HLA Antigens ; Hematopoietic Stem Cell Transplantation ; Humans ; Receptors, KIR ; Siblings

OBJECTIVETo detect the diversity of killer Ig-like receptor(KIR) gene content and the combination of haplotypes in Chinese Han population in Shanghai area.

METHODSDNA samples from 87 randomly unrelated healthy individuals in Shanghai Han population were genotyped with SSP/PCR method.

RESULTS(1) Frequencies of KIR genes: All of 18 known KIRs genes, such as 2DL1-5, 2DS1-5, 3DL1-3, 3DS1, KIR1D and the pseudogenes X, Xv and Z(KIR2DP1) were observed in Shanghai Hans. All individuals contain 3DL3, 2DL4, 3DL2 and 3DL1; the most common genes were 2DL3, Z, 2DL1 and X; the following were 2DS4, 1D, 2DL5, 2DS1, 3DS1 and 2DS5; the next were 2DS2, 2DL2, 2DS3 and Xv. (2) Frequencies of KIR gene haplotypes; there were 13 haplotypes detected in 87 Han individuals, among them, the most frequent one was type 2 (haplotypeA-2DS4). (3) Frequencies of KIR genotypes: 18 kinds of the combinations of the haplotypes were observed; the most frequent ones were AJ(2,2), AF (1,2). Also, In this study were identified five new genotypes FZ1 2 9 , FZ2 1 16 , FZ3 6 17 , FZ4 4 13 and FZ5 2 6 ,which had not been observed in Caucasians so far.

CONCLUSIONThese findings suggest that there are distinctive frequencies of KIR gene content, haplotype as well as genotype in Chinese Han population in Shanghai area.

China ; Gene Frequency ; Genetic Variation ; Genotype ; Haplotypes ; genetics ; Humans ; Receptors, Immunologic ; genetics ; Receptors, KIR ; Receptors, KIR2DL1 ; Receptors, KIR2DL3 ; Receptors, KIR2DL4 ; Receptors, KIR3DL1 ; Receptors, KIR3DL2 ; Receptors, KIR3DS1

The aim of study was to clarify the repertoire of killer immunoglobulin-like receptor (KIR) at the level of DNA and RNA in healthy persons and to compare KIR on genotypes and expression patterns. KIR genotypes including KIR2DL1, 2DL2, 2DL3, 2DL4, 2DL5, 2DS1, 2DS2, 2DS3, 2DS4, 2DS5, 3DL1, 3DL2, 3DL3, 3DS1, 2DP1 and 3DP1 were analyzed by PCR. The phenotypes including KIR2DL1, KIR2DL2, KIR2DL3, KIR3DL1, KIR2DS1, KIR2DS3, KIR2DS2/4 gene were determined by RT-PCR. The results showed that the genes of KIR2DL1, KIR2DL3, KIR2DL4, KIR3DL2, KIR3DL3 and KIR3DP1 could be detected in all healthy persons, NK-92MI cells and Molt4 cells, but all corresponding receptors were not expressed by Molt4 T cells. Only partial transcripts of KIR2DL1, KIR2DL3 and KIR2DS2/4 were detectable in NK-92MI cells. If genotypes of KIR2DL1, 2DL2, 2DL3, 2DS1, 2DS2, 2DS3 and 2DS4 were detected in healthy persons, almost all transcripts of corresponding receptors were expressed in peripheral blood NK cells. The expression levels of KIR were different. In conclusion, the repertoires of KIR are individually specific. The expression pattern of KIR is also specific, the expression levels of different KIRs are different in one individual, and the expression levels of the same KIR are also different in different individuals.
Cell Line ; Gene Frequency ; Genotype ; Haplotypes ; Humans ; Phenotype ; Receptors, Immunologic ; genetics ; Receptors, KIR ; classification ; genetics ; Receptors, KIR2DL4 ; genetics ; Receptors, KIR3DL1 ; genetics ; Receptors, KIR3DL2 ; genetics ; Receptors, KIR3DS1 ; genetics

PURPOSE OF REVIEW: Rheumatoid arthritis (RA) is characterized by a chronic T-cell response that has escaped normal control mechanisms. This review summarizes recent insights in pathways that are functional in RA and that favor continuous and pathogenic T-cell activation. RECENT FINDINGS: T-cell activation is ultimately determined by positive signals from costimulatory molecules and negative signals from regulatory T cells. Blockade of the classic costimulatory pathway, CD28-CD80 or CD86, is beneficial in RA. Additional pathways that predominantly control the activation of memory and effector T cells are functionally important in synovial inflammation. Some of these costimulatory molecules(such as stimulatory killer cell immunoglobulin-like receptors and NKG2D) appear to be relatively specific for RA and not to play a role in normal immune responses. In addition to this predominance of positive signals, age-disproportionate decline in thymic activity in RA may lead to a diminution of regulatory T cells and loss of their negative signals. SUMMARY: The successful treatment trial of RA with CTLA-4Ig clearly documents the importance of T-cell costimulation in RA disease activity. Novel costimulatory pathways may be of even greater significance than CD28 in RA and may represent promising new therapeutic targets. The finding of reduced thymic activity in RA is exciting and will stimulate further studies of T-cell homeostasis and the function of regulatory cells.
Arthritis, Rheumatoid* ; Autoimmunity ; Homeostasis ; Inflammation ; Lymphocytes* ; Memory ; Receptors, KIR ; T-Lymphocytes ; T-Lymphocytes, Regulatory ; United Nations

OBJECTIVETo categorize ambiguous allelic combinations encountered in genotyping of functional KIR genes by sequencing-based typing of the entire coding sequence and develop an efficient approach to identify such ambiguities.

METHODSFourteen KIR functional framework genes from 306 ethnic Chinese individuals were genotyped for the entire encoding sequence. The number of ambiguities was directly counted. Based on the differences within each ambiguous allelic combination, group-specific PCR primers and sequencing primers for amplifying the target allelic sequence were designed. The PCR products were then subjected to sequencing in order to identify the ambiguities.

RESULTSThe 14 functional KIR genes were subjected to sequencing-based typing (SBT) for the entire encoding sequence. Six ambiguous allelic combinations were identified. The most common ambiguity 3DL2*(002, 007/010, 015) has accounted for 12.09% of the 306 tested samples. The remaining 5 ambiguities were (2DL5A*001, 2DL5B*006/2DL5A*012, 2DL5B*008), 3DL3*(001, 010/009, 048), 3DL2*(007, 008/016, 027), 3DL3*(00801, 048/01001, 026) and 3DL3*(00802, 048/01002, 026) have accounted for 5.88% (18/306), 3.59% (11/306), 2.29% (7/306), 1.31% (4/306) and 1.31% (4/306) of all samples, respectively. For two ambiguities (2DL5A*001, 2DL5B*006/2DL5A*012, 2DL5B*008) and 3DL2*(007, 008/016, 027) subjected to group-specific PCR and re-sequencing, only one demonstrable genotype was identified, While for in each of other four ambiguities subjected to group-specific PCR and re-sequencing, two different genotypes were identified.

CONCLUSIONAn efficient approach by group-specific PCR and sequencing retest has been established to clarify the ambiguities during SBT testing for functional KIR framework genes, which may have a broad application in KIR sequencing-based typing.

Alleles ; Asian Continental Ancestry Group ; genetics ; Base Sequence ; China ; Genotype ; Humans ; Receptors, KIR ; genetics

To analyze killer immunoglobulin (Ig)-like receptor (KIR) gene content and allelic polymorphism in Zhejiang Han population, samples were genotyped by polymerase chain reaction sequence-specific primers (PCR-SSP). The results demonstrated that all 17 KIR genes could be observed in the population. All individuals contained 2DL4, 3DL2 and 3DL3 genes. The frequencies of these genes was 1.00. 2DL1, 2DL3, 2DP1, 3DP1*003, 3DL1, 2DS4*001/002 loci were more common, their frequencies were 0.902, 0.902, 0.902, 0.902, 0.7598, 0.5615 respectively, while the frequencies of 2DL2, 2DL5A, 2DL5B, 2DS1, 2DS2, 2DS3, 2DS4*003, 2DS5, 3DS1 and 3DP1*001/002 were relatively lower. The A KIR haplotype was the most prevalent (74.7%) in Zhejiang Han population and there were 12 different KIR haplotypes in total, the most common was 2 (53.0%). Twenty six different genotypes have been found in the population, AJ (2, 2) and AF (1, 2) showed higher frequencies, followed by AH (2, 5), NN2 (2, 6), AI (1, 5) and AG (1, 1). Fifteen of these genotypes have not been found in Caucasians so far and four new KIR profiles could not be assigned to the haplotypes according to standard assign method. In conclusion, there are distinctive frequencies of KIR gene content, haplotype as well as genotype in Zhejiang Han population.
China ; Gene Frequency ; Genotype ; Haplotypes ; Humans ; Polymerase Chain Reaction ; methods ; Polymorphism, Genetic ; Receptors, KIR ; genetics

OBJECTIVETo detect the frequencies of KIR2DS4 alleles in Chinese Han population and to study the impact of KR2DS4 alleles on clinical outcomes of HLA identical unrelated allo-HSCT.

METHODSA Sequence-Based Testing (SBT) and TOPO TA cloning system for identifying and distinguishing alleles of the KIR2DS4 gene were established. A total of 150 Chinese-Han individuals, including 75 leukemia patients who received allo-HSCT and their HLA high-resolution typing identical unrelated donors (URD) were entered this study. The patients underwent transplantation for CML (n = 24), AML (n = 19), ALL (n = 29) and other malignancies (n = 3).

RESULTSThe majority (139) of the 150 samples (92.7%) were positive for KIR2DS4. Sequencing of the whole length coding region of this gene identified four of the 12 known KIR2DS4 alleles, KIR2DS4*00101, *003, *004, and *007. 2DS4*00101 was the most frequent, being found in 109 of the 139 individuals (78.4%). The ratio of deleted to non-deleted versions of KIR2DS4 was approximately 1:2. Three novel KIR2DS4 alleles were identified. Transplantations from KIR haplotype B/x donors showed significantly higher overall survival rates than those from KIR haplotype A/A donors [RR 3.1 (95%CI 1.1 - 8.6), P = 0.007]. There was a lower overall survival rates in recipients when their donors carried two 2DS4 full-length allele (2DS4*001) than those carried less (0 or 1) 2DS4*001 allele (P = 0.031). In the haplotype A/A group, a higher risk of acute GVHD (aGVHD) [RR 9.0 (95%CI 1.2 - 66.9), P = 0.01], especially grade III-IV aGVHD (P = 0.006), was seen when the donor was homozygous for the full-length KIR2DS4*00101 allele.

CONCLUSIONThe development and application of the described SBT 2DS4 allele typing method highlights the diverse nature of the KIR gene family and displays the existence of KIR polymorphism that remains uncharacterized. Our findings suggest that KIR typing for 2DS4 be beneficial for selecting suitable donors.

Alleles ; Graft vs Host Disease ; genetics ; Haplotypes ; Hematopoietic Stem Cell Transplantation ; Humans ; Receptors, KIR ; genetics

OBJECTIVETo explore the relationship between CMV reactivation and KIR haplotype or HLA-Cw genotype in patients after unrelated-donor hematopoietic stem cell transplantation (HSCT).

METHODSFrom January 2003 to December 2008 the HLA-Cw/KIR genotype of 48 patient-donor pairs were determined by polymerase chain reaction with sequence specific primers (PCR-SSP) and sequence specific nucleotide (PCR-SSOP). Posttransplant CMV reactivation was performed by immune histochemically assay.

RESULTSOf 48 patients, 15 were transplanted from unrelated donors with an antigen mismatch for HLA Cw and 33 patient-donor pairs were matched for HLA-Cw. The CMV reaction rate was 66.7% for HLA-Cw mismatch group and 48.5% for HLA-Cw match group (chi(2) = 1.39, P = 0.2375). Thirty-seven donor-patients pairs belonged to group C1 and 11 to group C2, and CMV reaction rate was 64.9% in group C1 and 18.2% in group C2 (chi(2) = 18.13, P < 0.0001). Twenty-six patients received graft from KIR haplotype A (group A donor) and 22 from KIR haplotype B donors (group B donor) and CMV reaction rate was 57.7% in group A donor and 50.0% in group B donor (chi(2) = 0.28, P = 0.5941). The number of donor activating KIRs (aKIRs) was less than that of recipient aKIRs in 34 patient-donor pairs in which the CMV reaction rate was 70.6%, and the number of donor aKIRs was more than that of recipient aKIRs in 14 patient-donor pairs in which the CMV reactivation was 14.3%. There was a significan difference between the two group (chi(2) = 12.44, P = 0.0004).

CONCLUSIONKIR and HLA-Cw genotypes influence the rate of CMV reactivation following non-T cell deleted unrelated donor hematopoietic cell transplantation.

Genotype ; HLA-C Antigens ; genetics ; Haplotypes ; Hematopoietic Stem Cell Transplantation ; Humans ; Receptors, KIR ; genetics

This study was purposed to investigate the distribution of killer cell immunoglobulin-like receptor(KIR)in Jiangsu Han population of China. KIR genomic typing were analyzed by PCR-SSP typing methods in 269 randomly unrelated healthy individuals. The results showed that all 16 known KIR genes were found in Jiangsu Han population, the total KIR genes found in Jiangsu Han population were 34 genes. Out of 34 KIR found genotypes, 15 genotypes (AA1, BX2, BX3, BX4, BX5, BX6, BX8, BX9, BX11, BX13, BX33, BX68, BX69, BX70, BX75) had also been identified in Zhejiang and Hong Kong populations, otherwise, 19 genotypes (BX10, BX12, BX17, BX23, BX26, BX27, BX28, BX31, BX35, BX42, BX47, BX57, BX72, BX74, BX79, BX154, BX188, BX231, BX370) had never been observed in Zhejiang and Hong Kong populations. The rare allele BX42 (detected only in Greece population) and BX231 (detected only in Sweden population) and BX370 (detected only in Macedonia population) were identified in Jiangsu Han population. In conclusion, all 16 known KIR genes were detected in Jiangsu Han population, the total 34 KIR genotypes are found in Jiangsu province, among them the BX42, BX231 and BX370 are rare KIR genotypes.
Asian Continental Ancestry Group ; genetics ; China ; Gene Frequency ; Genotype ; Haplotypes ; Humans ; Polymorphism, Genetic ; Receptors, KIR ; genetics

OBJECTIVETo investigate the killer cell immunoglobulin-like receptor (KIR) gene frequencies and genotypes distributions in the Inner Mongolian population.

METHODSNinety genomic DNA samples were extracted from blood samples of randomly chosen Mongolian individuals. Gene-specific PCR amplification was used to identify genes present or absent for 16 KIR loci. KIR genotype distributions were obtained and compared to that of 24 populations published in literatures using principal component analysis by SAS8.0 software. Genetic tree was constructed by the calculate Nei's genetic distance.

RESULTS(1) The frequency of KIR 2DL2, 2DS2 in Mongolian individual is higher than that in north Mongoloid and less than that in Caucasian. (2) Haplotype AA was identified in 37.78% of individuals, which is higher than that in north Mongoloid and lower than that in Caucasian. (3) Mongolian was considered between north Mongoloid and Caucasian by principal component and genetic tree analysis.

CONCLUSIONMongolian might be affected by the north Mongoloid and Caucasian, and showed intermediate between the two populations.

Asian Continental Ancestry Group ; genetics ; China ; Genotype ; Humans ; Phylogeny ; Polymorphism, Genetic ; Receptors, KIR ; genetics