beta-defensins
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The surrounding environment contains plenty of pathogens, which represent a danger of infection. The simplest way for the pathological microorganism to enter the organism is the upper airways. Inflammation of the upper airways is among the most common and frequent diseases. This category includes nasal polyposis and chronic tonsillitis. In many cases it is associated with disorders in relation to the immune response. An inflammatory infiltration of mononuclears, eosinophils, plasma and mast cells can be found in the histological structure of the polypous as well as tonsillar mucosa. One aim of this study was to determine the expression of beta-defensins and various proteins, with a possible potential role in relation to the rise and development of those changes. Another aim was to determine the relationship between the inflammatory and malignant processes in the tonsils. The samples of nasal polyps were obtained during clinically indicated endonasal surgery from patients diagnosed with nasal polyposis (n=50). The samples of tonsils were collected during surgery from patients suffering from chronic tonsillitis (n=11) or tonsillar carcinoma (n=17). Immunohistochemical procedures for the detection of human beta-defensin 1, 2, 3 (HBD-1, 2, 3), Ki- 67, endothelial nitric oxide synthase (eNOS) and cleaved caspase 3 were performed on cryostate and paraffin sections. It was proven that HBD are secreted in fairly large amounts in cases of chronic inflammation. Their secretion during the malignant transformation is limited. This is a very probable fact that plays a role in malignant transformation in tonsillar tissue. The crucial role in the development of chronic inflammation, and maybe that of malignant transformation, is played by eNOS and its product NO molecule. eNOS and the NO molecule are involved in cell cycle regulation, in the apoptotic processes and cell proliferation, as well as in the angiogenesis and vasculogenesis. Our result confirmed that eNOS is presented in the tissues of the upper airways in both chronic inflammation and carcinomatous processes. Ki-67 and cleaved caspase 3 were used as markers of cell proliferation and apoptosis.
- MeSH
- antigen Ki-67 metabolismus MeSH
- apoptóza MeSH
- beta-defensiny metabolismus MeSH
- chronická nemoc MeSH
- fluorescenční protilátková technika nepřímá MeSH
- kaspasa 3 metabolismus MeSH
- lidé MeSH
- nádorová transformace buněk MeSH
- nosní polypy imunologie patologie MeSH
- nosní sliznice imunologie metabolismus MeSH
- proliferace buněk MeSH
- synthasa oxidu dusnatého, typ III metabolismus MeSH
- tonzilární nádory enzymologie patologie MeSH
- tonzilitida enzymologie patologie MeSH
- zánět etiologie patologie MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- práce podpořená grantem MeSH
Defensiny jsou antimikrobiální a imunomodulační peptidy, které jsou důležitou součástí přirozené imunity. Autoři stručně charakterizují přirozenou imunitu, chemickou stavbu a funkci defensinů. Zabývají se především lidskými defensiny na respiračních sliznicích a jejich účinkem na bakterie, a to hlavně na grampozitivní bakterii Staphylococcus aureus. Defensiny mají přímý destrukční účinek na bakteriální stěnu, ale mají i imunomodulační účinek. Lidské defensiny dělíme podle struktury na alfa a beta 1, beta 2, beta 3 a beta 4. Alfa jsou v granulocytech, ve sliznici urogenitálního a intestinálního traktu. Beta defensiny jsou ve všech epiteliálních tkáních. V epitelu dýchacích cest jsou beta defensiny 1 produkovány konstitučně a beta defensiny 2 a 3 jsou indukovány jako odpověď na infekční agens. Některé bakterie mohou modifikovat svoji stěnu tak, aby byly méně citlivé na defensiny. U bakterie Staphylococcus aureus byly zjištěny geny Dlt a MprF, které jsou zodpovědné za modifikaci kyseliny teichoové a phosphatidylglycerolu v membráně takovým způsobem, že se sníží atraktivita bakterie pro pozitivně nabité defensiny. Poruchy těchto genů způsobují zvýšenou senzitivitu této bakterie na defensiny. Nové poznatky o antimikrobiálních peptidech nám umožňují lépe rozumět infekčním onemocněním a nabízejí nové perspektivy v terapii těchto onemocnění.
Defensins are antimicrobial and immunomodulation peptides, which are the important part of natural immunity. The authors briefly characterize natural immunity, chemical structure and function of defensins. The review deals preferentially with human defensins on respiratory mucous membranes and their effects on bacteria, mainly on gram-negative Staphylococcus aureus. Defensins exert a direct destructive effect on bacterial wall, but they possess an immunomodulation effect as well. Human defensins are divided according to their structure to alpha, beta 1, beta 2, beta 3 and beta 4. Alpha defensins are in granulocytes, in mucosa of urogenital and intestinal tract. Beta defensins are in all epithelial tissues. In the epithelium of respiratory pathways beta defensins 1 are produced constitutively and beta defensins 2 and 3 are induced as a response to infection agents. Some bacteria can modify their wall and thereby become less sensitive to defensins. In the bacteria Staphylococcus aureus the genes Dlt and Mrpf were found responsible for modification of teichoic acid and phosphatidylglycerol in the membrane in such a way that it decreases attractiveness of the bacteria for the positively charged defensins. Defects in these genes caused increased sensitivity of this bacterium to defensins. New knowledge of antibacterial peptides helps us to understand better infectious diseases and offers new perspectives in the therapy of these diseases.
- MeSH
- alfa-defensiny genetika imunologie klasifikace MeSH
- beta-defensiny genetika imunologie klasifikace MeSH
- defensiny imunologie klasifikace terapeutické užití MeSH
- financování organizované MeSH
- imunologické faktory genetika imunologie MeSH
- kationické antimikrobiální peptidy genetika imunologie MeSH
- lidé MeSH
- rezistence dýchacích cest fyziologie genetika imunologie MeSH
- Staphylococcus aureus genetika imunologie MeSH
- Check Tag
- lidé MeSH
The nasal and tonsillar mucosa are exposed to massive incursions of pathological microorganisms. One of the mechanisms known to prevent an invasion of pathogens is an endogenous synthesis of antimicrobial peptides, which include human beta-defensins-1, 2, 3 (HBD-1, 2, 3). The aim of this study was to demonstrate the occurrence of HBD-1, 2 and 3 in the human nasal mucosa and palatine tonsils in healthy tissues and during chronic inflammation (nasal polyposis with and without the colonization of Staphylococcus aureus and chronic tonsillitis) and to evaluate their incidence under varying conditions. Another target was to compare the occurrence of human beta-defensins in these two different entities; that is, in the nasal mucosa and in the palatine tonsil. It was assumed that the incidence of HBD-1, 2, 3 was lower in tonsils than in nasal mucosa; however, inflamed samples of tonsils and nasal mucosa showed no difference in the production of HBD-1, 2, 3. The presence of all three subfamilies of HBD was significantly lower in nasal polyps with Staphylococcus aureus positive than in the negative control.
- Klíčová slova
- beta-defensins, nasal mucosa, palatine tonsil, chronic inflammation,
- MeSH
- beta-defensiny imunologie izolace a purifikace MeSH
- financování organizované MeSH
- imunohistochemie metody využití MeSH
- kationické antimikrobiální peptidy imunologie izolace a purifikace MeSH
- krční mandle fyziologie mikrobiologie patologie MeSH
- kultivační techniky metody využití MeSH
- lidé MeSH
- nosní polypy imunologie mikrobiologie MeSH
- nosní sliznice fyziologie mikrobiologie patologie MeSH
- Staphylococcus aureus imunologie izolace a purifikace MeSH
- studie případů a kontrol MeSH
- tonzilitida diagnóza imunologie mikrobiologie MeSH
- Check Tag
- lidé MeSH
Studies of the human defensins have been hampered by the lack of a simple expression system allowing for rapid production of functional peptide forms. Here, we describe a Saccharomyces cerevisiae AH22 expression system that meets that condition. The 42 amino acid form of human beta-defensin-1 was expressed under the control of the ADH1 promoter. The optimum conditions for expression were determined and the stable maintenance of the pVT103L-hBD-1 chimeric vector in the yeast population was confirmed. Expressed hBD-1 was secreted into the medium (approximately 55 microg l(-1)) and purified using cation-exchange chromatography. Isolated defensin exhibited strong bactericidal effect on Escherichia coli ML-35p. We conclude that the expression system described here will be a useful tool where readily prepared and active forms of the human defensins are needed.
- MeSH
- antibakteriální látky biosyntéza farmakologie izolace a purifikace MeSH
- beta-defensiny biosyntéza farmakologie genetika chemie MeSH
- Escherichia coli cytologie účinky léků MeSH
- genetické vektory genetika MeSH
- lidé MeSH
- molekulární sekvence - údaje MeSH
- proliferace buněk účinky léků MeSH
- rekombinantní proteiny biosyntéza farmakologie chemie izolace a purifikace MeSH
- Saccharomyces cerevisiae genetika MeSH
- sekvence aminokyselin MeSH
- Check Tag
- lidé MeSH
BACKGROUND: There have been conflicting reports in the literature on association of gene copy number with disease, including CCL3L1 and HIV susceptibility, and β-defensins and Crohn's disease. Quantification of precise gene copy numbers is important in order to define any association of gene copy number with disease. At present, real-time quantitative PCR (QPCR) is the most commonly used method to determine gene copy number, however the Paralogue Ratio Test (PRT) is being used in more and more laboratories. FINDINGS: In this study we compare a Pyrosequencing-based Paralogue Ratio Test (PPRT) for determining beta-defensin gene copy number with two currently used methods for gene copy number determination, QPCR and triplex PRT by typing five different cohorts (UK, Danish, Portuguese, Ghanaian and Czech) of DNA from a total of 576 healthy individuals. We found a systematic measurement bias between DNA cohorts revealed by QPCR, but not by the PRT-based methods. Using PRT, copy number ranged from 2 to 9 copies, with a modal copy number of 4 in all populations. CONCLUSIONS: QPCR is very sensitive to quality of the template DNA, generating systematic biases that could produce false-positive or negative disease associations. Both triplex PRT and PPRT do not show this systematic bias, and type copy number within the correct range, although triplex PRT appears to be a more precise and accurate method to type beta-defensin copy number.
- MeSH
- beta-defensiny genetika MeSH
- genetická predispozice k nemoci MeSH
- genom lidský genetika MeSH
- genová dávka * MeSH
- kohortové studie MeSH
- lidé MeSH
- mapování chromozomů metody MeSH
- molekulární sekvence - údaje MeSH
- populace MeSH
- populační genetika metody MeSH
- sekvence nukleotidů MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- srovnávací studie MeSH
- Geografické názvy
- Česká republika MeSH
- Dánsko MeSH
- Ghana MeSH
- Portugalsko MeSH
- Spojené království MeSH
Východiska: Pokroky v medicíně, zejména v oblasti imunologie v onkologii, přispívají k vývoji nových způsobů a metod léčby karcinomu ústní dutiny a orofaryngu. Jsou prováděny preklinické a klinické studie terapie cytokiny a dendritickými buňkami, blokády imunitních kontrolních bodů, vychytávání T lymfocytů, ošetření monoklonálními protilátkami a peptidovými vakcínami. Některé z těchto imunoterapeutických léčiv jsou zaváděny do lékařské praxe. Změny v imunitním systému pacientů s karcinomem ústní dutiny a orofaryngu odůvodňují proveditelnost dalšího cíleného výzkumu nových imunoterapeutických účinků na maligní nádory. Materiály a metody: Článek přináší data z analýzy protinádorové imunity 61 pacientů s karcinomem ústní dutiny a orofaryngu, kteří absolvovali (chemo) radioterapii na pozadí imunoterapie přípravkem s alfa/beta defenziny. Bylo provedeno srovnání laboratorních parametrů s klinickým pozorováním regrese tumoru pacientů. Vliv imunomodulačního přípravku s alfa/beta defenziny byl určen analýzou změn absolutního a relativního počtu lymfocytů, počtu CD3+ T lymfocytů, přirozených zabíječů (natural killers – NK) a NKT lymfocytů. Výsledky: Bylo zjištěno, že imunomodulační přípravek s alfa/beta difenziny zvyšuje cytostatický protinádorový efekt (chemo) radioterapie, má účinek závislý na dávce a cytoprotektivní účinek na NK buňky, takže imunitní odpověď na vývoj nádoru je použitím tohoto přípravku zvýšena. Data laboratorního vyšetření stavu imunity odpovídají přímým výsledkům regrese tumoru u pacientů s karcinomem ústní dutiny a orofaryngu. Klinicky vyšší míra regrese byla zaznamenána u pacientů léčených radioterapií a imunoterapií v dávkách 40 a 60 mg. Závěr: Stanovili jsme protinádorovou účinnost imunomodulačního přípravku s alfa/beta defenziny při léčbě pacientů se spinocelulárním karcinomem ústní dutiny a orofaryngu a její proveditelnost na klinice.
Background: Advances in medicine, especially in the field of immunology in oncology, contribute to the development of new ways and methods of treating cancer of the oral cavity and oropharynx. Preclinical and clinical studies of cytokine and dendritic cell therapy, blockade of immune checkpoints, T cell uptake, treatment with monoclonal antibodies and peptide vaccines are performed. Some of these immunotherapeutic drugs are introduced into medical practice. Changes in the immune system of patients with cancer of the oral cavity and oropharynx justify the feasibility of further targeted research of new immunotherapeutic effects on malignant tumors. Materials and methods: The article represents the data of the analysis of antitumor immunity in 61 patients with cancer of the oral cavity and oropharynx who received (chemo) radiotherapy against the background of immunotherapy with the immune agent containing alpha/beta-defensins, and compares laboratory indices with clinical observation of tumor regression in patients. The influence of the immune agent containing alpha/beta-defensins was determined by analyzing the changes in the absolute and relative numbers of lymphocytes, the number of CD3+ T cells, natural killers (NK) and natural killer T cells (NKT cells). Results: The preparation containing alpha/beta-defensins has been found to enhance cytostatic antitumor effect of (chemo) radiotherapy, having a dose-dependent and cytoprotective effects considering NK cells, so the immune response to the tumor development is enhanced with the use of this agent. The data of laboratory examination of immune status correspond to the direct results of tumor regression in patients with cancer of the oral cavity and oropharynx. Clinically higher regression indices are in patients receiving radiotherapy with immunotherapy at doses of 40 and 60 mg. Conclusion: We confirmed the antitumor efficacy of the immunomodulatory agent containing alpha/beta-defensins in the treatment of patients with squamous cell carcinoma of the oral cavity and oropharynx and the reasonability of its use on the clinic.
- MeSH
- beta-defensiny imunologie MeSH
- hojení ran fyziologie imunologie MeSH
- keratinocyty imunologie metabolismus MeSH
- lidé MeSH
- Check Tag
- lidé MeSH
