PhD Scientific Days 2026

Poster Session 1.A - Molecular Medicine

Respiratory burst and degranulation of neutrophil-like cells differentiated from CRISPR/Cas9 modified conditionally immortalized myeloid progenitors

Előadó neve

Jex, Emiliána

Neptune code

ED67G7

Előadó munkahelye

Department of Physiology, Semmelweis University

Előadó telefonszáma

+36205492501

Előadó e-mail címe

jex.emiliana@semmelweis.hu

Az előadás címe

Respiratory burst and degranulation of neutrophil-like cells differentiated from CRISPR/Cas9 modified conditionally immortalized myeloid progenitors

Szerző(k) neve és munkahelye

Emiliána Jex¹, Áron Pánczél¹, Attila Mócsai¹

1: Department of Physiology, Semmelweis University

Bemutatás módja

Poszter

Szekció

Poster Session 1.A - Molecular Medicine

Language of the presentation

English

Preferred session

Molecular Medicine

Összefoglaló szövege

Neutrophils are difficult to be manipulated by genetic approaches. This has been overcome by conditional immortalization of mouse myeloid progenitors (HoxB8 progenitors), from which HoxB8 neutrophil like cells (NLCs) can be differentiated. We have set up a system for the CRISPR/Cas9-based genetic manipulation of HoxB8 progenitors and were able to delete several isoforms of a protein family, including the PKC-α (Prkca CRKO), the PKC-β (Prkcb CRKO), the PKC-δ (Prkcd CRKO), and the PKC-ε (Prkce CRKO).
The aim of our experiments was to investigate the respiratory burst and the degranulation of the various CRKO HoxB8 NLCs.
HoxB8 NLCs were generated by estrogen withdrawal along with G-CSF treatment for 4 days. For the respiratory burst experiments, the cells were stimulated by immobilized IgG immune complexes, proinflammatory agonists on an integrin ligand surface, or by PMA stimulus. Superoxide production was measured by a cytochrome c reduction assay. For the other respiratory burst assay and degranulation experiments, the cells were stimulated by fMLP, or by PMA stimulus. Reactive oxygen species (ROS) production and degranulation was measured by flow cytometry.
Wild-type HoxB8 NLCs were able to produce superoxide upon various stimulation. Upon these stimulation, Prkcb CRKO and Prkcd CRKO NLCs showed reduced superoxide production. Wild-type, Prkca CRKO, Prkce CRKO NLCs were able to produce ROS upon PMA stimulation, but in case of Prkcb CRKO and Prkcd CRKO HoxB8 NLCs the ROS production was decreased. The wild-type, Prkca CRKO, and Prkce CRKO NLCs exhibited a normal degranulation response, whereas the degranulation reaction was reduced in Prkcb CRKO and Prkcd CRKO NLCs.
Our results indicate that PKC-β and PKC-δ may play a role in the respiratory burst and degranulation response of HoxB8 NLCs. Therefore, our CRISPR-based targeting strategy is suitable for deleting functionally relevant proteins from NLCs.
Funding: Hungarian National Research, Development and Innovation Office (KKP-129954, K-146160 and TKP2021-EGA-24), the HUN-REN Hungarian Research Network (0207007) and the Hungarian Academy of Sciences (LP2024-16/2024).

University

Semmelweis University

Supervisor

Prof. Dr. Attila Mócsai

Publication of my abstract

I give consent to the publication of my abstract on the website of the congress.

phd.section.field

in doctoral studies after complex exam (PhD)

Kind

Szabad

Status

elfogadva

Accepted presentation method

poszter

Előadás fájl jóváhagyás

nem rendelkezett róla

Előadó

8097

Start

17:12

End

17:15