Genes Polymorphism Study Of KIR And HLA Ligands In Hebei Han Population

Objective: Killer cell immunoglobulin-like receptor(KIR)is a group of protein receptors which expressed on the surface of natural killer cells(NK cell)and certain T cell,regulate NK cell activity through specific binding the human leukocyte antigen(HLA)-I molecules.Studies have shown that playing the role of KIR molecules specifically recognize its target cells HLA-I molecules,participat in the occurrence of graft-versus-host disease(GVHD)and affect the effect of graft-versus-leukemia(GVL),resulting in the difference of the effect of organ transplantation.Therefore,accurate detection of KIR molecules and HLA-I molecules are identical consistency,have very important significance for clinical transplantation and prognostic evaluation.According to the structure of the protein,KIR molecules are divided into inhibitory KIR receptor and activated KIR receptor.Since activated KIR receptor transduced activating signal is weak,even undetectable,therefore,NK cell mainly through inhibitory KIR receptor specific binding to the MHC-? molecules to conduct inhibitory signal to prevent the killing effect of their own cellsKIR and HLA genes are highly polymorphic and their characteristic of independent genetic,and specificity recognition of them,resulting in different receptor/ligand combinations among individuals.At present,There is no systematic study of KIR gene in Hebei area,and the genetic background in Hebei Han population has its specificity.The current domestic and foreign research found that different genetic background and race traceability makes the KIR/HLA-I combination of different characteristics.The purpose of this study is to research of KIR and HLA ligand gene polymorphism in Hebei Han population applying the PCR-SSP method,and statistics of its gene frequency.Method: 1 Preparation of genomic DNASamples come from a random sample of 325 Chinese hematopoietic stem cell donors database in Hebei depots of the Han nationality healthy unrelated volunteers(perfect HLA typing data).Using the TIANGEN blood DNA purification to prepare the genomic DNA.The extracted DNA should meet OD260 / OD280 ratio between 1.7-1.9,the concentration of 30 ng / ?l or so.2 GenotypingThe detecting of KIR genes adopts polymerase chain reaction-sequence specific primers(PCR-SSP).Log in Chinese bone marrow bank website and lookup the genotyping results of HLA-A,HLA-B.3 PCR product was analyzed by gel electrophoresisTransfer the PCR products to agarose gel electrophoresis,observe the results in the UV / gel imaging system,shoot imaging,and interpret the genotyping results.4 Statistical analysisGene frequency of all KIR genes and HLA genes is calculated by direct counting method.KIR and HLA gene frequencies of 325 Han population in Hebei are compared with other parts of the Han population and foreign populations in other regions by ?2 testResults: 1 The gene frequency of 16 knowm KIR genotypes obtained from 325 hematopoietic stem cell donors in Hebei Han population as follows:99.38%for KIR2DL1,18.77%for KIR2DL2,99.38%forKIR2DL3,100.00%for KIR2DL4,44.00%for KIR2DL5,94.15%for KIR3DL1,100.00%for KIR3DL2,100.00% for KIR3DL3,34.15%for KIR2DS1,17.85%for KIR2DS2,18.46%for KIR2 D S3,93.23%for KIR2DS4,23.08%for KIR2DS5,37.85%for KIR3DS1,99.38% for KIR2DP1,100.00%for KIR3DP1.2 The test result of HLA-A Bw4?B Bw4,HLA-A?B alleles corresp-onding to,obtained from 325 hematopoietic stem cell donors in Hebei Han population as follows:0.15%for HLA-A* 2301;14.31%for HLA-A* 2402,0.15%for HLA-A* 2403,0.31%for HLA-A* 2420;1.69%for HLA-A* 3201.3.85%for HLA-B* 1301,6.31%for HLA-B* 1302,0.15%for HLA-B* 13VJ;0.46%for HLA-B* 1517;0.15%for HLA-B* 2702,0.62%for HLA-B* 2704,0.92%for HLA-B* 2705,0.15%for HLA-B* 2707,0.15%for HLA-B* 2736,0.15%for HLA-B*27YNER;2.15%for HLA-B* 3701,0.46%for HLA-B* 37AD;0.77%for HLA-B* 3801,2.00%for HLA-B* 3802,0.62%for HLA-B* 38 XV,0.15%for HLA-B* 38 ACNUA,0.15%for HLA-B* 38ACNUB;1.85%for HLA-B* 4402,3.08%for HLA-B* 4403;5.23%for HLA-B* 5201,0.31%for HLA-B* 52DFT;0.92%for HLA-B* 5701;2.31%for HLA-B* 5801,0.15%for HLA-B* 58 VE.3 KIR/Bw4 pairing statistics of 325 hematopoietic stem cell donors in Hebei Han population as follow:24.62%for no KIR/HLA Bw4,46.77%for KIR3DL1/HLA Bw4,4.62%for KIR3DS1/HLA Bw4,24.00%for both KIR3 DL1/HLA Bw4 and KIR3DS1/HLA Bw4.Conclusion:1 In this study,statistical analysis of the KIR gene distribution obtained from 325 hematopoietic stem cell donors in Hebei Han population,found that framework 2DL4,3DL2,3DL3,3DP1 exist in almost all individuals,the high frequency of 2DL1,2DL3,3DL1,2DS4,2DP1 were> 93.00%,while the frequency of 2DL2,2DS2,2DS3,2DS5 are lower.2 The distribution of inhibitory KIR/HLA in Hebei Han population is more extensive.This is basically in line with the distribution characteristics of Chinese Han population,but also has its own characteristics.3 In this study,we master first-hand information of KIR gene distribution and KIR/HLA pairing in Hebei Han population,laying the foundation for further exploration of relationship between KIR/HLA and diseases.