Opredelitev specifičnega T celičnega odziva proti citomegalovirusu z določevanjem znotrajceličnih citokinov

Čelofiga, Nadja (author), Kopitar, Andreja Nataša (mentor)

Humani citomegalovirus (CMV) spada v družino Herpesviridae. Pri osebah z normalnim imunskim odzivom primarna okužba povzroča infekcijski mononulkeozi podobno bolezen ali poteka asimptomatsko. Pri imunokompromitiranih posameznikih pa zaradi razširjenosti virusa in latentne narave okužbe CMV predstavlja tveganje za hujši potek okužbe ali smrtnost bolnika. Okužba s CMV predstavlja velik problem po zaključku protivirusne profilakse pri bolnikih s presaditvijo organov. Z in vitro določanjem T celičnega imunskega odziva proti virusu CMV lažje ocenimo tveganje za okužbo s CMV po presaditvi organov in na podlagi tega individualno prilagodimo protivirusno trapijo ter preprečimo kasnejše zaplete. Namen naše raziskave je bil uvedba metode znotrajceličnega določevanja proizvodnje interferona gama (IFN-γ) v limfocitih T pri spodbujanju z različnimi antigeni CMV. V študiji je bilo vključenih 8 zdravih prostovoljcev, ki so že prišli v stik z virusom. V krvi bolnikov in zdravih oseb smo določili koncentracijo protiteles IgG in IgM proti CMV in na pretočnem citometru izmerili proizvodnjo IFN-γ po stimulaciji z različnimi antigeni. V naši študiji smo testirali različne CMV peptide, s katerimi smo stimulirali limfocite T in ugotovili, da US3 najbolje stimulira CD4+ in CD8+ populacijo limfocitov T. Merjenje specifičnega T celičnega odziva na CMV z merjenjem znotrajceličnega IFN-γ v limfocitih T je pomembno pri odločanju o zdravljenju bolnikov predvsem v primeru, ko je koncentracija limfocitov zmanjšana in IFN-γ ni mogoče določevati s komercialnim QuantiFERON-CMV testom.

• Language: Slovenian
• Keywords: virusi, humani citomegalovirus, imunski odziv, T celični imunski odziv, molekularna diagnostika, interferon gama, znotrajcelično označevanje citokinov
• Work type: Master's thesis/paper
• Typology: 2.09 - Master's Thesis
• Organization: BF - Biotechnical Faculty
• Publisher: [N. Čelofiga]
• Year: 2020
• PID: 20.500.12556/RUL-117312
• UDC: 578.083.3:577.27
• COBISS.SI-ID: 22146051
• Publication date in RUL: 05.07.2020

ČELOFIGA, Nadja, 2020, Opredelitev specifičnega T celičnega odziva proti citomegalovirusu z določevanjem znotrajceličnih citokinov [online]. Master’s thesis. N. Čelofiga. [Accessed 2 April 2025]. Retrieved from: https://repozitorij.uni-lj.si/IzpisGradiva.php?lang=eng&id=117312

Secondary language

• Language: English
• Title: Asessment of specific t-cell response to cytomegalovirus by determination of intracellular cytokines

Abstract

Human cytomegalovirus (CMV) is a part of Herpesviridae family. In subjects with a normal immune response, the primary infection is asymptomatic or causes a disease, similar to mononucleosis. With immunocompromised individuals, due to the latent nature and the prevalence of the virus, CMV presents a high risk for severe infection or even death of the patient. CMV infection poses a major risk in patients with solid-organ transplantation after the end of antiviral prophylaxis. With in vitro determination of the T cell immune response to CMV we can assess the risk of late onset of CMV infection and adapt the treatment individually so that we can avoid subsequent complications. The purpose of our study was to introduce an intracellular method for determining interferon gamma (IFN-γ) production in T lymphocytes when stimulated with various CMV antigens. The study included 8 healthy volunteers who had already come into contact with the virus. The levels of IgG and IgM antibodies against CMV were determined in the blood of patients and healthy subjects and the production of IFN-γ after stimulation with different antigens was measured on a flow cytometer. In our study, we tested various of CMV peptides that were stimulated by T lymphocytes and found that US3 is the best stimulating peptide for CD4 + and CD8 + population of T lymphocytes. Measurement of a specific T cell response to CMV by measuring intracellular IFN-γ in T lymphocytes is important in deciding treatment of patients especially when lymphocyte concentration is decreased and IFN-γ cannot be determined by the commercial QuantiFERON-CMV assay.

• Keywords: viruses, human cytomegalovirus, immune response, T-cell immune response, molecular diagnostics, interferon gamma, intracellular cytokine staining