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HLA class II sequence variation and disease susceptibility
62
Abstracts SUCCESSFUL
PREVENTION R.B.
Department of
ELLIOTT,
ANI W.
Paediatrics. Private
CUE
OF
OIABElES
CUTFIELO.
School
L.
of Medicine,
Bag, Auckland.
IN
ESCOBAR,
THE N.
N.O.D.
MOUSE
BIEBY
University
of Auckland,
New Zealand.
Pancreata with 15% Injected diabetic
from neonatal C.D. mice (6-e/litter) were diced and cultured in RPM1 1640 buffer fetal calf serum and cultured in 5% CO2 in air at 37’C for 1 week. The islets were intraparitonally either fresh or after cryo preservationintoNOD mice which had been for 3-10 days. The recipients were treated with Nicotinamide (0.5% solution as drinking water alone, or together with an hypoallergenic diet which contains no intact protein (Pregestimil @ ). Suitable controls were included. The results of animals maintained on insulin indefinitely or until death or regression of the diabetes occurred are shown. #
transplant
IO 7 10 IO 8
a
Nicotinamide
10 0 0 0
0 0 10 10
0 7 0 10
a
0
a a
Nicotinamide
Pregestimil@
a alone
or
in combination
a
Number subsequently non-diabetic 1 0 2 4 6 7
with Pregestimilm
reversed
Duration of non-diabetic state. p=permansnt 94
(3)~
P (1)
12 days
P P diabetes
permanently
in 5/20
animals. Animals given transplants with Nicotinamide alone or in combination with Either Pregestimip or Nicotinamide given Pregestimil@ reversed diabetes in 13/16 animals. from weaning to female NOD mice prevents diabetes (l/42(2%). O/23(0%) respectively. [30% A single subdiabetogenic dose of alloxan (lOOmg/Kg I.P.) (n=37) given controls (n=522)1. It is possible that free radicals at day 60 increases the diabetes rate to 80% in these mice. generated from dietary or other sources may initiate and perpetuate diabetes. including transplanted
tissues
in genetically
susceptible
animals.
HLA CLASS II SEQUENCE VARIATION AND DISEASE SUSCEPTIBILITY. B. Erlich, T. Bugawan and S. Scharf. Human Genetics Department, Cetus Corporation, 1400 Fifty-Third Street, Emeryville, CA 94608, USA. We have used the polymerase chain reaction (PCR) amplification method to sequence the polymorphic second exon of the HLA-DRH, DQa, DQS loci from a variety of haplotypes characterized by serologic and cellular typing, revealing, thus far, 24 allelic variants for the DRSI locus, 4 alleles for the DRHIII locus, 8 alleles for DQa, 14 alleles for DQD, 2 alleles for DPa, and 15 alleles at the DPD locus. Since some of the haplotypes characterized by sequence analysis were derived from IDDM, Pemphigus vulgaris (samples provided by Drs C Brautbar and A. Friedmann) and celiac disease (samples provided by Drs G.B. Ferrara and G. Angelini) patients, we have been able to compare the distribution of class II alleles in patients and controls by using oligonucleotide probe typing. For all these diseases, specific class II alleles were significantly increased among the patients. The DR4-linked susceptibility to IDDM is associated with a specific combination of DQS(3.2) and DRHI (Dw4 and DwlO) alleles. For some DR3 haplotypes, specific DPS alleles are also associated with IDDM. We have found that specific DPB alleles are also associated with celiac disease and juvenile rheumatoid athritis. For Pemphigus vulgaris, the DR4 susceptibility is associated with the DRDI allele (DwlO) and the DRw6 susceptibility with a rare DQB allele (DQDl.3) which differs from a common allele by only a Val to Asp substitution at position 57. In general, these studies have implicated the nature of the polymorphic residues at position 57 of the DQD and around position 70 of the DRBI and DP#3chains as playing a crucial role in the autoimmmune response. However, the nature of the residue at 57 does not per se predict predisposition. Thus, it is the allele rather than an individual &no acid residue which appears to confer suceptibility. The potential contributions of specific class II epitopes, alleles, haplotypes and genotypes to disease susceptibility will be discussed.
Abstracts SUCCESSFUL
PREVENTION R.B.
Department of
ELLIOTT,
ANI W.
Paediatrics. Private
CUE
OF
OIABElES
CUTFIELO.
School
L.
of Medicine,
Bag, Auckland.
IN
ESCOBAR,
THE N.
N.O.D.
MOUSE
BIEBY
University
of Auckland,
New Zealand.
Pancreata with 15% Injected diabetic
from neonatal C.D. mice (6-e/litter) were diced and cultured in RPM1 1640 buffer fetal calf serum and cultured in 5% CO2 in air at 37’C for 1 week. The islets were intraparitonally either fresh or after cryo preservationintoNOD mice which had been for 3-10 days. The recipients were treated with Nicotinamide (0.5% solution as drinking water alone, or together with an hypoallergenic diet which contains no intact protein (Pregestimil @ ). Suitable controls were included. The results of animals maintained on insulin indefinitely or until death or regression of the diabetes occurred are shown. #
transplant
IO 7 10 IO 8
a
Nicotinamide
10 0 0 0
0 0 10 10
0 7 0 10
a
0
a a
Nicotinamide
Pregestimil@
a alone
or
in combination
a
Number subsequently non-diabetic 1 0 2 4 6 7
with Pregestimilm
reversed
Duration of non-diabetic state. p=permansnt 94
(3)~
P (1)
12 days
P P diabetes
permanently
in 5/20
animals. Animals given transplants with Nicotinamide alone or in combination with Either Pregestimip or Nicotinamide given Pregestimil@ reversed diabetes in 13/16 animals. from weaning to female NOD mice prevents diabetes (l/42(2%). O/23(0%) respectively. [30% A single subdiabetogenic dose of alloxan (lOOmg/Kg I.P.) (n=37) given controls (n=522)1. It is possible that free radicals at day 60 increases the diabetes rate to 80% in these mice. generated from dietary or other sources may initiate and perpetuate diabetes. including transplanted
tissues
in genetically
susceptible
animals.
HLA CLASS II SEQUENCE VARIATION AND DISEASE SUSCEPTIBILITY. B. Erlich, T. Bugawan and S. Scharf. Human Genetics Department, Cetus Corporation, 1400 Fifty-Third Street, Emeryville, CA 94608, USA. We have used the polymerase chain reaction (PCR) amplification method to sequence the polymorphic second exon of the HLA-DRH, DQa, DQS loci from a variety of haplotypes characterized by serologic and cellular typing, revealing, thus far, 24 allelic variants for the DRSI locus, 4 alleles for the DRHIII locus, 8 alleles for DQa, 14 alleles for DQD, 2 alleles for DPa, and 15 alleles at the DPD locus. Since some of the haplotypes characterized by sequence analysis were derived from IDDM, Pemphigus vulgaris (samples provided by Drs C Brautbar and A. Friedmann) and celiac disease (samples provided by Drs G.B. Ferrara and G. Angelini) patients, we have been able to compare the distribution of class II alleles in patients and controls by using oligonucleotide probe typing. For all these diseases, specific class II alleles were significantly increased among the patients. The DR4-linked susceptibility to IDDM is associated with a specific combination of DQS(3.2) and DRHI (Dw4 and DwlO) alleles. For some DR3 haplotypes, specific DPS alleles are also associated with IDDM. We have found that specific DPB alleles are also associated with celiac disease and juvenile rheumatoid athritis. For Pemphigus vulgaris, the DR4 susceptibility is associated with the DRDI allele (DwlO) and the DRw6 susceptibility with a rare DQB allele (DQDl.3) which differs from a common allele by only a Val to Asp substitution at position 57. In general, these studies have implicated the nature of the polymorphic residues at position 57 of the DQD and around position 70 of the DRBI and DP#3chains as playing a crucial role in the autoimmmune response. However, the nature of the residue at 57 does not per se predict predisposition. Thus, it is the allele rather than an individual &no acid residue which appears to confer suceptibility. The potential contributions of specific class II epitopes, alleles, haplotypes and genotypes to disease susceptibility will be discussed.