吉林省汉族154例正常人HLA-Cw及KIR基因型频率的分析

目的:了解中国人HLA-Cw与KIR的识别方式,为今后研究其在疾病中的作用提供理论基础.方法:对154例吉林汉族无偿献血者,以PCR-SSP技术进行KIR和HLA-Cw基因分型.根据HLA-Cw与KIR识别方式,分析个体HLA-Cw与相应激活性或抑制性KIR受体的识别情况.结果:吉林汉族人群中,HLA-C2Lys80+ 2DL1的组合频率最高为27.27％,其次分别HLA-C1Asn80+2DL2/2DL3为68.83％,2DS2+HLA-C1Asn80为9.74％,2DS1+HLA-C2Lys80为9.74％,分别有72.08％和21.43％盼观察对象表达KIR2DL1、KIR2DS1而无相应HLA-Cw表达;21.43％的个体表达KIR2DS1而不表达HLA-Cwlys-KIR2DL1配对.2.60％的个体表达KIR2DS2而不表达HLA-Cwasn-KIR2DL2/L3配对.结论:在吉林汉族人群中,抑制性HLA-Cw-KIR受配体对表达高于活化性受配体对,约20％的个体单独表达激活性HLA-Cw-KIR受配体对.     

江苏汉族人群杀伤细胞免疫球蛋白样受体及其特异性配体HLA分析

目的:分析杀伤细胞免疫球蛋白样受体(KIR)及其特异性配体HLAⅠ类分子在江苏地区汉族人群的分布特点。方法:分别利用real-time PCR法和PCR-SSP方法对173例无血缘关系的江苏汉族健康人群进行KIR和HLA-Cw,HLA-Bw4分型,并分析KIR/HLA配对组合的类型和数量。结果:江苏汉族人群中,>92%的个体同时表达四种抑制性KIR(iKIR)(2DL1,2DL2/3,3DL1,3DL2)。2DL2/HLA-C1,2DL3/HLA-C1,2DL1/HLA-C2,3DL1/Bw4的频率分别依次分别为0.243,0.971,0.457,0.590;2DS1/HLA-C2,2DS2/HLA-C1的频率为0.162,0.231。54.3%的个体只表达2DL1而不表达相应的配体HLA-C2,32.9%的个体只表达3DL1而缺乏配体HLA-Bw4,另有5.8%的个体只表达HLA-Bw4而不表达3DL1。27.7%的个体同时表达3种iKIR/HLA,26%的个体同时表达两种iKIR/HLA,25.4%的个体只遗传一个iKIR/HLA配对,未发现3种iKIR/HLA同时缺失的个体。结论:在江苏汉族人群中,存在KIR与HLA表型分离现象,抑制性KIR/HLA配对表达高于刺激性KIR/HLA配对,约1/4个体只表达单个iKIR/HLA配对。     

广东汉族79例中iKIR及其HLA配体表达的初步分析

目的　探索杀伤细胞免疫球蛋白样抑制性受体 (iKIR)及其人白细胞抗原 (HLA)配体在中国广东汉族人群中的分布。方法　对 79例广东汉族人 ,采用引物序列特异性扩增法检测其iKIR表型 ,HLA A、B、Cw表型。HLA A、B分型采用BiotestHLA分型 (SSP)试剂盒 ,HLA C分型采用半量全自动PCR SSO分型。结果　KIR2DL1、2DL2、2DL3、2DL4、2DL5、3DL1、3DL2、3DL3的表型频率分别为0 .87、0 .13、0 .5 2、1、0 .18、0 .94、1和 0 .99。个体 86 .1%表达 1种以上iKIR HLA配对 ,其中 3DL1 HLA Bw4配对表型频率为 6 9.6 % ,2DL2 C2为 4 3.0 % ,2DL1 C1为 16 .5 %。由于不表达HLA A3,该组人群中无 3DL2 HLA A3受体配体对。 13.9%人群缺乏iKIR HLA配对。结论　约 7/ 8的广东汉族个体表达1种以上iKIR HLA配对 ,以 3DL1 HLA Bw4配对为主 ;约 1/ 8个体缺乏已知的iKIR HLA配对。     

中国人群HLA-Cw、KIR2D受体基因多态性分析

目的 分析中国南、北方两个汉族人群、一个蒙古族人群的人类白细胞抗原-Cw(human leucocyte antigen-Cw,HLA-Cw)遗传多态性;进一步分析南、北方汉族人群杀伤细胞免疫球蛋白样受体2D(killer immunloglobulin-like receptor 2D,KIR2D)基因的遗传多态性、与HLA-Cw的组合特点.方法 采用聚合酶链反应-序列特异性引物技术(polymerase chain reaction-sequence specific primer,PCR-SSP)检测湖南地区112名汉族人群、内蒙古地区98名汉族人群、内蒙古地区83名蒙古族人群HLA-Cw基因、第80位密码子(Lys80、Ash80)多态性;检测两个汉族人群KIR2DL 1/2/3、KIR2DS 1/2基因分布.结果 (1)湖南地区汉族人群与内蒙古地区汉族、蒙占族人群在HLA-Cw等位基因、第80位密码子的频率差异均有非常显著的统计学意义(P＜0.001),而内蒙古地区汉族、蒙古族人群之间上述频率差异无统计学意义(P＞0.05).(2)南、北方两个汉族人群间,5个KIR2D基因频率、各基因型频率差异无统计学意义(P＞0.05).(3)两个汉族人群均以Asn80/Asn80,2DL1+/2DL2-/2DL3+/2DS1-/2DS2-组合模式最为常见(45/112、29/98);其次为Asn80/Asn80,2DL1+/2DL2-/2DL3+/2DS1+/2DS2-(18/112,16/98)和Asn80/Lys80,2DL1+/2DL2-/2DL3+/2DS1-/2DS2-(11/112,17/98).Lys80/Lys80,2DL1+/2DL2-/2DL3+/2DS1-/2DS2-组合模式的频率差异有统计学意义(1/112,8/98;Fisher's P=0.0134),其余11种组合模式在两个人群间的频率差异均无统计学意义(P＞0.05).结论 提供了中国南方湖南地区、北方内蒙古地区正常汉族人群的HLA-Cw、5个KIR2D受体基因多态性数据、蒙古族人群HLA-Cw DNA分型数据;提示我国南、北方汉族人群在HLA-Cw第80位密码子、KIR2D受体基因的组合层面上可能存在着以抑制性信号通路为优势的共同特点.     

广东汉族恶性血液病患者KIR及其与HLA配伍频率分析

目的分析KIR-HLA基因在汉族恶性血液病患者中的分布。方法对81例广东汉族恶性血液病患者和83例正常人行KIR和HLA分型,进行统计分析。结果与正常人群相比,恶性血液病患者KIR频率、抑制性和活化性KIR-HLA配对频率、个体抑制性和活化性KIR-HLA配对数目均无显著性差异。结论广东汉族恶性血液病与KIR频率及KIR-HLA配对无关联。     

KIR受配体模式在治疗急性淋巴细胞白血病中的研究

目的 研究在无关供体造血干细胞移植中急性淋巴细胞白血病(ALL)的杀伤细胞免疫球蛋白样受体(KIR)受配体模式对自然杀伤(NK)细胞的异源反应性活性预示造血干细胞移植的影响.方法 采用基因测序和序列特异性引物聚合酶链反应(PCR-SSP)的方法,对中国造血干细胞捐献者资料库中提供的23对HLA全相合供受者进行KIR及HLA高分辨基因分型;流式缃胞术动态随访CD158分子表达水平;患者均为ALL.结果 23对供受者中17例供者KIR2D12/L3有相应的患者配体HLACw1、3、7、8、12、14;6例供者KIR2DL1有相应的患者配体HLA-Cw6、15;16例供者KIR3DL1有相应的患者配体HLA-Bw4;12例供者3DL2有相应的患者配体HLA-A11.23对供受者中有19对接受了造血干细胞移植,供受者KIR基因完全相同或宿主抗移植物(HVG)方向移植相关死亡率高,分别为33.3%和40.0%;移植物抗宿主(GVH)方向移植卡甘关死亡率低,为12.5%.供受者在GVH方向时,移植物抗宿主病(GVHD)发生率高(50.0%)且有多种激活性(aKIR)的组合;而HVG方向GVHD发生率低(20.0%).19对供受者有5对均为KIR基因A单体型,其中2对供受者为KIR2DS4*001/002亚型,移植后死亡;3对供受者KIR2DS4为KIR2DS4*003-007亚型,1年后无病生存.移植后随访无GVHD发生时,CD158a的表达逐渐下降;有GVHD发生时,CD158a的表达逐渐增高;移植后早期受者NK细胞百分比为(23.4±3.8)%,高于正常人水平[(2.04±0.58)%,P<0.05],差异有统计学意义.结论 供者的KIR2DL1、KIR3DL1是引起NK细胞异源反应活性的重要抑制性KIR.KIR受配体模式不仅能预示无关供体异基因造血于细胞移植的预后,更能帮助临床提高ALL异基凶造血干细胞移植的总生存率及无病生存率,降低移植后相关死亡率和防止白血病复发.     

杀伤细胞免疫球蛋白样受体基因多态性与不明原因复发性早期自然流产的关联性研究

目的探讨杀伤细胞免疫球蛋白样受体（killer cell immunoglobulin-like receptor，KIR）基因多态性与不明原因的复发性早期自然流产的关联性。方法采用序列特异性引物聚合酶链反应（PCR-SSP）法，分析我院就诊的100例不明原因的复发性早期自然流产（unexplained recurrent spontaneous abortions，URSA）患者和112例无血缘关系的正常妇女KIR基因位点的多态性。结果URSA病例组KIR2DS1（P=0．003）、KIR2DS2（P=0．008）、KIR2DL5（P=0．003）基因的阳性率较随机对照组显著升高，URSA病例组活化性KIR基因数目较对照组显著增多（P=0．002）。具有3个或3个以下活化性KIR基因的个体在病例组中占40．00％，在对照组中占58．04％；而具有3个以上活化性KIR基因的个体在病例组中占60．00％，在对照组中占41．96％，两者差异具有统计学意义（P=0．009）。结论活化性KIR基因数目增多可能参与不明原因复发性早期自然流产的发病。KIR2DS1、KIR2DS2、KIR2DL5基因频率升高可能是不明原因复发性早期自然流产的原因之一。     

HLA-Cw在广东汉族人群中的分布频率及其意义的初步分析

目的 :检测HLA Cw在广东汉族人群的分布频率 ,初步分析该人群中KIR与HLA Cw之间识别方式的特点及意义。方法 :骨髓移植供者 12 2例 ,ACD抗凝血提取DNA ,半量全自动PCR RSSO分型检测Cw。结果 :广东汉族人群HLA Cw基因频率分布由高至低依次为 :Cw 0 3(0 2 371) >Cw 0 7(0 2 15 9) >Cw 0 1(0 175 2 ) >Cw 0 8(0 112 9) >Cw 0 4 (0 0 5 0 5 ) >Cw 14、15(0 0 4 19) >Cw 12 (0 0 376 ) >Cw 0 6 (0 0 333) >Cw 0 5 (0 0 0 82 ) >Cw 16 (0 0 0 4 1)。第一组Cw 0 2 ,0 4 ,0 5 ,0 6识别KIR分子中2DL 2DSI;第二组Cw 0 1,0 3,0 7,0 8识别KIR分子中 2DL2 2DL3;2DS2 2DS3。两组分布频率相比有显著性差异 (P <0 0 1)。结论 :广东汉族人群HLA Cw 0 1,0 3,0 7,0 8出现频率较高 ,其与KIR的识别方式均属第二组。     

膜型/分泌型MICA对NK细胞受体NKG2D的相反调节效应及其对NK细胞受体谱的影响

目的　观察膜型和分泌型MICA对NK细胞受体表达的影响 ,以探讨NK细胞抗肿瘤活化机制及肿瘤细胞表达MICA分子的意义。方法　用MTT法测定人NK细胞系 (NK92 )的细胞毒活性 ;用RT PCR或FACS检测NK细胞受体 (NKG2D ,NKG2A B ,KIR2DL1,KIR2DS1)及NKG2D的识别配体MICA的表达。结果　肿瘤细胞表面的MICA分子可上调NKG2D的表达 ,下调抑制性受体NKG2A B和KIR2DL1的表达 ;而分泌型MICA (sMICA)分子对NKG2D及抑制性受体的表达均有抑制作用。结论　膜型MICA分子可上调NKG2D的表达 ,激发NK细胞对肿瘤细胞的细胞毒效应 ;分泌型MICA分子则通过降低NKG2D的表达下调机体的抗肿瘤免疫效应 ,肿瘤细胞分泌sMICA分子为肿瘤发生免疫逃逸的机制之一。     

青光眼睫状体炎综合征与杀伤细胞免疫球蛋白样受体-人类白细胞抗原受配体复合物遗传多态性的相关性研究

目的 研究和分析南方汉族人群青光眼睫状体炎综合征（PSS）与杀伤细胞免疫球蛋白样受体（KIR）及其人类白细胞抗原（HLA）配体基因多态性的相关性.方法 随机选择100例无关健康个体作为对照组,收集97例临床诊断为PSS的患者血样,采用KIR PCR-SSP方法检测KIR框架基因的有无,PCR-SBT法进行HLA-A、B、C基因的测序分型;统计分析健康对照组与患者的KIR框架基因频率、KIR基因单倍体组合型的频率、HLA配体的频率、单个KIR-HLA配体复合物的频率之间的差异.结果 与正常人群相比,PSS患者KIR框架基因频率及单倍体组合型频率差异无统计学意义（P均＞0.05）;HLA配体中的HLA-Bw4-80T在PSS患者及正常人群中的比例分别为22.1％和41.0％,差异有统计学意义（P＜0.01）;KIR-HLA配体组合中的3DL1＋ HLA-Bw4-80T在PSS患者及正常人群中的比例分别为22.1％和40.0％,差异有统计学意义（P＜0.05）,患者组与正常人群组相比都显著降低.结论 PSS与KIR框架基因及单倍型频率无关,但可能与HLA配体或KIR-HLA配体组合相关,这对研究PSS的发病机制及治疗方法具有重要意义.     

Gene for the activating natural killer cell receptor, KIR2DS1, is associated with susceptibility to psoriasis vulgaris

Psoriasis vulgaris, particularly its juvenile form, is strongly associated with the HLA-Cw*06 allele encoding the HLA-Cw6 molecule. This molecule is recognized by the inhibitory receptor KIR2DL1 and the activatory receptor KIR2DS1, which are expressed on natural killer cells and subpopulations of T lymphocytes. Humans differ by the presence or absence of particular KIR genes. We hypothesized that either activatory KIR2DS1 or inhibitory KIR2DL1 gene frequencies might be different in psoriatic patients from a control population. Therefore, we compared the frequencies of KIR2D inhibitory (L) and activatory (S) genes in 116 psoriasis vulgaris patients and in 123 healthy controls. Fourteen novel gene combinations were found. KIR2DS1 was present in 85% of the patients, but only in 51% of the controls (corrected p [pc] < 0.0009). Similarly, HLA-Cw*06 was much more frequent in patients (77%) than in controls (17%; pc < 0.00002). Statistical analysis suggests that, although the contribution of these two factors to psoriasis is partially independent, they interact nevertheless. This result strongly speaks for a role of KIR2DS1 on recognition of HLA-Cw6 in susceptibility to psoriasis.     

A role for KIR gene variants other than KIR2DS1 in conferring susceptibility to psoriasis

Recently we described an association between psoriasis and KIR2DS1, a gene for a stimulatory natural killer cell receptor, in a Polish population. The association was independently reported among Japanese and confirmed in a U.S. population. Prompted by these findings, we reanalyzed data by a multivariate approach in search of possible effects of KIR genes other than KIR2DS1 (non-KIR2DS1). The methodology was based on a stratified analysis and multiple logistic regression. We found that the non-KIR2DS1 genes had joint effects comparable to or stronger than the effects of KIR2DS1 in both the fraction of explained variance (0.174 vs 0.204, respectively, for KIR2DS1 and non-KIR2DS1) and the statistical significance (p = 0.000008 vs p = 0.000001, respectively). When individual genes were considered, a decrease in KIR2DS5 among patients vs controls (OR = 0.2, pcor = 0.0005) and a decrease in KIR2DS3 restricted to KIR2DS1-positive individuals (OR = 0.2, pcor = 0.005) were evident. We also performed a multivariate analysis of the HLA-Cw genotypes but failed to demonstrate any effects in addition to the known association with HLA-Cw*06. We conclude that the effect of the KIR genes on psoriasis susceptibility is complex, extending beyond the association with KIR2DS1 and involving protective effects and interactions.     

Association of killer cell immunoglobulin-like receptor and human leucocyte antigen-Cw gene combinations with systemic lupus erythematosus

Killer cell immunoglobulin-like receptors (KIRs) are a diverse family of activating and inhibitory receptors expressed on natural killer (NK) cells and T cells, the genes of which show extreme polymorphism. Some KIRs bind to human leucocyte antigen (HLA) class I subgroups, and genetic interactions between KIR genes and their ligand HLA have been shown to be associated with several autoimmune diseases. The present study aimed to investigate whether the combinations of KIR genes and HLA-Cw ligands associate with the susceptibility of systemic lupus erythematosus (SLE). Polymerase chain reaction using sequence-specific primers was used to determine the genotypes of KIR genes and HLA-Cw alleles. We found that the frequencies of HLA-Cw07 were statistically significantly higher in the patient group than those in the control group (P = 0·009). KIR2DS1⁺HLA⁻Cw^Lys was more common in subjects with SLE compared to control subjects (P = 0·015). In addition, the frequency of KIR2DS1 was increased in SLE when KIR2DL1/HLA-Cw are absent, and the difference was significant (P = 0·001). KIR genotype and HLA ligand interaction may potentially influence the threshold for NK (and/or T) cell activation mediated through activating receptors, thereby contributing to the pathogenesis of SLE.     

Polymorphisms of KIRs Gene and HLA-C Alleles in Patients with Ankylosing Spondylitis: Possible Association with Susceptibility to the Disease

INTRODUCTION: An emerging body of evidence is accumulating to suggest that killer cell immunoglobulin-like receptors (KIRs) and human leukocyte antigen (HLA) class I ligands contribute to the pathogenesis of diverse kinds of autoimmune diseases. However, the functional effects of their polymorphism remain largely unknown to date. Thus, the present study was undertaken to determine the association of the polymorphisms KIRs gene and HLA-C alleles with the susceptibility to ankylosing spondylitis (AS) by means of polymerase chain reaction/sequence-specific primers for genotyping KIRs from genomic DNA of 119 patients with AS together with 128 healthy donors as a control group. RESULTS AND DISCUSSION: We found that the frequencies of KIR3DS1 and KIR2DL5 were statistically significantly higher in the patient group than those in the control group (P = 0.016 and P = 0.003, respectively). Meanwhile, the percentage of patients, who were carrying two or more of the activating KIRs, was higher than that of control group. With respect to HLA-C alleles, individuals with AS showed an increased frequency of HLA-Cw02. If HLA-C was divided into group 1 or group 2 based on whether there was an asparagine or lysine present at position 80 of the alpha-chain, HLA-C group 2 was more common in subjects with AS compared to control subjects. The genotype 2DS1+/HLA-C lys(80)+ was more common in subjects with AS. Moreover, the CD69 expression, a NK activation marker, remarkably increased in patient with AS. CONCLUSION: In conclusions, this study suggests that KIR3DS1 may severe as AS susceptive genes to trigger continuous injury of arthrosis. The imbalance of activating and inhibitory KIR as well as HLA-C group 1 and group 2 may be the key factor, which influences the pathogenesis of AS. Moreover, KIR2DS1 might associate with the susceptibility of AS by influencing NK cell activity once group 2 HLA-C ligands are present.     

Crystal structure of the human natural killer cell inhibitory receptor KIR2DL1-HLA-Cw4 complex

Inhibitory natural killer (NK) cell receptors down-regulate the cytotoxicity of NK cells upon recognition of specific class I major histocompatibility complex (MHC) molecules on target cells. We report here the crystal structure of the inhibitory human killer cell immunoglobulin-like receptor 2DL1 (KIR2DL1) bound to its class I MHC ligand, HLA-Cw4. The KIR2DL1-HLA-Cw4 interface exhibits charge and shape complementarity. Specificity is mediated by a pocket in KIR2DL1 that hosts the Lys80 residue of HLA-Cw4. Many residues conserved in HLA-C and in KIR2DL receptors make different interactions in KIR2DL1-HLA-Cw4 and in a previously reported KIR2DL2-HLA-Cw3 complex. A dimeric aggregate of KIR-HLA-C complexes was observed in one KIR2DL1-HLA-Cw4 crystal. Most of the amino acids that differ between human and chimpanzee KIRs with HLA-C specificities form solvent-accessible clusters outside the KIR-HLA interface, which suggests undiscovered interactions by KIRs.