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.
Cathelicidins ; metabolism ; Chronic Disease ; Humans ; Sinusitis ; immunology ; metabolism ; pathology

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.
Cathelicidins ; biosynthesis ; Chromatography, Affinity ; Escherichia coli ; Genetic Vectors ; Recombinant Fusion Proteins ; biosynthesis ; Thioredoxins ; genetics

OBJECTIVETo investigate the effect of antibacterial peptide LL-37 on the integrity of Acinetobacter baumannii biofilm.

METHODSA 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.

RESULTSIn 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.

CONCLUSIONLL-37 even at a concentration far below its minimal inhibitory concentration can cause structural damage of Acinetobacter baumannii biofilm in vitro.

Acinetobacter baumannii ; drug effects ; physiology ; Biofilms ; drug effects ; Cathelicidins ; pharmacology ; Microbial Sensitivity Tests

BACKGROUND AND OBJECTIVES: Salivary secretions and the secreted IgA in the secretions play a critical role in maintaining oral health via innate host defense mechanism. Cathelicidins are a family of peptides thought to provide an innate defensive barrier against a variety of potential microbial pathogens. LL-37, an antimicrobial peptide, is the only Cathelicidin protein so far identified in humans. The purpose of this study was to examine the expression of Cathelicidin in human salivary glands and to investigate upregulation of Cathelicidin in inflammatory conditions. MATERIALS AND METHOD: Reverse transcriptase-polymerase chain reaction (RT-PCR) and immunohistochemical staining were performed on 20 salivary gland tissues, of which 10 were normal and 10 were chronic sialadenitis. RESULTS: Cathelicidin mRNA transcripts were detected in the normal salivary glands and chronic sialadenitis. The level of Cathelicidin mRNA in chronic sialadenitis was significantly increased compared with that in the normal salivary gland. Cathelicidin protein was expressed in the glandular epithelium of the normal salivary gland and chronic sialadenitis. CONCLUSION: The results indicate that Cathelicidin might play an important role in the innate host defense of human salivary glands.
Cathelicidins ; Epithelium ; Humans ; Immunoglobulin A ; Oral Health ; Peptides ; RNA, Messenger ; Salivary Glands ; Sialadenitis* ; Up-Regulation*

BACKGROUND AND OBJECTIVES: Antimicrobial peptides and proteins play an important role in innate host defense, and this may be particularly important at mucosal surfaces that form the initial barrier between the host and the external environment. Epithelial cells are the first line of defense mechanism against microorganisms, where antimicrobial peptides are the major participants. Cathelicidins are the precursors of potent antimicrobial peptides that have been identified in several mammalian species. Cathelin-related antimicrobial peptide (CRAMP) is one of the antimicrobial peptides and is the only member of cathelicidin family identified so far in mouse. The present study was undertaken to investigate the expression of CRAMP in the eustachian tube epithelium of mouse. MATERIALS AND METHOD: Tissue samples from the mouse eustachian tube were recovered from its pharyngeal, middle, and distal segments. CRAMP was localized by immunohistochemical staining and mRNA expression was determined by reverse transcriptase-polymerase chain reaction (RT-PCR). RESULTS: By immunohistochemical study, CRAMP was detected in epithelial cells and submucosal glands of the eustachian tube, but not in the negative control. Using RT-PCR, CRAMP mRNA was detected in the eustachian tube epithelium. CONCLUSION: The expression and localization of CRAMP in the epithelial cells and submucosal glands of the eustachian tube of mouse were defined. We found that CRAMP is one of the antimicrobial peptides found in the eustachian tube epithelium of mouse, and that it participates in the innate immune system of the eustachian tube.
Animals ; Cathelicidins ; Epithelial Cells ; Epithelium* ; Eustachian Tube* ; Humans ; Immune System ; Mice* ; Muscle Cramp ; Peptides ; RNA, Messenger

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.
Animals ; Cathelicidins ; Dermatitis, Atopic ; Epidermis ; Intercellular Signaling Peptides and Proteins ; Keratinocytes ; Mice ; Oxazolone ; Peptides ; Permeability ; Phenotype ; Stem Cells

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.
Humans ; Titanium/chemistry* ; Antimicrobial Peptides ; Cathelicidins ; Sincalide ; Anti-Bacterial Agents/pharmacology* ; Nanotubes/chemistry* ; Dental Materials ; Bacteria ; Keratinocytes ; Surface Properties

BACKGROUND: The large surfaces of the skin are often initial site of contact between microorganism and human. The skin are coated with epidermis and epithelial cells can recognize microorganism and mount a fast defense through the production of various inducible antibiotic peptides. This leads to chracteristic broad spectrum of antimicrobial activity against bacteria, fungi, and viruses. Recent studies introduce us new peptides with antimicrobial activity such as P,-defensins and cathelicidins. They are expressed on the epithelia and polymorphonuclear leukocytes, which are first lines of defence from various invasive environments. Futhermore, they are considered very interesting and important endogenous antibiotics. Our previous study has shown that the expression of human defensin(hBD-2) mRNA, which is potent antibiotic peptide against Gram-negative bacteria(P. aeruginosa), was upregulated with ultraviolet(UV) irradiation, tumor necrosis factor-α(TNF-α) and lipopolysaccharide(LPS) in HaCaT cells. A novel hBD-3, 5-kDa, nonhemolytic antimicrobial peptide, was demonstrated a salt-insensitive broad spectrum of potent antimicrobial activity against many potentially pathogenic microbes in especially, multiresistant S. aureus. We have analyzed the expression patterns of hBD-3 in HaCaT cell lines. OBJECTIVE: This research have done in order to evaluate the expression and regulation of hBD-3 mRNA in human keratinocyte cell lines. METHODS: HaCaT cell lines were used to all culture experiments. Cultured human keratinocytes were stimulated with UV irradiation or TNF-α or LPS to determine whether hBD-3 mRNA production occurred. Reverse transcription-polymerase chain reaction (RT-PCR) was per-formed to amplify hBD-3 cDNA from stimulated keratinocytes in a time dependant manner, and densitometry was used to verify the specificity of RT-PCR amplication products. RESULTS: Expression of hBD-3 was upregulated with UV irradiation, TNF-α and LPS in Ha-CaT cells compared to control CONCLUSIONS: Human keratinocytes are capable to induce hBD-3 mRNA, as well as hBD-2, in response to UV irradiation, TNF-α and LPS. suggesting that these cells could play an important role against the bacterial infection and UV light damage in human skin.
Anti-Bacterial Agents ; Bacteria ; Bacterial Infections ; Cathelicidins ; Cell Line* ; Densitometry ; DNA, Complementary ; Epidermis ; Epithelial Cells ; Fungi ; Humans* ; Keratinocytes ; Necrosis ; Neutrophils ; Peptides ; RNA, Messenger ; Sensitivity and Specificity ; Skin ; Ultraviolet Rays

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.
Animals ; Antimicrobial Cationic Peptides ; chemistry ; genetics ; physiology ; Blood Proteins ; chemistry ; genetics ; physiology ; Cathelicidins ; chemistry ; genetics ; physiology ; Drug Design ; Recombinant Proteins ; chemistry ; genetics ; Sheep

OBJECTIVETo investigate the core functional region of antimicrobial peptide LL-37, which inhibites RSV replication and could be developed for theraputic aplication.

METHODSA 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.

RESULTSN-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.

CONCLUSIONThe 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.

Cathelicidins ; pharmacology ; Cell Line ; Cytokines ; genetics ; immunology ; Humans ; Respiratory Syncytial Virus Infections ; genetics ; immunology ; virology ; Respiratory Syncytial Viruses ; drug effects ; physiology ; Virus Replication ; drug effects