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Gene cluster polymorphisms autoimmunity

• IMM430H1S March 2005
• Zenon Harley

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The Paper
1. Summary
2. DNA sleuthing
3. Strain analysis
4. Lupus model
5. Discussion
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Summary

• 1 in 2,000 Canadians have lupus (10x more in
  women)
• The cause is unknown
• Diagnosis is difficult

1. Summary
2. DNA sleuthing
3. Strain analysis
4. Lupus model
5. Discussion
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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.

1. Summary
2. DNA sleuthing
3. Strain analysis
4. Lupus model
5. Discussion
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DNA sleuthing

• Lupus-prone mouse NZ2410
• Non-lupus-prone mouse B6

1. Summary
2. DNA sleuthing
3. Strain analysis
4. Lupus model
5. Discussion
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DNA sleuthing

• Known causative interval Sle1

1. Summary
2. DNA sleuthing
Sle1a,b,c
3. Strain analysis
4. Lupus model
5. Discussion
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DNA sleuthing
1. Summary
2. DNA sleuthing
3. Strain analysis
4. Lupus model
5. Discussion
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DNA sleuthing

• Two congenic strains B6.Sle1b and B6.FcR

1. Summary
2. DNA sleuthing
3. Strain analysis
4. Lupus model
5. Discussion
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DNA sleuthing

• Gender-biased anti-nuclear antibody production.

1. Summary
2. DNA sleuthing
3. Strain analysis
4. Lupus model
5. Discussion
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DNA sleuthing

• Molecular map 0.9Mb
• Sequencing 24 expressed genes, 19 in spleen
• SLAM/CD2 family

1. Summary
2. DNA sleuthing
3. Strain analysis
4. Lupus model
5. Discussion
Telomeric FcR
Telomeric Sle1b
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DNA sleuthing

• Structural polymorphisms

1. Summary
2. DNA sleuthing
3. Strain analysis
4. Lupus model
5. Discussion
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DNA sleuthing

• RT-PCR analysis of expression levels in
  lymphocytes.

1. Summary
2. DNA sleuthing
3. Strain analysis
4. Lupus model
5. Discussion
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DNA sleuthing
1. Summary
2. DNA sleuthing
3. Strain analysis
4. Lupus model
5. Discussion
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Strain analysis

• Does this polymorphism contribute to autoimmunity
  in other mouse strains?

1. Summary
2. DNA sleuthing
3. Strain analysis
4. Lupus model
5. Discussion
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Strain analysis

• ANA production compared to B6 mice.

1. Summary
2. DNA sleuthing
3. Strain analysis
4. Lupus model
5. Discussion
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Lupus model

• How do polymorphisms among different strains
  contribute to autoimmunity?

1. Summary
2. DNA sleuthing
3. Strain analysis
4. Lupus model
5. Discussion
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Lupus model

• Ly108 is known to have alternative splice forms

1. Summary
2. DNA sleuthing
3. Strain analysis
4. Lupus model
5. Discussion
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Lupus model

• Ly108-1 is favoured in Lupus-prone congenic
  strains

1. Summary
2. DNA sleuthing
3. Strain analysis
4. Lupus model
5. Discussion
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Lupus model

• RT-PCR using isoform-specific primers B6 and
  B6.Sle1b

1. Summary
2. DNA sleuthing
3. Strain analysis
4. Lupus model
5. Discussion
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Lupus model

• Additional possible isoform in B6.Sle1b and NZW

1. Summary
2. DNA sleuthing
3. Strain analysis
4. Lupus model
5. Discussion
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Lupus model

• T-cell stimulation

1. Summary
2. DNA sleuthing
3. Strain analysis
4. Lupus model
5. Discussion
(1) T-cell stimulus, (2) Streptavidin, (3)
Ionomycin, (4) MnCl2
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Lupus model

• T-cell stimulation

1. Summary
2. DNA sleuthing
3. Strain analysis
4. Lupus model
5. Discussion
(1) T-cell stimulus, (2) Streptavidin, (3)
Ionomycin, (4) MnCl2
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Lupus model
1. Summary
2. DNA sleuthing
3. Strain analysis
4. Lupus model
5. Discussion
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Discussion

• Epistatic interactions

1. Summary
BB black Bb black bb brown CC normal Cc
normal cc albino
2. DNA sleuthing
3. Strain analysis
4. Lupus model
5. Discussion
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Discussion

• Polymorphisms diversify the immune response.
• Favoured by natural selection due to diversity of
  pathogens.

1. Summary
2. DNA sleuthing
3. Strain analysis
4. Lupus model
5. Discussion
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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.

1. Summary
2. DNA sleuthing
3. Strain analysis
4. Lupus model
5. Discussion