- Email: [email protected]
HLA variation in two populations of the Iberian Peninsula
Abstracts
53
P275 NOVEL Ill.A-DQB1 ALLELE AND CLASS II HAPWTYPE Fontaine Pierre, Fajardy Isabelle, Weill Jacques, Dame P.M. LaboratoireHLA, CHRU ULLE, France
Using a PCR-RFLP strategy, we studied the HLA-DQAl, DQBl and DRBI polymorphism to generate the HLA genotype of a family and found a novel HLA-DQBl allele constituting a novel DRBI-DQAI-DQBI extended haplotype. We studied a nuclear family: the father, the mother and their son; the father is originated from Martinique, the mother from the North of France. The father and his son bear a HLA-DQBl *0201 like allele found by irregular restriction pattern and confirmed by sequencing. This new allele named DQBI*0203 (WHO nomenclature) shows identical exoo2 sequence to DQBI *0201 except for a single nucleotide substitution at codon 57 (GCC (Alanine) -> GAC (Aspartate)). The candidate haplotype in which this substitution must have occured is DRBI*0301-DQAI*0501-DQBI*020l, haplotype described with a significant linkage disequilibrium in different ethnic groups. Surprisingly, the observed haplotype is DRB I *0302-DQA I *0501-DQB I *0203 instead of the DRBI*0302-*DQAI*040I-DQBI*0402 combination usually described in black american populations . The unexpected haplotype we observe allows us to propose a sheme of evolution which involves an interlocus crossing over event followed by a ponctual substitution generating both the american black specific haplotype and the novel DQBl*0203 allele.
P276
C LOCUS: XII WORKSHOP SERA AND DNA TYP1NG Longo A. Delfino L. Morabito A, Ferrara GB. Immunogenetics Lab,. CBA/IST. Genova, Italy In the frame of the XII Workshop Anthropological study 101 samples of a Northern Italian Population have been typed. As far as the C locus is concerned, DNA typing has been performed for 17 alleles. At serological level 190 As. and Moab anti locus C sent by the Workshop have been studied. Only 49 samples out of 190 have an r. higher than .700 and among these only 13 samples have an r. higher than .850. The splits typed on DNA Cw303/304 and Cw70l+02+03/704 are not serologically identified with any sera. Evenly, the new alleles typed on DNA (Cw12/l203/l4/l5/l60l/l602 and 1701) covering all together the 18,1% of frequence in this population are not serologically identified. Two significant differences of frequencies between the data from the XII Workshop,Italian Population, (a) and data from this Northern Italian Population typing (b) have been identified for: Cwl~ 3.0 in (a) and 7.4 in (b) and Cw3~ 3.6 in (a) and 6.0 in (b). The data of the eight alleles typed either at DNA and serologic level are identical.
P277 HLA CW ARMS-PCRTYPING IN THE SARDINIAN POPULAnON
P278
Cantu Licinio, Floris Lauretta, Carcassi Carlo, Deidda Donatella, Alba Francesco, Gessa 5./ Boero Roberta, Mulargia Marina, La Nasa Giorgio, Cirillo Daniela. Cattedra di Genetica Medica, Universita di Cagliari, Cagliari, Italy.
David Comas, Eva Mateu, Francese Calafell. Jaume Bertnnpetit, laboratori d'Antropologia. Facultat de Biologia. Universitat de Barcelona. Diagonal 645. 08028 Barcelona, Catalonia, Spain.
HLA-C alleles have always been the least well characterized of the HLA class I genes and the genetic nature of HLA-C gene products is of increasing interest in the field of immunology for their involvement in their recognition of natural killer cells as well as for their importance in transplantation, disease susceptibility and anthropological studies. In order to identify serologically detectable and undetectable HLA-Cw antigens we used a sequence-specific primer (peR-SSP) system (23 primer
mixes) to detect all sequenced HLA-Cw alleles in 119 random individuals belonging to the Sardinian population. The results are shown in the following
table. Cw allele '01 '02 '0302 '0303 '04 '0501 '0602
Frequency 0.050 0.050 0.004 0.021 0.176 0.184 0.088
Cw allele '07 '08 '1201/2 '1203 '15 '1601 '1602 'POI
Frequency 0.21 0.071 0.008 0.071 0.046 0.008 0.004 0.004
HLA VARJATlON IN TWO POPULATIONS OF THE IBERIAN PENINSULA
The genetic differentiation of the Basques has long been recognized although results are not the same when analyzing allele frequencies or mtDNA sequences. In order to analyze the level of differentiation we ha...e srudied the HLA system comparing Basques with Catalans, a hardly diferentiated European population as shown by classical markers. The two populations are found at the edges of the Pyrenees and previous evidence show a sharp genetic cba.nge. The high polymorphism of the system and 111e high resolution at the DNA level of the typing allow to distinguish between both Iberian Peninsula populations and discuss the genetic level of differentiation as HLA may be both treated as allele frequencies or DNA sequences. One sample of 100 autocthonous individuals from the Basque country (Guipuzkoa) end one sample of 88 autocthcncus individuals from Calalonia (North of Girona) have been typed for HLA Class [ and Class D. High resolution DNA typing has been used in both populations. Samples have been typed for HLA Class I alleles (HLA-A, HLA-B and HLA-C) by an Amplification Refractory Mutation System (ARMS)·PCR assay: thirty-two pairs of primers have been used for HLA-A, twenty-seven for HLA-B and twenty-three for HLA-e. The 12th lntemational Histocompatibility workshop SSO-PeR technique has been used 10 type at a high allelic resolution level the loci of the Class n HLA antigens (HLA-DRB, HLA-DQA, HLA-DQB, HLAl>PA and IRA-DPB) using ll. large number of specific probes. The allele and the haplotype frequencies of HLA Class I and Class II have been compared to the anthropological data collected for the 11th International Histocompatibility Workshop. The large amount of data produced for the 12th International Histocompatibility Workshop will allow to compare the present populations with other populations around the world using the same high resolution DNA techniques.
As was expected we observed a lower level of Cw homozygosity (homozygous
+ heterozygous/blank) by ARMS-peR when compared to serology: 17.64%vs 62.18%.
P279
P280
HLA ALLELE AND HAPLOlYPE FREQUENCIES IN ALGERIANS: RELATEDNESS TO SPANIARDS AND BASQUES.
SURNAME AND HLA GENE FREQUENCIESTO REVEAL RECENT AND ANCIENT MIGRATION
Antonio Arnaiz-Villena, Djamal Benmamar, Nieves Diaz-Campos, Eduardo Gomez-Casado, Miguel Alvarez, Pilar Varela, Jorge Martinez-Laso Department of Immunology, Hospital 12 de Ocrubre, Universidad Complutense, Madrid, Spain.
C.R. Guglielmino 1, A. De Silvestrll , M. Martinetti2, L. Spada3• A. Rossi3, A. Rossi3, and M. Cuccial 1 Dipartimento di Genetica e Microbiologia-Universita di Pavia-Italy 2 Servizio di Immunoematologia e Trasfusione IRCCS Policlinico S. Matteo-Pavia-Italy 3 Servizio Immunoematologico Az. Osp. USSL2-Piacenza-ltaly
The HLA system has been used as a powerful genetic polymorphism in order to identify individuals and populations. Ethnic groups may be characterized byspecificHLA allele frequencies and particular extended HLA haplotypes; also, genetic relationships among these groups may be deduced. In the present study,seroiogyand DNA typinghave been used to detect HLAA, -B, -C, -DR and -DQ alleles in each individual and to calculate characteristic haplotypes in Algerians. These results bave been compared to the previously obtained in other populations, particularly Northern Meciterraneans: genetic distances and their respective dendrograms place Basques and Spaniards closer to Algerians than to other Europeans. Also,
characteristic Basque and/or Spanish haplotypes are found in Algerians, i.e.: A30-B18-Cw5-DR3-DQ2, AI-B57-Cw7.DR7-DQ2. This supports that the Algerian population, mainly its paleo-North African component (Berbers), have a common descent with Basques and Spaniards, probably reflecting a preneolithic relationship between Iberians and paleo-North Africans.
The combined use of HLA genes and surnames reveals genetic barriers, migration and can enlight the historical origin of populations even in a small area. We studied Pavia and Piacenza provinces situated in Lombardy and in Emilia respectively (Northern Italy). Both provinces share a mountainous region. while the remaining part is flat. With principal component analysis, both kind of data show that the mountainous region of the two province are separated from the plain areas and closed one to each other. The separation is even more clear with the HLA data; since HLA genes reflect more ancient features of populations, we can speculate that the mountainous populations of the two provinces share a common ancestor. There is archaeological evidence that the mountainous area was inhabited from prehistorical time till Middle Age by the Ligurians, one of the most ancient Italic population.
53
P275 NOVEL Ill.A-DQB1 ALLELE AND CLASS II HAPWTYPE Fontaine Pierre, Fajardy Isabelle, Weill Jacques, Dame P.M. LaboratoireHLA, CHRU ULLE, France
Using a PCR-RFLP strategy, we studied the HLA-DQAl, DQBl and DRBI polymorphism to generate the HLA genotype of a family and found a novel HLA-DQBl allele constituting a novel DRBI-DQAI-DQBI extended haplotype. We studied a nuclear family: the father, the mother and their son; the father is originated from Martinique, the mother from the North of France. The father and his son bear a HLA-DQBl *0201 like allele found by irregular restriction pattern and confirmed by sequencing. This new allele named DQBI*0203 (WHO nomenclature) shows identical exoo2 sequence to DQBI *0201 except for a single nucleotide substitution at codon 57 (GCC (Alanine) -> GAC (Aspartate)). The candidate haplotype in which this substitution must have occured is DRBI*0301-DQAI*0501-DQBI*020l, haplotype described with a significant linkage disequilibrium in different ethnic groups. Surprisingly, the observed haplotype is DRB I *0302-DQA I *0501-DQB I *0203 instead of the DRBI*0302-*DQAI*040I-DQBI*0402 combination usually described in black american populations . The unexpected haplotype we observe allows us to propose a sheme of evolution which involves an interlocus crossing over event followed by a ponctual substitution generating both the american black specific haplotype and the novel DQBl*0203 allele.
P276
C LOCUS: XII WORKSHOP SERA AND DNA TYP1NG Longo A. Delfino L. Morabito A, Ferrara GB. Immunogenetics Lab,. CBA/IST. Genova, Italy In the frame of the XII Workshop Anthropological study 101 samples of a Northern Italian Population have been typed. As far as the C locus is concerned, DNA typing has been performed for 17 alleles. At serological level 190 As. and Moab anti locus C sent by the Workshop have been studied. Only 49 samples out of 190 have an r. higher than .700 and among these only 13 samples have an r. higher than .850. The splits typed on DNA Cw303/304 and Cw70l+02+03/704 are not serologically identified with any sera. Evenly, the new alleles typed on DNA (Cw12/l203/l4/l5/l60l/l602 and 1701) covering all together the 18,1% of frequence in this population are not serologically identified. Two significant differences of frequencies between the data from the XII Workshop,Italian Population, (a) and data from this Northern Italian Population typing (b) have been identified for: Cwl~ 3.0 in (a) and 7.4 in (b) and Cw3~ 3.6 in (a) and 6.0 in (b). The data of the eight alleles typed either at DNA and serologic level are identical.
P277 HLA CW ARMS-PCRTYPING IN THE SARDINIAN POPULAnON
P278
Cantu Licinio, Floris Lauretta, Carcassi Carlo, Deidda Donatella, Alba Francesco, Gessa 5./ Boero Roberta, Mulargia Marina, La Nasa Giorgio, Cirillo Daniela. Cattedra di Genetica Medica, Universita di Cagliari, Cagliari, Italy.
David Comas, Eva Mateu, Francese Calafell. Jaume Bertnnpetit, laboratori d'Antropologia. Facultat de Biologia. Universitat de Barcelona. Diagonal 645. 08028 Barcelona, Catalonia, Spain.
HLA-C alleles have always been the least well characterized of the HLA class I genes and the genetic nature of HLA-C gene products is of increasing interest in the field of immunology for their involvement in their recognition of natural killer cells as well as for their importance in transplantation, disease susceptibility and anthropological studies. In order to identify serologically detectable and undetectable HLA-Cw antigens we used a sequence-specific primer (peR-SSP) system (23 primer
mixes) to detect all sequenced HLA-Cw alleles in 119 random individuals belonging to the Sardinian population. The results are shown in the following
table. Cw allele '01 '02 '0302 '0303 '04 '0501 '0602
Frequency 0.050 0.050 0.004 0.021 0.176 0.184 0.088
Cw allele '07 '08 '1201/2 '1203 '15 '1601 '1602 'POI
Frequency 0.21 0.071 0.008 0.071 0.046 0.008 0.004 0.004
HLA VARJATlON IN TWO POPULATIONS OF THE IBERIAN PENINSULA
The genetic differentiation of the Basques has long been recognized although results are not the same when analyzing allele frequencies or mtDNA sequences. In order to analyze the level of differentiation we ha...e srudied the HLA system comparing Basques with Catalans, a hardly diferentiated European population as shown by classical markers. The two populations are found at the edges of the Pyrenees and previous evidence show a sharp genetic cba.nge. The high polymorphism of the system and 111e high resolution at the DNA level of the typing allow to distinguish between both Iberian Peninsula populations and discuss the genetic level of differentiation as HLA may be both treated as allele frequencies or DNA sequences. One sample of 100 autocthonous individuals from the Basque country (Guipuzkoa) end one sample of 88 autocthcncus individuals from Calalonia (North of Girona) have been typed for HLA Class [ and Class D. High resolution DNA typing has been used in both populations. Samples have been typed for HLA Class I alleles (HLA-A, HLA-B and HLA-C) by an Amplification Refractory Mutation System (ARMS)·PCR assay: thirty-two pairs of primers have been used for HLA-A, twenty-seven for HLA-B and twenty-three for HLA-e. The 12th lntemational Histocompatibility workshop SSO-PeR technique has been used 10 type at a high allelic resolution level the loci of the Class n HLA antigens (HLA-DRB, HLA-DQA, HLA-DQB, HLAl>PA and IRA-DPB) using ll. large number of specific probes. The allele and the haplotype frequencies of HLA Class I and Class II have been compared to the anthropological data collected for the 11th International Histocompatibility Workshop. The large amount of data produced for the 12th International Histocompatibility Workshop will allow to compare the present populations with other populations around the world using the same high resolution DNA techniques.
As was expected we observed a lower level of Cw homozygosity (homozygous
+ heterozygous/blank) by ARMS-peR when compared to serology: 17.64%vs 62.18%.
P279
P280
HLA ALLELE AND HAPLOlYPE FREQUENCIES IN ALGERIANS: RELATEDNESS TO SPANIARDS AND BASQUES.
SURNAME AND HLA GENE FREQUENCIESTO REVEAL RECENT AND ANCIENT MIGRATION
Antonio Arnaiz-Villena, Djamal Benmamar, Nieves Diaz-Campos, Eduardo Gomez-Casado, Miguel Alvarez, Pilar Varela, Jorge Martinez-Laso Department of Immunology, Hospital 12 de Ocrubre, Universidad Complutense, Madrid, Spain.
C.R. Guglielmino 1, A. De Silvestrll , M. Martinetti2, L. Spada3• A. Rossi3, A. Rossi3, and M. Cuccial 1 Dipartimento di Genetica e Microbiologia-Universita di Pavia-Italy 2 Servizio di Immunoematologia e Trasfusione IRCCS Policlinico S. Matteo-Pavia-Italy 3 Servizio Immunoematologico Az. Osp. USSL2-Piacenza-ltaly
The HLA system has been used as a powerful genetic polymorphism in order to identify individuals and populations. Ethnic groups may be characterized byspecificHLA allele frequencies and particular extended HLA haplotypes; also, genetic relationships among these groups may be deduced. In the present study,seroiogyand DNA typinghave been used to detect HLAA, -B, -C, -DR and -DQ alleles in each individual and to calculate characteristic haplotypes in Algerians. These results bave been compared to the previously obtained in other populations, particularly Northern Meciterraneans: genetic distances and their respective dendrograms place Basques and Spaniards closer to Algerians than to other Europeans. Also,
characteristic Basque and/or Spanish haplotypes are found in Algerians, i.e.: A30-B18-Cw5-DR3-DQ2, AI-B57-Cw7.DR7-DQ2. This supports that the Algerian population, mainly its paleo-North African component (Berbers), have a common descent with Basques and Spaniards, probably reflecting a preneolithic relationship between Iberians and paleo-North Africans.
The combined use of HLA genes and surnames reveals genetic barriers, migration and can enlight the historical origin of populations even in a small area. We studied Pavia and Piacenza provinces situated in Lombardy and in Emilia respectively (Northern Italy). Both provinces share a mountainous region. while the remaining part is flat. With principal component analysis, both kind of data show that the mountainous region of the two province are separated from the plain areas and closed one to each other. The separation is even more clear with the HLA data; since HLA genes reflect more ancient features of populations, we can speculate that the mountainous populations of the two provinces share a common ancestor. There is archaeological evidence that the mountainous area was inhabited from prehistorical time till Middle Age by the Ligurians, one of the most ancient Italic population.