Zhejiang Han Population And Killer Cell Immunoglobulin-like Receptor Genetic Polymorphism Research

Killer cell immunoglobulin-like receptors (KIRs) on natural killer cells recognize groups of HLA class I alleles. Seventeen KIR genes have been identified at present, and two kinds of KIR haplotypes (group A and B) have been described based on their gene contents. Immunogenetic analysis of different ethnic populations shows significant differences in terms of the distribution of group A and B haplotypes. Here, genomic DNA from 104 healthy unrelated Chinese Han individuals was typed for the presence or absence of KIR genes. All 17 KIR genes were observed in the population, and framework genes 3DL3, 3DP1, 2DL4, and 3DL2 were present in all individuals. Twentysix different genotypes were found, four of which could not be assigned to haplotypes according to the model of Hsu et al.. Group A haplotypes outnumbered group B haplotypes in frequency by approximately 3:1, with individuals having two group A haplotypes accounting for 58.7%. Analysis indicated that some pairs of KIR genes showed remarkable linkage disequilibrium. Our data demonstrated that the Chinese Han population is distinct in KIR gene frequencies and putative KIR haplotypes in comparison to some other populations.We established a PCR-SSP method to detect all KIR genes. 19 pairs of primers were designed to detect the different KIR genes, with human growth hormone specific primer added as control, and the KIR PCR-SSP Kit was used for confirmation. The PCR-SSP method we established could detect all KIR genes, and the PCR products show perfect specificity. The genotypes observed had reach 35, of which AJ(2, 2) was the most popular one. The genotype that had not been observed in Caucasians had reach 21. The genotypes that could not be assign to haplotypes had reach 7, with NN16 been observed 3 times.We established polymerase chain reaction sequence based typing (PCR-SSOP) procedure to identify alleles of KIR3DL2 gene. The method was designed around the specific amplification of exon 3 to exon 4 and exon 8 to exon 9 of the KIR3DL2 gene, and 21 digoxigenin-labelled probes were used for the SSOP analysis. Genomic DNA from 104 healthy unrelated Chinese Han individuals was typed for KIR3DL2 alleles.Each sample was assigned to the putative allele combination according to the probe profiles of all KIR3DL2 alleles. We observed 18 different genotypes and 8 KIR3DL2 alleles in the population, with KIR3DL2*002 having the highest frequency, 0.558, and confirmed a new KIR3DL2 allele. Our data demonstrated that the established PCR-SSOP methods for KIR3DL2 allele typing were reliable, and Chinese Han population is distinct in KIR3DL2 allele frequencies.We established polymerase chain reaction sequence based typing (PCR-SSOP) procedure to identify alleles of KIR2DL3 gene. The method was designed around the specific amplification of exon 4 to exon 5, exon 6 and exon 7 to exon 9 of. the KIR2DL3 gene, and 13 digoxigenin-labelled probes were used for the SSOP analysis. Genomic DNA from 96 healthy unrelated Chinese Han individuals was typed for KIR2DL3 alleles. Each sample was assigned to the putative allele combination according to the probe profiles of all KIR2DL3 alleles. 95 of the 96 samples were KIR2DL3 gene positive. 6 genotypes were observed, and two alleles were confirmed, which were KIR2DL3*001 and KIR2DL3*002, with KIR2DL3*001 having the higher frequency, 0.78. One sample have two copies of KIR2DL3 gene and one KIR2DL2 gene.Polymerase chain reaction sequence-based typing (PCR-SBT) procedures identifying alleles of the KIR2DL4 gene have been established. The method was designed around the specific amplification of exon 3 to exon 5 and exon 7 to exon 9 of the KIR2DL4 gene and produce discrimination of KIR2DL4 alleles. Genomic DNAs from 83 healthy unrelated Chinese Han individuals were typed for KIR2DL4 alleles by this method. Each sample was assigned to the putative KIR2DL4 allele combination according to the nucleotide polymorphism profiles of all KIR2DL4 alleles. Twenty-one different genotypes and seven KIR2DL4 alleles were observed in the population, with KIR2DL4*00102 having the highest frequency, 0.5. Five individuals bear a recombinant allele KIR3DP*004 that associated with three putative KIR2DL4 alleles. Our data demonstrated that the established PCR-SBT method for KIR2DL4 allele typing was reliable, and Chinese Han population is distinct in KJR2DL4 allele frequencies in comparison to some other populations.Polymerase chain reaction sequence-based typing (PCR-SBT) procedures identifying alleles of the KIR2DS4 gene have been established. The method was designed around the specific amplification of exon 4 to exon 5 of the KIR2DS4 gene and produce discrimination of KIR2DS4 alleles. Genomic DNAs from 105 healthy unrelated Chinese Han individuals were typed for KIR2DS4 alleles by this method. Each sample was assigned to the putative KIR2DS4 allele combination according to the nucleotide polymorphism profiles of all KIR2DL4 alleles. 11 different genotypes and 4 KIR2DL4 alleles were observed in the population, with KIR2DS4*00101 having the highest frequency, 0.576. The newly defined KIR2DS4 allele KIR2DS4*007 also have a relative high frequency, 0.114.