- Email: [email protected]
Polymorphism in the promoter region of DQA1 gene: A study in celiac disease patients
48
Abstracts
II-21
II-22
F,Petronzelli, A.Kimura'. C,Tassone. P.Ferrante, D.Arma and M.C.Mazzilli
PRIMARY INVOLVEMENT IN C E L I A C D I S E A S E
T,Sasazuki',
Dipsrtimento di Medicsna Sperimentale, Universita' "La Sapienza", Viale Regina Elena 324. 0 0 1 6 1 R o m a , Italy snd •Department of Genetics, Medical Institute of Bioregulation. Kyushu University, Fukuoka, Japan
IN THE PROMOTER REGION OF STUDY IN CELIAC DISEASE PATIENTS POLYMORPIIISM
DOAI
GENE:
A
It is known that Celiac Disease (CD) is strongly associated to the DQ(aI'0S01.~I'0201) heterodimer. Furthermore many studies indicate that the quantitative expression of HLA class II antigens may play a role in the development of the disease. Since a polymorpbism in the promoter region of the IX)AI gene (QAP) as been described (Kimura and Sasazuki. "HLA 1991", in press) we investigated its contribution to CD. Sixty-four Italian patients oligotyped for DQAI, DQBI and DRBI genes, were studied for ten QAP alleles using the chemiluminescent PCR-SSO method and the results compared to those of 61 healthy controls. QAP-DQAI relations follow a very complex pattern: few combinations seem fixed while, in other cases, the same QAP allele could be associated to different DQAI or, similarly, one DQAI allele shows different promoters, according to the DRBI polymorphism. Focusing on the DQAI alleles positively and negatively associated to CD. we found no significant differences in the patients versus HLA-matched controls. In fact. all DQAI'0501 subjects (patients and controls) resulted QAP4.1 positives irrespective of the DR allele (DR3, DRII, DRI2, DRI303) and the DQAl*0101,*0102. and '0103 variants, negatively associated to CD, showed a complex pattern of QAP-DQAI combinations, but not significantly different in the two populations, It could be ef some interest that the DQAI'0501 allele at risk of CD, is always associated to the QAP4.1 promoter.
II-23 Br0nnler G., Yao Z.. Bettmotti M. diP.. Keller E., Bartova A.*, Knick A.*, and Albert E. D. Labormr Imm..n..w~k ~odet~iktimkdcr Umv*~ttitmfit~hen.Pemmkolerltr.~, * H L A - L a ~ . Hoeptuflof the UmvenayOtomou¢.CSFR
8000 M~chen2.
OF HL~ D Q A I * 0 S 0 1 - D Q B I * 0 2 0 1
HETERODIMER
Canossi A., Di Rocco M., Contasta I., Liberatore G., Romano P., Papola F.,*Mazzetti da Pietralata M.,*Sandri G., Adorno D. and Casciani C.U. Istituto CNR di Tipizzazione Tissutale e Problemi della Dialisi -p/le Collemaggio, 67100 L'Aquila *Ambulatorio MalassorDimento Intestinale e Celiachia, Roma The gluten is the etiologic factor of celiac disease (CD) but genetic factors are clearly present in this pathology. It involves a T-cell mediated response to the lamina propria of jejunum. In our previous studies we evidenced an association of CD with HLA antigens B8 (Pcorr<0.001, RR=6.894), DR3 (Pcorr
This work is supported in part by CNR grants "Ingegneria Genetica" and "Biotecnologie e Biostrumentazioni".
Ge~ny
DRBI-DQA1-DQB1-DPB1 Polymorphism in Patients with Celiac Desease Celiac Desease (CD) is a glutensansitive entempathy associated with DR3 and DR7. The hypothesis was put forward, that the primary association is with the DQAI*0501, DQBI*0201 comhinatton in cis- or trans- position. There is controversy over a possible involvement of the DPBI poiymorphism in CD. 102 patients and 142 unrelated, healthy control-persons were collected at the childrens hospital and tissue-typing-laboratory Olomouc, Czechoslovakia. They were tested for DRB 1-, DQA 1-, DQB l- and DPB l-alleles by nonradzactive otigotyping.
II-24 IGLESIAS-CASARRUBIOSP.', S£GURADO O.G.'*, MORALESIt.. MARTINEZ-LASOJ.",PARTANEN J.*, CAMPBELL R.D.*, ARNA/Z-VIIJ.ENAA'.
"De!~mn~t t~ I m m ~ , Hospital 12 de Octubre.~pain *lnstit~e ol Immunoio~, Unt~rsiw~ Munich. FRC "Tmsuel~ing I~horatuW,Hehtn~, Fmland aHRC Immunochera~,txy Urat. O,lord. UK
We obtained the following results: DRB1
PQAI
DOBI
Pat Con
RR
3 0501 0201 3 0501 0201 17 0 28 .......................................................... 3
0501
n-value < 0,0001
A SINGLE C4 GENE HAPLOTYPES
MIGHT
BE A MARKER
OF DIABETOGENIC
0201
7 0201 0201 35 7 I0 < 0,0001 ................................................................ 3 0501 0201 5 0501 0301 4 4 1,4 n.s. ............................................................... 5 0501 0301 7 0201 0201 9 8 1,6 n.s. ................................................... 3 0501 0201 X X X 21 14 2,4 < 0,05 ............................................................ From this we conclude, that the DQAl-/DQBl-hypothesis must be amended to incorporate the high frequency of DR 3/7 heterozygotes first descrihed by our group in 1977. There is a quantitative element of susceptibility associated with homozygosity for DQBl*0201 and not for DQA 1"0501. The quesnon of an independent conmbution of DPBI*0101, which is found significantly increased in patients (p < 0.0001) was answered in the negative: DPBI*0101 in the absence of DR3 is not more common in patients than in controls.
A novel "low frequency" diabetogenic haplotype (HLA-B49,DR4.DQw8) has been found in the Spanish population. It carries the SC01 complotype (BFS, C2C, CAAQ0 (deleted gene) and CAB1). usually present in the HLA-BS,DR3,DQw2 haplotype assonated with insulin-dependent diabetes meli/tus ( I D D M ) in Caucasoids. Family studies of CA Chido and Rodgers antigenic determinants and their specific C4d nucleotide sequences showed that this novel hapfotype bears Chido -3,-6. Thus, it may not be due to a recent recombination from the common HLA-BS,DR3 hapfotype bearing Ch/do 3,6; furthermore our data show that the TNFB allele which goes together with HLA-B49,SCOLDR4 is TNFB2 (or "long") and HLA-B8,SC01,I:)R3 is TNFBI (or "short"). These data support the notion that many structural differences exist between both haplotypes, although the serofogical SC01 markers have been maintained by both of them. This is the first DR4-bearing [DDM-susceptible haplotype with a deleted CA gene described so far and the only DR4-bearmg hapfotype found in the Spanish population. This report raises the intriguing fact that extended hapfotypes with deleted (or not duplicated) genes in the class 1|1 region, also including the HI.AB18,FIC30,1:)R3,DQw2 hapiotype, contain IDDM-suscoptibil/ty factors more often than non-deleted (or duplicated) haplotypes in Caucasoid and especially Mediterranean populations.
Abstracts
II-21
II-22
F,Petronzelli, A.Kimura'. C,Tassone. P.Ferrante, D.Arma and M.C.Mazzilli
PRIMARY INVOLVEMENT IN C E L I A C D I S E A S E
T,Sasazuki',
Dipsrtimento di Medicsna Sperimentale, Universita' "La Sapienza", Viale Regina Elena 324. 0 0 1 6 1 R o m a , Italy snd •Department of Genetics, Medical Institute of Bioregulation. Kyushu University, Fukuoka, Japan
IN THE PROMOTER REGION OF STUDY IN CELIAC DISEASE PATIENTS POLYMORPIIISM
DOAI
GENE:
A
It is known that Celiac Disease (CD) is strongly associated to the DQ(aI'0S01.~I'0201) heterodimer. Furthermore many studies indicate that the quantitative expression of HLA class II antigens may play a role in the development of the disease. Since a polymorpbism in the promoter region of the IX)AI gene (QAP) as been described (Kimura and Sasazuki. "HLA 1991", in press) we investigated its contribution to CD. Sixty-four Italian patients oligotyped for DQAI, DQBI and DRBI genes, were studied for ten QAP alleles using the chemiluminescent PCR-SSO method and the results compared to those of 61 healthy controls. QAP-DQAI relations follow a very complex pattern: few combinations seem fixed while, in other cases, the same QAP allele could be associated to different DQAI or, similarly, one DQAI allele shows different promoters, according to the DRBI polymorphism. Focusing on the DQAI alleles positively and negatively associated to CD. we found no significant differences in the patients versus HLA-matched controls. In fact. all DQAI'0501 subjects (patients and controls) resulted QAP4.1 positives irrespective of the DR allele (DR3, DRII, DRI2, DRI303) and the DQAl*0101,*0102. and '0103 variants, negatively associated to CD, showed a complex pattern of QAP-DQAI combinations, but not significantly different in the two populations, It could be ef some interest that the DQAI'0501 allele at risk of CD, is always associated to the QAP4.1 promoter.
II-23 Br0nnler G., Yao Z.. Bettmotti M. diP.. Keller E., Bartova A.*, Knick A.*, and Albert E. D. Labormr Imm..n..w~k ~odet~iktimkdcr Umv*~ttitmfit~hen.Pemmkolerltr.~, * H L A - L a ~ . Hoeptuflof the UmvenayOtomou¢.CSFR
8000 M~chen2.
OF HL~ D Q A I * 0 S 0 1 - D Q B I * 0 2 0 1
HETERODIMER
Canossi A., Di Rocco M., Contasta I., Liberatore G., Romano P., Papola F.,*Mazzetti da Pietralata M.,*Sandri G., Adorno D. and Casciani C.U. Istituto CNR di Tipizzazione Tissutale e Problemi della Dialisi -p/le Collemaggio, 67100 L'Aquila *Ambulatorio MalassorDimento Intestinale e Celiachia, Roma The gluten is the etiologic factor of celiac disease (CD) but genetic factors are clearly present in this pathology. It involves a T-cell mediated response to the lamina propria of jejunum. In our previous studies we evidenced an association of CD with HLA antigens B8 (Pcorr<0.001, RR=6.894), DR3 (Pcorr
This work is supported in part by CNR grants "Ingegneria Genetica" and "Biotecnologie e Biostrumentazioni".
Ge~ny
DRBI-DQA1-DQB1-DPB1 Polymorphism in Patients with Celiac Desease Celiac Desease (CD) is a glutensansitive entempathy associated with DR3 and DR7. The hypothesis was put forward, that the primary association is with the DQAI*0501, DQBI*0201 comhinatton in cis- or trans- position. There is controversy over a possible involvement of the DPBI poiymorphism in CD. 102 patients and 142 unrelated, healthy control-persons were collected at the childrens hospital and tissue-typing-laboratory Olomouc, Czechoslovakia. They were tested for DRB 1-, DQA 1-, DQB l- and DPB l-alleles by nonradzactive otigotyping.
II-24 IGLESIAS-CASARRUBIOSP.', S£GURADO O.G.'*, MORALESIt.. MARTINEZ-LASOJ.",PARTANEN J.*, CAMPBELL R.D.*, ARNA/Z-VIIJ.ENAA'.
"De!~mn~t t~ I m m ~ , Hospital 12 de Octubre.~pain *lnstit~e ol Immunoio~, Unt~rsiw~ Munich. FRC "Tmsuel~ing I~horatuW,Hehtn~, Fmland aHRC Immunochera~,txy Urat. O,lord. UK
We obtained the following results: DRB1
PQAI
DOBI
Pat Con
RR
3 0501 0201 3 0501 0201 17 0 28 .......................................................... 3
0501
n-value < 0,0001
A SINGLE C4 GENE HAPLOTYPES
MIGHT
BE A MARKER
OF DIABETOGENIC
0201
7 0201 0201 35 7 I0 < 0,0001 ................................................................ 3 0501 0201 5 0501 0301 4 4 1,4 n.s. ............................................................... 5 0501 0301 7 0201 0201 9 8 1,6 n.s. ................................................... 3 0501 0201 X X X 21 14 2,4 < 0,05 ............................................................ From this we conclude, that the DQAl-/DQBl-hypothesis must be amended to incorporate the high frequency of DR 3/7 heterozygotes first descrihed by our group in 1977. There is a quantitative element of susceptibility associated with homozygosity for DQBl*0201 and not for DQA 1"0501. The quesnon of an independent conmbution of DPBI*0101, which is found significantly increased in patients (p < 0.0001) was answered in the negative: DPBI*0101 in the absence of DR3 is not more common in patients than in controls.
A novel "low frequency" diabetogenic haplotype (HLA-B49,DR4.DQw8) has been found in the Spanish population. It carries the SC01 complotype (BFS, C2C, CAAQ0 (deleted gene) and CAB1). usually present in the HLA-BS,DR3,DQw2 haplotype assonated with insulin-dependent diabetes meli/tus ( I D D M ) in Caucasoids. Family studies of CA Chido and Rodgers antigenic determinants and their specific C4d nucleotide sequences showed that this novel hapfotype bears Chido -3,-6. Thus, it may not be due to a recent recombination from the common HLA-BS,DR3 hapfotype bearing Ch/do 3,6; furthermore our data show that the TNFB allele which goes together with HLA-B49,SCOLDR4 is TNFB2 (or "long") and HLA-B8,SC01,I:)R3 is TNFBI (or "short"). These data support the notion that many structural differences exist between both haplotypes, although the serofogical SC01 markers have been maintained by both of them. This is the first DR4-bearing [DDM-susceptible haplotype with a deleted CA gene described so far and the only DR4-bearmg hapfotype found in the Spanish population. This report raises the intriguing fact that extended hapfotypes with deleted (or not duplicated) genes in the class 1|1 region, also including the HI.AB18,FIC30,1:)R3,DQw2 hapiotype, contain IDDM-suscoptibil/ty factors more often than non-deleted (or duplicated) haplotypes in Caucasoid and especially Mediterranean populations.