The Gene Polymorphism And Expression Of Key Receptors On NK Cell And The Ligands In Autoimmune Diabetes

Objective:To characterize the gene polymorphism, haplotypes, and genotypes of killer cell immunoglobin-like receptor (KIR), the gene polymorphism of HLA-C and the KIR/HLA-C gene compatibility in Hunan Han population. The obtained results will be used as basic data for further study in its linkage with disease and transplantation immunity.Design:Cross-sectional study.Methods:The method of polymerase chain reaction-sequence specific primer (PCR-SSP) was used to detect KIR and HLA-C genotypes in 429 healthy individuals of Hunan Han population. Then the distribution and characteristics of the haplotypes, genotypes of KIR genes and the KIR/HLA-C gene compatibility were analyzed.Results:(1)16 KIR genes were detected in Hunan Han population, of which the frequencies of framework genes KIR2DL4,3DL2,3DL3, and 3DP1 were 100%. KIR2DL1,2DL3,3DL1,2DS4 and 2DP1 genes were common with frequencies more than 90%, while the frequencies of KIR2DS1,2DL5,3DS1,2DL2,2DS2,2DS3,2DS5 were lower than 40%. (2) In Hunan Han population, the distribution of KIR genes frequencies were similar to Japanese, but the frequencies of KIR2DL2, 2DS2,2DS3 and 2DL5 were lower than Caucasian, the frequencies of KIR2DL3 was higher than Caucasian. (3) The KIR genes gave priority to A haplotype in Hunan Han population, the frequencies of A and B haplotype were 69.6% and 30.4% with a ratio of 2:1. (4) 49 KIR genotypes were found in the Hunan Han population,4 of which had not been reported. AA was the main genotype of KIR gene (44.8%) (5) HLA-C1 was dominant in Hunan Han population, with a frequency of 98.1%. (6)HLA-C1(+)/HLA-C2(-), KIR2DL1(+)/2DL2(-)/2DL3(+)/2DS1 (-)/2DS2(-) was the most common KIR2D/HLA-C gene compatibility in Hunan Han population (23.9%).Conclusion:Distrubution of KIR genes, HLA-C genes and KIR/ HLA-C gene compatibility show rich polymorphism, with distinctive KIR genotypes in Hunan Han population. Objective:To investigate the gene polymorphisms of Killer cell immunoglobin-like receptor (KIR) and HLA-C in autoimmune diabetes.Design:Cross-sectional and case control study.Methods:The method of polymerase chain reaction-sequence specific primer (PCR-SSP) was used to detect the gene polymorphisms of KIR and HLA-C in 387 T1ADM patients [180 acute onset autoimmune diabetes(T1ADM) patients and 207 latent autoimmune diabetes in adults (LADA)patients] and 429 healthy controls (199 control A group and 230 control B group) who matched for sex, age in Hunan Han population.Results:(1)Compared with the corresponding controls, the frequencies of KIR2DL1 (98.9% vs.92.0%,OR=7.781, P<0.01), 3DL1(94.5% vs.86.4%, OR=2.669, P<0.01) and 2DS4 (83.9% vs. 70.9%, OR=2.142, P<0.01) increased in T1ADM patients; the frequency of KIR2DL3 (99.5% vs.96.1%, OR= 8.389, P<0.05) increased in LADA patients; The genetic susceptibility of KIR genes in Hunan Han patients with autoimmune diabetes was different from other races. (2) The total number of activating KIR genes in T1ADM patients was higher than their controls (2.02 vs.1.78, P<0.05). (3) The frequencies of A and B haplotype of KIR gene were 69.7% and 30.3% in T1ADM patients, 75.4% and 24.6% in LADA patients. The ratio between A and B haplotype were about 2:1 and 3:1 respectively. (4)There are no differences in frequencies of HLA-C genes (HLA-C1, HLA-C2) between T1ADM or LADA patients and their corresponding controls. (5) The frequencies of KIR2DL1(-)/HLA-C2(-) (0.6% vs.6.0%, OR= 0.087, P< 0.01) in T1ADM patients and KIR2DL3(-)/HLA-C1(+) (0.5% vs.3.9%, OR=0.119,P<0.05) in LADA patients were lower than their corres-ponding controls. (6) Compared with the controls, the frequency of HLA-C1(+)/HLA-C2(-),KIR2DL1(+)/2DL2(-)/2DL3(+)/2DS1(+)/2DS2(-) (24.4% vs.15.6%, OR=1.753, P<0.05) increased in T1ADM patients.Conclusion:The KIR gene polymorphism and KIR/HLA-C gene compatibility are associated with the susceptivity of autoimmune diabetes. Objective:To analyze the interactive effects between KIR gene and HLA-DQ or MICA gene in acute onset autoimmune diabetes (T1ADM).Design:Cross-sectional and case control study.Methods:All DNA samples of 180 T1ADM patients and 199 healthy controls of Hunan Han were typed for the polymorphisms of HLA-DQ and MICA genes with PCR sequencing-base typing (PCR-SBT) method. After analyzed the susceptible alleles, haplotypes or genotypes of HLA-DQ and MICA gene, Svejgaard& Ryder test was performed to investigate the interactive effects between susceptible KIR gene (KIR 2DL1,3DL1 or 2DS4) and susceptible HLA-DQ haplotypes or suscep-tible MICA alleles.Results:(1) The susceptible alleles of HLA-DQ gene in Hunan Han were DQA1*03, DQA1*0501/0503, DQB1* 0201, DQB1*0303and DQB1*0401. The protective alleles were DQA1*0101/0104, DQA1 *0102, DQA1* 0103, DQ A1*0601, DQB1* 0301, DQB1*0502, DQB1 *0601 and DQB1*0602. The susceptible haplotypes were DQA1*03-DQB1*0303, DQA1*03-DQB1*0401 and DQA1*05-DQB1*0201. The protective haplotypes were DQA1*0601-DQB1*0301, DQA1*0102-DQB1*0601 and DQA1*0102-DQB1*0602. (2) The susceptible allele of MICA in Hunan Han was MICA9 and the protective was MICA5.1. The susceptible genotypes were MICA4/9 and MICA9/9. (3) The risk of T1ADM increased in ones who carried susceptible KIR gene (KIR2DL1, 3DL1 or 2DS4) and HLA-DQ haplotypes or MICA9 (OR=1.077-21.111). (4) Susceptible HLA-DQ haplotypes or MICA9 increased the risk of TIADM in ones who carried KIR2DL1 or KIR3DL1 (OR= 2.929-7.222). (5) The susceptible effects of HLA-DQ haplotypes was stronger than that of KIR2DS4 (OR=3.150). (6) KIR3DL1 or KIR2DS4 increased the risk of T1ADM in ones who without MICA9 (OR= 3.792 or 2.424).Conclusion:KIR genes have a synergistic action on the susceptible effects of HLA-DQ or MICA gene in T1ADM. Objective:To observe the variances of NK cells and its surface receptors in autoimmune diabetes patients, and to explore the role of NK cells in the pathogenesis of T1ADM, in order to offer some ideas and means for the forecast, monitor and therapy of autoimmune diabetes.Design:Cross-sectional and case control study.Methods:53 T1ADM patients (35T1ADM patients and 18 LADA patients) from the patients admitted to the department of endocrinology in our hospital,28 healthy controls from health-check screening were included in the study. The percentages of T cell subsets and NK cells and the expression of surface receptors (KIR and NKG2D) of NK cell were detected by flow cytometry. The activities of NK cells before and after incubated with IL-2 were detected by the assay of MTT. We compared and analyzed the variances of NK cells and its surface receptor in autoimmune patients.Results:(1) Compared with the corresponding controls, the propor-tion of CD3+ T cells increased (70.5% vs.64.6%, P<0.05), while the proportion of NK cell decreased (13.8% vs.20.1%, P<0.01) in T1ADM patients.(2)The NK cell activity of T1ADM patients and LADA patients was lower than the corresponding controls (30.9% vs.47.1%, P<0.01; 39.8% vs.49.2%, P<0.05). The NK cell activity of T1ADM patients (30.9% vs.39.8%, P<0.01), whose recent-onset subgroup patients (33.2% vs.40.9%, P<0.05) and long course subgroup patients (28.1% vs. 38.6%, P<0.05) were lower than the corresponding LADA patients; (3) After incubated with IL-2, the NK cell activities of T1ADM patients and LADA patients were enhanced (43.2% vs.30.9%,47.4% vs.39.8%, P< 0.05). The growth of NK cell activity in T1ADM patients and its recent-onset subgroup patients were higher than the corresponding LADA patients(9.9% vs.7.4%, P<0.05,12.9% vs.8.0%, P<0.05). (4) Compared with the control A group, the expression of KIR2DL1 (10.2% vs.6.2%, P <0.05), NKG2D(96.9% vs.95.2%, P<0.05) increased in T1ADM patients.Conclusion:(1) The proportion of CD3+ T cells increase, while the proportion and the activity of NK cells decrease in the peripheral blood of autoimmune diabetes patients. The NK cell-mediated immunity abnormality of T1ADM patients is more severe than LADA patients. (2)IL-2 can improve the NK cell activity of autoimmune diabetes patients, and the effect is more obvious in the early stage of T1ADM than LADA. (3)The expression of the surface receptors of NK cell (KIR2DL1 andNKG2D) increase in T1ADM, which may play a role in the patho-genesis of T1ADM.