This study was purposed to investigate the killer immunoglobulin-like receptor (KIR) genotype in patients with hematological malignancies. The sequence specific primer-polymerase chain reaction (PCR-SSP) technique was performed for the amplification of six inhibitory KIR genes (KIR2DL1-2DL4, 3DL1-3DL2) and six activating KIR genes (KIR2DS1-S5, 3DS1). The methods of KIR-SSP was used to determine the KIR genotypes of 54 leukemia patients, including 14 patients with acute myeloid leukemia (AML), 16 with acute lymphoblastic leukemia (ALL), 20 with chronic myeloid leukemia (CML), 3 with myelodysplastic syndrome (MDS) and 1 with acute myeloid-lymphoblast leukemia (AMLL). 54 patients were classified as high risk group (n = 27) and standard risk group (n = 27). The expression of KIR in NK cells and T cells was detected by flow cytometry. The frequencies of activating KIR genes in standard risk group were higher than those in high risk group, especially 2DS1 (p = 0.014), or 2DS2 (p = 0.046), or 3DS1 (p = 0.027). However, the frequencies of inhibitory KIR genes in standard risk group were similar to those in high risk group (p > 0.05). The frequencies of activating KIR genes were also higher in standard risk patients with acute AML, as compared with those in high risk patients with acute AML, particularly 2DS1 (66.7% vs 29.4%, p = 0.022), 2DS2 (57.6% vs 17.6%, p = 0.013), and 2DS3 (33.3% vs 5.9%, p = 0.039). The percentages of patients in high-risk group who expressed more than two kinds of activating KIRs were lower that those in standard-risk group (p = 0.035). There was no difference in the expressions of CD158a, CD158b, and CD158e on NK cells and T cells between high-risk group and standard-risk group (p > 0.05). In conclusions, different expressions of activating KIR genes were found in patients between high-risk group and standard-risk group.
Genotype ; Hematologic Neoplasms ; genetics ; immunology ; Humans ; Killer Cells, Natural ; immunology ; metabolism ; Receptors, KIR ; genetics ; Risk Factors ; T-Lymphocytes ; immunology ; metabolism

The interaction of killer cell immunoglobin-like receptors (KIR) with HLA molecules has particular relevance to the genetics, immune responses and allogeneic stem cell transplantation. The genes of KIR and HLA are located in different chromosomes and segregate independently. The repertoire of KIR molecules varies among NK cells and is determined by the KIR genotype. The HLA genotype has only subtle impact on the KIR phenotype. Three major HLA specificity groups are recognized by KIRs. Donor versus recipient NK-cell alloreactivity, when recipients lack HLA ligand for their donor inhibitory KIR, can benefit allogeneic stem cell transplantation, especially the HLA haploidentical hematopoietic stem cell transplantation. The outcome of stem cell transplantation can be best predicted by the presence of KIRs on the donor's NK cells and the absence of corresponding KIR ligand in the recipient's HLA repertoire-a receptor-ligand model. In this paper the interaction of KIR and HLA in hematopoietic stem transplantation, the genetic basis of KIR and HLA, the relation of KIR expression on NK cells with HLA and the role of KIR and HLA in immune responses were reviewed.
Hematopoietic Stem Cell Transplantation ; Histocompatibility Antigens Class I ; genetics ; immunology ; Humans ; Killer Cells, Natural ; cytology ; immunology ; Receptors, Immunologic ; genetics ; immunology ; Receptors, KIR ; Transplantation Immunology

The aim of this study was to investigate how the killer immune globulin-like inhibition receptor (KIR) in match with HLA-Cw impacts on NK cell activity. Mononuclear cells were isolated in 20 ml peripheral blood from 27 healthy persons by Ficoll-Hypaque and purified by NK cell isolation kit. Target cells were mononuclear cells isolated from bone marrow of 30 de novo AML patients. The KIR expression were detected by flow cytometry with antibodies against CD158a, CD158b. The 2 ml of peripheral blood from healthy persons and AML patients were collected, the DNA was extracted by using PROTRANS method, the HLA-Cw and KIR gene were detected by PCR-SSP typing with sequence specific primers. The NK cell cytotoxicity against AML cells was determined by MTT after combination of KIR with HLA-Cw gene. The results indicated that the purity of NK cells was (90.8 +/- 6.08)%. The less the KIR/HLA-Cw matched, the more activity was shown in NK cells. When no match of NK cell/target cell (KIR/HLA-Cw) there was, the cytotoxicity was (50.66 +/- 8.40)%, 1 or 2 matches showed cytotoxicity of (38.28 +/- 6.71)% and (19.74 +/- 4.15)% (p < 0.001). Expression level of KIRs on NK cells also was related with cytotoxicity level (p < 0.001). It is concluded that the interaction between inhibitory KIR and HLA ligands, and also expression level of KIRs on NK cells both impact significantly on NK cell function, that the less match of KIR/HLA-Cw, and the less expression of KIRs on NK cells result in the stronger NK cell cytotoxicity.
Adult ; Female ; Genotype ; HLA-C Antigens ; genetics ; Humans ; Killer Cells, Natural ; immunology ; metabolism ; Male ; Receptors, KIR ; genetics

The aim of study was to analyze natural killer immunoglobulin (Ig)-like receptor (KIR) gene content in HLA-identical sibling and to investigate the possibility of their KIR match. Samples were genotyped for HLA by Luminex method and polymerase chain reaction sequence based typing, the KIR gene was detected by polymerase chain reaction sequence-specific primers. The results showed that 17 KIR genes could be observed in the 27 pairs HLA-A, -B, -Cw and -DRB1 locus identical sibling samples. All individuals contained KIR3DL3, KIR3DP1, KIR2DL4 and KIR3DL2.; 20 different KIR genotypes and 12 haplotypes have been found, the most common KIR genotypes was 2,2 with frequency 29.6% and KIR haplotype was 2 with frequency 53.0%. The A KIR haplotype was the most prevalent with frequency 67.2%; 12 pairs (44.4%) HLA identical sibling donor-recipients showed KIR match in genotype and haplotype, 13 pairs (48.1%) with one KIR haplotype mismatch and 2 pairs (7.4%) with two KIR haplotype mismatch; 1 pair was matched between donor KIR2DL1 and patient HLA-Cw (Lys80) ligand, 17 pairs were matched between KIR2DL2/KIR2DL3 and HLA-Cw (Asn80) ligand, 5 pairs were matched between KIR3DL1 and HLA-Bw4 ligand. It is suggested that the probability of KIR mismatch is high in HLA-identical sibling.
HLA Antigens ; genetics ; immunology ; Haplotypes ; Hematopoietic Stem Cell Transplantation ; methods ; Histocompatibility Testing ; Humans ; Multigene Family ; Receptors, KIR ; genetics ; Siblings

HLA-Cw belongs to classic HLA-I gene, HLA-Cw molecules have high polymorphism like HLA-A and B molecules. They distribute extensively on the surfaces of karyote, not only presenting endogenetic antigen to CD8+ T cells to induce specific killing effect, but also participating in immunologic reaction as the ligands of killer cell immunoglobulin-like receptor (KIR). Thus it has been valued for their relations to diseases and the functions in transplantation immunity, anti virus and anti-tumor immunity.
Autoimmune Diseases ; immunology ; Graft vs Host Disease ; etiology ; HLA-C Antigens ; genetics ; physiology ; Humans ; Neoplasms ; immunology ; Receptors, Immunologic ; metabolism ; Receptors, KIR ; Virus Diseases ; immunology

The profound graft-versus-leukemia (GVL) effect presented after allogeneic hematopoietic cell transplantation (allo-HSCT) has been evidenced. In contrast to T cell mediated GVL, natural killer (NK) cells recognize target cells and introduce GVL effect by using an integration of activating and inhibitory receptors. This review has summarized current literatures from 2001 - 2003 on human killer cell immunoglobulin receptors (KIR) and other NK cell receptors involved in recognition of tumor targets and the polymorphism of KIR genes of donor/recipient pairs of related and unrelated Allo-HSCT. KIR epitope mismatch may facilitate engraftment and reduce leukemia relapse post transplant by mediating lysis of recipient's cells and introducing GVL effect under the condition of KIR epitope mismatch. Clinical roles of KIR in Allo-HSCT and immunotherapy are discussed. Technologic approach in allogeneic reactive NK cells introduction, identification and selection in vitro, development of inhibitory receptor blockade as well as up-regulation of activating NK cells may significantly enhance GVL immune response. Further investigation on the regulation of KIR inhibitory receptors enables to design novel strategy in cancer immunotherapy over the forthcoming decade.
Graft vs Host Disease ; immunology ; Hematopoietic Stem Cell Transplantation ; Humans ; Immune Tolerance ; Killer Cells, Natural ; immunology ; Major Histocompatibility Complex ; Receptors, Immunologic ; chemistry ; genetics ; physiology ; Receptors, KIR ; Transplantation, Homologous

Natural killer (NK) cells play an important role in innate immunity, especially in the response to viral infections, such as hepatitis C virus (HCV). Killer cell immunoglobulin-like receptors (KIRs) are the primary receptors of NK cells that mediate innate immunity. KIRs are also involved in acquired immunity, because some KIRs are expressed on the surface of certain subsets of T cells. In this study, the frequency of KIR genes, HLA-C allotypes, and combinations of KIR genes with their HLA-C ligands were evaluated in two different groups of the Korean population: controls and patients with chronic HCV infection. The study population consisted of 147 Korean patients with chronic HCV infection. The frequency of KIR2DS2 in patients with chronic HCV infection was 9.5% which was significantly lower than 19.5% of the control (P < 0.01). However, there were no significant differences in the frequency of other KIR genes, HLA-C allotypes or different combinations of KIR genes with their HLA-C ligands. This study can contribute to the further prospective study with a larger scale, suggesting the assumption that KIR2DS2 might aid in HCV clearance by enhancing both the innate and acquired immune responses of people in Korea.
Adult ; Aged ; Female ; Genes, MHC Class I ; Genotype ; HLA-C Antigens/genetics ; Hepacivirus/immunology ; Hepatitis C, Chronic/*genetics/immunology ; Humans ; Killer Cells, Natural/immunology/virology ; Male ; Middle Aged ; Receptors, KIR/*genetics/immunology ; Republic of Korea ; T-Lymphocyte Subsets/immunology

OBJECTIVETo investigate the genotypes of natural killer cell immunoglobulin-like receptor (KIR) genes and their frequencies in Chinese subjects and explore the mechanism of the actions of nature killer cells.

METHODSThe DNA samples were obtained from 67 randomly selected unrelated Chinese Han individuals for genotyping of the KIR genes using PCR with sequence-specific primers (PCR-SSP), and the frequencies of the KIR genes in these Chinese subjects were compared with the reported frequencies in populations of other nationalities.

RESULTSSixteen KIR genes were identified in these Chinese subjects, and 87.5% of these genes were expressed at frequencies above 0.35. Fourteen functional KIR genes combined into 25 KIR genotypes, among which the most frequent genotype KIR-2DL1-2DL3-2DL4-3DL1-3DL2-3DL3-2DS4 showed a frequency of 0.373, while the frequencies of all the other genotypes were no greater than 0.09. Comparison of the KIR combinations in Chinese Han population with those of Japanese, Korean, and Caucasians populations identified 8.93% of the KIR combinations shared by all these populations; the Chinese, Koreans and Caucasians shared 5.36% common KIR combinations, whereas only 1.79% common combinations were found in Chinese and Caucasians. In this study, 16 new gene combinations were identified (25.28%).

CONCLUSIONThis study shows the high-frequency distribution of a single KIR gene polymorphism. The KIR combination KIR-2DL1-2DL3-2DL4-3DL1-3DL2-3DL3-2DS4 has the highest frequency in Chinese, Japanese, Korean and Caucasian populations, indicating that inhibitory signal transduction pathway plays an important role in the function of the natural killer cells. This study provide clues for new approaches for improving the prognosis of kidney transplantation by enhancing or inhibiting the function of the natural killer cells instead of life-time usage of immunosuppressive agents.

Asian Continental Ancestry Group ; ethnology ; genetics ; Gene Frequency ; Genotype ; Humans ; Killer Cells, Natural ; immunology ; Polymorphism, Genetic ; Receptors, KIR ; genetics ; Sequence Analysis, DNA

OBJECTIVETo study the biological function of killer cell immunoglobulin-like receptor (KIR) and the role of donor inhibitory KIR and recipient genetic background in HLA matched unrelated hematopoietic stem cell transplantation (HSCT).

METHODSHLA genotype of 51 patients (ALL 18 cases, CML 15 cases, AML 10 cases and others 8 cases) and their respective matched unrelated donors from Database of China Marrow Registration was determined by polymerase chain reaction sequence oligonucleotide probes (PCR-SSOP) and sequence specific primers (PCR-SSP). The KIR genotype was determined by PCR-SSP.

RESULTSAll the patients and the donors expressed KIR2DL1, KIR2DL2/L3, KIR2DL4, KIR3DL2 and KIR3DL3. 96.7% individuals expressed KIR3DL1. Among them, 21.57% of KIR was completely identical, while 78.43% was not. Of the non-identical KIRs, 25.49% were the recipient's KIR genotype containing the donor's ones, and 27.45% was the donor's containing the recipient's. 74.62% of donor's KIR2DL1 lacked recipient's C2 ligand, 5.91% of donor's KIR2DL2/L3 lacked recipient's C1 ligand, 19.74% of donor's KIR3DL1 lacked recipient's Bw4 ligand and 54.91% of donor's KIR3DL2 lacked recipient's A3, A11 ligand.

CONCLUSIONKIR genotype and HLA class I antigen are inherited independently. KIR2DLI and KIR3DL2 of donors may cause alloreactivity of NK cell. The mismatch of KIR/HLA in donor-recipient plays a very important role in matched unrelated allo-HSCT. The outcome of HSCT can be better predicted by the model of the presence of KIRs on the donor' sNK cells and the absence of corresponding KIR ligand in the recipient's HLA.

Adolescent ; Adult ; Child ; Child, Preschool ; Female ; Gene Frequency ; Genotype ; HLA Antigens ; genetics ; Hematopoietic Stem Cell Transplantation ; Humans ; Infant ; Killer Cells, Natural ; immunology ; Male ; Receptors, KIR ; genetics ; Transplantation, Homologous

OBJECTIVETo study the effects of small interfering RNA (siRNA)-mediated silencing of CD158b expression on the efficiency of the natural killer (NK) cells in killing allogeneic dendritic cells.

METHODSAfter the knockdown of CD158b by CD158b -SiRNA, the CD158b mRNA expression in natural killer cells was examined by qRT-PCR and the CD158b protein expression by flow cytometer. The cytotoxic activity of RNAi-NK cells and normal NK cells against the allogeneic dendritic cells was detected by LDH release assay.

RESULTSThe CD158b mRNA expression and its protein expression were decreased significantly in the NK cells by CD158b siRNA (P/0.05). The cytotoxic activities of alloreactive NK cells generated by RNAi CD158b expression against allogeneic dendritic cells were increased significantly.

CONCLUSIONSilencing CD158b gene can inhibit the NK cell CD158B mRNA and protein expression. Alloreactive NK cells generated by RNAi CD158b expression have the potential for use in interventions of GVHD.

Bone Marrow Transplantation ; immunology ; methods ; Cytotoxicity, Immunologic ; immunology ; Dendritic Cells ; cytology ; immunology ; Flow Cytometry ; Gene Silencing ; Graft vs Host Disease ; immunology ; Humans ; Killer Cells, Natural ; cytology ; immunology ; RNA Interference ; RNA, Messenger ; biosynthesis ; genetics ; RNA, Small Interfering ; genetics ; Receptors, KIR ; metabolism ; Receptors, KIR2DL3 ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction