Transcript Lengeling_MRC Mouse Network

Macrophage Biology
and Innate Immunity
Andreas Lengeling
Consortium members will be drawn from the following
organizations.
•
•
•
•
•
•
•
•
•
The Roslin Institute (www.roslin.ed.ac.uk; Hume, Lengeling, Mabbott, Manson,
Freeman),
The Royal (Dick) School of Veterinary Studies Pathology Unit (Philbey, Harrison)
Wellcome Trust Centre for Immunity, Infection and Evolution (http://ciie.bio.ed.ac.uk/;
Allen)
Centre for Molecular Medicine (www.mmc.med.ed.ac.uk; Rheumatic Diseases Unit
(Ralston), Gastrointestinal Diseases Unit (Satsangi)
Edinburgh Cancer Research UK Centre (www.ecrc.ed.ac.uk; Frame)
MRC Centre for Inflammation Research (www.cir.med.ed.ac.uk; Dransfield, Rossi,
Iredale, Forbes, Gregory)
MRC Human Genetics Unit (www.hgu.mrc.ac.uk, Dorin)
MRC Centre for Regenerative Medicine (www.crm.ed.ac.uk; ffrench-Constant)
Edinburgh Division of Pathway Medicine (www.ed.ac.uk/schoolsdepartments/pathway-medicine; Ghazal)
Why study macrophages!
Macrophage Activation
Innate immune defence
Wound repair
Homeostasis
Chronic inflammation
Multiple organ failure
Tumour growth
Fibrosis/Atherosclerosis
Obesity
We aim to establish an in vitro
screening panel to examine
the impact of knockouts on
myeloid differentiation, innate
immunity and antigen
presentation.
Gene Prioritisation
More than 70% of the genes in the genome are
expressed in macrophages in some state of
activation
Knockouts of macrophage-specific genes, or
conditional knockouts of essential genes in
macrophages, are unlikely to be lethal.
Condition Knockouts can use lysM-cre, CD11b/ccre, Csf1r-cre
Biogps.org
Clustering of the BioGPS data using Biolayout
The phagocyte cluster
Genes in the phagocyte cluster
Research Plan
Bone marrow cells can be frozen, recovered, and cultured in
appropriate colony-stimulating factors to generate all of the major
myeloid lineages. We propose that IMPC routinely freeze marrow from
all viable lines. Foetal liver is an alternative for late gestational or
postnatal lethals.
Transcriptomic profiling of culture-derived macrophages
Measurements of differentiation (expression of markers), proliferation
and apoptosis
Assay of the outcome of interactions of macrophages with a range of
pathogens including mycobacteria, salmonella, listeria, group A
streptococcus, candida, leishmania, influenza, adenovirus and
cytomegalovirus.
Assay of classical (interferon gamma) and alternative (IL-4) activation
based on induction of standard marker genes
Assay of antigen-presentation based upon activation of T cells from
the OT-II transgenic mouse line in response to ovalbumin.
Funding/Research Strategy
• The principal of phenotyping using frozen marrow has been
confirmed by ourselves and others. We are developing to the
pipeline to test the phenotype of an ENU mutant in the Mpeg1
macrophage-specific gene.
• We proposed to develop a programme grant application to the
MRC that would include provision for funding of a dedicated
technician to collect and freeze bone marrow at Harwell.
• Although we have prioritised a set of macrophage-expressed
genes of specific interest to our own research, we would also be
interesting in participating in other proposals. A priori, we can
provide information on whether any gene is expressed in an
informative manner in macrophages generated from bone
marrow.
• We also propose to establish a major Centre of excellence in
comparative pathology at DickVet/Roslin that would be please to
be named and involved in any phenotyping activity.