PhD Scientific Days 2024

Molecular Medicine I.

Identification of Cell Types Involved in the Uptake and Expression of Nucleoside-modified mRNA-LNP Complexes

Előadó neve

Ms. Sági, Stella Márta

Neptun code

G6NO54

Előadó munkahelye

Department of Physiology, Semmelweis University, Budapest, Hungary

Előadó telefonszáma

+36307009163

Előadó e-mail címe

sagi.stella98@gmail.com

Az előadás címe

Identification of Cell Types Involved in the Uptake and Expression of Nucleoside-modified mRNA-LNP Complexes

Szerző(k) neve és munkahelye

Stella Márta Sági¹, Gábor Kovács¹, Kornél Molnár¹, Éva Kemecsei¹, Norbert Pardi², Zoltán Lipinszki³, Zoltán Jakus¹

1: Department of Physiology, Semmelweis University, Budapest, Hungary
2: University of Pennsylvania, Perelman School of Medicine, Philadelphia, PA, USA
3: Biological Research Centre of the Hungarian Academy of Sciences, Szeged, Hungary

Bemutatás módja

Szóbeli

Szekció

Molecular Medicine I.

Language of the presentation

English

Preferred session

Molecular Medicine

Összefoglaló szövege

Introduction: Vaccines based on the lipid nanoparticle-encapsulated nucleoside-modified messenger RNA (mRNA-LNP) technology recently gained major scientific interest, as two vaccines against SARS-CoV-2 accepted for emergency use utilize this technology. It has been shown that mRNA-LNP vaccines are able to induce potent humoral immune response, however the underlying molecular and cellular mechanisms involved are still not fully understood. The lymphatics are involved in several immunological processes, but the role of lymphatics in the mRNA-LNP vaccine induced immune response requires further investigation.
Aims: We aimed to characterize tissue-specific cell types involved in the uptake and expression of mRNA-LNP complexes.
Methods: Dil fluorophore-labelled LNP and mRNA-LNP encoding eGFP were injected into the hind paws of wild-type mice, following we have collected histological samples from the injection site and regional lymph nodes, which were subsequently either paraffin-embedded and sectioned, then examined by fluorescent immunostaining; or cell suspensions were made from which we quantified the cell types uptaking mRNA-LNPs by flow cytometry measurements.
Results: Histological sections were examined, and eGFP-expressing cells were detected at the injection site. eGFP expression was observed in certain structures labeled with the LYVE1 lymphatic endothelial cell specific marker and extensive neutrophil infiltration was observed following the injection of mRNA-LNP. Flow cytometry measurements revealed that LNPs were taken up primarily by cells bearing markers specific for subcapsular sinusoidal macrophages, neutrophil granulocytes, and dendritic cells. Furthermore, our results to date indicate that macrophages express eGFPs, whereas neutrophil granulocytes do not.
Conclusions: Our results suggest that certain populations of the lymphatics are able to take up and express mRNA-LNP complexes, raising the possibility that the lymphatics may be involved in the immune response induced by mRNA-LNP-based vaccines. In lymph nodes, LNPs are mostly taken up by cells bearing cell surface markers of subcapsular sinusoidal macrophages, suggesting that LNPs are firstly solubilized in lymph and then reach lymph nodes. Our results suggest that neutrophils take up mRNA-LNP complexes, however, do not express it.
Funding: K139165, TKP2021-EGA-29,TKP2021-EGA-24,SE250+ fund

University

Semmelweis University

Supervisor

Dr. Zoltán Jakus

Publication of my abstract

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

Kind

Szabad

Status

elfogadva

Accepted presentation method

szóbeli

Előadás fájl jóváhagyás

nem rendelkezett róla

Előadó

8047

Start

11:45

End

11:55