MHC-Ⅰ链相关基因A在免疫性疾病中的研究进展

MHC-Ⅰ链相关基因A（MICA）位于人第6号染色体的MHC—Ⅰ类区域中,该基因包含6个外显子,编码的蛋白分子结构域与经典的Ⅰ类分子α链相似,包括胞外结构域（α1、α2与α3）、跨膜区和胞质区,但不含β2微球蛋白。MICA表达于内皮细胞、上皮细胞、单核细胞、成纤维细胞以及大多数上皮性肿瘤细胞表面,但在CD4^＋T细胞、CD8^＋T细胞和CD19^＋细胞不表达。MICA在肾脏等器官移植排斥反应和移植物存活率有重要的相关性,其抗体阳性者预后较差。MICA在肿瘤、自身免疫性疾病、感染性疾病的发生、发展过程中起着独特的作用。     

MICA基因研究进展及与疾病的关系

MICA基因在人类位于6P HLA-Ⅰ类基因区HLA-B位点着丝粒端的46kb左右，其组成、表达和产物与HLA-Ⅰ类基因不同。该基因包含6个外显子，编码的蛋白分子结构域与经典的Ⅰ类分子α链相似，包括3个胞外结构域：α1、α2和α3，加上跨膜区和胞质区，不含β2m。MICA大量表达于上皮细胞和纤维母细胞的表面，在T、B细胞表面不表达，但在大多数上皮性肿瘤细胞表面都有表达。MICA分子与其配体NKG2D受体结合后可以激活NK细胞、Vδ1γδT细胞等，参与天然免疫应答。现已发现MICA与某些肿瘤和免疫相关性疾病（如白塞氏病、1型糖尿病）存在关联，并且在肿瘤和某些疾病的发展中起着重要作用。     

中国湖北汉族HLA—Ⅱ类等痊基因频率的群体调查

调查中国湖北汉族人群ＨＬＡ－Ⅱ类基因频率。方法,用聚合酶链反应／序列特异性引物和聚合酶链反应／限制性片段长度多态性技术,对中国湖北汉族１６８名正常个体进行了ＨＬＡ－ＤＲＢ１（ｎ＝１６８）、ＨＬＡ－ＤＱＢ１（ｎ＝１６０）、ＨＬＡ－ＤＰＢ１（ｎ＝９３）基因的多态性检测。结果共检出３９种ＤＲＢ１、１５种ＤＱＢ１和１７种ＤＰＢ１等位基因型别,等位基因频率较高的分别是：ＤＲＢ１＊０９０１（ｇｅｎｅｆｒｅｑｕ     

Real-time PCR分型法检测苏南地区汉族人群KIR基因多态性

目的利用SYBR Green I Real-time PCR分型方法检测杀伤细胞免疫球蛋白样受体(killer cell immunoglobulin-likereceptor,KIR)基因,探讨苏南地区汉族人群KIR基因的分布特点。方法应用SYBR Green I Real-time PCR法对191名苏南地区汉族非亲缘健康人群进行KIR基因分型。结果 SYBR Green I Real-time PCR法有效地进行了KIR基因分型。已知的16种KIR基因在苏南地区汉族人群均被检出。框架基因2DL4、3DL2、3DL3和假基因3DP1存在于所有受检个体中。最常见的非框架基因为2DL1、2DL3、3DL1、2DS4以及假基因2DP1。共检出33种KIR基因型,最常见的为AA1(39.27%),其次为BX2、BX4和BX8。发现仅在新加坡华人报道的罕见基因型BX331和BX337,及仅在墨西哥人群罕见的基因型BX427。结论苏南汉族人群中检测出已知的16种KIR基因,共发现33种基因型,最常见的为AA1,并见到3个罕见基因型BX331、BX337和BX427。     

广西地区汉族系统性红斑狼疮与HLA-DRB1等位基因相关性研究

系统性红斑狼疮 (SLE)是典型的自身免疫性疾病 ,由于因人种、民族、地理环境等因素不同 ,HLA抗原分布可出现较大的差异 ,其与SLE相关的抗原型与基因型亦存在较大差异。我们应用聚合酶链式反应 序列特异性引物 (PCR SSP)技术 ,对广西汉族SLE的HLA DRB1基因进行分型研究。基金项目 :广西科学基金项目 ( 9912 0 41)作者单位 :广西医科大学第一附属医院皮肤科 (郑文军、梁伶、苏家光、刘栋华 ) ,内分泌科 (罗佐杰 ) ;广西医科大学病理生理学教研室 (潘尚领 )通讯作者 :苏家光 ,5 3 0 0 2 1南宁 ,广西医科大学第一附属医院皮肤科 ,Ema…     

上海地区汉族人群杀伤细胞免疫球蛋白样受体基因多样性及单倍型组合研究

目的 分析上海地区汉族人群杀伤细胞免疫球蛋白样受体(killer Ig-like receptors，KIR)基因多态性及单倍型，为进一步研究不同人群KIRs与某些疾病的相关性提供线索。方法 采用序列特异性引物PCR法对87名无关汉族健康志愿者进行KIR基因低分辨率检测和单倍型分析。结果 (1)共检出Xv、KIRlD在内的18个KIRs基因。3DL3、2DL4、3DL2和3DLl存在于所有个体；较常见为2DL3、Z、2DLl、X；其次为2DS4、1D、2DL5、2DSl、3DSl、2DS5；2DS2、2DL2、2DS3、Xv频率较低。(2)共检出13种单倍型，最常见的是单倍型2。(3)共检出18种基因型．以AJ(2．2)、AF(1．2)型最常见，其次为AH(5，2)、AI(1，5)和AG(1，1)。另外有5种基因型FZl(2，9或6，16)、FZ2(1，16或2，15)、FZ3(6，17)、FZ4(4，13)和FZ5(2，6)，目前在白人中尚未发现。结论 上海地区汉族人群有其独特的KIRs基因频率、单倍型频率和基因型频率分布。     

KIR2DL4基因的多态性与南方汉族白血病的相关性研究CSCD

目的探讨南方汉族人群白血病与KIR2DL4等位基因多态性的相关性。方法提取南方汉族急性淋巴细胞白血病（acute lymphoblastic leukemia,ALL）（n=283）、急性髓系白血病（acutemyeloid[eukemia,AML）（n=227）、慢性髓系白血病（chronic myeloid leukemia,CMI。）（n=80）患者和306名南方汉族随机正常志愿捐血者的DNA基因组,应用本实验室自行设计的引物对KIR2DL4基因的全部8个外显子进行测序分型,用Assign4．7分析软件判定基因型。用SPSS13．0统计软件分别对检出的每个KIR2DIA等位基因、10A型等位基因和9A型等位基因的检出比例分别进行差异显著性分析,并进行P值校准（Pc）。结果ALL、AML、CMI,患者和正常对照均检出5种等位基因：KIR2DIA＊001、＊005、＊006、＊008、＊011。ALL病例组与对照组的KIR2DI．4＊011等位基因检出比例、10A型KIR2DL4等位基因检出比例的差异有统计学意义（KIR2DL4＊011：OR=1．66,P=0．01;10A型K琥2DIA：OR=0．42,P=0．03）,但尸值经过校准后,差异均无统计学意义（Pc≥O．05）。AML、CML病例组分别与对照组检出的各个KIR2DL4等位基因、10A型等位基因和9A型等位基因的检出比例进行比较,组间比较差异均无统计学意义（P〉0．05、Pc〉0．05）。结论南方汉族ALL、AML、CML白血病与KIR2DL4等位基因多态性无显著关联。     

中国南方汉族群体HLA-A2基因亚型分析

ＨＬＡ Ａ2是一种主要ＨＬＡＩ类抗原 ,正式命名的等位基因 (截止 1997年 )达 2 2种[1] ,本文初步调查了中国南方汉族群体ＨＬＡ Ａ2等位基因亚型分布。1　材料与方法1.1　研究对象　 4 0名随机选择、无血缘关系的健康汉族个体来自湖南长沙地区。1.2　主要试剂　ＨＬＡ Ａ2亚型     

中国汉族群体RhD阴性个体D基因外显子多态性研究

目的 研究中国汉族RhD阴性[RhD(-)]个体Rh血型系统D基因8个外显子的构成情况。方法 应用聚合酶链反应－序列特异性引物（PCR－SSP）方法,对50名汉族人RhD(-)个体RHD基因及RHCE基因进行了研究。扩增RHD基因的第2,3,4,5,6,7,9,10外显子和内含子4,以及RHCE基因的第1,2,4,5外显子和内含子4。结果 50名RhD(－）供者的Rh表型为ccdee 22人,ccdEe 3 w ,CE3w ,在表型为ccdee或ccdEe的25人中,D基因8个外显子全部缺失,在表型为Ccdee或CcdEe的25人中,RHD基因则表现出多态性：9人存在全部基因8个外显子,7人缺失全部D基因8个外显子,5人存在D基因第2外显子,3人存在D基因第6外显子,1人存在D基因第2,6,10外显子,结论 中国汉族RhD(-)个体D基因外显子具有多态性,Rh表型为cc的个体D基因8个外显子全部缺失,在表型为Cc的个体中观察到5种D基因多态性,中国汉族人D基因结构不同于高加索人,故在应用RHD基因定型结果时,应慎重对待。     

Human leucocyte antigen but not KIR alleles and haplotypes associated with chronic HCV infection in a Chinese Han population

The host immune system plays a key role in the elimination of infected cells which depend on killer‐cell immunoglobulin‐like receptors (KIR), human leucocyte antigen (HLA) class I molecules and their combinations. To evaluate the roles of HLAclass I, KIR genes and their combination in Chronic hepatitis C virus (HCV) infection (CHC), a total of 301 CHCs and 239 controls in a Chinese Han population were included for HLA and KIR genotyping using next‐generation sequencing and multiplex PCR sequence‐specific priming, respectively. The allele frequency of HLA‐C*08:01 was significantly higher in the CHCs than that of the controls (0.088 vs. 0.040, OR = 2.332, 95%CI: 1.361–3.996, p = 0.022), while the frequencies of B*13:01 (0.032 vs. 0.084, OR = 0.357, 95%CI: 0.204–0.625, p = 0.009) and C*08:04 (0.008 vs. 0.038, OR = 0.214, 95%CI: 0.079–0.581, p = 0.022) were significantly lower in the CHCs. The frequencies of haplotype A*11:01‐C*08:01 were higher in the CHCs (0.058 vs. 0.019, OR = 3.096, 95%CI: 1.486–6.452, p = 0.026), while haplotype B*13:01‐C*03:04 were lower in the CHCs compared to the controls (0.028 vs. 0.071, OR = 0.377, 95%CI: 0.207–0.685, p = 0.012). No association of CHC with KIR genes, genotypes, or haplotypes, as well as HLA/KIR combinations was observed. Our results indicated that HLA‐C*08:01 was a risk factor for CHC, while HLA‐C*08:04 and HLA‐B*13:01 were protective factors against CHC. Haplotypes HLA‐A*11:01‐C*08:01 could increase susceptibility to CHC, while HLA‐B*13:01‐C*03:04 could be protective against CHC in the Chinese Han population.     

Distribution of KIR genes and KIR2DS4 gene variants in two Mexican Mestizo populations

Killer immunoglobulin-like receptors (KIR) are transmembrane proteins that regulate NK and T cell subsets by recognizing HLA-I molecules as ligands. The KIR gene family consists of 16 genes, located at chromosome 19q13.4. KIR gene frequencies vary among populations. In Mexico, HLA and genetic ancestry studies show that Mestizo populations have different genetic backgrounds based on admixture with European, African, and Asian ancestry. This study aimed to evaluate the frequencies of KIR genes and genotypes in Guerrero and Jalisco, two Mexican Mestizo populations located in the south and the west of the country, respectively, and to compare these frequencies with those of other populations. KIR genotyping was performed by SSP-PCR. We observed that KIR gene frequencies were similar in both populations. There were 24 genotypes observed in Guerrero, 38 genotypes observed in Jalisco, 15 genotypes shared in both populations and 32 genotypes unique to one population or the other. In 10 individuals, nine novel genotypes were identified. KIR2DS4 gene variants showed significant differences: The KIR2DS4full gene was more common in Guerrero (p < 0.0001), and the KIR2DS4del variant was more common in Jalisco (p < 0.05). Differences in KIR2DS4 gene variants and genotypic profiles could be influenced by the genetic admixture in both regions.     

KIR genes polymorphism in Argentinean Caucasoid and Amerindian populations

In natural killer cells, killer immunoglobulin-like receptors (KIRs) loci code for either inhibitory or activating receptors, and according to the number of genes present in each individual, it is possible to identify a high rate of polymorphism in the populations. We performed KIR typing by polymerase chain reaction-sequence-specific oligonucleotide probing in 402 Argentinean Caucasoid and in two Amerindian populations (101 Wichis and 54 Chiriguanos) from the North of Argentina. KIR2DL4, KIR3DL2, KIR3DL3 and KIR3DP1 were always present, whereas the frequencies of KIR2DL1, KIR2DL3, KIR2DS4, KIR3DL1 and KIR2DP1 ranged between 84% and 96%. The frequencies of KIR2DS2, KIR2DL2, KIR2DL5, KIR2DS5, KIR2DS1 and KIR3DS1 ranged between 41% and 62%. The KIR2DS3 with a frequency of 29% in Argentinean Caucasoid population was present at a very low frequency in Amerindian populations. Haplotype segregation studies performed in 10 Wichi families showed the presence of only three haplotypes: A, B5 and B1. The Amerindian populations showed several similarities to Asian but not to Caucasoid populations with regard to the frequency of KIR2DS3, full-length KIR2DS4 gene and KIR2DL4 alleles.     

Diversity distributions of killer cell immunoglobulin-like receptor genes and their ligands in the Chinese Shaanxi Han population

In the present study, 17 killer cell immunoglobulin-like receptors (KIR) genes and KIR ligands (human leukocyte antigen [HLA] -A and -B) were detected by using a polymerase chain reaction-sequence-specific primer (PCR-SSP) method in 104 unrelated healthy Han individuals living in Shaanxi province, China. The observed carrier frequencies of the 12 KIR genes ranged from 0.14 to 0.96. KIR2DL4, 3DL2, 3DL3, 2DP1 and 3DP1 were found to be present in every individual. A total of 51 different KIR gene profiles were identified, in which 11 gene profiles exclusively belonged to the study population. Neighbor-joining phylogenetic tree between the studing population and its neighboring ethnic groups was constructed using the observed carrier frequencies of 13 KIR loci. The phylogenetic tree shows that the Shaanxi Han population, Han populations in different regions, Yi, Japanese, and Koreans were in the same cluster. KIR/HLA relationships show that KIR3DS1(-)/3DL1(+)/Bw4(+) was the most common association in the population. In conclusion, the present study findings reveal the high polymorphism of KIRs in the Shaanxi Han population, demonstrate the KIR/HLA association in the study population, and enrich the KIR and HLA gene resources. The obtained KIR data will further the understanding of genetic relationships among populations in different geographic areas, and assist in answering questions regarding KIR/HLA relationships.     

Association of KIR Genotypes and Haplotypes with Susceptibility to Chronic Hepatitis B Virus Infection in Chinese Han Population

Killer immunoglobulin-like receptor （KIR） genes can regulate the activation of NK and T cells upon interaction with HLA class I molecules. Hepatitis B virus （HBV） infection has been regarded as a multi-factorial disorder disease. Previous studies revealed that KIRs were involved in HCV and HIV infection or clearance. The aim of this study was to explore the possibility of the inheritance of KIR genotypes and haplotypes as a candidate for susceptibility to persistent HBV infection or HBV clearance. The sequence specific primer polymerase chain reaction （SSP-PCR） was employed to identify the KIR genes and pseudogenes in 150 chronic hepatitis B （CHB） patients, 251 spontaneously recovered （SR） controls, and 412 healthy controls. The frequencies of genotype G7 M, FZ1 increased in CHB patients compared with healthy control subjects. The frequency of genotype AH was higher in SR controls than that in both CHB patients and healthy controls. The carriage frequencies of genotype G and AH were higher; while, the frequencies of AF and AJ were lower in SR controls than those in healthy control subjects. The frequency of A haplotype was lower, whereas, the frequency of B haplotype was higher in CHB patients and SR controls than those in healthy controls. In healthy controls, haplotype 4 was found lower compared with that in CHB patients and SR controls and the frequency of haplotype 5 was higher in SR controls than that in other two groups. Based on these findings, it seems that the genotypes M and FZ1 are HBV susceptive genotypes; AH, on the other hand, may be protective genotypes that facilitate the clearance of HBV. It appears that the haplotype 4 is HBV susceptive haplotype, whereas, haplotype 5 may be the protective haplotype that facilitates the clearance of HBV. Cellular ＆ Molecular Immunology. 2008;5（6）：457-463.     

上海地区汉族人群KIR2DL5基因多态性及单元型分析

KIR2DL5基因是杀伤细胞免疫球蛋白样受体 (KIR )单元型B的特征性标志 ,至少有 4个变异体 ,其中 2DL5A 0 0 1和2DL5B 0 0 3是表达型 ,统称为KIR2DL5 1;2DL5B 0 0 2和 2DL5B 0 0 4是不表达型 ,统称为KIR2DL5 2。本文用序列特异性引物PCR法 (PCR SSP )分析了KIR2DL5表达型KIR2DL5 1和不表达型KIR2DL5 2在上海地区正常汉族人群的分布及其在KIR单元型的组成。结果发现 (1)在 87名健康汉族人和 14个家族中表达型KIR2DL5 1和不表达型KIR2DL5 2基因频率分别为0 1977和 0 0 4 11;(2 )KIR2DL5基因变异体在不同单元型中分布不同 ,有 5种不同组合 ,其中单元型 1、 2、 3、 4、 11、 12、15和 16不含KIR2DL5及其变异体 ;单元型 5和 14只含有KIR2DL5 1;单元型 13和 17只含有KIR2DL5 2 ;单元型 6含有单一KIR2DL5 1或二者兼有 ;单元型 8和 9同时含有KIR2DL5 1和KIR2DL5 2。     

Comparison of KIR gene content profiles revealed a difference between northern and southern Persians in the distribution of KIR2DS5 and its linked loci

Killer cell immunoglobulin-like receptors (KIR) are the key receptors of human natural killer (NK) cells that mount an early immune response against infection and tumors. The number and type of KIR genes are substantially variable between individuals and populations. Recently we reported KIR gene content diversity in a Persian population living in the southern province of Fars, which is comparable to that of European Caucasians. These results are consistent with the ethnic ancestry and affinity between Persians and Caucasians. Herein we analyzed another Persian population living in the northern province of Tehran and discovered an unexpected increase in the distribution of KIR2DS5 and its linked loci KIR3DS1, -2DS1, and -2DL5 in northern Persians compared with that reported in the southern Persian population. Although the geographic barriers may have limited the gene flow, the impact of the local environment on the natural selection of KIR2DS5 and its linked loci in the northern Persians cannot be completely ruled out. The difference in northern and southern populations in activating KIR gene content creates an appealing hypothesis that KIR2DS5-enriched northern Persians are more resistant to developing clinical conditions demonstrated to be associated with KIR2DS5, such as psoriasis vulgaris, endometriosis, ankylosing spondylitis, and acute rejection of kidney grafts, compared with those living in the southern part of the country.     

A genome-wide association study combining pathway analysis for typical sporadic amyotrophic lateral sclerosis in Chinese Han populations

Sporadic amyotrophic lateral sclerosis (sALS) is a severe neurodegenerative disease that causes progressive motor neuron death. Although the etiology of sALS remains unknown, genetic variants are thought to predispose individuals to the disease. Several recent genome-wide association studies have identified a number of loci that increase sALS susceptibility, but these only explain a small proportion of the disease. To extend the current genetic evidence and to identify novel candidates of sALS, we performed a pooling genome-wide association study by 859,311 autosomal single-nucleotide polymorphisms of IlluminaHumanOmniZhongHua-8 combining pathway analysis in 250 typical sALS cases precluding age, clinical course, and phenotype interference and 250 control subjects from Chinese Han populations (CHP). The results revealed that 8 novel loci of 1p34.3, 3p21.1, 3p22.2, 10p15.2, 22q12.1, 3q13.11, 11q25, 12q24.33, and 5 previously reported loci of CNTN4 (kgp11325216), ATXN1 (kgp8327591), C9orf72 (kgp6016770), ITPR2 (kgp3041552), and SOD1 (kgp10760302) were associated with sALS from CHP. Furthermore, the pathway analysis based on the Gene Set Analysis Toolkit V2 showed that 10 top pathways were strongly associated with sALS from CHP, and among them, the 7 most potentially candidate pathways were phosphatidylinositol signaling system, Wnt signaling pathway, axon guidance, MAPK signaling pathway, neurotrophin signaling pathway, arachidonic acid metabolism, and T-cell receptor signaling pathway, a total of 39 significantly associate genes in 7 candidate pathways was suggested to involve in the pathogenesis of sALS from CHP. In conclusion, our results revealed several new loci and pathways related to sALS from CHP and extend the association evidence for partial loci, genes, and pathways, which were previously identified in other populations. Thus, our data provided new clues for exploring the pathogenesis of sALS.     

Allelic diversity of KIR3DL1/3DS1 in a southern Chinese population

The inhibitory KIR3DL1 and the activating KIR3DS1 segregate as alleles of the same locus. KIR3DL1 is highly diversified at the allele level and KIR3DL1 alleles exhibit varied levels of expression and ligand binding affinity resulting in varied degrees of NK cell inhibition. Previous studies have shown that the KIR3DL1/3DS1 polymorphism associated with viral infection, cancer and transplantation. However, little is known about the population distribution of KIR3DL1/3DS1 alleles in Chinese. The present study examined allelic diversity of KIR3DL1/3DS1 in a southern Chinese population (N = 306) using PCR-SSP and sequencing based typing. The presence of KIR3DL1 and KIR3DS1 were detected in 97.1% and 34.0% of the tested individuals respectively. A total of 10 KIR3DL1 alleles (including 2 novel ones) and 6 KIR3DS1 alleles (including 5 novel ones) were identified. Common KIR3DL1 alleles (>10%) were KIR3DL1*01502 (74.8%), KIR3DL1*00501 (23.9%) and KIR3DL1*00701 (15.7%). KIR3DS1*01301 was the predominant KIR3DS1 allele with other KIR3DS1 alleles only sporadically observed. The knowledge of the allelic polymorphism of KIR3DL1/3DS1 may help to better understand the role played by KIR3DL1/3DS1 in associated diseases and clinical transplantation in southern Chinese.