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HLA class II allele frequencies in unrelated donor bone marrow transplantation
105
Abstracts
P573
P574
THE FREQUENCIES AND DISTRIBUTION OF HLA CLASS II ALLELES IN FOUR SOUTHERNAFRICAN POPULATIONS B Arendse, G Harms, M Jacons, J Glaser, K Meehan, V Pokorny & J Rousseau
A COMPARISON OFHLA CLASS II ALLELE FREQUENCIES WlTHlN AFRICA: IMPLlCAnONS FORVACCINEDEVELOPMENT
ProvincialLaboratory for Tissue Immunology, Cape Town, Soutfl Africa
AB Begcvich. JD Chang, LL Steiner, CA Aldrich,and PA Zimmerman'. Roche Molecular Systems, Alameda, CA and"National Institutes of Allergyand Infectious Diseases, NIH, Bethesda, MD USA
The HLA class II allele distribution in 4 Southem African populations was analysed by amplification of the second exon sequences from the DRB1, DQB1, DOA1 and DPBl genes in unrelated Xhosa, Cape Coloured, Khoi and San individuals. The Xhosa group are representative of the Ngunispeaking tribes; the Cape Coloureds possess Westem European, Southem African and Asian genes; the Khoi and San are descendant from people of the latter Stone Age. The most commonly occurring DR81 alleles were: Xhosa, DRB1'0302 (f=0.14); Cape Coloured, DRB1'0701 (f=0.12); Khoi, DRB1'1501 (f=0.30) and San, DRB1'0401 (f=0.41). Although the DRB1'0701 allele occurred most frequently in the Cape Coloureds, the DRB1'11 (f=0.1ff) and DRB1'15 (f=0.16) groups had higher frequencies. There appeared to be an almost total absence of DRB1'16 alleles in all 4 populations. The most commonly occurring D081 alleles were: Xhosa, DQB1'0602 (f=0.21); Cape Coloured, D081'0301 (f=0.20); Khoi, DOB1'0602 (f=0.26) and San, DQB1'0302 (f=0.47). The most commonly occurring DQA1 alleles were: Xhosa, DOA1'0102 (f=0.28); Cape Coloured, DQA1'0501 (f=0.27), Khoi, DOA1'0102 (f=0.43) and San, DQA1'0301 (f=0.62). No deviations from Hardy-Weinberg eqUilibrium were seen, except for the San DQA1'0301 allele, which is probably due to its association with the DRB1'04 and DRB1'07 alleles which account for more than 60% of the allele complement. The most common haplotypes were: Xhosa, DRB1'0302, DQB1'0402, DQA1'0401; Cape Coloured, DRB1'0701, DOB1'0201, DQA1'0101; Khoi, DRB1'1501, DQB1'0602, DQA1'0102 and San, DRB1'0401, DQB1'0302, DQA1'0301. No association was seen between the DRB1 and DPB1 alleles in the Khoi or the San.
P575 HLA CLASS 11 ALLELEFREQUENCIES IN UNRELATED DONOR BONE MARROW TRANSPLANTAnON Yunis E, Begovich A, Awdeh Z, Baxter-Lowe LA, Noreen H, Schmeckpeper B,
Spalding T, Williams TM. Hegland J, Salazar M National Marrow Dooor Program (NMDP), Minneapolis, Minnesota, USA The allele frequencies of class IJ loci of the major histocompatibility complex were assigned in 1,302 donors-recipient pairs from unrelated-donor bone marrow transplants
facilitared bytheNMOP between 1988 and 1994. Seventy-three percent of the patients and 87% of'the donors were of Caucasoid origin. The most commonly observed allele frequencies, illustrated below, are not surprising considering this predominantly Caucasian sample set.
Locus
Allele Frequency %
DRBI 0701 (15.0) 1501 (14.6) 0301 (13.7) 0401 (10.1)
DQBI 0301 (16.2) 0002 (15.9) 0201 (12.1) 0202 (10.8)
DQAI 0501 (23.1) 0102 (21.8) 0201 (15.9) 0101 (11.0)
DPBI 0401 (32.6) 0201 (14.3) 0402 (13.4) 0301 (11.7)
OPAl 0103 (80.7) 0201 (14.6) 0202 (3.7) 0104 (0.6)
Although there is a high level of poLymorphism at each class II locus, the four most frequently observed alleles together constitute greater than 50% of the observed allele frequencies in this population. Allele frequencies observed in the patient and donor groups were virtually identical. Based on high resolution typing, there were new associations identified, such as: DRBl*1001 and *1401 with DQAl ·0104; DRBI ·0701
withOQBI*0202; and DRBI'0901 withDQAI'OI03,DQB1'0303. Haplotype frequencies will be calculated and confirmed in the patient population by obtaining
famlly studies fromparticipating transplant centers. This study was sponsored bythe NMDP (Office ofNaval Research Grant #N00014-93-1-0658).
P577
ANALYSIS OF SEGREGATION OF 'MHC BLOCK PROFILES WITH HLA LOCI IN RECOMBINANT HAPLOTYPES H Grosse-Wilde', G Tay", S Gauldieri', CS Witt', HOttinger', FT Ohrisfiansen" and RL Dawkins'. 'Institute for Immunology, Universitatsklinikum Essen, Essen, Germany and for Moleculary Immunology and Instrumentation, The University of Westem Australia, Perth, Australia.
"centre
Previously, we have demonstrated the value of non-HLA polymorphisms in the MHC ('MHC block matching') in selecting MHC identical siblings. In the recombinant family component of the 121HW, we studied families in which recombinant haplotypes were identified by HLA typing and family study. We hypothesised that recombination does not occur between the non-HLA polymorphism used in 'block matching' and the HLA loci due to suppression of recombination in these highly polymorphic regions. For example, recombination is rare between HLA-DR and -DO. We predicted that recombinationwould occur preferentially in less polymorphic regions between these polymorphic blocks of several hundred kllobases, The aim of this study was to determine whether the non-HLA polymorphism used by MHC block matching segregate with the HLA loci, even in recombinant haplotypes. In each family, the MHC alpha, beta and delta block profiles from the offspring with the recombinant haplotype were compared with other siblings. The nonHLA profiles were invariably as predicted by HLA typing, confirming that recombination rarely occurs between blocks. Block matching allowed the assignment of haplotypes in ambiguous cases. Studies using microsatellites are currently in progress to refine the location of recombination occurring between these blocks, (9542)
In order to understand hoththeorigins of African-American populations and the genetic relationships within sub-Saharan Africa, we have initiated a study of HLA class II allele frequencies in Africa.PCR-hased assays have been used to typethe HLA-DRBI, -DQAI,-DQBI,- DPAI and -DPBlloci in over450individuals representing 17villages in five WestAfricancountries (Senegal, Guinea, Sierra Leone, Ivory Coast, & Ghana) and I villagein theCameroon. Twenty-three DRB!, 8 DQA1, 12DQBI, 8 DPAI and 25DPBI alleles have been identified. There are significant differences between the allele frequencies found in West Africa relative to theCameroon. Forexample, three OPB I alleles arenewandunique to the Cameroon. Analysis of DR-DQhaplotypes reveals a great dealof diversity aswell as a number of novelhaplotypes notobserved in non-African populations. A minimum of 38 distinctDR-DQhaplotypes have been identified in these two populations some of which are unique to the Cameroon. There are also significant differences in thehaplotype frequencies between thetwo Africanlocations. These data suggest thatthere is notonlya great dealof diversity within Africa butalso between different Africanpopulations and have important implications for vaccine development. Theyalso provide thebeginnings of adatabase to studythe population structure of African-derived populations.
P576
TNF MICROSATELLITE POLYMORPHISMS: A COMPARlSON OF ALLELE FREQUENCIES IN UK AND OTHER EUROPEAN POPULA nONS Poulton K, Hajeer A, Worthington J, Fryer AA, StrangeR, Oilier WER University of Manchester,Manchester,UK. Allele frequencies for TNF microsatelliteswere investigatedin 2 UK control cohorts and compared with published data on other Europeanpopulations. UK Group I comprised of 109 laboratory workers and UK Group 2 comprised of 251 individuals who were admitted to hospital for minor routine surgery. Genomic DNA was amplified using fluorescently labelled primers for 4 TNF microsatellites (a.b.c.d). PCR products were pooled and analyzedby automated gene scanning. The resultswere comparedwith data from 56 Basques, 45 Greeks,75 Danish, controls. Therewere somedifferencesin TNFa allele frequencies betweenthe groups. Allele 2 was the most frequent allele in all groups and TNFa 6, 10 and II were generallyin high frequencies comparedto TNFa 3,8 and 9 which were at low frequency in all groups. TNF b allele frequencieswere similar in all but the Frenchgroup. TNFc is biallelic and its distribution was similar in all groups. TNFd datawere not available for the non-UK groups. The variation in allele frequencies observed betweenthe groups may reflect true ethnic variation or may result from the variable and often small sample sizes. Testing oflarger cohorts will be necessary to address this question. Selection of appropriate control groups for microsatellite based disease associationstudieswill obviously be important.
P578 ALLELIC FREQUENCIES OF CLASS
U ( DR, DQ, Dp) AND OF NEW GENES (TAPl-2, DMA, DMB, LMP2) MARKERS IN A PANEL OF 100 FRENCH INDIVIDUALS, Cheneau ML, Chevrier D, Pinson MI, Cesbron A. MullerN. andBignon lD. HLALaboratory,Blood bank, 44011 Nantes cedex 01,France.
If classical class II alleles namely DR,DQand OPare commonly assessed and validated in numerous populations, new markers encoded by new genes located in MIlCnamely TAPI, TAP2, DMA, DMB and LMP2 are no! yet weD validated in ethnically homogeneous populations. Frequency evaluations of thesenew alleles remains veryusefulnotonly from ethnical point of view, but also for liLA and disease studies. Therefore, using PeR technics(pcR-SSO, ARMS-PCR), we typed for our panel of 100 bealtbyindividuals. DR, DQandOPallele freqnencies were not dilIeren! from oar previousdata or otherpublisbed data from freech caucasoids. Newgene allele frequencies (Afq) are summarised in the foUowing table :
TAP2 A.fq DMA A.fq DMB TAP1 A.fq *0101 0.895 ["810110103 0.520 "0101 0.81 101 *0201 0.095 1"0102 0.015 "0102 0.12 102 *0301 0.010 1"0201 0.330 *0103 0.02 "0103 0.135 *0104 0.05 "0104 ·New • includingTAP2Cand TAP2D.
ru
1'0
A.fq LMP2 A.fq 0.72 0.750 A 0.030 H 0.28 0.195 0.025
Weare currently assessing linkage disequilibriabetween classical class II genes and newly identified genesby nsing selected familie'
Abstracts
P573
P574
THE FREQUENCIES AND DISTRIBUTION OF HLA CLASS II ALLELES IN FOUR SOUTHERNAFRICAN POPULATIONS B Arendse, G Harms, M Jacons, J Glaser, K Meehan, V Pokorny & J Rousseau
A COMPARISON OFHLA CLASS II ALLELE FREQUENCIES WlTHlN AFRICA: IMPLlCAnONS FORVACCINEDEVELOPMENT
ProvincialLaboratory for Tissue Immunology, Cape Town, Soutfl Africa
AB Begcvich. JD Chang, LL Steiner, CA Aldrich,and PA Zimmerman'. Roche Molecular Systems, Alameda, CA and"National Institutes of Allergyand Infectious Diseases, NIH, Bethesda, MD USA
The HLA class II allele distribution in 4 Southem African populations was analysed by amplification of the second exon sequences from the DRB1, DQB1, DOA1 and DPBl genes in unrelated Xhosa, Cape Coloured, Khoi and San individuals. The Xhosa group are representative of the Ngunispeaking tribes; the Cape Coloureds possess Westem European, Southem African and Asian genes; the Khoi and San are descendant from people of the latter Stone Age. The most commonly occurring DR81 alleles were: Xhosa, DRB1'0302 (f=0.14); Cape Coloured, DRB1'0701 (f=0.12); Khoi, DRB1'1501 (f=0.30) and San, DRB1'0401 (f=0.41). Although the DRB1'0701 allele occurred most frequently in the Cape Coloureds, the DRB1'11 (f=0.1ff) and DRB1'15 (f=0.16) groups had higher frequencies. There appeared to be an almost total absence of DRB1'16 alleles in all 4 populations. The most commonly occurring D081 alleles were: Xhosa, DQB1'0602 (f=0.21); Cape Coloured, D081'0301 (f=0.20); Khoi, DOB1'0602 (f=0.26) and San, DQB1'0302 (f=0.47). The most commonly occurring DQA1 alleles were: Xhosa, DOA1'0102 (f=0.28); Cape Coloured, DQA1'0501 (f=0.27), Khoi, DOA1'0102 (f=0.43) and San, DQA1'0301 (f=0.62). No deviations from Hardy-Weinberg eqUilibrium were seen, except for the San DQA1'0301 allele, which is probably due to its association with the DRB1'04 and DRB1'07 alleles which account for more than 60% of the allele complement. The most common haplotypes were: Xhosa, DRB1'0302, DQB1'0402, DQA1'0401; Cape Coloured, DRB1'0701, DOB1'0201, DQA1'0101; Khoi, DRB1'1501, DQB1'0602, DQA1'0102 and San, DRB1'0401, DQB1'0302, DQA1'0301. No association was seen between the DRB1 and DPB1 alleles in the Khoi or the San.
P575 HLA CLASS 11 ALLELEFREQUENCIES IN UNRELATED DONOR BONE MARROW TRANSPLANTAnON Yunis E, Begovich A, Awdeh Z, Baxter-Lowe LA, Noreen H, Schmeckpeper B,
Spalding T, Williams TM. Hegland J, Salazar M National Marrow Dooor Program (NMDP), Minneapolis, Minnesota, USA The allele frequencies of class IJ loci of the major histocompatibility complex were assigned in 1,302 donors-recipient pairs from unrelated-donor bone marrow transplants
facilitared bytheNMOP between 1988 and 1994. Seventy-three percent of the patients and 87% of'the donors were of Caucasoid origin. The most commonly observed allele frequencies, illustrated below, are not surprising considering this predominantly Caucasian sample set.
Locus
Allele Frequency %
DRBI 0701 (15.0) 1501 (14.6) 0301 (13.7) 0401 (10.1)
DQBI 0301 (16.2) 0002 (15.9) 0201 (12.1) 0202 (10.8)
DQAI 0501 (23.1) 0102 (21.8) 0201 (15.9) 0101 (11.0)
DPBI 0401 (32.6) 0201 (14.3) 0402 (13.4) 0301 (11.7)
OPAl 0103 (80.7) 0201 (14.6) 0202 (3.7) 0104 (0.6)
Although there is a high level of poLymorphism at each class II locus, the four most frequently observed alleles together constitute greater than 50% of the observed allele frequencies in this population. Allele frequencies observed in the patient and donor groups were virtually identical. Based on high resolution typing, there were new associations identified, such as: DRBl*1001 and *1401 with DQAl ·0104; DRBI ·0701
withOQBI*0202; and DRBI'0901 withDQAI'OI03,DQB1'0303. Haplotype frequencies will be calculated and confirmed in the patient population by obtaining
famlly studies fromparticipating transplant centers. This study was sponsored bythe NMDP (Office ofNaval Research Grant #N00014-93-1-0658).
P577
ANALYSIS OF SEGREGATION OF 'MHC BLOCK PROFILES WITH HLA LOCI IN RECOMBINANT HAPLOTYPES H Grosse-Wilde', G Tay", S Gauldieri', CS Witt', HOttinger', FT Ohrisfiansen" and RL Dawkins'. 'Institute for Immunology, Universitatsklinikum Essen, Essen, Germany and for Moleculary Immunology and Instrumentation, The University of Westem Australia, Perth, Australia.
"centre
Previously, we have demonstrated the value of non-HLA polymorphisms in the MHC ('MHC block matching') in selecting MHC identical siblings. In the recombinant family component of the 121HW, we studied families in which recombinant haplotypes were identified by HLA typing and family study. We hypothesised that recombination does not occur between the non-HLA polymorphism used in 'block matching' and the HLA loci due to suppression of recombination in these highly polymorphic regions. For example, recombination is rare between HLA-DR and -DO. We predicted that recombinationwould occur preferentially in less polymorphic regions between these polymorphic blocks of several hundred kllobases, The aim of this study was to determine whether the non-HLA polymorphism used by MHC block matching segregate with the HLA loci, even in recombinant haplotypes. In each family, the MHC alpha, beta and delta block profiles from the offspring with the recombinant haplotype were compared with other siblings. The nonHLA profiles were invariably as predicted by HLA typing, confirming that recombination rarely occurs between blocks. Block matching allowed the assignment of haplotypes in ambiguous cases. Studies using microsatellites are currently in progress to refine the location of recombination occurring between these blocks, (9542)
In order to understand hoththeorigins of African-American populations and the genetic relationships within sub-Saharan Africa, we have initiated a study of HLA class II allele frequencies in Africa.PCR-hased assays have been used to typethe HLA-DRBI, -DQAI,-DQBI,- DPAI and -DPBlloci in over450individuals representing 17villages in five WestAfricancountries (Senegal, Guinea, Sierra Leone, Ivory Coast, & Ghana) and I villagein theCameroon. Twenty-three DRB!, 8 DQA1, 12DQBI, 8 DPAI and 25DPBI alleles have been identified. There are significant differences between the allele frequencies found in West Africa relative to theCameroon. Forexample, three OPB I alleles arenewandunique to the Cameroon. Analysis of DR-DQhaplotypes reveals a great dealof diversity aswell as a number of novelhaplotypes notobserved in non-African populations. A minimum of 38 distinctDR-DQhaplotypes have been identified in these two populations some of which are unique to the Cameroon. There are also significant differences in thehaplotype frequencies between thetwo Africanlocations. These data suggest thatthere is notonlya great dealof diversity within Africa butalso between different Africanpopulations and have important implications for vaccine development. Theyalso provide thebeginnings of adatabase to studythe population structure of African-derived populations.
P576
TNF MICROSATELLITE POLYMORPHISMS: A COMPARlSON OF ALLELE FREQUENCIES IN UK AND OTHER EUROPEAN POPULA nONS Poulton K, Hajeer A, Worthington J, Fryer AA, StrangeR, Oilier WER University of Manchester,Manchester,UK. Allele frequencies for TNF microsatelliteswere investigatedin 2 UK control cohorts and compared with published data on other Europeanpopulations. UK Group I comprised of 109 laboratory workers and UK Group 2 comprised of 251 individuals who were admitted to hospital for minor routine surgery. Genomic DNA was amplified using fluorescently labelled primers for 4 TNF microsatellites (a.b.c.d). PCR products were pooled and analyzedby automated gene scanning. The resultswere comparedwith data from 56 Basques, 45 Greeks,75 Danish, controls. Therewere somedifferencesin TNFa allele frequencies betweenthe groups. Allele 2 was the most frequent allele in all groups and TNFa 6, 10 and II were generallyin high frequencies comparedto TNFa 3,8 and 9 which were at low frequency in all groups. TNF b allele frequencieswere similar in all but the Frenchgroup. TNFc is biallelic and its distribution was similar in all groups. TNFd datawere not available for the non-UK groups. The variation in allele frequencies observed betweenthe groups may reflect true ethnic variation or may result from the variable and often small sample sizes. Testing oflarger cohorts will be necessary to address this question. Selection of appropriate control groups for microsatellite based disease associationstudieswill obviously be important.
P578 ALLELIC FREQUENCIES OF CLASS
U ( DR, DQ, Dp) AND OF NEW GENES (TAPl-2, DMA, DMB, LMP2) MARKERS IN A PANEL OF 100 FRENCH INDIVIDUALS, Cheneau ML, Chevrier D, Pinson MI, Cesbron A. MullerN. andBignon lD. HLALaboratory,Blood bank, 44011 Nantes cedex 01,France.
If classical class II alleles namely DR,DQand OPare commonly assessed and validated in numerous populations, new markers encoded by new genes located in MIlCnamely TAPI, TAP2, DMA, DMB and LMP2 are no! yet weD validated in ethnically homogeneous populations. Frequency evaluations of thesenew alleles remains veryusefulnotonly from ethnical point of view, but also for liLA and disease studies. Therefore, using PeR technics(pcR-SSO, ARMS-PCR), we typed for our panel of 100 bealtbyindividuals. DR, DQandOPallele freqnencies were not dilIeren! from oar previousdata or otherpublisbed data from freech caucasoids. Newgene allele frequencies (Afq) are summarised in the foUowing table :
TAP2 A.fq DMA A.fq DMB TAP1 A.fq *0101 0.895 ["810110103 0.520 "0101 0.81 101 *0201 0.095 1"0102 0.015 "0102 0.12 102 *0301 0.010 1"0201 0.330 *0103 0.02 "0103 0.135 *0104 0.05 "0104 ·New • includingTAP2Cand TAP2D.
ru
1'0
A.fq LMP2 A.fq 0.72 0.750 A 0.030 H 0.28 0.195 0.025
Weare currently assessing linkage disequilibriabetween classical class II genes and newly identified genesby nsing selected familie'