engleski

Microsatellite markers in association with HLA-B alleles in the Croatian population

HLA region contains a large amount of microsatellite loci which can serve as highly informative polymorphic markers. We have investigated polymorphism of two microsatellite loci (D6S265 and D6S273), HLA-B locus and their associations. A panel of 156 healthy, unrelated individuals from our capital city, Zagreb was analysed for 2 of the above microsatellites and HLA-B alleles. DNA was isolated from lymphocites using standard salting out method. Molecular typing of HLA-B alleles was performed using Reverse Line Strip method according to 13thInternational Histocompatibility Workshop protocols. Microsatellites were analysed using electrophoresis on polyacrilamide gel (ALFexpress, Pharmacia-Amersham). Their alleles were determined using Fragment Manager software. Our findings showed that the most frequent alleles at D6S265 were: 130bp (35.1%), 134bp (25.7%) and 138bp (15.7%). At D6S273 locus three most present alleles were: 134bp (37.7%), 136bp (27.9%) and 132bp (17.8%). Results gained for our population concerning the frequencies of alleles for a given locus showed that there is no significant difference between our and other populations so far tested. Analysis of HLA-B polymorphism showed that three most frequent alleles were: B*5101 (11.9%), B*1801 (11.2%) B*0702 (9.9%). Haplotypic associations analyses of these 3 most frequent HLA-B alleles with D6S273 alleles showed that B*1801 allele was always associated with D6S273 130bp allele, but the other two HLA-B alleles did not show such exclusive associations. Allele B*5101 occurred with 7 different D6S273 alleles, out of which association with allele 130bp was most frequent (43.2%). Similar findings were observed for B*0702 allele, which associated with allele 132bp in most cases (29.0%). In summary, this study will have important consequences for association and linkage studies using polymorphism within HLA region. These results should provide a useful reference for further anthropological studies, as well as for investigation of the role of microsatellites in transplantation.

D6S265; D6S273; HLA-B; haplotypic associations; Croatian population

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