congenic mice infected with trypanosoma congolense harry noyes university of liverpool

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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

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200

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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

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