Izražanje izbranih genov v periferni krvi bolnikov med anafilaksijo in po stimulaciji in vitro

Svetina, Manca (author), Rijavec, Matija (mentor), Koren, Ana (co-mentor)

Anafilaksija je resna sistemska preobčutljivostna reakcija, ki se razvije v nekaj minutah po stiku z alergenom. Za potrjevanje klinične diagnoze anafilaksije se trenutno uporablja določanje triptaze v serumu. Biooznačevalec, ki bi omogočal nedvoumno diagnozo anafilaksije, še ni znan. V predhodnih raziskavah se je izkazalo, da bi lahko imeli pomembno vlogo pri bioloških procesih med anafilaksijo tudi citokin onkostatin M (OSM), matrična metaloproteinaza 9 (MMP9) in receptor za interlevkin 1 tipa II (IL1R2). Zaradi tega smo zasnovali raziskavo, v kateri smo s pomočjo verižne reakcije s polimerazo v realnem času (RT-qPCR) merili izražanje genov IL1R2, MMP9 in OSM na ravni mRNA v krvi bolnikov med anafilaksijo. Izražanje izbranih genov se je med akutno fazo anafilaksije statistično značilno povišalo. S krivuljami ROC smo prikazali, da sta občutljivost in specifičnost oblikovanih diagnostičnih testov celo boljši kot pri diagnostičnem testu triptaze. Za eksperimentalno primerjavo smo v drugem delu naloge iz krvi bolnikov s potrjeno preobčutljivostjo za strup čebele ali ose izolirali periferne mononuklearne celice (PMNC), jih in vitro stimulirali z alergenom ter preverili izražanje prej omenjenih genov.

• Language: Slovenian
• Keywords: anafilaksija, RT-qPCR, izražanje genov, označevalci, IL1R2, MMP9, OSM
• Work type: Master's thesis/paper
• Typology: 2.09 - Master's Thesis
• Organization: BF - Biotechnical Faculty
• Place of publishing: Ljubljana
• Publisher: [M. Svetina]
• Year: 2021
• PID: 20.500.12556/RUL-133356
• UDC: 606:57.083.32:601.4:577.21(043.2)
• COBISS.SI-ID: 88805891
• Publication date in RUL: 24.11.2021

Secondary language

• Language: English
• Title: Peripheral blood gene expression in patients during anaphylaxis and after stimulation in vitro

Abstract

Anaphylaxis is a serious systemic hypersensitivity reaction that develops within minutes after exposure to a specific allergen. Serum tryptase concentration is the most used laboratory test to confirm the clinical diagnosis of anaphylaxis. A biomarker that would allow an unambiguous diagnosis of anaphylaxis is still not recognized. Previous research has shown that cytokine oncostatin M (OSM), matrix metalloproteinase 9 (MMP9) and interleukin 1 type II receptor (IL1R2) could play an important role in biological processes during anaphylaxis. For this reason, we designed a study where we measured the relative mRNA expression of IL1R2, MMP9 and OSM genes in the blood of patients during anaphylaxis using real-time polymerase chain reaction (RT-qPCR). The mRNA expression of selected genes increased during the acute phase of anaphylaxis. ROC curves showed that the sensitivity and specificity of the designed diagnostic tests are even better than tryptase. For experimental comparison, peripheral mononuclear cells (PMNCs) were isolated from the blood of patients with confirmed hypersensitivity to bee or wasp venom, in vitro stimulated with an allergen, and mRNA expression of the aforementioned genes was measured.

• Keywords: anaphylaxis, RT-qPCR, gene expression, biomarkers, IL1R2, MMP9, OSM