PhD Scientific Days 2020

Molecular Sciences II. Lectures

Characterization of Neutrophil Granulocytes Generated In Vitro From HoxB8-Transduced Myeloid Progenitor Cells

Előadó neve

Anita, Orosz

Előadó munkahelye

Semmelweis University, Department of Physiology

Előadó telefonszáma

+36302852921

Előadó e-mail címe

orosz.anita@med.semmelweis-univ.hu

Az előadás címe

Characterization of Neutrophil Granulocytes Generated In Vitro From HoxB8-Transduced Myeloid Progenitor Cells

Szerző(k) neve és munkahelye

1. Anita Orosz, Department of Physiology, Budapest
2. Attila Mócsai, Department of Physiology, Budapest

Szekció

Molecular Sciences II. Lectures

Language of the presentation

English

Section, first choice

Molecular Sciences

Section, second choice

Theoretical and Translational Medicine

Összefoglaló szövege

Neutrophils play a critical role in the innate immunity. However, deeply understanding their biology has been challenging, as they are short-lived, terminally differentiated cells.
Our aim is to overcome this obstacle using the HoxB8-driven, immortalized myeloid progenitor cell line. This allows us to generate unlimited amounts of neutrophils, followed by detailed analysis of various cell functions.
HoxB8 progenitors were cultured in medium containing β-estrogen. Neutrophils were grown in estrogen free medium supplemented with G-CSF. Reactive oxygen species (ROS) production was monitored via cytochrome-c reduction. In vitro phagocytosis was measured using green fluorescent Staphylococcus aureus (USA300). HoxB8 chimeras were generated with the adoptive transfer of HoxB8 progenitors. Migration and phagocytosis assays were performed in vivo. KBxN serum transfer arthritis model was used to monitor HoxB8 neutrophils’ role in autoantibody-induced inflammation.
Hoxb8 progenitors differentiated into neutrophils in 5 days, in vitro. These HoxB8 neutrophils could produce ROS upon various stimuli comparable to WT bone marrow- derived neutrophils. They could carry out phagocytosis of opsonized bacteria in vitro. Upon adoptive transfer of HoxB8 progenitors, HoxB8 neutrophils soon appeared in the circulation of the recipients’. These neutrophils were able to migrate into the inflamed peripheral tissues, where they carried out phagocytosis of heat inactivated Candida albicans particles. However, in 2 days HoxB8 neutrophils dissappeared from the recipients, possibly because the progenitors were unable to colonize their bone marrow permanently. Upon arthritogenic serum treatment, chimeras having only WT Hoxb8 neutrophils developed a systemic joint inflammation, comparable to the WT animals. Meanwhile Syk (Spleen Tyrosine Kinase) KO HoxB8 chimeras seemed to be completely protected in the same circumstance.
The HoxB8 progenitor cell line is a robust tool to generate neutrophils in vitro. HoxB8 neutrophils, both in vitro and in vivo, are quite similar to their BM derived counterparts as far as their effector functions are concerned. Their role in acute inflammation further proves the cell line’s utility in wide-ranging studies of neutrophils. Moreover, the in vitro cultured HoxB8 progenitors can act as good targets for genetic modifications, manifesting on the neutrophil level.

Additional Information

Attila Mócsai
mocsai.attila@med.semmelweis-univ.hu

Bemutatás módja

Szóbeli

Kind

Szabad

Status

elfogadva

Accepted presentation method

szóbeli

Előadás fájl jóváhagyás

nem rendelkezett róla

Előadó

4716

Start

12:20

End

12:35

Authors (legacy)

1. Anita Orosz, Department of Physiology, Budapest
2. Attila Mócsai, Department of Physiology, Budapest