Article
Reference
HLA class II DRB1, DQA1 and DQB1 polymorphisms in the Polish population from Wielkopolska
JUNGERMAN, M, et al.
Abstract
HLA DRB1, DQA1 and DQB1 alleles were determined by DNA PCR-SSO typing in a sample of 99 individuals originating from Wielkopolska (midwestern Poland). A high number of alleles (38 DRB1, 8 DQA1 and 14 DQB1) was detected at each locus, many of them presenting notable frequencies in this population. The three HLA loci are thus characterized by very high heterozygosity levels (93% for DRB1, 85% for DQA1, and 88% for DQB1), which confirms the results found for other European populations. A total of 6 DRB1-DQA1-DQB1 haplotypes are detected with an estimated frequency higher than 5%, namely, DRB1*1501-DQA1*0102-DQB1*0602, DRB1*0701-DQA1*0201-DQB1*0201, DRB1*0101-DQA1*0101-DQB1*0501, DRB1*1101-DQA1*0501-DQB1*0301, DRB1*03011-DQA1*0501-DQB1*0201, and DRB1*1301-DQA1*0103-DQB1*0603. A genetic distance analysis between the Polish and other world populations tested for HLA class II indicates that the Wielkopolska community is close to geographically close, rather than linguistically related populations from Europe. More generally, a good agreement between genetics and geography is found for DRB1 and DQB1 polymorphisms in Europe, [...]
JUNGERMAN, M, et al. HLA class II DRB1, DQA1 and DQB1 polymorphisms in the Polish population from Wielkopolska. Tissue Antigens, 1997, vol. 49, no. 6, p. 624-628
DOI : 10.1111/j.1399-0039.1997.tb02810.x PMID : 9234484
Available at:
http://archive-ouverte.unige.ch/unige:17064
Disclaimer: layout of this document may differ from the published version.
HLA class I1 DRBl, DQAl and DQBl
polymorphsms in the Polish population from Wielko polska
M. Jungerman, A. Sanchez-Mazas, l? Fichna, R. Ivanova, D. Charron, J. Hors, S. Djoulah. HLA class 11 DRBl, DQAl and DQBl
polymorphisms in the Polish population from Wielkopolska.
Tissue Antigens 1997: 49: 624-628. 6 Munksgaard, 1997
HLA DRB1, DQAl and DQBl alleles were determined by DNA PCR- SSO typing in a sample of 99 individuals originating from Wielkopolska (midwestern Poland). A high number of alleles (38 DRBl, 8 DQAl and 14 DQBl) was detected at each locus, many of them presenting notable frequencies in this population. The three HLA loci are thus characterized by very high heterozygosity levels (93% for DRB1, 85% for DQAI, and 88% for DQBl), which confirms the results found for other European populations. A total of 6 DRBI-DQAl -DQB1 haplotypes are detected with an estimated frequency higher than 5%, namely, DRB1*1501- DQA1*0102-DQB1*0602,
DRB 1 *010 l-DQAl*O 1 01-DQB 1 *0501,
DRBl*0701-DQA 1 *0201-DQB 1 *0201,
D R B 1
*
1 101-DQA 1 *050 I - DQB 1 *030 I , DRBl*03011-DQA l*050f -DQBl*O20 I , and DRB 1*
130 1- DQAl*0103-DQB1*0603. A genetic distance analysis between the Polish and other world populations tested for HLA class I1 indicates that the Wielkopolska community is close to geographically close, rather than linguistically related populations from Europe. More generally, a good agreement between genetics and geography is found for DRBl and DQBl polymorphisms in Europe, suggesting that these two loci are highly in- formative for assessing historical relationships among humans.Until now, the study of HLA genetic diversity in the European Polish population was limited to the serological typing of class I loci (XJth Interna- tional Histocompatibility Workshop and Confer- ence (IHWC), Yokohama 199 1). Previous genetic distance analyses of HLA-A and -B polymorph- isms suggest that European populations differen- tiate into a northern and a southern group (1).
According to these data, the Polish appear to be genetically close to their geographic neighbors the Slovaks and Ukrainians, who also speak an Indo- European Slavic language (2).
A more limited number of populations were tested for HLA class I1 loci during the XIth IHWC, and Poland was not represented. We thus selected a random sample of Polish individuals to analyze the SSP polymorphism of DRB I , DQA I and DQBl loci. We present here the allele and haplotype frequencies estimated for this popula-
M. Jungermanl, A. Sanchez=Mazas*, P. Fichna3, R. Ivanova4,
D. Charron4s5, J. H0rs4 and S. Djoulah4
'Institute of Human Genetics, Palish Academy of Sciences, Poznan, Poland, 2Laboratory of Genetics and Biometry, Department of Anthropology and Ecology, University of Geneva.
Switzerland, 31nstitute of Pediatrics, Academy of Medicine, Poznan, Poland, 4Department of Immunology, Saint-Louis Hospital, Paris, 51NSERM U396, Paris, France
Key words: DNA typing - Europe - HLA-DR31, DQA1, DQBI - PCR-SSO - Poland
Received 9 September 1996, revised, accepted for pubrication 6 February 1997
tion, as well as a preliminary comparison to other Europeans.
Material and methods Subject studies
DNA was isolated for a total of 99 healthy individ- uals originating from Wielkopolska (5 1"3'-53"
North, 15"-18"30' East), which is a historic prov- ince located in mid-western Poland. I t covers an area of approximately 45,000 squiire kilometcrs in- habited by 5.5 million people, which represent about 14% of the whole area and population of Poland. The largest city of the province is Poznan.
It was Wielkopolska when the state of Poland was brought to life, on the turn of the 9th century.
Since then, some migrations took place, especially from neighboring Germany.
HLA class I1 DNA typing in Poznan, Poland
H
LA ty p i ngMLA DRBI and DQBl alleles were determined by INNO-LIPA HLA typing tests (Innogenetics N.V., Zwijnaarde, Belgium) based on the reverse hybridi- zation principle. Amplified biotinylated DNA ma- ttria1 was hybridized with specific oligonucleotide probes immobilized as parallel lines on membrane- based strips. After hybridization, streptavidin labeled with alkaline phosphatase was added and bound to any biotinylated hybrid previously formed. Incubation with BCIPNBT chromogen resulted in a purple/brown precipitate.
For HLA-DQAl typing, we used the DNA PCR-SSO method. The SSO probes were 3' end labeled enzymatically by DNA deoxynucleotidyl transferase (Tdt, Boehringer Mannheim GmbH, Germany) with one digoxigenin molecule, Digoxig- enin di-deoxy-Uridine-Triphosphate (DIG- 1 1
-
ddUTP) (3).
Statistical analysis
Allele and haplotype frequencies were estimated by a maximum likelihood method aIgorithm using the program package ARLEQUIN (Excoffier et al.. , personal communication, Department of Ant hropoIogy and Ecology, University of Geneva, Switzerland). Genetic distances, defined as
k
D,=0.5x
C
Ix,--.Y,J ,i= I
where xi and y i are the frequencies of the ith al- lele in populations x and y, respectively, and k is the size of the frequency vector (4), were com- puted as for previous HLA data analyses (5-8).
A set of 16 population samples from all parts of Europe was considered (see legend for Fig. 1).
The following subset of currently known HLA alleles was taken into account for the genetic dis- tance analysis:
DRBl (44 frequencies): 0101, 0102, 0103, 1501, 1502, 1601, 1602, 0301, 0302, 0401, 0402, 0403, 0404, 0405, 0406, 0407, 0408, 0409, 0410, 041 1, 1101, 1102, 1103, 1104, 1201, 1202, 1301, 1302, 1303, 1304, 1305, 1401, 1402, 1403, 1404, 1405, 07, 0801, 0802, 0803, 0804, 0901, 1001, "other".
DQAI (9 frequencies): 0101, 0102, 0103, 0201, 03, 0401, 0501, 0601, "other".
DQBl (16 frequencies): 0501, 0502, 0503, 0504, 0601, 0602, 0603, 0604, 0605, 0201, 0301. 0302, 0303, 040 I , 0402, "other".
The mean genetic distance matrix computed over loci D R B l , DQA1. a n d DQBi was used for a
principal coordinate analysis, using the local pro- gram ANCOO (Roessli et al., personal comrnuni- cat ion).
Results
Allele frequencies
HLA-DRB 1, DQA 1 and DQB 1 allele frequencies estimated in the Polish population are presented in Table 1. As a high number of alleles (38 DRBl, 8 DQAl and 14 DQB1) is found for each locus, and no allele exhibits a very high frequency, the ob- served (obs.) hetexozygosities almost reach the maximum values expected (in brackets) for each allelic set DRBI: obs. 0.93 (0.975); DQA1: obs.
0.847 (0.875); DQBl: obs. 0.879 (0.929). The sub- typing of DRBl"0301 further reveals the presence of both 0301 1 and 03012 alleles. No blank is found
O 1
I
GYP
1
ITA-.4.1 1
"'I
BAS O."
1
SPA '*
'* 0
I
01 SAR GYP.
OAN & FRE + 'o'o'
i'*"
CRO E UL.
-0.1
J
Figzrre 1. Principal coordinate analysis for 16 European popula- tion samples tested For D R R I , DQAl, and D Q U l polymorph- isms. Top: axis 1 (horizontal) and 11 (vertical); bottom: axes 1 (horizontal) and 111 (vertical). POL: 99 Polish from Wielkopol- ska (present study); BUL: 100-1 16 Bulgarian (10); CRO: 141 Croatian ( ] I ) , CZE: 99 Czech (12). DAN: 53-55 Danes (1).
FRE: 171-180 French ( I ) . GER: SS-90 Gernian ( t ) , ITA: 284- 492 Itiilinn (1) and 99-100 ItAiun (13). R U M : 51-73 Rumanian ( 1 ) iind 83-76 Rumunian (14). SAR: 91 Sardinian ( I ) . SPA:
154-166 Spanish ( I ) titid 176 Spunish ( 1 5 ) . BAS: 80 Spanish B;isLlues ( 15); GYP: Spiinish Gypsies ( 16).
Table 1.
DQ81, DQA1 and ORBl allele frequencies in the Polish population from Wid- kopolska (n=99)
. ~~~
Allele Frequency Allele Frequency
DQBI '0201 DQBt '0602 DQBl'0501 OQB1'0301 DQBl'O302 OQBl'0603 OQBl'0402 OQB1'0601 OQBl'O303 OQBl"O304 OIlBl'O502 DOB1'0305 DQBI '0503 BLANK DQA1'0501 DQAl'0102 OQA1'0101 DQA1'0301 DQAl'O201 DQAl"OlO3 DQAl'O401 DQAl"O502 BLANK o m 1 '0604
0.1 786 0.1693 0.1 526 0.1 268 0.0992 0.0797 0.0454 0,0404 0.0303 0.0202 0.01 51 0.0051 0.0051 0.0051 0.0271 0.2171 0.1 877 0.1 707 0.1 206 0.1173 0.0997 0.0454 0.0051 0.0364
DRB1'1501 DRB1'0701 ORB1 '0101 DRB1'0301' ORB1 '1301 ORB1 '1 101 ORB1 '0104 ORB1 '0401 ORBl '0806 DRBl'1303 O R B l '1601 ORB1 *1307 ORB1 '0406 ORB1 *0413 DR61'0801 DRB1'1 I 0 4 DR61'1502 DRB1'0103 DR61'0403 ORB1 '0407 ORB1 '0414 OR 8 1 '0901 ORB1 '1001 ORB1 '1 103 ORB1 '1 106 DRBl'1201 ORBl '1503 DRBl'1504 ORBl '0304 ORBl '0402 ORB1 '041 9 ORB1 '0805 DRBl'1102 ORB1 '1107 ORB1 '1202 ORB1'1203 DRB1'1401 ORBl '1407 BLANK
0.1616 0.1 212 0.0959 0.0757 0.0707 0.0657 0.0404 0.0354 0.0252 0.0252 0.0252 0.0202 0.0151 0.0151 0.01 51 0.0151 0.0151 0.0101 0.0101 0.0101 0.0101 0.0101 0.0101 0.0101 0.0101 0.0101 0.0101 0.01 01 0.0051 0.005 1 0.0051 0.0051 0.0051 0.0051 0.0051 0.0051 0.005 1 0 0.005 1
a ORB1 '0301 1: 0.0657; ORB1 '03012: 0.0051; DRBl '0301 not subtyped:
0.0051.
for DRBI, and only 3 4 % for the other two loci, indicating a high resolution power of SSO methods.
Some relevant allelic comparisons between the Polish and other Europeans may be done for the DREH and DQBl polymorphisms. DRB1*1501 and DRB1*0101, which are among the most fre- quent alleles observed in the Polish (16.Bk and 9.6% rcspcctively), also exhibit high frequencies in Danes (17.4% and 12.7) and Czech (10.7% and 10.7'%). A striking result is that these two alLeles are likely frequent in Spanish Basques (16.9% and I2.3'1/;1), whiie in Sardinian and Spanish Gypsies, they show very low frequencies. A comparable fre- quency pattern is found for DQB1*0602. Danes and Czech are also close to the Polish for
DRB 1
*
130 1 (7.lD/o, 8.2% and 6.8% respectively), also frequent in Bulgarians (9%). The frequency of DRB1*0303 (7.6%) observed in the Polish is simi- lar to the one found in other Europeans, except Basques and Sardinians where it is much higher.DRB1*07 (12.1% in Polish) is also generally com- mon in Europe, although relevant differences are found in Spanish, Spanish Basques and Czech (higher frequencies), on one hand, and Sardinians, Bulgarians, and Spanish Gypsies (lower frequen- cies), on the other hand.
Haplotype frequencies
The most frequent DQB1-DQAl-DRBl haplo- types found in the Polish population are shown in Table 2. All these haplotypes are usually common in other Europeans.
DQBl*0602-DQA1*0102-DRBl* 1501 (1 3%) also shows a high frequency in Danes (17%), while the frequency is generally lower than 8%
in other Europeans.
shows very close frequencies in other Euro- peans, except Sardinian (less than 5'3'0).
DQB 1 *0101-DQAl*0101-DRB 1'0501 (9740) ex- hibits a similar frequency in French. The higher frequency, among Europeans, is found in Danes (12%), while it is less than 7% in other popula- tions.
presents heterogeneous frequencies in Europe, al- though generally comprised between 8 and 12%.
DQB1*0201-DQA1*0501-DRB1*0301 (7%) is common in Europe, but is especially frequent in S a rd i n ia n (24%).
DQB 1 *0603-DQAl*O 103-DRB 1
*
130 1 (6%) iscommonly observed with a low frequency in Europeans.
DQB l"020 1 -DQA 1 *02O 1 -DRB 1 *070 1 (10%)
DQB 1 *03O 1 -DQA 1 *050 1 -1DRB 1
*
1 101 (7%)Genetic distances
The mean genetic distances computed over loci DRB1, DQAl and DQBl indicate that the Polish
Table 2.
Three-locus OW-DQA1 -ORB1 haplotype frequencies in the Polish population from Wielkopolska (n=99)
Haplotype Frequencya
DQB1'0602 DI2A1'0102 DR81'1501 0.1 303 DQB1'0201 DQA1*0201 ORB1 '0701 0.1010 0.0888
OQBl'O501 OQAf'0101 ORB1 '0101
OQB1'0301 OQAI'0501 DR81'1101 0.0657 DQBl'O201 DQAl'O501 DRBl '0301 0.0707 0.0556 DOBl "0603 00A1'0103 ORB1 '1301
Only haplotypes whose frequency is higher than 5% are listed.
HLA class 11 DNA typing in Poznan, Poland population from Wielkopolska is close to the
French and the Danes, followed by the Czech, the Busques, the German and the Spanish.
The results of the principal coordinate analysis is presented in Figure 1. The first 3 principal axes represent 69% of the total variance. The first axis ( WYO) mainly differentiates the northern and west- ern European populations (Polish, Danes, Ger- man, French, Czech, Spanish and Basques) from the south-eastern European populations (Italian, Rumanian, Croatian, Bulgarian and Sardinian) and the Gypsies. According to the second axis (1 80/0), a main differentiation is observed for Sardi- nians, on one hand, and Gypsies, on the other hand. The peculiar segregation of the Gypsies sup- ports the hypothesis of their eastern (probably In- dian) origin. The second axis also segregates Polish and Danes from the other western and northern populations. The third axis (12%) separates Basques and Spanish, on one hand, and Germans,
00 the other hand, the latter being related to the French and Polish.
The results obtained for each locus taken separ- ately (personal analyses not shown) are in close a8reement with this overall analysis.
'
Di$cussion
The present results show that the Polish popula- ticm from Wielkopoiska is genetically related to geog rap h ical 1 y n e i g h bo r i n g PO p 11 1 at i o n s ( n or t he r n aqd central Europeans), rather than to other Bal- to-Slavic speakers (Czech, Bulgarian, Croatian).
Thus, the close relationship found between Polish, S l ~ v a k , and Ukrainian, for HLA class I ( I ) , is pmbably not (only) due to their linguistic affin- ities, but (also) to their geographic proximity.
More generally, the HLA class I1 multivariate aqdysis reveals that European populations are merely distributed according to geography, rather than to linguistics. The highest variance compon- ent (39%) of our principal coordinate analysis (Figure 1) seems to be related to a southeastern/
northwestern European differentiation, as it has been observed for many other genetic systems, and explained by the demic diffusion of Neolithic farmers, some 10,000 years ago (9). On the other hand, the populations belonging to a same linguis- tic: family (either Italic family including French, Spanish, Italian, Sardinian, and Rumanian or Ger- mimic family including German and Danish or S 1 iiv ic f a n il y i ncl Lid i ng Po 1 is h, B i i I ga ria n , Czech , Croatian) appear to be genetically heterogeneous.
Tliese distinct linguistic families (all belonging to the Indo-European phylum, except the Basque) owrlitp. The BXL~LKS are a linguistic isolate poss- ibly related to North Caucasians (9), showing :i
number of genetic differences from other Euro- pean populations. According to our results, how- ever, they are not as distant, genetically, as the geo- graphically isolated Sardinians are from other Europeans. Thus, geographic rather than linguistic boundaries are detectable by HLA class I1 data analysis in Europe.
This preliminary study of HLA class I1 DNA data in Europe includes a set of populations which are not tested for completely equivalent alIelic series. For example, 14 alleles found in the Polish have been included in the category "DRB 1 -other"
for genetic distance analysis. Although this constraint probably does not introduce an import- ant bias, due to the fact that all these alleles are very rare (the highest frequency is 4%, for DRBl*OZ04), the new data collected within the scope of the XlIth IHWC (Saint Malo and Paris 1996) will permit to update the present analysis, based on equivalent data sets. An important chal- lenge will be to compare numerous European populations, geographicalIy widely distributed, and linguistically diverse, to test the relative contri- bution of geography and language in the human migration history of Europe.
Acknowledgments
We would like to thank the Poznan volunteer do- nors for their participation in this work. This study was supported by FNRS (Switzerland) grant 3 100- 039847.93, INSERM and GREG. S. Djoulah is the recipient of a grant BFA/95/6748 from CMEP
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