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Objective:Establish the decision threshold value of mean fluorescence intensity of anti-human leukocyte antigen(HLA)antibody through statistical analyzing the results of international proficiency testing(PT)organized by American Society for Histocompatibility and Immunogenetics(ASHI).Methods:Single antigen reagent and liquid chip(Luminex)technique were used to detect anti-HLA antibody. A retrospective analysis of the HLA antibody PT results of 55 quality control samples from 11 times organized by ASHI from 2012 to 2019 was reviewed.Results:Among 79 kinds of HLA-I antibodies, 21, 43 and 15 types of HLA-A, B and Cw antibodies were detected respectively, while among 44 kinds of HLA-Ⅱ antibodies, 18, 7 and 19 types of HLA-DRB1, DQB1 and DPB1 antibodies were detected respectively. After analyzing the MFI detection value of different specific antibodies in each PT samples at our laboratory and the coincidence rate of the negative / positive results judged by ASHI through summarizing the results of multicenter participating in the same period, MFI values of HLA antibody were arranged from high to low into the intervals of possible saturation value, positive decision value, positive judgment threshold value, suspicious positive reference value and suspicious negative reference value , according to the coincidence rate of 95%, 90%, 80%, 79%~50% and <50%.Thus, the decision limit value table of HLA specific antibody at our laboratory was established. And 42 kinds of HLA antibody types were detected with complete data.When the MFI values of various HLA-I or HLA-Ⅱ antibodies are found to be 80% or more in the table, it can be used to judge the detection of HLA antibodies. When HLA antibody MFI value reaches the positive decision value, it may have a certain guiding significance for clinical diagnosis and treatment. And when antibody MFI value reaches the saturation value and lies in the suspicious positive or suspicious negative reference threshold, it just suggests that the clinical need for dynamic follow-up of anti-HLA antibody detection.Conclusions:The decision limit value of MFI of laboratory HLA antibody is established based on the international PT experimental results, which is of reference value for the interpretation of experimental results and clinical diagnosis and treatment. A transplant ation center should pay attention to the quality control of comparison test between laboratories in the detection of HLA antibodies.
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Objective@#To establish a real-time PCR (RT-PCR) assay for detecting mRNA expression of killer cell immunoglobulin-like receptor (KIR) 2DS1 gene( KIR2DS1 ) on the surface of natural killer (NK) cells, and evaluate its performance. @*Methods@#A total of 57 recipient-donor pairs of allogeneic hematopoietic stem cell transplantation (Allo-HSCT) were enrolled in this study. The specific primers and probe of KIR2DS1 gene were designed for Taqman-MGB fluorescence quantitative PCR detection system. The performance parameters of the detecting system, such as coincidence rate, repeatability, sensitivity, scope of application of the instrument and reproducibility of operation technicians were evaluated and validated. @*Results@#The KIR-SSO Genotyping Test was used as the gold standard. The results of 35 samples showed the accuracies of self-built method were all 100% for both of positive and negative KIR2DS1 . Three samples with high, median and low value of Ct values were used to verify the repeatability. The coefficients of variation of intra-assay and inter-assay were ranged from 0.09% to 0.46% and 0.71% to 1.13% respectively. The sensitivity of the established method was up to 10 2 copies/μL at least. The coefficients of variation of the three samples with sensitivity of 10 2 copies/μL were 5.37%, 2.71% and 5.51% in five repeated tests respectively. The regression analysis for the samples measured by ABI-7500 and LC-480 fluorescence quantitative PCR instrument showed regression equation was Y=0.973 6X+0.118 3 (R 2 =0.961 9, R 2 ＞0.95). The reproducibility of 10 samples with positive KIR2DS1 operated by two technicians showed that the biases were all less than ±5%. @*Conclusion@#A TaqMan-MGB real-time PCR assay for detection of mRNA expression of KIR2DS1 gene was established successfully with fine performance.
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Objective@#To analyze family-based haplotype frequencies of HLA-A, -B, -C, -DRB1 and -DQB1 genes and their clinical significance.@*Methods@#The data of HLA genotyping in 3568 families undergoing related haploidentical transplantation between 2012 and 2017 at the First Affiliated Hospital of Soochow University were retrospectively evaluated. The HLA genotyping was performed by PCR amplification with sequence-based typing (PCR-SBT) and sequence-specific oligonucleotide probe (PCR-SSOP) methods. The family genetic analysis and haplotype frequencies were also investigated.@*Results@#All the families were divided into 3 groups, including group1 of 1 422 entire families; group2 of 1 310 patients and either of their parents or one of their children; group3 of 836 patients and their HLA≥5/10 matched sibling donors. In the haplotypes with frequencies greater than 0.1% in group1+ group2, the frequency of A*11∶01-B*40∶01-C*03∶04-DRB1*11∶01-DQB1*03∶01, A*02∶07-B*51∶01-C*14∶02-DRB1*09:01-DQB1*03∶03 were significantly different between group1 and group2 (P=0.029, 0.033) . The frequency of A*11∶01-B*46∶01-C*01∶02∶01G-DRB1*09∶01-DQB1*03∶03 was significantly different between group1 and group3 (P=0.035) . The frequency of A*02∶01-B*40∶01-C*07∶02-DRB1*09∶01-DQB1*03∶03 was significantly different between group1 and group2 (P=0.034) , or group1 and group3 (P=0.034) . The frequency of A*24∶02-B*13∶01-C*03∶04-DRB1*12∶02-DQB1*03:01 was significantly different between group2 and group3 (P=0.046) .@*Conclusion@#In this study, we summarize the prevalence of haplotype frequencies in terms of HLA-A, -B, -C, -DRB1 and-DQB1. Based on the database of family haplotype analysis, patients and donor candidates are sorted with matched HLA genotype while unmatched HLA haplotype. Even in patients without entire family information, HLA haplotype analysis assists in choosing the optimal related or unrelated donors.
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Objective@#To investigate the immune reconstruct regularity profile of KIR2DL1 and KIR3DL1 in unrelated-donor allogeneic hematopoietic stem cell transplantation (allo-HSCT) with KIR-AA genotype.@*Method@#75 donor-recipient pairs were performed by KIR genotying using PCR-SSP, and all donors were identified with KIR-AA genotype. Dynamic detections (including unrelated-donor on the day of transplantation and the recipient each month post allo-HSCT) of the expression of KIR2DL1/3DL1 on NK cell and mRNA level were performed in 291 cases using flow cytometry (FCM) and real-time fluorescent quantitation PCR (RT-qPCR) .@*Result@#①The median expression of KIR2DL1 in unrelated-donor on transplant’s day was 21.60%, the median expression of KIR2DL1 in recipient 1M, 2M, 3M and 3-6M after transplantation were 7.40%, 12.00%, 16.92%, 17.64% respectively. The median expression of KIR2DL1 in unrelated-donor on transplant’s day was 265.14 copies/10 000abl copies, the median expression of KIR2DL1 in recipient 1M, 2M, 3M, 3-6M, 6-9M, 9-12M after transplantation were 332.17, 438.31, 723.25, 414.17, 180.76 and 234.67 copies/10 000abl copies respectively. The median expression of KIR2DL1 on NK cells and mRNA level gradually increased at all time points after transplantation, and reached the highest expression at 3 months after transplantation. But mRNA expression levels increased earlier than NK cell membrane proteins. ②The median expression of KIR3DL1 in unrelated-donors on transplant’s day was 18.56%, the median expression of KIR3DL1 in recipient 1M, 2M, 3M, 3-6M after transplantation were 23.83%, 22.57%, 23.02%, 21.60% respectively. The median expression of KIR3DL1 in unrelated-donor on transplant’s day was 572.29 copies/10 000abl copies, the median expression of KIR3DL1 in recipient 1M, 2M, 3M, 3-6M, 6-9M, 9-12M after transplantation were 1 233.74, 1 140.42, 876.73, 1 057.07, 739.02 and 514.43 copies/10 000abl copies respectively. The median expression of KIR3DL1 on NK cells and mRNA level were higher than donors at 1 month after transplantation, and stable expression at all time points after transplantation, so mRNA and NK cell membrane proteins expression increased at the same time.@*Conclusion@#The immune reconstruct regularity of KIR2DL1 and KIR3DL1 gene were different, which provided an experimental basis for selecting the best time to detect the expressions of KIR2DL1 and 3DL1 after transplantation.
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Objective@#To analyze the distribution and proportion of donor-specific activated killer cell immunoglobulin like receptor (aKIR) genes and their clinical application values in unrelated allogeneic hematopoietic stem cell transplantation (allo-HSCT) .@*Methods@#Retrospective analyses of KIR genotyping using polymerase chain reaction with sequence specific primers (PCR-SSP) were performed in 216 pairs of donors and recipients.@*Results@#The frequency of donor-specific KIR genes was 53.7% (116/216) in 216 patients receiving unrelated allo-HSCT, with the frequency of 78.3% (112/143) in the KIR genes mismatched group and 5.5% (4/73) in matched group. Of the 116 patients with detectable donor-specific KIR genes, 99.1% (115/116) patients had various donor-specific aKIR genes. Among 55 pairs of donors’ KIR-Bx genotype and patients’ KIR-AA genotype group, the most commonly observed genotypes were Bx1, Bx2, Bx3, Bx4, in which the donor-specific KIR genes were respectively KIR 3DS1, 2DL5A, 2DS5, 2DS1; KIR 3DS1, 2DL5A, 2DS3, 2DS1; KIR 2DS2, 2DL2; KIR 2DS2, 2DL2, 3DS1, 2DL5A, 2DS5, 2DS1. Of 44 pairs of donors’ KIR-AA genotype and patients’ KIR-Bx (AB) genotype group, 36.4% (16/44) recipients had donor-specific KIR2DS4 (FUL) gene. In 143 pairs of KIR mismatched group, the frequencies of donor-specific KIR genes were KIR2DS1 (35.7%) , KIR3DS1 (32.9%) , KIR2DS5 (29.4%) , KIR2DS4 (FUL) (25.9%) , KIR2DL2 (25.2%) , KIR2DS2 (24.5%) , KIR2DS3 (21.7%) and KIR3DL1 (8.4%) , respectively.@*Conclusion@#The donor-specific aKIR genes mainly existed in KIR mismatched group after unrelated allo-HSCT, and the different pairs of donors’ and patients’ KIR genotypes led to the diverse donor-specific aKIR. But there were higher specific aKIR genes in higher frequency of KIR AA, Bx1, Bx2, Bx3, Bx4 genotypes. All these can provide the experimental basis for studying the role of the donor-specific aKIR genes on the prognosis of HSCT.
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<p><b>OBJECTIVE</b>To investigate the role of different immunoglobulin- like receptor (KIR)haplotypes in haplo- identical hematopoietic stem cell transplantation (HSCT).</p><p><b>METHOD</b>Killer cell KIR genotyping was performed on 468 individuals from 156 unrelated families by PCR-SSP. A total of 624 KIR haplotypes from the parents were used for haplotype analysis. Ninety-two patients received haplo-identical HSCT from one of the parents.</p><p><b>RESULTS</b>The family study showed segregation of one A haplotype and at least 20 unique B haplotypes. The frequency of haplotype A was 72.92% (455/624). The most commonly observed haplotypes in group B were B1, B2, and B3, present at a frequency of 10.26%, 5.77%, and 4.48%, respectively. Compared to KIR gene matched donors (n=17), grafts from KIR gene mismatched donors (n= 14) had a positive effect on survival after haplo- identical HSCT for AML/MDS patients (OS: 88.2%vs 42.9%,P=0.015; RFS: 88.2%vs 35.7%,P=0.007). No effect was observed for ALL/NHL patients (OS: 76.0%vs 75.0%,P=0.727; RFS: 68.0%vs 65.0%,P=0.866). A significantly lower survival rate was observed for transplants from AA (n=52) and AB1/AB2 donors (n=15), compared to other group Bx donors (n=25) (OS: 53.3%vs 96.0%,P=0.017; RFS: 53.3%vs 92.0%,P=0.019). Meanwhile, the risk of relapse was much higher in AA group (n=52) compared to Bx group (n=40) (25.0%vs 5.0%,P=0.009). A higher risk of TRM was observed in AB1/AB2 group (P=0.012). In addition, transplant from donors carried Cen-B was associated with an increased survival compared with Cen-A homozygous donors (OS: 94.7%vs 68.5%,P=0.036; RFS: 89.5%vs 64.4%,P=0.045).</p><p><b>CONCLUSION</b>Overall, KIR genotyping and haplotype analyses should be useful for selection of the most optimal donors with favorable KIR gene grafts. KIR gene mismatch donors should be preferred for AML/MDS patients. Selecting donors carried Cen- B and avoiding the selection of donors of KIR genotype AA/AB1/AB2 was strongly advisable for haplo-identical HSCT.</p>
Subject(s)
Humans , Chronic Disease , Genotype , Haplotypes , Hematopoietic Stem Cell Transplantation , Killer Cells, Natural , Leukemia, Myeloid, Acute , Therapeutics , Neoplasm Recurrence, Local , Receptors, KIR , Genetics , Survival Rate , Tissue DonorsABSTRACT
<p><b>OBJECTIVE</b>To study KIR3DL1 expression level on NK cell surface of normal donors for hematopoietic stem cell transplantation（HSCT）.</p><p><b>METHODS</b>Ninety- two donors were performed by using of KIR genotyping, HLA high resolution genotyping and KIR3DL1 expression level using sequencebased testing（SBT）, PCR- sequence specific primer（SSP）and flow cytometry methods.</p><p><b>RESULTS</b>In 92 donors, the frequencies of KIR-A/A, Bx1, Bx2 for common genotypes were 46.74%（43/92）, 18.48% （17/92）and 9.78%（9/92）respectively（P<0.001）; KIR-A, B1, B2, B3 for common KIR haplo-type were 70.33%（128/182）, 10.99%（20/182）, 7.14%（13/182） and 4.39%（8/182） respectively（P<0.001）; the frequencies of HLA-BW4/BW4, HLA-BW4/BW6, BW6/BW6 ligands were 13.79%, 67.81% and 18.39% respectively（P<0.001）. KIR3DL1 middle expression level among haplo- type KIR- A/A and KIR- Bx, KIR-B/B were 18.77%（3.11%-49.24%）, 13.14%（1.70%-63.32%）and 0.37%（0.20%-2.60%）respectively （P<0.05）. KIR3DL1 expression level［18.77%（3.11%-49.24%）］in haplo-type KIR-A/A was higher than haplo-type KIR-Bx at the same time did not express 2DL2 group［11.20%（3.50%-36.08%）］（P=0.019）. KIR3DL1 expression level in recognition group（HLA-BW4 positive group）［17.61%（1.40%-49.24%）］ was higher than KIR3DL1 unrecognized group（HLA-BW4 negative group）［10.60%（3.50%-18.56%）］ （P=0.006）.</p><p><b>CONCLUSION</b>The expression levels of KIR3DL1 in different KIR genotypes, haplotypes and HLA ligands were statistically significance.</p>
Subject(s)
Humans , Genotype , HLA-B Antigens , Genetics , Haplotypes , Hematopoietic Stem Cell Transplantation , Hematopoietic Stem Cells , Killer Cells, Natural , Metabolism , Ligands , Receptors, KIR3DL1 , Genetics , Tissue DonorsABSTRACT
Objective To explore the efficiency and safety of fondaparinux and low molecular weight heparin(LMWH) on the elder patients with non-ST-segnent elevation acute coronary syndromes (NSTE-ACS).Methods One hundred and forty patients over 75 years old with NSTE-ACS were randomly divided into treatment group(n =70) and control group (n =70).Patients in treatment group were given the conventional treatment combined with fondaparinux,and in control group were given the basis of conventional treatment combined with LMWH.The therapeutic efficacy,the cardiovascular events at 4 d,7 d and 30 d during the treatment and bleeding incidence rate were observed.Reslts There was no statistically significant difference between the treatment group and control group in the total effective rate (x2 =0.475,P ＞ 0.05.Meanwhile,no significant differences were found between the two groups in cardiovascular events at 4 d,7 d and 30 d (x2 =0.257,0.475 and 0.317,P ＞0.05).The incidence rate of bleeding in treatment group was obviously lower than that in control group and there was statistically significant difference (2.9％ vs.31.4％ ; x2 =20.115,P ＜0.01).Conclusion The effectiveness of fondaparinux used in elderly patients with non-ST-segment elevation acute coronary syndromes is similar with LMWH,but the incidence rate of bleeding is lower than LMWH.
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Objective To research the consistency of testing results with three different antimajor histocompatibility complex class Ⅰ-related chain A(MICA) specific antibody reagents in order to evaluate their clinical application's value.Method An collaborative study of 18 laboratories was undertaken at the 16th International HLA and Irnmunogenetics Workshop.Total of 16 sera(4 batchs)were tested for anti-MICA antibodies by Luminex method with three different reagents (Kit-A,-B and -C).Result Anti-MICA antibodies were found in 15 sera,except one sera(no.S04) ; No.S10 sera showed positive results in all the laboratories.The anti-MICA antibodies were divided into MICA-G1 group (MICA01,02,07,12,17 and 18) and MICA-G2 group (MICA 04,06,08/27,09 and 19).MICA-G1 group specific antibodies were detected in 5 sera with Kit-A and-B reagent; but there were false-positive results of anti-MICA08/27 and MICA19 antibodies in this 5 sera with Kit-C.MICA-G2group specific antibodies can be detected in other 5 sera with Kit-A and-B,But the MICA specific antibodies testing gave different results with Kit-A,-B and-C in all the last 5 sera samples.Testing of MICA08/27 showed highest consistency results (86.67％,13/15) with Kit-A,-B and-C; and testing of MICA19 showed lowest consistency results (40％,6/15) with this 3 reagents.There were 80％ consistency results of anti-MICA specific antibodies in 13 sera with Kit-B.Conclusion There are the same effect to judgment positive or negative result for anti-MICA antibodies with 3 different reagents,but the results of anti-MICA specific antibodies are not the same.Therefore,it's better to use two or more reagents to test anti-MICA specific antibodies,or choose reagent with wide detection range.
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Objective To investigate whether soluble lectin-like oxidized low-density lipoprotein receptor-1 (sLOX-1) as the early diagnostic biomarker of acute ST elevation myocardial infarction (STEAMI).Methods Sixty-five patients with STEAMI and 30 patients with stable coranary heart disease or other heart disease(control group) were enrolled as our subjects.Serum sLOX-1 levels were measured.Results The median(P25,P75) of Serum sLOX-1 in the patients with STEMI were 210.0 (130.0,356.0) ng/L,significantly higher than that of control group(65.5 (55.2,85.2) ng/L,Z =6.17,P ＜ 0.001).Logistic regression analysis revealed that sLOX-1 alone was an independent factor associated with STEAMI (B =0.036,P ＜ 0.001).The area under the ROC curve of sLOX-1 for detecting STEMI was 0.895,and 95％ CI was 0.831-0.959 (P＜0.001).Taking sLOX-1 =87.5 ng/L as cut-off value,the sensitivity was 89.6％ and specificity was 82.4％for the diagnosis of STEAMI.Conclusion Serum sLOX-1 was significantly higher in the STEAMI and it might served as the early diagnostic marker for STEAMI.
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Objective To compare the short-term prognostic value of glycohemoglobin (HbA1c) and admission plasma glucose in non-diabetic patients with acute ST-elevation myocardial infarction.Methods Eighty-four non-diabetic patients with acute ST-elevation myocardial infarction from January 2010 to June 2011 were included.Both HbA1c and plasma glucose was measured on admission.Cardiovascular event was followed up in 30 days.Results The average of HbA1c and admission plasma glucose was as cut-off point.The patients were divided into HbA1c ＜ 5.5％ group (40 cases) and HbA1c ≥5.5％ group (44 cases) according to HbA1c level.The patients were divided into admission plasma glucose ≤ 8.6 mmol/L group (42 cases) and admission plasma glucose ＞ 8.6 mmol/L group (42 cases) according to admission plasma glucose.The incidence of cardiovascular event in 30 days in admission plasma glucose ＞ 8.6 mmol/L group was higher than that in admission plasma glucose ≤ 8.6 mmol/L group [19.0％ (8/42) vs.2.4％ (1/42)],and there was significant difference(P＜ 0.05).There was no significant difference in the incidence of cardiovascular event in 30 days between HbA1c ≥5.5％ group and HbA1c ＜ 5.5％ group (P ＞ 0.05).Admission plasma glucose showed weak correlation with blood creatine kinase isoenzyme MB peak (r =0.233,P ＜0.05).Conclusion In non-diabetic patients with acute ST-elevation myocardial infarction,elevated admission plasma glucose levels are associated with higher cardiovascular event in 30 days.
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Objective To explore the influence of the killer cell immunoglobulin like receptor (KIR) gene polymorphism on cytomegalovirus (CMV) infection and pathogenesis after hematopoietic stem cell transplantation (HSCT).Methods The KIR genotype was determined by sequence-specific primer polymerase chain reaction (PCR-SSP) in 138 pairs of donors and recipients before HSCT during October,2005 and May,2011.Posttransplant monitoring for CMVpp65 antigen was performed by indirect immune histochemically assays since week 2 after transplantation.The differences between CMV positive group and negative group,inhibitive and active KIR of donors and recipients,and KIR haplotype frequency of donors and recipients were analyzed.Results There were no significant differences in frequency of KIR gene and haplotype AA,AB,BB between the donors and recipients.The frequencies of 2DS2 and 2DS4 * 003-007 of donors in CMV positive group were obviously lower than those in CMV negative group with significant differences(8％ vs 16％,P =0.0420;3％ vs 13％,P =0.0050).There was no significant difference in KIR gene between CMV positive group and CMV negative group.The CMV infection rates of haplotype AA,BB,AB donors were 64.38％,36.84％ and 50.00％,while CMV infection rates of haplotype AA,BB,AB recipients were 53.73％,46.15％ and 51.72％,respectively.The CMV infection rate was higher in the patients received KIR haplotype AA donor than in those received KIR haplotype BB donor (36.84％ vs 64.38％,P =0.0299).2DS4 * 003-007 and haplotype BB of donor were found associated with CMV infection in multifactor analysis.Conclusion KIR genotypes of donors are associated with CMV infection after HSCT.
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Objective To investigate the prediction of anti-human leukocyte antigen antibodies (HLA) and anti-major histocompatibility complex class I-related chain A antibodies (MICA) to the development of acute rejection (AR) and kidney allograft function. Methods Forty-one kidney transplant patients were prospectively tested for anti-HLA and anti-MICA. Thirty-seven patients were screened using Luminex/single-antigen beads to determine the HLA and MICA-specific antibody levels at 0,30,90, 180,360,720 and 1080 days post-transplantation. The patients and donors of HLA and MICA allele typing were determined by PCR-SSOP, and donor specific antibody (DSA) and non-donor specific antibody (NDSA) were identified.Simultaneously,their serum creatinine (SCr) levels and clinical data were analyzed. Results Nine patients (21.95 % ,9/41 ) had pre-existing anti-HLA and(or) anti-MICA, including 6 cases of anti-MICA,2 cases of anti-HLA, and one case of anti-MICA and anti-HLA. Nine patients had pre-existing DSA and NDSA. In the 37 patients, 6 patients (16.2% ) developed de novo anti-HLA, and 3 (8.1%) developed de novo antiMICA. In patients positive for de novo anti-HLA, the titer of antibody was gradually increased during the follow-up of three years. Four patients out of 9 patients with pre-existing antibodies were suffered from AR (44.4%); In 6 patients positive for de novo anti-HLA,three cases (50.0%) were suffered from AR; In three patients positive for de novo anti-MICA,no AR occurred (P＜0.05). In two patients positive for DSA of HLAⅡ antibody detected at the third and seventh day after transplantation, the renal grafts were renovecd due to rejection. The Scr levels in patients positive for pre-existing MICA with AR were higher than in those positive for pre-existing MICA without AR at each scheduled time point during the follow-up period (P＜0.05). The Scr levels in patients negative for antibodies pre-transplantation and having AR were higher than in those having no AR at each scheduled time point during the follow-up period (P＜0. 01 ). The Scr levels in patients positive for de novo HLA and MICA and having AR one month following transplantation were higher than in those negative for antibodies and having no AR (P＜0.01 ). Conclusion Pre-existing and de novo anti-HLA were the irnportant factors for the development of AR, but the mismatch of HLA and MICA alleles in donors and patients was primary causes for generation of de novo antibodies.
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Objective To investigate the effect of KIR-HLA receptor-ligand model on the unrelated allo-hematopoietic stem cell transplantation (Allo-HSCT) of acute lymphoblastic leukemia (ALL). Methods The KIR genotype of 23 pairs of ALL patients and their HLA-matched unrelated donors obtained from the Database of China Marrow Donor Program. KIR genotype was determined using PCR-SSP. The expression of inhibitory KIR(iKIR) was determined by flow cytometry analysis on recipients after HSCT. Results Among all 23 donor/recipient pairs, 17 donors with KIR2DL2/L3 could find corresponding HLA-Cw1, 3, 7, 8, 12, 14 ligands in their recipients. Six donors with KIR2DL1 could match with HLA-Cw6, 15 in recipients. Sixteen donors with KIR3DL1 could recognize HLA-Bw4 and 12 donors with 3DL2 could find HLA-AI1 in their corresponding recipients, respectively. Ninteen patients were successfully transplanted, and the death rate of transplantation were 33.3% (2/6)and 40.0% (2/5) in KIR receptor-ligand matched model and the graft versus leukemia(HVG) KIR ligand-mismatching pattern. The frequency of acute graft versus host disease(GVHD) was 50.0% and death rate was 12.5% (1/8) in GVH KIR ligand-mismatching. The incidence rate of activated GVHD(aGVHD) was 20.0% in the HVG KIR ligand-mismatching. Five donor/recipient pairs of KIR gene typing were the KIR-haplotype A, 2 donor/recipient pairs with KIR2DS4 * 001/002 were died, 3 donor/recipient pairs with KIR2DS4 * 003-007 were obtained the disease free survival. The expression of CD158a/2DL1 was low when the patient had no aGVHD, but became much higher when aGVHD occurred. The percentage of NK cell of the patients was decreasing since transplantation, but still higher than normal after HSCT[ (23.4 ± 3.8 ) % vs (2.04 ± 0.58) %, P < 0.05 ]. Conclusion Analysis on KIR-HLA gene loci pattern may provide a useful parameter in predicting the clinical outcome of HLA-matched unrelated allogeneic hematopoietic stem cell transplantation for leukemia patients. Moreover, it may help to increase overall survival and disease free survival after HSCT by preventing the development of GVHD.
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Objective To study the influence of human leucocyte antigen(HLA) and major his-tocompatibility complex class Ⅰ chain-related gene A (MICA) specific antibodies on renal allograft function and graft rejective reaction by monitoring their changes from preoperative to postoperative pe-riods. Methods Twenty-seven patients with renal aliografts were tested with the specificity of anti-HLA antibodies (anti-HLA class Ⅰ and anti-HLA class Ⅱ) and anti-MICA antibodies and their posi-tive value changes by flow PRATM beads. The HLA genotype was integrated to distinguish donor specific antibody(DSA) and non-donor specific antibody(NDSA). Their serum creatinine levels and clinical data were analyzed simultaneously. Results Of the 27 patients, 22 cases accepted renal transplantation from dead bodies and 5 eases accepted from live donors. Except 1 failed patient, the other 26 patients had good functional renal allografts. Twenty-four survival patients were followed up on month 1, 3, 6 and 12 after transplantation. Seven out of 27 patients had pre-exist antibody before transplantation. Among them, 2 patients had anti-HLA antibody; 3 patients had anti-MICA antibody; 2 patients had both anti-HLA and anti-MICA antibody. Three patients with no anti-HLA and anti-MICA antibodies before transplantation created antibodies after transplantation from 3 to 6 months. One patient created NDSA after transplantation and appeared chronic rejection. There were 3 patients who had anti-MICA antibodies before transplantation. The expression levels of antibodies had changed from high to low, but the specific anti-MICA antibody had not changed during the follow-up on month 1, 3, 6 and 12 after transplantation. The patient with pre-transplantation low level of anti-HLA class Ⅱ antibody appeared acute rejection with fever and his CMV was positive as well. The patient's SCr levels changed from 171 μmol/L to 236 μmol/L after I to 3 months post-transplantation. Twenty-four patients were divided into positive and negative groups according to the specific antibody. There was significant difference of SCr levels between the 2 groups 1 month and 1 year after transplantation(P= 0.03, 0.05). Conclusions It is important to detect the specificity and positive value of anti-HLA antibodies and anti-MICA antibody regularly during the post transplantation follow-up. This will make an effective therapy for decreasing the occurrenee and development of acute or chronic rejection and hy-pofunction on renal allograft.