# Presentation: 666106

## Chemicals and Immunologic Lung Disease:Gene and Environment

- Meinir G Jones PhD
- NHLI @ ICSM
- London

## Only a minority of subjects exposed  develop disease
Cases tend to develop within the first two years of exposure
Exposure response - disease develops at low exposures
Suggestive of immune response gene effects

- Cases tend to develop within the first two years of exposure
- Exposure response - disease develops at low exposures
- Suggestive of immune response gene effects

## Sensitisation to acid anhydrides and association with HLA-DR3

- Young et al 1995

## Isocyanate asthma and HLA-DQB1

- Bignon et al 1994

## Balboni et al 1996

## DQB1*0503 and *0501 differ by one amino acid at position 57

DQB1*0503 has aspartic acid at position 57

DQB1*0501 has valine at position 57

- DQB1*0503 has aspartic acid at position 57
- DQB1*0501 has valine at position 57

## Possible role for Aspartic acid at position 57 - HLA-DQB1

- Balboni et al 1996

## Chronic beryllium diseaseHLA-DPB1

- Richeldi et al 1993

## HLA-DPB1*0201 and *0401 differ at three positions

position 36

position 55 and 56 - aspartic acid, glutamic acid /alanine, alanine

position 69 - glutamic acid /valine

- position 36
- position 55 and 56 - aspartic acid, glutamic acid /alanine, alanine
- position 69 - glutamic acid /valine

## HLA-DPB1 Glutamate 69

- DPB1*0201 - glutamate at position 69
- DPB1*0401 - valine at position 69
- Glu69 variant - 97% CBD cases	                                                       			  27% referents

- Richeldi et al 1993

## Position 57 DQB1 associated with insulin dependent diabetes mellitus and isocyanate induced asthma

Position 71 DRB1 associated with rheumatoid arthritis

The side chains of DP Glu69 and Asp55 are projected towards the peptide binding cleft

- Position 71 DRB1 associated with rheumatoid arthritis
- The side chains of DP Glu69 and Asp55 are projected towards the peptide binding cleft

## Slide 12

## 95% of CBD Glu69

30-45% referents Glu69 

~10% develop CBD

- 30-45% referents Glu69
- ~10% develop CBD

## HLA-DPB1 Glu69

- Glu69/Glu69		- 6/20 (30%) CBD			                 	-1/75  (1%) control

- Wang et al 1999

## Phenotypic frequency of Glu69 carriers in CBD and referents

- Wang et al 1999

## CBD carried predominantly non-*0201 allele (84%)
Controls carried predominantly *0201  allele (68%)

- Controls carried predominantly *0201  allele (68%)

## Specific Glu69 alleles and copy numbers may confer greatest susceptibility on CBD

## Association of cobalt withHLA-DPB1Glu69

- Potolcchio et al 1997

## T cell responses to beryllium

- 25 T cell clones raised from 3 CBD patients
- All proliferated to Be, but not Co or Ni

- Lombardi et al 2001

## Panel of homozygous cell lines

- EBV-transformed B lymphoblastoid cell lines (Xth IHW)
- 02012
- 0402
- 1001

## All were restricted by HLA-DP alleles with Glu69

T cell response to Be completely inhibited by anti-DP mAb

- T cell response to Be completely inhibited by anti-DP mAb

## Is Glu69 critically important in T cell recognition of Be?

- Murine DAP.3-DP2 transfectants
- only 3/12 T cell clones able to respond to DAP.3DP2 transfectants
- DPB1*0201 recognised, but not *0402

## T cell response to beryllium

- T cell lines raised from lungs of CBD patients
- T cell response inhibited by anti-DP
- DP alleles which presented beryllium matched those implicated in disease association studies

- Fontenot et al 2000

## Beryllium binding to HLA-DP 

- Soluble HLA-DP2 molecule
- Glu69 variant
- mutated lysine at position 69

- Amicosante et al 2001

## Beryllium could compete out from DP Glu69 molecule the biotinylated-CLIP at low concentration

Be bound at pH 5 and 7.5 - suggesting Be bound in absence of antigen processing

Be altered the binding of a mAb, thought to recognise epitopes within the binding groove

- Be bound at pH 5 and 7.5 - suggesting Be bound in absence of antigen processing
- Be altered the binding of a mAb, thought to recognise epitopes within the binding groove

## Cobalt binding to DP

- HLA-DP but not -DR bound cobalt
- DP*0201 bound cobalt at least three times more efficiently than *0401 (Asp55/Glu69)
- DP*0201 binds more cobalt than *0402 (Glu69)
- DP*0402 binds more cobalt than *0401 (Asp55)

- Potolicchio et al 1999

## Strong evidence from disease association and binding studies of genetic susceptibility

Is there a gene environment interaction?

- Is there a gene environment interaction?

## Association of HLA-DR3 with exposure to anhydrides in the development of specific IgE 

## Platinum study - subject characteristics

## HLA-DR3 and Platinum sensitivity

## HLA-DR6 and platinum sensitivity

## Strength of HLA-DR3 and DR6 association with sensitisation to platinum varies with intensity of exposure

## CBD, HLA-DPB1 Glu69 and Be exposure

- Richeldi et al 1997

## Genetic susceptibility increased risk at high exposure of beryllium

## Conclusions

- Identified genetic susceptibility
- Gene-environment interaction