In vitro differentiation of near-unlimited numbers of functional mouse basophils using conditional Hoxb8

Gurzeler, Ursina; Rabachini de Almeida, Tatiana; Dahinden, Clemens A.; Salmanidis, Marika; Brumati, Gabriela; Ekert, Paul G.; Echeverry, Nohemy; Bachmann, Daniel; Simon, Hans-Uwe; Kaufmann, Thomas (2013). In vitro differentiation of near-unlimited numbers of functional mouse basophils using conditional Hoxb8. Allergy, 5(68), pp. 604-613. Wiley-Blackwell 10.1111/all.12140

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Background: Basophils constitute a rare leukocyte population known for their effector functions in inflammation and allergy, as well as more recently described immunoregulatory roles. Besides their low frequency, functional analysis of basophils is hindered by a short life span, inefficient ex vivo differentiation protocols, and lack of suitable cell models. A method to produce large quantities of basophils in vitro would facilitate basophil research and constitute a sought-after tool for diagnostic and drug testing purposes. Methods: A method is described to massively expand bone marrow–derived basophils in vitro. Myeloid progenitors are conditionally immortalized using Hoxb8 in the presence of interleukin-3 (IL-3) and outgrowing cell lines selected for their potential to differentiate into basophils upon shutdown of Hoxb8 expression. Results: IL-3-dependent, conditional Hoxb8-immortalized progenitor cell lines can be expanded and maintained in culture for prolonged periods. Upon shutdown of Hoxb8 expression, near-unlimited numbers of mature functional basophils can be differentiated in vitro within six days. The cells are end-differentiated and short-lived and express basophil-specific surface markers and proteases. Upon IgE- as well as C5a-mediated activation, differentiated basophils release granule enzymes and histamine and secrete Th2-type cytokines (IL-4, IL-13) and leukotriene C4. IL-3-deprivation induces apoptosis correlating with upregulation of the BH3-only proteins BCL-2-interacting mediator of cell death (BIM) and p53 upregulated modulator of apoptosis (PUMA) and downregulation of proviral integration site for Moloney murine leukemia virus 1 kinase (PIM-1). Conclusion: A novel method is presented to generate quantitative amounts of mouse basophils in vitro, which moreover allows genetic manipulation of conditionally immortalized progenitors. This approach may represent a useful alternative method to isolating primary basophils.

Item Type:

Journal Article (Original Article)

Division/Institute:

04 Faculty of Medicine > Pre-clinic Human Medicine > Institute of Pharmacology
04 Faculty of Medicine > Department of Haematology, Oncology, Infectious Diseases, Laboratory Medicine and Hospital Pharmacy (DOLS) > Institute for Immunology (discontinued)

UniBE Contributor:

Gurzeler, Ursina; Rabachini de Almeida, Tatiana; Dahinden, Clemens A.; Echeverry, Nohemy; Bachmann, Daniel; Simon, Hans-Uwe and Kaufmann, Thomas

Subjects:

600 Technology > 610 Medicine & health

ISSN:

0105-4538

Publisher:

Wiley-Blackwell

Funders:

[4] Swiss National Science Foundation
[8] 3R Research Foundation Switzerland

Language:

English

Submitter:

Anita Dähler

Date Deposited:

06 Dec 2013 03:21

Last Modified:

05 Jun 2015 08:51

Publisher DOI:

10.1111/all.12140

PubMed ID:

23590216

Uncontrolled Keywords:

apoptosis; basophils; Hoxb8; IL-3; mast cells

URI:

https://boris.unibe.ch/id/eprint/39530

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