Chemicals and Immunologic Lung Disease:Gene and

EnvironmentMeinir G Jones PhD

NHLI @ ICSMLondon

• 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

Sensitisation to acid anhydrides and association with HLA-DR3

Cases Refs P value OR

TMA 8/11 2/14 0.004 16

PA 2/12 2/14 NS 1.2

TCPA 5/7 0/0

Total 15/30 4/28

Young et al 1995

Isocyanate asthma and HLA-DQB1

DQB1 Isocyanateasthman=56

Refs

n=32

OR

*0501 1 (2%) 5 (16%) 0.14

*0503 7 (13%) 0 (0%) 9.85

Bignon et al 1994

DQB1 Isocyanateasthmatic

n=30

referentsn=12

*0503 30% 0%

*0501 0% 25%

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

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

DQB1* TDI asthman=30

Referentsn=138

Asp57++ 17 (57%) 44 (32%)

Asp57+ 12 (40%) 73 (53%)

Asp57- 1(3%) 21 (15%)

Balboni et al 1996

Chronic beryllium diseaseHLA-DPB1

DPB1 CBDn=33

Controlsn=44

Gen pop

*0201 30% 10% 10%

*0401 14% 48% 52%

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

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

• 95% of CBD Glu69

• 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

HLA-DPB1 CBDn=19

Refsn=34

02010202

Glu69+

9(42%)

23(67%)

non0201

Glu69+

16(84%)

12(35%)

Wang et al 1999

• CBD carried predominantly non-*0201 allele (84%)

• Controls carried predominantly *0201 allele (68%)

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

Association of cobalt withHLA-DPB1Glu69

DPB1* Cobalt casesn=20

Referentsn=35

Glu69 + 19/20 (95%) 17/35 (48%)

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

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

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?

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

Exposure Cases Refs OR

Low 1/1(100%)

2/34(6%)

39(1.2,1277)

Med 3/5(60%)

3/15(20%)

6(0.7,54)

High 1/1(100%)

1/3(33%)

5(0.1,220)

Platinum study - subject characteristics

Casesn=44

Referentsn=57

Age 30 (24-55) 29 (23-50)Duration ofemployment

64 (6-256) 69 (19-176)

Currentsmoking

29 (66%) 19 (33%)

Race, blackExposure

(high)

21 (48%)33 (75%)

31 (54%)39 (68%)

HLA-DR3 and Platinum sensitivity

Cases Refs Case +Ref -

Case –Ref +

OR

All 18(41%)

15(26%)

14 6 2.3(1.0,5.6)

Low 6(55%)

4(22%)

4 0 Infinite

High 12(36%)

11(28%)

10 6 1.6(0.6,4.1)

HLA-DR6 and platinum sensitivity

Cases Refs Case +Ref -

Case –Ref +

OR

All 16(36%)

34(60%)

8 19 0.4(0.2,0.8)

Low 2(18%)

12(67%)

8 19 0.1(0.02,1.1)

High 14(42%)

22(56%)

7 11 0.5(0.2, 1.3)

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

CBD, HLA-DPB1 Glu69 and Be exposure

Nonmachinist

Machinist Total

Glu69 –ve 0/55(0%)

1/31(3.2%)

1/86(1.2%)

Glu69 +ve 1/25(4%)

4/16(25%)

5/41(12%)

Total 1/80(1%)

5/47(10%)

6/127(5%)

Richeldi et al 1997

• Genetic susceptibility increased risk at high exposure of beryllium

Conclusions

• Identified genetic susceptibility

• Gene-environment interaction