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Slide 1 of 30
Gene cluster polymorphisms &
autoimmunity
IMM430H1S: March 2005Zenon Harley
Slide 2 of 30
The Paper
5. Discussion
4. Lupus model
3. Strain analysis
2. DNA sleuthing
1. Summary
Slide 3 of 30
Summary
• 1 in 2,000 Canadians have lupus (10x more in women)
• The cause is unknown
• Diagnosis is difficult
5. Discussion
4. Lupus model
3. Strain analysis
2. DNA sleuthing
1. Summary
Slide 4 of 30
Summary
• Lupus studied in a mouse model because of congenics.
• Isolate lupus-causing genes from a lupus-prone mouse and cause lupus in a healthy mouse.
5. Discussion
4. Lupus model
3. Strain analysis
2. DNA sleuthing
1. Summary
Slide 5 of 30
DNA sleuthing
• Lupus-prone mouse: NZ2410
• Non-lupus-prone mouse: B6
2. DNA sleuthing
1. Summary
5. Discussion
4. Lupus model
3. Strain analysis
Slide 6 of 30
DNA sleuthing
2. DNA sleuthing
1. Summary
5. Discussion
4. Lupus model
3. Strain analysis Sle1a,b,c
• Known causative interval: Sle1
Slide 7 of 30
DNA sleuthing
2. DNA sleuthing
1. Summary
5. Discussion
4. Lupus model
3. Strain analysis
Slide 8 of 30
DNA sleuthing
2. DNA sleuthing
1. Summary
5. Discussion
4. Lupus model
3. Strain analysis
• Two congenic strains: B6.Sle1b and B6.FcR
Slide 9 of 30
DNA sleuthing
2. DNA sleuthing
1. Summary
5. Discussion
4. Lupus model
3. Strain analysis
• Gender-biased anti-nuclear antibody production.
Slide 10 of 30
DNA sleuthing
2. DNA sleuthing
1. Summary
5. Discussion
4. Lupus model
3. Strain analysis
• Molecular map: 0.9Mb
• Sequencing: 24 expressed genes, 19 in spleen
• SLAM/CD2 family
Telomeric FcR
Telomeric Sle1b
Slide 11 of 30
DNA sleuthing
2. DNA sleuthing
1. Summary
5. Discussion
4. Lupus model
3. Strain analysis
• Structural polymorphisms
Slide 12 of 30
DNA sleuthing
2. DNA sleuthing
1. Summary
5. Discussion
4. Lupus model
3. Strain analysis
• RT-PCR analysis of expression levels in lymphocytes.
Slide 13 of 30
Slide 14 of 30
DNA sleuthing
2. DNA sleuthing
1. Summary
5. Discussion
4. Lupus model
3. Strain analysis
Slide 15 of 30
Strain analysis
• Does this polymorphism contribute to autoimmunity in other mouse strains?
5. Discussion
4. Lupus model
2. DNA sleuthing
1. Summary
3. Strain analysis
Slide 16 of 30
Slide 17 of 30
Strain analysis
5. Discussion
4. Lupus model
2. DNA sleuthing
1. Summary
3. Strain analysis
• ANA production compared to B6 mice.
Slide 18 of 30
Slide 19 of 30
Lupus model
• How do polymorphisms among different strains contribute to autoimmunity?
5. Discussion
3. Strain analysis
2. DNA sleuthing
1. Summary
4. Lupus model
Slide 20 of 30
Lupus model
5. Discussion
3. Strain analysis
2. DNA sleuthing
1. Summary
4. Lupus model
• Ly108 is known to have alternative splice forms
Slide 21 of 30
Lupus model
5. Discussion
3. Strain analysis
2. DNA sleuthing
1. Summary
4. Lupus model
• Ly108-1 is favoured in Lupus-prone congenic strains
Slide 22 of 30
Lupus model
5. Discussion
3. Strain analysis
2. DNA sleuthing
1. Summary
4. Lupus model
• RT-PCR using isoform-specific primers: B6 and B6.Sle1b
Slide 23 of 30
Lupus model
5. Discussion
3. Strain analysis
2. DNA sleuthing
1. Summary
4. Lupus model
• Additional possible isoform in B6.Sle1b and NZW
Slide 24 of 30
Lupus model
• T-cell stimulation
5. Discussion
3. Strain analysis
2. DNA sleuthing
1. Summary
4. Lupus model
(1) T-cell stimulus, (2) Streptavidin, (3) Ionomycin, (4) MnCl2
Slide 25 of 30
Lupus model
• T-cell stimulation
5. Discussion
3. Strain analysis
2. DNA sleuthing
1. Summary
4. Lupus model
(1) T-cell stimulus, (2) Streptavidin, (3) Ionomycin, (4) MnCl2
Slide 26 of 30
Slide 27 of 30
Lupus model
5. Discussion
3. Strain analysis
2. DNA sleuthing
1. Summary
4. Lupus model
Slide 28 of 30
Discussion
• Epistatic interactions
4. Lupus model
3. Strain analysis
2. DNA sleuthing
1. Summary
5. Discussion
BB = black
Bb = black
bb = brown
CC = normal
Cc = normal
cc = albino
Slide 29 of 30
Discussion
• Polymorphisms diversify the immune response.
• Favoured by natural selection due to diversity of pathogens.
4. Lupus model
3. Strain analysis
2. DNA sleuthing
1. Summary
5. Discussion
Slide 30 of 30
Discussion
• Disease not always caused by a “bad” allele.
• Immunity and autoimmunity involve a number of complex and inter-dependent pathways.
• This presents a major therapeutic challenge.
4. Lupus model
3. Strain analysis
2. DNA sleuthing
1. Summary
5. Discussion