PhD Scientific Days 2025

Poster Session II. - G: Pharmaceutical Sciences and Health Technologies

Lineage-Specific Analysis of Basal Tyrosine Phosphorylation in Circulating Leukocytes of Hck–/–Fgr–/–Lyn–/– Mice

Előadó neve

Deli Dorottya, MSc

Neptun code

KLF695

Előadó munkahelye

Department of Physiology, Semmelweis University, Budapest, Hungary

Előadó telefonszáma

+36-1-459-1500 x60-408

Előadó e-mail címe

deli.dorottya@phd.semmelweis.hu

Az előadás címe

Lineage-Specific Analysis of Basal Tyrosine Phosphorylation in Circulating Leukocytes of Hck–/–Fgr–/–Lyn–/– Mice

Szerző(k) neve és munkahelye

Dorottya Deli¹, Dorottya Markó¹, Krisztina Futosi¹, Attila Mócsai¹

1: Department of Physiology, Semmelweis University, Budapest, Hungary

Bemutatás módja

Poszter

Szekció

Poster Session II. - G: Pharmaceutical Sciences and Health Technologies

Language of the presentation

English

Preferred session

Pharmaceutical Sciences and Health Technologies

Összefoglaló szövege

Introduction: Tyrosine kinases are central regulators of immune cell signaling and are key drug targets in inflammatory diseases. We recently developed a rapid, quantitative flow cytometry–based assay to assess basal tyrosine phosphorylation of circulating neutrophils in mice.

Aims: We aimed to extend this method to allow parallel analysis of multiple leukocyte subsets and enable cell lineage–specific resolution in complex in vivo models.

Methods: Peripheral blood was collected from wild type and Hck–/–Fgr–/–Lyn–/– triple knockout mice, as well as from mixed bone marrow chimeras generated using knockout donor cells. Samples were stained with fluorescent antibodies specific for neutrophils, monocytes, B- or T-cells. After fixation and permeabilization, tyrosine-phosphorylated proteins were detected intracellularly using fluorescently labeled anti-phosphotyrosine or isotype control antibodies. For specificity control, anti-phosphotyrosine antibodies were preincubated with 1 mM soluble phosphotyrosine. Fluorescence signals reflecting basal phosphorylation were analyzed by flow cytometry.

Results: The assay revealed high basal tyrosine phosphorylation in all leukocyte populations. In Hck–/–Fgr–/–Lyn–/– mice, myeloid cells showed a marked reduction in phosphorylation, while lymphocytes remained unaffected. In mixed bone marrow chimeras, knockout and wild-type myeloid cells were distinguishable based on differential phosphorylation, demonstrating the method’s suitability for lineage-specific in vivo analysis.

Conclusions: This optimized assay enables rapid, quantitative, cell-type–resolved analysis of basal tyrosine phosphorylation in vivo. Its use in mixed chimeras allows within-animal comparison of wild-type and kinase-deficient myeloid cells, supporting its utility in lineage-specific functional studies and translational signaling research.

Funding: Funded by the Hungarian National Research, Development and Innovation Office (KKP-129954, FK-146729 and TKP2021-EGA-24) and the HUN-REN Hungarian Research Network (0207007)

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 before complex exam (PhD)

Kind

Szabad

Status

elfogadva

Accepted presentation method

poszter

Előadás fájl jóváhagyás

nem rendelkezett róla

Előadó

9117

Start

18:30

End

18:36