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Vzpostavitev modelnega sistema za določanje imunomodulatornih lastnosti mlečnokislinskih bakterij na celični liniji THP-1
ID Repar, Lara (Author), ID Lunder, Mojca (Mentor) More about this mentor... This link opens in a new window

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Abstract
Mlečnokislinske bakterije so grampozitivne bakterije, ki imajo podobne fiziološke in metabolne lastnosti. Uporabljajo se predvsem v živilski industriji, pri proizvodnji fermentiranih izdelkov, ter v farmacevtski industriji kot dostavni in ekspresijski sistemi. Zaradi ugodnih učinkov na zdravje gostitelja številne mlečnokislinske bakterije uvrščamo med probiotike. Različne študije so pokazale, da mlečnokislinske bakterije sprožijo aktivacijo monocitov, spodbudijo izločanje citokinov in vplivajo na delovanje drugih imunskih celic. Učinki mlečnokislinskih bakterij so zelo raznoliki in odvisni od vrste ter seva bakterije, zato je izbira ustrezne bakterije ključnega pomena za uspešnost študije. Teste in vitro uporabljamo za iskanje kandidatov z ustreznimi imunomodulatornimi lastnostmi. V okviru diplomske naloge smo določali porast izločanja vnetnega citokina interlevkina 8 po izpostavitvi celic THP-1 inaktiviranim modelnim mlečnokislinskim bakterijam. Citokine izločajo številne imunske celice pod vplivom različnih dražljajev, kot so bakterijski endotoksini. Kot modelno mlečnokislinsko bakterijo smo uporabili Lactococcus lactis, ki smo jo inaktivirali z izpostavitvijo ultravijoličnemu valovanju. Porast izločanja citokina smo merili z encimsko-imunskim testom (ELISA) in količino izločenega interlevkina 8 primerjali z izločanjem po tretiranju celic THP-1 z bakterijo Escherichia coli, ki nam je služila za kontrolo. Kot model za primarne monocite smo uporabili celično linijo THP-1, ki se običajno uporablja za določanja dejavnosti monocitov in makrofagov v študijah in vitro. Vzpostavili smo test za določanje vpliva mlečnokislinskih bakterij na izločanje interlevkina 8 iz celične linije THP-1. Potrdili smo, da inaktivirana Lactococcu lactis modulira izločanje interlevkina 8 iz monocitne celične linije THP-1.

Language:Slovenian
Keywords:mlečnokislinske bakterije, Lactococcus lactis, interlevkin 8, citokini, celična linija THP-1
Work type:Bachelor thesis/paper
Organization:FFA - Faculty of Pharmacy
Year:2021
PID:20.500.12556/RUL-129912 This link opens in a new window
Publication date in RUL:09.09.2021
Views:1259
Downloads:69
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Secondary language

Language:English
Title:A THP-1 cell line-based model system setup for testing immunomodulatory properties of lactic acid bacteria
Abstract:
Lactic acids bacteria are gram positive bacteria with similar physiologic and metabolic properties. They are primarily used in the food industry in the production of fermented items and in the pharmaceutical industry as delivery and expression systems. Due to their positive effects on human health, they are classified as probiotics. Many studies have concluded that lactic acid bacteria trigger the activation of monocytes, induce cytokine secretion and affect the activity of other immune cells. The effects of lactic acid bacteria are very diverse and dependent on the sort and the strain of bacteria, therefore the selection of a suitable strain is crucial to the success of a study. In vitro tests are used for searching candidates that possess suitable immunomodulatory properties. In our research, we established a model for testing the effect of inactive lactic acid bacteria on the release of the pro-inflammatory cytokine interleukin 8 in THP-1 cells. The cytokines are secreted by several immune cells, while under the influence of a variety of stimuli, such as bacterial endotoxins. We used Lactococcus lactis as our model bacteria. The bacteria were deactivated by exposure to ultraviolet light. The increase of cytokine secretion was measured with enzyme-linked immunosorbent assay (ELISA). The quantity of the secreted interleukin 8 was then compared with the secretion after treatment of THP-1 cells with Escherichia coli, which served as a control. We used the THP-1 cell line as a model for primary monocytes, which is usually used for defining monocyte and macrophage activity in in vitro studies. We established a test to determine the effect of lactic acid bacteria on the secretion of interleukin 8 from the THP-1 cell line. We confirmed that inactivated Lactococcus lactis modulates the secretion of interleukin 8 from the monocyte cell line THP-1.

Keywords:lactic acid bacteria, Lactococcus lactis, interleukin 8, cytokines, THP-1 cell line

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