congenic mice infected with trypanosoma congolense harry noyes university of liverpool
Post on 15-Jan-2016
220 views
TRANSCRIPT
Congenic mice infected with Trypanosoma congolense
Harry Noyes
University of Liverpool
Participants• ILRI, Nairobi• Morris Agaba• John Gibson• Olivier Hanotte• Steve Kemp• Hassan Musa• Joel Mwakya• Daniel Mwanga• Jan Naessens• Joseph Nganga• Moises Ogugu• John Wambugu
• University of Manchester
• Andy Brass• Helen Hulme• Leo Zeef• Leanne Wardlesworth
• University of Liverpool
• Anthea Broadhead• Derek Daly• Harry Noyes• Katie Rennie• Luke Thomas
• Roslin Institute• Alan Archibald• Susan Anderson• Laurence Hall
Mouse models of trypanotolerance
.
Survival of F6 and parental strains
0
10
20
30
40
50
60
70
80
90
100
1 21 41 61 81 101 121
Days Post Challenge
% Survival
F6
AJ
C57BL
Survival of F6 and parental populations
Trypanosoma infection response (Tir) loci
C57/BL6 x AJ and C57/BL6 x BALB/C
Iraqi et al Mammalian Genome 2000 11:645-648 Kemp et al. Nature Genetics 1997 16:194-196
Microarray design for each condition
Resistant C57BL/6Susceptible AJ
High throughput tissue collection
Correlation of expression and SNP in parental inbred lines
Number of differentially expressed probes per 50 probe bin
Number of SNP in 1kb upstream of genes in each bin
r2 = 0.1
Number of SNP in 1kb upstream region
0 2 4 6 8 10 12
Relative Risk
0
2
4
6
8
10
12
Rel
ativ
e R
isk
of D
iffer
entia
l Exp
ress
ion
Only genes with tightly regulated expression (p<0.005) were associated
with SNP
Effect of copy number variation
Poster 30
Effect of copy number variation on Glyoxalase1 expression
C57BL/6 regions carried by congenics
C57BL/6 regions in
Tir 1 congenics
Tir 2 congenics
Tir 3 congenics
Survival of congenic mice
Response of Congenic mice to T. congolense infection
0
20
40
60
80
100
120
21-30 31-40 41-50 51-60 61-70 71-80 81-90 91-100
Days post intection
% survivingTir1AA (n = 20)Tir1CC (n = 25)Tir2AA (n = 60)Tir2CC (n = 120)Tir3AA (n = 20)Tir3CC (n = 90)TirnAA (n = 100)
p53
Networks in Congenic Spleen
QTL workflowGiven a species, chromosome, Affymetrix Array Name, Start and Stop position, Get back the EnsEMBL Gene idsGet EnsEMBL Gene ids
Retrieve a list of Information about each EnsEMBL Gene id
Extract the SwissProt ids from this Gene information list
Send gene ids and uniprot accesion numbers off to KEGG and retrieve a KEGG id
Remove any null values
Send swissprot ids to NCBI and get gene ids
Send kegg gene ids off to KEGG and retrieve a pathways
Merge pathways into a single text file
Merge pathways ids with gene ids
Fisher et al. NAR 2007
Polymorphism in Daxx in hyperacidic region
C57BL/6 ETDDDDDDDDDDDDEDNEESEEEEEEEEEE129/J .......D......................BALB/c .......-......................A/J .......-......................
Cholesterol metabolism
Total Cholesterol levelsCHOLESTEROL
0.00
0.50
1.00
1.50
2.00
2.50
3.00
3.50
4.00
0 8 21 4
DAYS POST INFECTION
AJ_HIGH_FAT
AJ_LOW_FAT
BALB_HIGH_FAT
BALB_LOW_FAT
c57_HIGH_FAT
C57_LOW_FAT
Survival of C57BL/6ByJ
C57BL/6ByJ spontaneous mutant with low cholesterol and low risk of atherosclerosis
Paul DarkInfection, Injury and Inflammation Research GroupHope Hospital Intensive Care UnitThe University of Manchester, UK
Total cholesterol responses in Intensive care n = 607 33% dead within 28 days
2
2.2
2.4
2.6
2.8
3
3.2
3.4
0 2 4 6 8 10ICU day
Total cholesterol (mmol/l)
Died Survived
Results
Analysis of early non-HDL cholesterol responses
-0.50
-0.40
-0.30
-0.20
-0.10
0.00
0.10
0.20
0.30
Day 1 to 2 Day 2 to 3
Change in non-HDL cholesterol
Died
Survived
NATURE MEDICINE • VOLUME 9 • NUMBER 2 • FEBRUARY 2003
LXR agonists lower cholesterol and inhibit NFkB mediated inflammatory signals
RXRb protein by Western BlotRxRb expression in congenic strain spleen - TOTAL
0
50
100
150
200
250
300
350
0 3 7 9 17
Day
Density (as % C57 d 0)
CCAA
Conclusions
• Glo1 gene duplication may be associated with differences in oxidative stress
• Daxx aspartate polymorphism may regulate apoptosis
• RXRb expression differences may be associated with regulation of cholesterol and inflammation