HLA DR-DQ disease associations with cervical carcinoma show human papillomavirus-type specificity

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CONTRIBUTIVE ROLE OF DRB l *0404/0405 TO IDDM SUSCEPTIBILITY M. Bois, E. Tron de Bouchonv. M. Guillot, M.C. Pasquet, R. Marechaud, D. Alcalay. CRTS, CHU de Poitie~rs, France It is known that among caucasians, HLA DRB 1 04- confers an increas~ risk Ibr developing insulin dependent diabetus m¢llitus (IDDM). A consensus has developed that certain related alleles at HLA DQ A and B loci are more closely associated with susceptibility to the disease (i.e. DQAI 0301, DQB1 0302). Studies in various ethnic groups with different HLA DR-DQ linkage desequilibrium gave different answers to the question of an independent role of DRBI 04- alleles as IDDM susceptibility marker. We report here on unbalanced distribution of DRB 1"04- alleles among 41 DRB 1"04- unrelated caucasians IDDM patients registered in our institution since 1989. Each subtyping and DQB 1 typing were made with 2 techniques : PCR ASP (HLA DRB 1"0401-0408) and PCR SSP (DRB 1"0401-0411), Xth IHW reference ceils for DRB 1"0401, DRB 1"0404, DRBI*0405 were used as internal controls. DRBI*04- subtype analysis : DRBI*0404 allele frequency was significantly decreased (1 out of 47 haplotypes : 2,1% [95 CI : 0-6%]) compared with 19 out of 63 haplotypes in 59 unrelated caucasian controls (30,1% [19-41%])(p<0,01, RR=2,17). DRB 1"0405 was increased (l 7 out of 47 haplotypes) (36,1% [23-49%]) compared with 6 out 0f63 (9,5% [2-16%]) control haplotypes (p<0,01, RR=5,35). Concerning DQB 1 alleles, DQBI*0302 allele frequency was increased. All of the 41 DRBI*04- IDDM patients were typed DQBl*0302 compared with 32 out of 50 DRB 1"04- healthy controls (64% [51-77%]) (p<0,01, RR=56,25). But in controls, DRB 1"0404 as well as DRB 1*0405 were constantly associated with DQB 1"0302 (and DQA 1"0301). Thus, despite the fact that DQB 1"0302 is confirmed as major determinant of IDDM susceptibility in these DRB 1*04- IDDM patients, a significant high allele frequency of DRB 1"0405 (and low frequency of DRB 1*0404) raises again the question of the contributive role of DRB 1"04- alleles to the IDDM suseeptibility conferred by DQAI and DQBI alleles.

TNF AND MHC HAPLOTYPES IN BASQUES : RELATION TO INSULIN DEPENDENT DIABETES MELLITUS (IDDM). A. Cam_bpn-Thomsen, B. CrouauRoy, N. Bouzekri, J. Doutreix, R. Jambou, E. Ohayon, M. Abbal. CNRS UPR 8291 and Histocompatibility laboratory, CHU Purpan, Toulouse, France Basques have been studied for their MHC genes : the major haplotypes found are : A30 (or A24), Cw5, B18, BFFI, C4A3, BQ0, DR3, DQ2 ; A29, Cw-. B44, BFF, C4A3, BI, DR7. DQ2 ; A1, Cw6 or 7, B57, BFS, C4A6, B l, DR7, DQ2. Microsatellite markers located in the TNF genes region have marked linkage disequilibrium with other alleles at ocighbouring loci. We studied these TNF microsatellites in the Basque population (N = 57) and found especially a charactedstic combination of alleles TNF ni, b5 which are highly specific of the BI8, DR3 haplotype. The BI8, DR3 haplotype is associated with IDDM in Southern Europe ; Sardinia which has the highest frequency of this haplotype has also the second highest incidence rate of IDDM in Europe ; apart from the association with HLA-DQ possible other genetic factors exist in the central part of the H L A region. We thus, in addition to the normal Basque population study, conducted a genetic epidemiology study of IDDM in the French Basque population ; 100 fanlilies with at least one case of IDDM accepted to participate, half of them being of strictly Basque origin. The study of HLA and TNF markers showed that DR3, DQ2 is present in 87.2 % of the padents vs 35.7% in the Basque controls (p<0.0005, odds ratio = 12.3 [4.8-31.2[, whereas DR4, DQ8 was also significantly but less strikingly increased among patients (I7% vs 8.9% ; p.~0.001, odds ratio = 5.3 [1.9-14.9]. The reverse is usually found in northern populations where DR4, DQ8 is more strongly associated with IDDM than DR3, DQ2. However, as in all caucasoid populations the highest odds ratio is for heterozygtes DR3, DQ2.rDR4, DQ8 : 44 [2.5-67.7]. Our present data show a significant association of the haplotypes TNF al, b5 and TNF a2, b3 with IDDM. Due to linkage disequilibrium it is difficult to show the presence of a specific genetic susceptibility factor related to TNF independently of class II alleles. However the odds ratio of DR3 associated to TNF al, b5 is 16.4 [5.44-9.7] compared to 6.5 [ 1.9-22.61 for DR3 in the absence of TNF at. b5 suggesting that the susceptibility genomic region could include TNF.

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P O L Y M O R P H I S M O F THE DQBI P R O M O T E R R E G I O N IN INSULIN DEPENDENT DIABETES MELLITUS. Idriss Djilali-Saiah, Sophie CaillatZucman, Jos4 Timsit, Christian Boitard, Roger Assan* and Jean-Franqois Bach. INSERM U25, Hopital Necker. and *Service de DiabrtoJogie, Hopital Bichat.

TAP2 GENE POLYMORPHISM CONTRIBUTES TO THE GENETIC SUSCEPTIBILITY T O M U L T I P L E S C L E R O S I S . H. M o i n s - T e i s s e r e n c I, M. A l i z a b e h 2 , P. L o i s e a u I , G. E d a n 3 , B. B i r e b e n t 2 , B. G e n e t e t 2 , O. S a b o u r a u d 3 , G. S e m a n a I, D. C h a r r o n 1 (1)Laboratoire d'In~unologle et d'Histocompatibilit6, H6pital SaintLouis, Paris, France. (2)Laboratoire d'Histocompatibilite, CRTS, Rennes, France. (3)Clinique Neurologique, CHRU, Rennes, France The aim of this study is to analyse TAP1 and TAP2 membrane transporter genes involvement in the susceptibility to m u l t i p l e s c l e r o s i s (MS). T h e s e t w o g e n e s are located within the HLA class II r e g i o n and their products a r e i n v o l v e d in e n d o g e n o u s antigen processing. A population of i 1 6 u n r e l a t e d MS was compared f i r s t to a control population of 46 s u b j e c t s a n d t h e n to a c o n t r o l population of 79 H L A D R 1 5 s u b j e c t s . T A P 1 a n d T A P 2 a l l e l e s (4 f o r T A P 1 and 8 for TAP2) were typed by the PCR-SSO technique.An additional silent mutation of TAP2 gene generating two variants (I/J} was also analysed. Comparison of T A P 1 a n d T A P 2 a l l e l e s f r e q u e n c i e s in M S a n d controls showed no significant alleles variations. But, analyse of TAP2 I and TAP2 J mutations revealed an i n c r e a s e d f r e q u e n c y of J v a r i a n t in M S p a t i e n t s n o t d u e to a linkage disequilibrium of TAP2 J with HLA class II polymorphism. This suggests that TAP2 J variant is a c t i n g as an additional susceptibility genetic marker of MS. S i n c e T A P 2 J is a s i l e n t m u t a t i o n , it s e e m s o f p a r t i c u l a r interest to study the coding regions flanking this mutation.

Paris. France

Expression of MHC class 11 genes is controlled by cis-acting sequences located in upstream regulatory regions (URR) of these genes. Allele-specific polymorphism of the DQB1 promoter region has been described, suggesting that nucleotide variations in transcriptional regulatory elements could lead to a differential expression of the various DQBI alleles. To adress the question of whether particular URR variants are involved in susceptibility to insulin dependent diabetes mellitus (IDDM), we compared DQBI promoter polymorphism among 130 Caucasian fDDM patients and 70 Caucasian normal controls, all DQBl*0201,0301 or 0302. PCR amplification of the DQB 1 URR located at -315 to - 10 bp upstream from exon 1 was followed by hybridization using 8 different sequence-specific oligonueleotidic probes. The distribution of the DQB l URR variants was not different between IDDM patients and matched controls. Thus, the same variants were observed among all subjects expressing the same DQBI allele. Four unusual DQBI URR variants, i.e unexpected on the basis of known polymorphism, were observed and confirmed by single strand conformation polymorphism (SSCP) analysis. However, their frequency was similar in IDDM and control subjects (15% versus 13% respectively). Our data indicate that predisposition to IDDM is not associated with particular sequence in the transcriptional regulatory region of the DQB1 ceDe. Whether differences in URR polymorphism correlates with differences in DQ expression and could thus regulate initiation of an autoimmune response is still unclear.

114 HLA DR-DQ DISEASE ASSOCIATIONS WITH CERVICAL CARCINOMA SHOW HUMAN PAPILLOMAVIRUS-TYPE SPECIFICITY. R.J. Apple', C.M. Wheeler~, W. Klitz2, M.M. Manos3, T.M Becket4, and H.A. Erlich~. 'Dept. of Human Genetics, Roche Molecular Systems, Alameda, CA; 2Dept. of Integrative Biology, University of California, Berkeley, CA.; aDept, of Immunology and Infectious Disease, Johns Hopkins School of Public Health, Baltimore, MD; 4UNM Cancer Center, New Mexico Tumor Registry, Albuquerque, NM Cervical carcinoma (CC) is now known to be associated with human papillomaviruses (HPV), but the evidence for a link with specific HLA loci is highly controversial. The role of genetic vadation at the HLA class II loci and among HPV types in CC was investigated by PCR DNA amplification and oligonucleotide probe typing of paraffin-embedded invasive CC tissue from 98 Hispanic patients and of cervical swabs from 220 Hispanic controls. HPV was detected in 79/89 ampnfiable tumor biopsies; 53 of these contained HPV16. Frequencies of all DQB1 alleles, including the DQB1 alleles encoding the DQw3 specificity, were similar among patients and controls. However, patient and control DR-DQ haplotype frequencies differed significantly (p<0.01), and specific haplotypes were associated with either CC susceptibility or resistance. These DR-DQ assocMtions were HPV type-specific. The class II haplotype DRB1*1501DQBI*0602 was present in 13/53 HPV16-positive cancer cases and 14/220 controls (OR=4.78; 95%CI 1.90, 11.83; p=0.00007), while DR 13 haplotypes were negatively associated with HPV16-positive CC. These results suggest that specific HI.#, class II haplotypes may influence the immune response to specific HPV-encocled epitopes and affect the risk of cervical neoplasia.

115 COMP~qlSON

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T~MIOR

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SERUM

HLA

CLASS l

EXPRESSION IN

PATZ~S WITK B ~ S T CANCER. T. Kleint S. Warchaizer, I. Levin, A. Nyska, R. Narinsky, n. Kfir, E. Livni and B. Klein, Departments of Oneology and Pathology, Golda Medical Center, and Tissue Typing Laboratory, neilinson Medical Center, Petah Tiqva, Israel The proper expression of class antigens is contingent upon association of the class I heavy chain with the light chain B-2microglobuin (B-2M). When the HLA complex is metabolised, B-2M is shed into the serum. A large variety of human and experimental tumors have altered MHC class I expression. In a previous study (Cancer 67:2295, 1991) we observed elevated mean B-2M serum levels in B.C. patients (pts) compared to controls. To study the relationship between tumor expression and serum levels, we examined 54 pts with B.C. Tumo~ B-2M was determined with iramunohistochemistry and serum levels by ELISA technique. Of the 54 pts, 33 had low and 21 had high B-2M expression on the tumor. Of the 36 infiltrating ductal tumors, 70% were low expressors. When tumor expression was compared to B-2M serum level 95% of pts with high tumor B-2M expression had high serum levels of B-2M, in contrast only, 53% Of pts with low tumor B-2M expression had low serum levels of B-2M (
responses.