This research aims to establish a continuously growing cell line of alternatively activated mouse macrophages to replace the use of cells isolated from mouse organs or bone marrow.
Macrophages are phagocytes and antigen-presenting cells which perform key roles in innate and adaptive immunity. Activated macrophages are classified as M1 or M2. M1 macrophages have an acute inflammatory phenotype and are highly important for fighting bacteria. The alternatively activated anti-inflammatory M2 macrophages on the other hand promote cell proliferation and tissue repair, and are thought to have a role in allergic diseases, such as asthma.
There is increasing interest in the role of M2 macrophages in both allergy and infection. Currently, the only way of obtaining these cells for study relies on harvesting directly from animals. These cells have a limited life-span, thus requiring the repeated use of animals – it is estimated that around 300,000 mice are used for this purpose worldwide each year. Establishing a permanently growing M2 macrophage cell line would replace the number of animals currently used, and potentially provide a screening platform for drugs which modify M2 macrophage function to treat diseases.
Research details and methods
Recently a method to obtain continuously growing non-transformed primary macrophages (MPI cells) from mice without chemical or viral mutagenesis or genetic manipulation has been developed.
The research will establish if MPI cells, when treated with the cytokines interleukin 4 or interleukin 13, can be induced to form an M2-like phenotype. The cells will be characterised and compared to parental MPI cells and bone marrow-derived M2 macrophages using a number of approaches, including gene expression patterns and the induction of inflammatory mediating molecules in response to challenge with pathogens and other innate immune system stimulators.
In addition, the ability of MPI-derived M2 macrophages to control mouse cytomegalovirus infection and replication of some bacterial agents will also be compared to parental MPI cells and bone marrow-derived macrophages.
Stichling N et al. (2018). Lung macrophage scavenger receptor SR-A6 (MARCO) is an adenovirus type-specific virus entry receptor. PLoS pathogens 14(3):e1006914. doi: 10.1371/journal.ppat.1006914
Maler MD et al. (2017). Key Role of the Scavenger Receptor MARCO in Mediating Adenovirus Infection and Subsequent Innate Responses of Macrophages. mBio 8(4). doi: 10.1128/mBio.00670-17
Fejer G et al. (2015). Self-renewing macrophages--a new line of enquiries in mononuclear phagocytes. Immunobiology 220(2):169-74. doi: 10.1016/j.imbio.2014.11.005
Rawson FJ et al. (2015). Fast, Ultrasensitive Detection of Reactive Oxygen Species Using a Carbon Nanotube Based-Electrocatalytic Intracellular Sensor. ACS Applied Materials & Interfaces 7(42):23527-37. doi: 10.1021/acsami.5b06493
- Awards and Prizes: NC3Rs 3Rs Prize Highly Commended, 2014
- NC3Rs blog: Immune cells grown in lab could significantly reduce animal use in research
Principal investigatorDr Gyorgy Fejer
InstitutionUniversity of Southampton
Co-InvestigatorProfessor Simon Jackson
Professor Nigel Silman
Dr Lars Dolken