Immunogenomics Laboratory: Current Research Projects
Oxford-ANU Wellcome Trust Programme: Dr C Vinuesa,
Dr R Cornall, Prof J Bell, Prof C Goodnow
This project is producing and screening large libraries of mutagenized
C57BL/6 or TCR transgenic mice to identify genes controlling the immune
response. Dr Carola Vinuesa heads a large effort screening for defects
in most components of the humoral immune response by immunization
and serological analysis. Mapping, gene identification, and detailed
biological analysis of mutants arising from the programme is a joint
effort at the ANU and Oxford University.
Different patterns of odies identifying putative new mutant strains by serological screening. Research of Dr C Vinuesa
Refs:
Nelms KA, Goodnow CC (2001) Genome-wide ENU mutagenesis to reveal
immune regulators. Immunity 15, 409-418.
Molecular Analysis of Pathways in Diabetes (MAPDB) Special Programme
of the Juvenile Diabetes Research Foundation and the National Health
and Medical Research Council: Dr G Hoyne, Dr K Nelms, Dr N Petrovsky,
Prof C Goodnow
This project is producing and screening large libraries of mutagenized
TCR:insHEL double transgenic mice to identify genes and mechanisms
controlling Type 1 diabetes and its complications. The starting strain
is genetically sensitised with subclinical insulitis and a trackable
population of islet-reactive helper T cells, to allow detection of
genes and mechanisms that modify different steps in this disease.
Homogeneous subclinical insulitis in double transgenic mice expressing hen egg lysozyme
in pancreatic islet beta cells and carrying anti-lysozyme antigen receptors on most CD4 T cells.
Genes in Immunity and Tolerance Consortium - Large RO1 of the National
Institutes of Health: Dr C Vinuesa, Dr G Hoyne, Dr J Cyster, Prof
L Lanier, Prof A Weiss, Prof C Goodnow.
This project began in Sept 2002 and is producing and screening
large libraries of mutagenized IgM/G:sHEL double transgenic mice to
identify genes and mechanisms controlling B cell tolerance and memory,
lymphocyte migration, leukocyte inhibitory receptors, and lymphocyte
signal transduction mechanisms. Mutagenesis, mapping and gene identification
are performed at the ANU, and screen development and detailed analysis
performed jointly at the ANU and the University of California San
Francisco.
Mechanisms controlling B cell responses to self and foreign antigens.
NHMRC Project Grant: Dr C Vinuesa, Mr J Jun, Dr M Blery, Prof C Goodnow
In this project we aim to understand how B lymphocyte responses
to antigen are regulated by plasticity in signalling by the B cell
antigen receptor (BCR). The project builds on our previous work in
three interconnected problems: 1. defining how changes in subcellular
distribution and trafficking of BCRs on anergic B cells selectively
reduces immunogenic responses and signalling to NFkB and JNK, while
leaving tolerogenic signalling via NFAT and ERK intact; 2. defining
how molecular changes in the membrane-spanning tail of the BCR enhance
the proliferative burst of memory B cells; and 3. understanding how
the changes in BCR signalling during tolerance or memory arise by
modulating the formation of immunogenic signalling complexes - a hypothesized
"immunon" - formed by a novel member of the membrane associated
guanylate kinase (MAGUK) family of proteins, Carma-1/Card-11. This
takes advantage of a mouse strain, unmodulated, arising from one of
our mutagenesis programs which carries a point mutation in Carma-1.
Healy JI, Dolmetsch RE, Timmerman LA, Cyster JG, Thomas
ML, Crabtree GR, Lewis RS, Goodnow CC (1997). Different nuclear signals
are activated by the B cell receptor during positive versus negative
signaling. Immunity 6, 419-428 .
Glynne R, Akkaraju S, Healy JI, Rayner J, Goodnow CC
& Mack D (2000) How self-tolerance and the immunosuppressive drug
FK506 prevent B cell mitogenesis. Nature 403, 72-676.
Martin SW, Goodnow CC (2002). Burst-enhancing role of
the IgG membrane tail as a molecular determinant of memory. Nat Immunol
3, 182-8.
Jun JE, Wilson LE, Vinuesa CG,
Lesage S, Blery M, Miosge LA, Cook MC, Kucharska EM, Hara H, Penninger
JM, Domashenz H, Hong NA, Glynne RJ, Nelms KA and Goodnow CC (2003).
Identifying the MAGUK-protein Carma-1 as a central regulator of humoral
immune responses and atopy by genome wide mouse mutagenesis. Immunity.
In press.
Mechanisms controlling CD4 T cell tolerance to pancreatic islet-specific
antigen. NHMRC Project Grant: Mr A Liston, Dr G Hoyne, Dr L Wicker,
Dr L Peltonen, Prof C Goodnow.
This project focuses on identifying how diabetes susceptibility
or resistance genes contributing to disease in the NOD mouse or in
human Autoimmune Polyendocrine Syndrome 1 (Aire) affect the regulation
of T cells that potentially could destroy insulin-producing islet
beta cells. The project builds on our recent findings that defects
in Aire and NOD genes affect different steps in a specialized mechanism
for eliminating "forbidden clones" of organ-specific T cells.
Germinal Centres (GC) producing anti-islet autoantibodies forming in the pancreatic islets of TCR insHEL
double transgenic mice as a result of NOD diabetes susceptibility genes
that allow islet-reactive CD4 T cells to escape detection in the thymus.
Refs:
Lesage S, Goodnow CC (2001) Organ-specific autoimmune disease: A deficiency
of tolerogenic stimulation. J Exp Med 194, F31-F36.
Prasad SJ, Goodnow CC (2002). Intrinsic in vitro abnormalities in
dendritic cell generation caused by non-MHC non-obese diabetic genes.
Immunol Cell Biol 80, 198-206.
Prasad SJ, Goodnow CC (2002). Cell-intrinsic effects of non-MHC NOD
genes on dendritic cell generation in vivo. Int Immunol 14, 677-684.
Lesage S, Hartley SB, Akkaraju S, Wilson J, Townsend M and Goodnow
CG (2002). Failure to censor forbidden clones of CD4 T cells in autoimmune
diabetes. J Exp Med 196, 1175-88.
Liston A, Lesage S, Wilson J, Peltonen L, Goodnow CC (2003). Organ-specific
T cells escape thymic censoring as a result of autoimmune polyendocrinopathy
syndrome 1 mutation. Nature Immunology (epub).
Genetic analysis of B and T cell homeostasis: L Miosge, A Loy,
P Papathanasiou, J Jun, L Rui, A Fahrer, G Hoyne, C Vinuesa, K Nelms,
C Goodnow.
Our first large scale pilot genome-wide mutagenesis library and
immunological screen of blood yielded a large number of mutant strains
with lymphopenia or lymphoma/leukemia. We have now mapped and identified
the mutation in almost all these strains, representing point mutations
at many critical points in the pathways controlling circulating T
and B cell numbers. Because many of the mutations inactivate discrete
domains of proteins such as Tap2, CD45, Zap70, Slp76, Carma-1, Jak3,
NFkB2, p53, Ikaros, rather than simply knocking out the protein altogether,
these strains provide opportunities to define how antigen receptor
and cytokine receptor signalling pathways intersect to regulate lymphocyte
counts, immunity, and leukemia/lymphoma.
Refs:
Nelms KA, Goodnow CC (2001) Genome-wide ENU mutagenesis to reveal
immune regulators. Immunity 15, 409-418.
Loy AL, Goodnow CC (2002). Novel approaches for identifying genes
regulating lymphocyte development and function. Curr Opin Immunol
14, 260-265.
Miosge LA, Blasioli J, Blery M and Goodnow C (2002). Analysis of an
ENU-generated mouse mutation defines a cell intrinsic role of NFkB2
in regulating circulating B cell numbers. J ExpMed 196, 1113-1119.
Jun JE, Wilson LE, Vinuesa CG, Lesage S, Blery
M, Miosge LA, Cook MC, Kucharska EM, Hara H, Penninger JM, Domashenz
H, Hong NA, Glynne RJ, Nelms KA and Goodnow CC (2003). Identifying
the MAGUK-protein Carma-1 as a central regulator of humoral immune
responses and atopy by genome wide mouse mutagenesis. Immunity. In
press.
Molecular analysis of memory B cell formation in germinal centres:
C Vinuesa, I MacLennan, M Cooke, C Goodnow.
Germinal centres are specialized sites of B cell proliferation
and receptor gene hypermutation, where high affinity foreign-antigen
specific B cells are stringently selected to form memory cells and
plasma cells. While these processes are critical for long-term immunity
to infection, and our previous work has shown they are also stringent
steps for deleting self-reactive B cells, little is known about the
underlying molecular and cellular mechanisms. By sorting germinal
centre B cells destined to form memory, and matched control germinal
centre cells that are blocked in differentiation to memory cells,
we have used Affymetrix microarrays to identify a set of memory-associated
genes. Remarkably, many of these have not previously been found to
be expressed in immune cells, but are known for their roles in synaptic
plasticity and memory in the nervous system. This project is pursuing
the immunological functions of these memory genes.
Immunohistochemical staining in blue reveals high expressing in germinal centre B cells
of a novel memory-associated gene product identified by gene expression
profiling on Assymetrix microarrays. Research of C Vinuesa
