Analysis Of Actual Mismatch Risk For KIR Genes And KIR Iigand Related Genes In Kidney Transplant Recipients

With the progress of the tissue matching technique in kidney transplantation,new immunosuppressive agent and biological agent used to clinic become normal, which keep the supper-acute rejection(AR)away on the whole, meanwhile, the rate and strength of the AR is cut down. But in the same condition of tissue matching and immunosuppressive agent used, some patient occur AR. The AR not only occur graft failure in earlier period, but the one of the most important factor of the graft long term survival. This make to reduce AR after kidney transplantation and to prolong the live time of the graft become a important topic. Beside the tissue matching, there may be other transplantation immunity access play a role. The relationship of killer cell immunoglobulin-like receptor which is the ligand of the HLA in the surface of nature killer cell and part of effector T cells with transplantation AR have much attention.KIR molecule belong to immunoglobulin superfamilly, express to the surface of the NK cell and part effector T cells. KIR Genes coding for them are found on chromosome19q13.4, express codominance, owned16KIR gene, contents14function genes and2silent genes. Its encoding proteinum KIR receptor is transmembrane glycoprotein, according to the side of cytoplasmic domain, KIR divide to L-type and S-type, L-type transmit inhibitory signal, named inhibitory KIR; S-type transmit activate signal, named activatory KIR. KIR receptor can recognite and blind HLA-I antigen specificity, so transmit and regulate NK cell and part effector T cells kill target cell. According to the numbers of the immunoglobulin like domain extracellular region, KIR gene cord protein individe as KIR2D and KIR3D, KIR2D consist of KIR2DL1-L5, KIR2DS1-S5:KIR3D consist of KIR3DLI-L3and KIR3DS1。For example:inhibitory receptor KIR3DL1have three immunoglobulin like domain and immunoreceptor tyrosine-based inhibitory motif (ITIM) activatory receptor KIR3DS1, its transmembrane domain has with positive charge residue that can bind specificity junct protein, no ITIM. inhibitory KIR receptor transmit inhibitor signal control NK cell kill target cell, activatory KIR receptor lack of ITIM, let activatory signal access form active NK cell and effective T cell kill target cell.A NK cell may express multitude KIR3D and KIR3D molecule alone or at equal pace, likewise, can express multitude activatory or inhibitory receptor. Generally, the ligand of KIR2D is HLA-C, and the ligand of KIR3D is HLA-A and HLA-B4. compared activatory receptor, inhibitory receptor have more closed affinity with HLA-I, when activatory or inhibitory receptor recognite and bind the same HLA-I antigen, inhibitory effect play the main role. KIR bind corresponding HLA-I may have four results followed:1、When inhibitory KIR bind HLA-I and there has no activatory KIR bind it, cell isn’t dissolve because inhibitory signal access was done;2、When activatory KIR bind corresponding HLA-I and there has no inhibitory KIR bind it, form activate signal, lead cell dissolve;3、If activatory or inhibitory receptor both bind HLA-I, both signal access is make done, when activatory signal play the main role, NK cell still actived, lead target cell dissolve;4.. If activatory or inhibitory receptor both bind HLA-I, both signal access is make done, when inhibitory signal play the main role, or both signal access don’t play, NK cell do not make effect. This is NK cell recognize normal tissue. At present, in clinic, tissue matching contains HLA-A、B、DR genotyping, PRA and CDC.Recent years, some foundation study of HLA biology and hemopoietic stem cell transplantation finding that HLA-C-KIR, HLA and its ligand KIR form differ immunity transmit signal and lead differ immunity regulate access in hemopoietic stem cell transplantation and tumor treatment. In addition, bone marrow transplants are already will KIR-ligand-HLA-Bw4180related gene (KIR ligand HLA-Bw4180related genes) as organization type-specific mismatch analysis in an independent analysis parameters.The frequencies of KIR gene in different race and different individual is different, so the KIR genotype in individual may different. KIR gene is express codominanced, KIR genotype differ between individual, so in vivo NK cell and part effect T cell surface KIR receptor type may differed, lead immunity transmit signal may differed. Kidney transplantation is recipient accept donor’s antigent, a series of immunity reaction aim to graft occurd necessarily.Whether or not HLA-I-KIR play the role at immune response in the AR on earlier period after kidney transplantation, with the deepening of the research to KIR, KIR and its ligands complicated relation between will be further clarify, make through the active intervention control KIR and its ligand of the interaction between the immune treatment to be, to improve the prognosis of patients with transplantation has important significance. The present study known and KIR genes associated mainly includes:KIR-L genes, KIR-S genes, KIR ligand HLA-C gene and KIR ligand HLA-Bw4180related gene these kinds of genes.This topic for kidney transplant surgery patients with uremia and corresponding donor as the research object. The Sequence Specific Primer PCR (Polymerase Chain Reaction Sequence Specific Primer, PCR-SSP) method to detect patients with uremia KIR-L genes, KIR-S genes, KIR ligand HLA-C gene and KIR ligand HLA-Bw4180related gene and analyzes the polymorphism, and compared with normal healthy people, KIR and its ligands way between recognition, this paper discusses the relationship between the two in uremic patients and significance. The application of retrospective analysis method, through the analysis of renal transplant recipients KIR ligand HLA-Bw4180related gene and its corresponding donor match has been discussed, and the acute renal transplant rejection and acute rejection reversal, the correlation between the results as a graft for clinical predictors for treatment to provide reference. This article contains two chapters followed:Chapter1Kidney transplant recipients for KIR genes, KIR gene ligand of the actual mismatch probability ObjectiveDetection KIR-L genes, KIR-S genes, KIR ligand HLA-C gene and KIR ligand HLA-Bw4180related genes in donor kidney transplantation recipients with the distribution of frequency, and discusses kidney transplant recipients for, this a few kinds of genes between actual mismatch probability.MethodSelect from January2007to June2012complete kidney transplant recipients of322cases (uremia group) and196cases of donor (normal group), extraction peripheral venous blood2ml, using the polymerase chain reaction sequence specific primer parting technology (PCR-SSP) analysis data, direct counting method for calculation of phenotype frequency (phenotype frequencies, PF). Gene frequency (gene frequencies, GF) application formula GF=1-(1-PF)1/2computation. At the same time calculation for recipients to322between these kinds of gene actual mismatch probability. Two groups of gene phenotype frequency, genotype frequency comparison using chi-square test. The comparison of the measurement data by using two independent sample t-test. To P<0.05for difference have statistical significance.ResultsThe normal group and uremia group KIR-L gene KIR2DL1,2DL2,2DL3,2DL4,2DL5,3DL1,3DL2and3DL3the phenotype frequency (%) were95.9/92.6,31.6/43.5,92.8/98.1,99/100,35.2/50.9,96.9/96.3,100/100,100/100. Of322kidney transplant recipients for, between KIR-L gene mismatch probability is0%-15.70%, and the average mismatch probability4.725%, among them KIR3DL2and3DL3mismatch in the probability is0%, KIR2DL5mismatch in the15.70%chance. The normal group and uremia group KIR-S gene KIR2DS1,2DS2,2DS3,2DS4and2DS5and3DS1the phenotype frequency (%) were31.6/43.5,32.6/43.5,21.4/19.9,90.3/92.6,18.4/29.2,39.3/56.5. Of322kidney transplant recipients for, between KIR-S gene mismatch probability is1.50%-17.20%, and the average mismatch probability9.10%, among them the chance to KIR2DS3mismatch is1.50%, KIR3DS1mismatch in the probability of17.20%. The normal group and uremia group KIR ligand HLA-C gene HLA-C01, C02, C03, C04, C05, C06, C07, C08, C12, C13, C14, C15, C16, C17and C18phenotypes frequency (%) were35.2/34.5,3.1/1.9,46.9/43.5,13.3/12.7,1.5/3.7,8.2/7.5,36.7/36.3,21.9/23.6,16.8/9.0,0/0,10.2/14.6,5.1/10.9,1.5/1.9,0/0,0/0. Of322kidney transplant recipients for, between KIR ligand HLA-C gene mismatch probability is0.00%-7.80%, and the average mismatch probability is1.95%, with HLA-C13, C17, C18mismatch probability is0%, HLA-industrial mismatch probability is7.80%. Of322kidney transplant recipients for, between KIR ligand HLA-Bw4180related genes actual mismatch risk was76.09%, among which supply and recipients have mismatch between245cases (76.09%), no mismatch77cases (23.91%), mismatch group is no mismatch group of3.18times. KIR ligand HLA-Bw4180related gene mismatch probability and other KIR genes mismatch is likely there were significant differences (P<0.05).ConclusionsIn all KIR related genes, KIR ligand HLA-Bw4180related genes in kidney transplant recipients for the mismatch between the highest rate of may and KIR ligand HLA-Bw4180related gene blood type allocation more relevant.Chapter2KIR ligand HLA-Bw4180related gene mismatch and acute rejection after transplantation (AR) and (AR) the incidence rate of reverse connectionObjectiveKidney transplant recipients to explore for, of KIR ligand HLA-Bw4180related gene mismatch in kidney transplant recipients postoperative acute rejection (acute rejection, AR) in the occurrence and the possible role of in reversal. Method322pairs of kidney transplantation recipients for, extraction peripheral venous blood, using PCR-SSP method to detect HLA-Bw4I80related gene. According to KIR ligand HLA-Bw4I80related genes in between for recipients whether mismatch is divided into mismatch group and no mismatch group, including mismatch group there were245cases of recipients, no mismatch group there were77cases of recipients, the analysis is two groups in the kidney transplantation recipients of acute rejection (AR) and the incidence of acute rejection reversal rate is consistent. The diagnosis of acute rejection standard according to the clinical manifestation, laboratory examination, transplant kidney doppler ultrasound, transplant kidney biopsy, and other comprehensive judgment. Using SPSS13.0statistical software for statistical analysis. Mismatch between the groups with and without mismatch between the group and the rate of incidence of AR reverse compares the chi-square test. The comparison of the measurement data by using two independent sample t-test. To P<0.05for difference have statistical significance.ResultsOf322kidney transplant recipients for the KIR ligand HLA-Bw4I80related gene happen mismatch with245cases of recipients, no mismatch of a total of77cases of recipients.322cases occurred in patients with AR recipients, there are26cases (all in accordance with the criteria for the diagnosis of AR in article1,2, no diagnosis renal biopsy).245cases of KIR ligand HLA-Bw4I80related genes mismatch group after transplantation recipients AR a rate of8.16%(twenty cases),77cases of no mismatch group after transplantation recipients AR a rate of7.79%(6cases), two groups of AR incidence is no statistical significance (P>0.05). Mismatch group of twenty cases of AR in recipients of17cases MP impact after treatment an average of15.4d reversal,2cases of MP impact after treatment is invalid to OKT3reversal,1cases of treatment is invalid after resection of kidney transplantation. No mismatch group of6patients with AR recipients in4cases of MP impact on average after23d reversal,2cases treatment is invalid after resection of kidney transplantation. Mismatch group AR recipients was10%(2cases) MP impact invalid after treatment by OKT3reversal, reverse rate is95.00%; No mismatch group only100%recipients with MP shock treatment, reverse rate is66.67%, two groups of OKT3utilization rate and AR reversal rate all have obvious difference, but no statistical significance (P>0.05).ConclusionsKIR ligand HLA-Bw4I80related gene mismatch in kidney transplant recipients AR happen and AR reversal of the different role, tip should adopt new supply and recipients compatibility between spectrum model research KIR genes and the relationship between the kidney transplantation, and large sample study to further discuss on foreign NK cells for renal damage effect the kidney transplant recipients benefit in the long run.