PhD Scientific Days 2026

Poster Session 3.U - Molecular Medicine

Development of Mouse Cytokine-expression Panel for Detection of Immunomodulation of Cellular Therapies

Előadó neve

Balázs, Sámuel

Neptune code

E5YNWZ

Előadó munkahelye

HUN-REN RCNS, Institute of Molecular Life Sciences

Előadó telefonszáma

+36307121385

Előadó e-mail címe

balazs.samuel@gmail.com

Az előadás címe

Development of Mouse Cytokine-expression Panel for Detection of Immunomodulation of Cellular Therapies

Szerző(k) neve és munkahelye

Sámuel Balázs¹, Ferenc Fekete¹, Károly Hegedűs^1,2,3, Katalin Monostory¹

1: HUN-REN RCNS, Institute of Molecular Life Sciences
2: Semmelweis University Doctoral School
3: Toxi-Coop Toxicological Research Center Plc.

Bemutatás módja

Poszter

Szekció

Poster Session 3.U - Molecular Medicine

Language of the presentation

English

Preferred session

Molecular Medicine

Összefoglaló szövege

Over the past 40 years, cellular therapies have revolutionized targeted immunomodulation, evidenced by over 10,000 clinical trials conducted worldwide. To assess the efficacy and to evaluate the safety of these products, it is crucial to obtain a comprehensive overview of their immunomodulatory effects. To achieve this, we measure cytokine mRNA expression in immunocompetent cells using high-throughput qPCR, which enables a reliable, rapid, and multiplexed workflow.
We aimed to design and validate qPCR methods for the analysis of a 23-component cytokine panel in mice, targeting genes previously reported to exhibit altered expression during immunomodulatory cell therapies.
The in silico primer design followed a strict pipeline, ensuring that their physicochemical properties (melting temperature, amplicon length, hairpin and dimer formation) matched those already established in our laboratory. Furthermore, either the forward or the reverse primer was designed to span an exon-exon junction, thereby preventing amplification of genomic DNA impurities during PCR. The amplification product was tested by melt curve analysis, while the amplicon length was verified using an automated gel electrophoresis system. Primer specificity was validated by Sanger sequencing, and amplification efficiency was determined using our high-throughput qPCR.
Regular melting curves with a single peak were obtained for all primer pairs. Gel electrophoresis showed an average deviation of 5% from the in silico calculated amplicon lengths. Sequencing results were cross referenced using the NCBI Nucleotide Blast, confirming that all amplicons matched their intended target genes. Finally, an average efficiency of 88% was observed across all assays.
The 23 PCR primer-pairs in the panel for measuring mouse cytokine expression met our predefined requirements. The single peaks observed in melt curve analysis confirmed that only a single product was amplified during PCR. Gel electrophoresis revealed only minor deviations between the calculated and measured amplicon lengths, and sequencing verified that all the amplicons matched their respective target genes. An average amplification efficiency of 88% supports consistent and reproducible measurements of cytokine expression.
This work was supported by the SE 250+ Excellence PhD Scholarship of Semmelweis University and RGH-151536 provided by the NRDIO

University

Semmelweis University

Supervisor

Katalin Monostory

Publication of my abstract

I do not give consent to the publication of my abstract on the website of the congress.

phd.section.field

in doctoral studies before complex exam (PhD)

Kind

Szabad

Status

elfogadva

Accepted presentation method

poszter

Előadás fájl jóváhagyás

nem rendelkezett róla

Előadó

9732

Start

14:24

End

14:27