive intermediates by these cells under the influence of AMP extract were assessed. As estimated, the addition of AMP extract into cultures of macrophages decreased superoxide anion generation after 5 days of incubation. Furthermore, AMP extract inhibited degranulation and respiratory burst in neutrophils, therefore in this regard it suppress proinflammatory effect of two studied populations of leukocytes.
Subject(s)
Cathelicidins , In Vitro Techniques , Intercellular Signaling Peptides and Proteins , Leukocytes , Macrophages , Methods , Neutrophils , Peptides , Platelet-Rich Plasma , Respiratory Burst , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization , SuperoxidesABSTRACT
The objective of this study was to construct an improved thioredoxin fusion protein expression system, and express the cathelicidin-derived peptide, Lf-CATH2. The improved fusion vector Lf-CATH2-pET32α(-TS) was successfully constructed by firstly deleting the thrombin site and S tag from the pET-32α vector, then inserting the Lf-CATH2 plus a thrombin site instead. Afterwards, Lf-CATH2 was expressed in Escherichia coli as fusion protein. After the cleavage by thrombin, Lf-CATH2 was released and subsequently separated using affinity chromatography. The antimicrobial activity of purified Lf-CATH2 was also examined. The improved expression vector significantly increased enzyme cleavage efficiency by 37%, and Lf-CATH2 could be expressed in high yield and maintain the biological activity. This novel thioredoxin fusion protein expression system enables a quick production of high-yield bioactive cationic peptides like cathelicidins.
Subject(s)
Cathelicidins , Chromatography, Affinity , Escherichia coli , Genetic Vectors , Recombinant Fusion Proteins , Thioredoxins , GeneticsABSTRACT
It has been reported that patients with progressive tuberculosis (TB) express abundant amounts of the antimicrobial peptides (AMPs) cathelicidin (LL-37) and human neutrophil peptide-1 (HNP-1) in circulating cells, whereas latent TB infected donors showed no differences when compared with purified protein derivative (PPD) and QuantiFERON®-TB Gold (QFT)-healthy individuals. The aim of this study was to determine whether LL-37 and HNP-1 production correlates with higher tuberculin skin test (TST) and QFT values in TB household contacts. Twenty-six TB household contact individuals between 26-58 years old TST and QFT positive with at last two years of latent TB infection were recruited. AMPs production by polymorphonuclear cells was determined by flow cytometry and correlation between TST and QFT values was analysed. Our results showed that there is a positive correlation between levels of HNP-1 and LL-37 production with reactivity to TST and/or QFT levels. This preliminary study suggests the potential use of the expression levels of these peptides as biomarkers for progression in latent infected individuals.
Subject(s)
Adult , Female , Humans , Male , Middle Aged , Blood Cells/chemistry , Cathelicidins/blood , Latent Tuberculosis/diagnosis , Mycobacterium tuberculosis/immunology , alpha-Defensins/blood , Biomarkers/blood , Contact Tracing , Cathelicidins/metabolism , Disease Progression , Gene Expression , Interferon-gamma Release Tests/methods , Latent Tuberculosis/metabolism , Neutrophils/metabolism , Tuberculin Test/methodsABSTRACT
<p><b>OBJECTIVE</b>To investigate the effect of antibacterial peptide LL-37 on the integrity of Acinetobacter baumannii biofilm.</p><p><b>METHODS</b>A model of Acinetobacter baumannii biofilm in vitro was constructed by plates and crystal violet staining method, and the minimal inhibitory concentration of LL-37 was determined by broth dilution. The biofilm morphology was observed by scanning electron microscopy and biofilm formation was analyzed by the crystal violet staining of the adherent biofilm in the presence of antibacterial peptide LL-37.</p><p><b>RESULTS</b>In the Acinetobacter baumannii biofilm model, the minimal inhibitory concentration of LL-37 was 64 µg/ml; LL-37 caused structural damage of the biofilm at a low concentration of 2.5 µg/ml. The biofilm was decreased gradually as the concentration of LL-37 increased.</p><p><b>CONCLUSION</b>LL-37 even at a concentration far below its minimal inhibitory concentration can cause structural damage of Acinetobacter baumannii biofilm in vitro.</p>
Subject(s)
Acinetobacter baumannii , Physiology , Biofilms , Cathelicidins , Pharmacology , Microbial Sensitivity TestsABSTRACT
The purpose of this review is to explain the function of LL-37 in the pathogenesis of chronic sinusitis. LL-37 is the only human cathelicidin identified so far. LL-37 is an integral part of the innate immune,the role of which in chronic sinusitis is attracting more and more s attention.
Subject(s)
Humans , Cathelicidins , Metabolism , Chronic Disease , Sinusitis , Allergy and Immunology , Metabolism , PathologyABSTRACT
<p><b>OBJECTIVE</b>The aim of this study was to investigate the role of macrophages in promotion of ovarian tumor cell proliferation mediated by over-expression of antimicrobial peptide LL-37.</p><p><b>METHODS</b>To co-culture ovarian tumor cells SKOV3, 3AO and HO-8910 with macrophages. The Transwell(®) inserts system was used in the co-culture model. The effect of macrophages promoted ovarian tumor cell proliferation was assessed by BrdU-ELISA and cell number counting. Expressions of mRNA and protein of LL-37 in the macrophages and SKOV3 cells were determined by RT-PCR and Western blot analysis. To observe that LL-37 is responsible for macrophage-promoted ovarian tumor cells growth, LL-37 neutralizing antibody was added to abrogate the LL-37 activation.</p><p><b>RESULTS</b>The cell number assay showed that after 4 days coincubation with macrophages in the proportion of 1:0.5, the number of SKOV3 cells increased from (6.0 ± 0.5)×10(4) to (11.8 ± 1.3)×10(4), showing a significant difference (P < 0.05). It also showed that the growth of the SKOV3 cells was dependent on the macrophage number (P < 0.05). The number variability of 3AO and HO-8910 cells was as the same as SKOV3 cells upon co-culture with macrophages. As determined by BrdU-ELISA, the resulted proliferation of ovarian tumor cells was similar to the result of cell number counting. RT-PCR and Western blot results showed that the expression of LL-37 mRNA and protein in the macrophages was remarkably enhanced in a time dependent manner upon coincubation with SKOV3 cells, but did not work in SKOV3 cells. BrdU-ELISA assay exhibited that treatment of cells with LL-37 significantly stimulated HO-8910 and 3AO cell proliferation. Addition of LL-37 neutralizing antibody markedly inhibited macrophage-promoted ovarian tumor cell (SKOV3, 3AO and HO-8910 cells) proliferation. The OD values of these three cells were decreased from 2.95 ± 0.11 to 1.45 ± 0.04, from 3.39 ± 0.36 to 1.32 ± 0.09 and from 3.93 ± 0.17 to 1.68 ± 0.23, respectively (P < 0.05).</p><p><b>CONCLUSIONS</b>Over-expression and release of LL-37 from macrophages is responsible for proliferation of ovarian tumor cells in co-culture condition. The data presented indicate that LL-37 may be critical for macrophage-induced tumor progression.</p>
Subject(s)
Female , Humans , Antibodies, Neutralizing , Pharmacology , Cathelicidins , Genetics , Metabolism , Pharmacology , Cell Line, Tumor , Cell Proliferation , Coculture Techniques , Macrophages , Cell Biology , Physiology , Ovarian Neoplasms , Metabolism , Pathology , RNA, Messenger , MetabolismABSTRACT
BACKGROUND: A stem cell is an undifferentiated cell that has the potential for self-renewal and differentiation. Adipose-derived stem cells (ADSCs) have advantages in accessibility and abundance compared to other kinds of stem cells and produce many growth factors and hormones. OBJECTIVE: We investigated whether ADSC cultured media could be used as a therapy for atopic dermatitis. METHODS: ADSC cultured media was topically applied twice daily for 5 days to oxazolone-treated atopic dermatitis-like hairless mice. RESULTS: Topical application of ADSC cultured media improved the epidermal permeability barrier and keratinocyte differentiation, and restored the predominant Th2 phenotype when compared to vehicle. ADSC cultured media-treated epidermis also showed an increase in the expression of antimicrobial peptides cathelin-related antimicrobial peptide, mouse beta-defensein 3. CONCLUSION: Topical ADSC cultured media could be useful in the treatment of atopic dermatitis.
Subject(s)
Animals , Mice , Cathelicidins , Dermatitis, Atopic , Epidermis , Intercellular Signaling Peptides and Proteins , Keratinocytes , Oxazolone , Peptides , Permeability , Phenotype , Stem CellsABSTRACT
<p><b>OBJECTIVE</b>To investigate the core functional region of antimicrobial peptide LL-37, which inhibites RSV replication and could be developed for theraputic aplication.</p><p><b>METHODS</b>A panel of 6 partial LL-37 peptides (referred to as P1 to P6) was synthesized according to LL-37 amino acide sequence. Hep-2 cells were infected with RSV, treated with LL-37 or partial peptides respectively. Cells were collected after 24 hours incubation at 37 degrees C, CO2 5%. Total RNA was obtained from the cells. Expression level of RSV N gene was quantified by real-time PCR. Meanwhile enzyme-linked immunosorbent assay (ELISA) was used to quantify the chemokines RANTES, IL-8, MCP1 in the supernatants of Hep-2 cultures after 24 h incubation with or without LL-37 and partial peptide P6.</p><p><b>RESULTS</b>N-terminal partial LL-37 peptide (corresponding to residues 1-12 of LL-37) had no significant effects on RSV replication (P > 0.05). In contrast, C-terminal (corresponding to residues 13-37) and a panel of 4 overlapping 22-mer partial peptides (from the peptide incorporating aa 13-34 through that spanning aa 16-37) showed significant inhibitory effect on RSV replication to some extent (P < 0.05 or P < 0.01). LL-37 induced significant expression of chemokine RANTES, IL-8 and MCP-1 in Hep-2 cells. In contrast, partial peptide P6 had no significant effect on expression of the chemokines in Hep-2 cells.</p><p><b>CONCLUSION</b>The LL-37 C-terminal 22-mer partial peptide P6 was putative core functional region for inhibition of RSV replication. The partial peptide didn't induce significant expression of chemokine RANTES, IL-8 and MCP-1.</p>
Subject(s)
Humans , Cathelicidins , Pharmacology , Cell Line , Cytokines , Genetics , Allergy and Immunology , Respiratory Syncytial Virus Infections , Genetics , Allergy and Immunology , Virology , Respiratory Syncytial Viruses , Physiology , Virus ReplicationABSTRACT
Antibacterial peptides are a family of host-defense peptides most of which are gene-encoded and produced by living organisms of all types. Antibacterial peptides are small molecular proteins with broad antimicrobial spectrum against bacteria, viruses, fungi and sometimes even as anticancer peptide. SMAP-29, a cathelicidin-like peptide derived from sheep myeloid, line alpha-helical Structure, exerts a powerful broad antimicrobial activity against different pathogens including Gram-positive and Gram-negative bacteria, fungi, viruses, parasites, spirochaetes, chlamydia and antiendotoxin activity, and particular antibacterial mechanism, rapidly to permeabilize membranes of susceptible organisms. This paper summarizes the lately research progress of SMAP-29 and Variants including the characteristics of structure, structure-activity relationships, mode of action, diverse biological functions, gene recombinant and expression. We put emphasis on the necessity of molecular design, and primary and secondary structure-based modification, to provides a strong foundation for further drug development and design of SMAP-29.
Subject(s)
Animals , Antimicrobial Cationic Peptides , Chemistry , Genetics , Physiology , Blood Proteins , Chemistry , Genetics , Physiology , Cathelicidins , Chemistry , Genetics , Physiology , Drug Design , Recombinant Proteins , Chemistry , Genetics , SheepABSTRACT
Antimicrobial peptides/proteins are ancient and naturallyoccurring antibiotics in innate immune responses in a variety of organisms. Additionally, these peptides have been recognized as important signaling molecules in regulation of both innate and adaptive immunity. During mycobacterial infection, antimicrobial peptides including cathelicidin, defensin, and hepcidin have antimicrobial activities against mycobacteria, making them promising candidates for future drug development. Additionally, antimicrobial peptides act as immunomodulators in infectious and inflammatory conditions. Multiple crucial functions of cathelicidins in antimycobacterial immune defense have been characterized not only in terms of direct killing of mycobacteria but also as innate immune regulators, i.e., in secretion of cytokines and chemokines, and mediating autophagy activation. Defensin families are also important during mycobacterial infection and contribute to antimycobacterial defense and inhibition of mycobacterial growth both in vitro and in vivo. Hepcidin, although its role in mycobacterial infection has not yet been characterized, exerts antimycobacterial effects in activated macrophages. The present review focuses on recent efforts to elucidate the roles of host defense peptides in innate immunity to mycobacteria.
Subject(s)
Humans , Adaptive Immunity , Anti-Bacterial Agents , Antimicrobial Cationic Peptides , Autophagy , Cathelicidins , Chemokines , Cytokines , Homicide , Immunity, Innate , Immunologic Factors , Macrophages , Negotiating , PeptidesABSTRACT
Antimicrobial peptides/proteins are ancient and naturallyoccurring antibiotics in innate immune responses in a variety of organisms. Additionally, these peptides have been recognized as important signaling molecules in regulation of both innate and adaptive immunity. During mycobacterial infection, antimicrobial peptides including cathelicidin, defensin, and hepcidin have antimicrobial activities against mycobacteria, making them promising candidates for future drug development. Additionally, antimicrobial peptides act as immunomodulators in infectious and inflammatory conditions. Multiple crucial functions of cathelicidins in antimycobacterial immune defense have been characterized not only in terms of direct killing of mycobacteria but also as innate immune regulators, i.e., in secretion of cytokines and chemokines, and mediating autophagy activation. Defensin families are also important during mycobacterial infection and contribute to antimycobacterial defense and inhibition of mycobacterial growth both in vitro and in vivo. Hepcidin, although its role in mycobacterial infection has not yet been characterized, exerts antimycobacterial effects in activated macrophages. The present review focuses on recent efforts to elucidate the roles of host defense peptides in innate immunity to mycobacteria.
Subject(s)
Humans , Adaptive Immunity , Anti-Bacterial Agents , Antimicrobial Cationic Peptides , Autophagy , Cathelicidins , Chemokines , Cytokines , Homicide , Immunity, Innate , Immunologic Factors , Macrophages , Negotiating , PeptidesABSTRACT
Los péptidos antimicrobianos son las moléculas efectoras del sistema inmune innato, cuyas familias se encuentran en casi todos los organismos, desde bacterias hasta mamíferos. Son una familia de sustancias polifacéticascon complejos mecanismos deacción relacionados con la interacción con el patógeno a través de su membrana, o afectando blancos internos, como la replicación del ADN y la síntesis de proteínas, e interactuando con el huésped con funciones inmunomoduladoras de la regulación delproceso inflamatorio y de la cicatrización. Aunque la generación de resistencia a los péptidos antimicrobianos es mucho menorsi se compara con la generada por losantibióticos convencionales, existen mecanismos de resistencia ya descritos, como la degradación por proteasas, la liberación de proteínas inhibidoras o los cambios en la conformación de la membrana externa del patógeno. El estudio de estas sustancias hapermitido evidenciar sus usos potenciales en el ámbito clínico para contrarrestar los inconvenientes de la resistencia a los antibióticos; sin embargo, a pesar de los grandesavances logrados en este campo, aún quedan puntos controversiales por dilucidar.
The antimicrobial peptides (AMP) are theeffectors molecules of the innate immunesystem, finding groups of this kind of substances in almost all living organisms from bacteria to mammals. They are a family of versatile substances with complexes action mechanisms in the pathogen they interact with membrane, DNA synthesis and protein synthesis and folding, and also with the hostshowing immunomodulatory functions inwound healing and inflammation process.Even though the generation of resistance to the AMP is lower compare with conventional antibiotics there are resistance mechanism already describe to this kind of substances like degradation by proteases, releasing ofinhibitory substances or conformationalchanges in the external membrane of thepathogen. Actually the study of this group of substances has make them see as potential tools for clinical use helping to coun-teract the problem of antibiotic resistance, but even great progress had been made in this field there still exist some controversial issues for future study.
Subject(s)
Cathelicidins , Cecropins , Antimicrobial Cationic Peptides , alpha-Defensins , Anti-Bacterial AgentsABSTRACT
OBJECTIVE@#To investigate the expressions of LL-37 and IL-8 in chronic sinusitis with nasal polyps.@*METHOD@#Semi-quantitative reverse transcription polymerase chain reaction (RT-PCR) and immunohistochemical staining were used to detect the expressions of LL-37 and IL-8 in nasal polyp tissues of 31 patients with chronic sinusitis and inferior turbinate tissues of 11 patients with chronic rhinitis.@*RESULT@#LL-37 and IL-8 mRNA were all positively expressed in all nasal polyps and inferior turbinate tissues. There were significant increases of LL-37 and IL-8 mRNA expressions in nasal polyps compared with the inferior turbinate tissues (P < 0.01). There were also significant increases of positive expression rates of LL-37 and IL-8 protein in nasal polyps, compared with the inferior turbinate tissues (P < 0.01). There was a positive relationship between the mRNA and protein expressions of LL37 and IL-8 (P < 0.01).@*CONCLUSION@#The expressions of LL-37 and IL-8 in nasal polyps suggest that they may play a role in the pathogenesis of chronic sinusitis. Besides its innate immune, LL-37 could enhance human body's anti-infected function by increasing acquired immune.
Subject(s)
Adult , Female , Humans , Male , Middle Aged , Young Adult , Antimicrobial Cationic Peptides , Cathelicidins , Metabolism , Chronic Disease , Interleukin-8 , Metabolism , Nasal Mucosa , Metabolism , Nasal Polyps , Metabolism , Sinusitis , MetabolismABSTRACT
<p><b>OBJECTIVE</b>To develop and investigate GLL-37, a substitution analogue of the human antimicrobial peptide LL-37 with anti-enzymatic degradation activity and improved efficacy.</p><p><b>METHODS</b>The bactericidal activities of LL-37 and newly developed GLL-37 against 6 Gram-negative and -positive bacteria were determined by Broth microdilution assays. The minimum inhibitory concentrations of LL-37 and GLL-37 against E.coli ATCC 25922 in different NaCl concentration medium were also detected. Both peptides were co-incubated with elastase, and then analyzed by PAGE electrophoresis and bactericidal activity determination.</p><p><b>RESULT</b>GLL-37 showed a stronger elastase resistance ability than LL-37, and was significantly more effective than LL-37 under high-salt condition.</p><p><b>CONCLUSION</b>The antimicrobial peptide GLL-37 derived form LL-37 has the potential as a new therapeutic agent for bacterial infections.</p>
Subject(s)
Animals , Female , Humans , Anti-Bacterial Agents , Pharmacology , Therapeutic Uses , Antimicrobial Cationic Peptides , Pharmacology , Therapeutic Uses , Blood Bactericidal Activity , Cathelicidins , Cell Membrane Permeability , Escherichia coli , Membrane Proteins , Metabolism , Monocytes , Pseudomonas Infections , Drug TherapyABSTRACT
In order to obtain a high activity antibacterial peptide, An expression vector pPICZalphaA-pl is constructed with a tandem of four antimicrobial peptides in the same direction,which includes Protegrin-1 (PG-1), Scorpion Defensin (SD), Metalnikowin-2A and Sheep Myeloid Antibacterial Peptide (SMAP-29) (serial number in GenBank are AAB27599, AAAB27538, P80409 and P49928 respectively). At the same time the expression vector pPICZalphaA-sd which express Scorpion Defensin was contructed. The expression vectors of pPICZalphaA-pl and pPICZalphaA-sd were linearized and transformed into the yeast host strain X-33 respectively. Under the control of the promoter AOX1 (alcohol oxidase1), the peptides PL and SD were secreted expressed. Their heat-stable property, acid-stable property and MIC were detected in vitro. The results suggest the peptides PL and SD have good heat-stable and acid-stable properties, and the combinant PL peptide showes higher antibacterial activity against several Gram-positive bacteria (G+) and Gram-negative bacteria (G-) than the peptide SD, especially against Escherichia coli. The antibacterial activity of combinant antimicrobial peptide PL shows its far exploiting perspective.