Study Of The Association Between KIR Genotypes And Acute Rejection After Kidney Transplantation

With the progress of the tissue matching technique in kidney transplantation, new immunosuppressive agent and biological agent used to clinic become nomal, 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 orther 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 immuneglobulin superfamilly,express to the surface of the NK cell and part effector T cells.KIR Genes coding for them are found on chromosome 19q13.4,express codominance,owned 16 KIR gene,cotents 14function genes and 2silent genes.Its encoding proteinum KIR receptor is transmembrane glycoprotein,according to the side of cytoplasmic domain,KIRdivide 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-Ⅰantigen 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 KIR3DL1～L3 and KIR3DS1。For example:inhibitory receptor KIR3DL1 have 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 NKcell 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-Cw,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-Ⅰantigen,inhibitory effect play the main role.KIR bind corresponding HLA-Ⅰmay hame four results followed:1,When inhibitory KIR bind HLA-Ⅰand there has no activatory KIR bind it,cell isn't dissolve because inhibitory signal access was done;2,When activatory KIR bind corresponding HLA-Ⅰand 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 recognice nomal 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-Cw-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. 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 sufface KIR receptor type may differed,lead immunity transmit signal may deffered.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,this research review the frequencies that KIR genes in population and the distribution of its genotypes,analysis the correlation that inhibitory genotypes of kidney donor-recipient pair with AR.This article contains three chapters followed:Chapter one The detection of killer cell immunoglobulin-like receptor KIR genesObjective To investigate the KIR gene frenquencies of killer cell immunoglobulin-like receptor of uremia patients.Methods Uremia patients were chosen to drawn venous blood According the KIR basic radical coded sequence that publication by IDP KIR sequence library(www.ebi.ac.uk/ipd/kir) on November 10,2006.Twenty-three pairs of primers were designed,whole were verification by BLAST software,which can recognize 16 KIR genes,contains 14 functional genes and 2 silent genes.In our unit,55 uremia patients who waiting for kidney transplantation.Analysis by polymerase chain recation-sequences specific primer,PCR-SSP,work out the KIR gene frenquencies of 55 uremia patients,compared with Korean,Japanese,Southern Asian.Results Sixteen genes were observed in uremia patients,contains 14 functional genes(2DL1,2DL2,2DL3,2DL4,2DL5,3DL1,3DL2,3DL3,2DS1,2DS2,2DS3,2DS4,2DS5,3DS1)and 2 silent genes(2DP1,3DP1).The frequencies(GF)of 2DL4,3DL2,3DL3 and 3DP1 are 1.00;2DL1,2DL3, 3DL1 and 2DS4 are 0.73～0.87;2DL2,2DS1,2DS2,2DS3 and 2DS5 are 0.11～ 0.25;2DL5,3DS1 and 2DP1 are 0.30～0.41.Compared the frequencies of uremia patients(GF 0.73～1.00 in KIR-2DL4,3DL1,3DL2,3DL3,2DS4) &(GF 0.11～0.34 in KIR-2DL2,2DL5,2DS1,2DS3,2DS5,3DS1)with Korean,Japanese, Southern Asian subjects,the difference were less than 0.20.In addition,the frequencies of KIR-2DL1 in uremia patients was lower than Japanese,2DS2 was lower than Southern Asian,and 2DL3 was higher than Southern Asain.Conclusions All 16 known genes observed in uremia patients indicated that the KIR genes are polymorphic.The most frequent genes were 2DL4,3DL2,3DL3 and 3DP1;followed by 2DL1,2DL3,3DL1 and 2DS4;rare of 2DL2,2DS1, 2DS2,2DS3 and 2DS5.This study shows that there is a high concordance but variability in the frequent distributions of KIR gene among the uremia patients and Asian populations.Chapter two Analysis of killer cell immunoglobulin-like receptor KIR genotypesObjective To investigate the killer cell immunoglobulin-like receptor KIR genetypes distribution of populationMethods DNA samples from 67 randomly unrelated individuals in Chinese Han population were genotyped with PCR-SSP method and a comparison of frequencies of KIR genes were done with other populations.According the KIR basic radical coded sequence that publication by IDP KIR sequence library (www.ebi.ac.uk/ipd/kir) on November 10,2006.Twenty-three pairs of primers were designed,whole were verification by BLAST software,which can recognize 16 KIR genes and alliles.Results 16 KIR genes were found in Chinese Han individual in this study, contains 14 functional genes(2DL1,2DL2,2DL3,2DL4,2DL5,3DL1,3DL2,3DL3,2DS1,2DS2,2DS3,2DS4,2DS5,3DS1)and 2 silent genes(2DP1,3DP1). There were 25 genetypes constitution by 14 functional genes were detected.among them the most frequent one was KIR-2DL1-2DL3-2DL4-3DL1-3DL2-3DL3- 2DS4(0.373),the other 24 were≤0.09.Comparing the Chinese Han KIR combinations with these of Japanese,Korean,Caucasians,there are 56 KIR genotypes summary. Analysis the KIR genotypes in the four populations,they are share 5 KIR genotypes, 8.93%of the KIR combinations were detected in all these populations;5.36%were detected in Chinese,Koreans and Caucasians;1.79%were only found in Chinese and Caucasians;In this study,16 kinds of new gene combinations were identified (28.58%).2 kinds of genotypes were only found in Japanese(3.57%);9 kinds of genotypes were only found in Korean(16.07%);and 16 kinds of genotypes were only found in Caucasians(28.58%).Conclusions This study shows that Among 25 KIR genotypes in the 67 Chinese Han,only the distribution frequency of KIR-2DL1-2DL3-2DL4-3DL1-3DL2-3DL3-2DS4 was 0.373(4%),which is an inhibitable signal transduction pathway.We found one kind of KIR combination(KIR-2DLl-2DL3-2DL4-3DL1-3DL2-3DL3-2DS4) is the most frequent in all these populations(Chinese,Japanese, Koreans and Caucasians).Chapter three Association between inhibitory KIR genotype and acute rejection after kidney transplantationObjective To investigate the effect of inhibitory KIR genotypes(L-KIR,AA) on acute rejection(AR) after cadaveric kidney transplantation.Mothods DNA samples from 24 donor-recipient pair were genotyped with PCR-SSP method and a comparison of frequencies of KIR genes were done with other populations.According the KIR basic radical coded sequence that publication by IDP KIR sequence library(www.ebi.ac.uk/ipd/kir) on November 10,2006. Twenty-three pairs of primers were designed,whole were verification by BLAST software,which can recognize 16 KIR that contains 14 functional genes(2DL1,2DL2,2DL3,2DL4,2DL5,3DL1,3DL2,3DL3,2DS1,2DS2,2DS3,2DS4,2DS5,3DS1)and 2 silent genes(2DP1,3DP1),reference to the report that make by Marsh in WHO-KIR Nomenklatur Kommission in 2002.It identify the 14 functional genes to inhibition genotype and non inhibition genotypes,inhibition genotype consist by some inhibitory KIR genes(2DL1,2DL3,2DL4,3DL1,3DL2,3DL3) and a activatory KIR gene(2DS4),named group AA.non inhibition genotypes content two group,group BB and AB.BB consist by some inhibitory KIR genes (2DL1,2DL3,2DL4,3DL1,3DL2,3DL3) and some activatory KIR genes(2DS1, 2DS2,2DS3,2DS5,3DS1)(no 2DS4);AB consist by some inhibitory KIR genes (2DL1,2DL3,2DL4,3DL1,3DL2,3DL3) and some activatory KIR genes(2DS1, 2DS2,2DS3,2SD4,2DS5,3DS1)(content 2DS4).RESULTS:Of the 24 pairs,AA KIR genotype was found in 9 donors and 10 recipients;BB KIR genotype was found in 3 donors and 4 recipients;AB KIR genotype was observed in 12 donors and 10 recipients.AR occurred in 10%(1/10) of the recipients with AA KIR genotype and in 64.29%(9/14) of the recipients with BB & AB KIR genotype.The difference between the two groups was significant (P＜0.05).11.11%(1/9) of the donors with AA KIR genotype had AR while 60% (9/15) of the donors with BB & AB KIR genotypes suffered AR,with significant difference between the two groups(P＜0.05).0%(0/4) of the donors/recipients with AA/AA match had AR while 50%(10/20) of the donors/recipients with non-AA/AA match suffered AR,without significant difference(P＞0.05).CONCLUSIONS:Recipients with AA KIR genotype tend to have a lower incidence of AR,while for recipients with BB & AB KIR genotypes,choice of a graft with AA KIR genotype may reduce the incidence of AR.These may be due to the possible mechanism that AA KIR genotype was characterized by inhibitory conducting signals after kidney transplantation early.