非HLA基因在异基因造血干细胞移植中的研究进展

GVHD和感染等移植后并发症仍是异基因造血干细胞移植中的主要困难。目前许多研究对GVHD、复发和死亡率进行了非HLA的基因预测。本文讨论了与异基因移植相关的非HLA基因组与造血干细胞移植后并发症的关系与临床意义。     

异基因造血干细胞移植研究进展

异基因造血干细胞移植已成为治愈血液系统恶性肿瘤、骨髓衰竭性疾病、某些先天性及代谢性疾病等的重要方法。本文就外周血干细胞移植（PBSCT）、脐血造血干细胞移植（CBSCT）、非清髓性干细胞移植（NST）、移植物抗宿主病（GVHD）与移植物抗白血病（GVL）效应以及间充质干细胞（MSC）在异基因造血干细胞移植中的应用等方面的新进展进行介绍。     

自体及非清髓异基因造血干细胞移植治疗多发性骨髓瘤一例

异基因造血干细胞移植是目前可以治愈多发性骨髓瘤(MM)的方法之一。为了降低治疗相关死亡率，减少复发，我们对1例多发性骨髓瘤患者进行了2次移植。第1次为自体外周血造血干细胞移植，第2次为非清髓性异基因外周血造血干细胞移植。现报告如下。     

高危急性髓系白血病异基因造血干细胞移植后复发的防治

急性髓系白血病（AML）是一组高度异质性的克隆性疾病,化学药物治疗和造血干细胞移植均为治疗AML的方法。对于高危AML患者而言,异基因造血干细胞移植为治疗该疾病的有效手段,但部分AML患者造血干细胞移植后仍可能面临疾病复发的问题,大多数复发患者再行化学药物治疗、二次移植等的效果不佳,是导致患者异基因造血干细胞移植后死亡的主要原因。因此,加强对异基因造血干细胞移植后AML患者的随访,并采取一些合适的手段预防移植后复发显得尤为重要。本文就高危AML患者异基因造血干细胞移植后复发的监测、药物治疗和细胞治疗进行综述,以期为改善高危AML患者异基因造血干细胞移植预后提供参考。     

用于异基因移植的造血干细胞来源

造血干细胞移植(hcmatopoiedc stem cell transplantation，HSCT)的临床应用已有近50年的历史，目前已成为治疗多种良恶性血液病、实体肿瘤、遗传性疾病和重症自身免疫性疾病等的有效手段。本文试就异基因造血干细胞移植中应用的不同来源造血干细胞(hematopoietic stemcell，HSC)的特点及其对移植效果的影响作一评述。     

异基因造血干细胞移植治疗慢性活动性EB病毒感染患者的护理

目的总结异基因造血干细胞移植治疗成人慢性活动性EB病毒感染的护理经验。方法回顾性分析7例成人慢性活动性EB病毒感染患者异基因造血干细胞移植的治疗及护理方法。结果 5例患者获得完全缓解,2例死亡。结论慢性活动性EB病毒感染患者病情严重,治疗难度大且预后不佳,异基因造血干细胞移植可改善患者预后。做好患者治疗期间的情绪管理、症状管理及并发症防护是护理的关键。     

肾细胞癌的异基因造血干细胞移植治疗进展

近年来研究发现,异基因造血干细胞移植后可以产生抗肿瘤作用,这种作用对耐细胞因子的肾细胞癌也有很好的疗效,为肾细胞癌的治疗提供了一种新的方法。本文就异基因造血干细胞移植治疗肾细胞癌的研究进展作一综述。     

异基因外周血造血干细胞移植治疗多发性骨髓瘤二例

多发性骨髓瘤(MM)为浆细胞恶性克隆增生性疾病,传统化疗和自体造血干细胞移植虽可使疾病得到暂时缓解,但最终不免复发和进展,患者的中位存活时间仅3～4年。异基因造血干细胞移植是目前惟一可治愈该病的方法,我院采用外周血造血干细胞移植治疗多发性骨髓瘤2例,报告如下。     

ABO血型不合异基因造血干细胞移植

目的 探讨ABO血型不合异基因造血干细胞移植(Allo-HSCT)的疗效及并发症。方法 6例患者中1例ABO血型主要不合，5例次要不合；2例脐血，1例外周血干细胞，3例骨髓联合外周血干细胞；对主要不合者骨髓用重力沉降法去除红细胞，对次要不合骨髓以离心分离方法去除血浆，外周血造血干细胞及脐血未经处理直接输注。结果 6例均成功植入，仅在输注干细胞时发生短暂轻微溶血，而无延迟性溶血反应及移植后红系造血延迟。结论 ABO血型不合可顺利进行Allo-HSCT，获得干细胞的植入，移植前后防止溶血发生至关重要。     

异基因造血干细胞移植患儿营养状况的纵向调查

目的了解异基因造血干细胞移植患儿营养状况的变化。方法分别于移植前、移植后30d、60d、100d对89例异基因造血干细胞移植患儿进行身高、体质量、体质指数(BMI)、三头肌皮褶厚度(TSF)、中上臂围(MUAC)、腰围(WC)等人体学测量,同时测量脂肪组织(FM)、体脂百分比(%BF)、去脂组织(FFM)及去脂组织百分比(%FFM)等人体成分,并进行生化指标白蛋白(ALB)、前白蛋白(PreALB)测量,比较异基因造血干细胞移植患儿营养状况的改变。结果人体学测量结果显示,在移植后100d内患儿体质量、BMI、TSF均呈现先降后升趋势,在移植后30d降至最低点,然后逐渐上升(P0.05,P0.01),WC在移植后100d内持续升高(P0.01);人体成分测量结果显示,%BF、FM在移植后60d内明显升高(均P0.05),%FFM在移植后60d内显著降低(P0.01);生化测量结果显示,ALB呈先升后降再上升的趋势,PreALB在移植后60d内显著上升(P0.01)。结论在异基因造血干细胞移植后30d内,多数人体学及人体成分测量指标下降;而在移植后60d则存在FFM丢失和FM不断蓄积的现象。不建议将ALB和PreALB作为评价造血干细胞移植患儿营养状况的可靠指标。     

Molecular genetic analysis of interleukin-1 promoter and receptor antagonist tandem repeat polymorphisms among HLA-identical renal transplant recipient and donor pairs

The cytokine gene polymorphisms as well as the minor histocompatibility antigen (mHAg) disparities have been thought to result in transplant graft rejection and graft-versus-host reaction between HLA-identical donor/recipient pairs. We detected interleukin (IL)-1 promoter and receptor antagonist tandem repeat polymorphisms among 21 HLA identical renal transplant recipient and donor pairs; of 21 pairs, 15 (71%) showed at least one locus difference between donor and recipient, and 73% of the pairs with the disparities at these loci showed either recipient to donor (R-->D) or donor to recipient (D-->R) positive CD4+ cell response measured by a modified Cylex assay developed in our laboratory. Allele 2 of IL-1 receptor antagonist genotype (IL-1ra VNTR), a gene associated with an increasing IL-1beta production, was detected in the three recipients who developed rejections. One HLA-identical pair that had variations on all four loci of IL-1 gene cluster consistently showed positive CD4+ cell immune responses between the donor and the recipient. This study may provide information of the role of non-HLA genes participating in allograft rejection; this demonstrates that in addition to the disparities of the mHAgs, the non-HLA antigens have to be considered as risk factors in HLA-identical solid organ transplantation.     

Effects of non-HLA gene polymorphisms on development of islet autoimmunity and type 1 diabetes in a population with high-risk HLA-DR,DQ genotypes

We assessed the effects of non-HLA gene polymorphisms on the risk of islet autoimmunity (IA) and progression to type 1 diabetes in the Diabetes Autoimmunity Study in the Young. A total of 1,743 non-Hispanic, white children were included: 861 first-degree relatives and 882 general population children identified as having high-risk HLA-DR/DQ genotypes for type 1 diabetes. Of those, 109 developed IA and 61 progressed to diabetes. Study participants were genotyped for 20 non-HLA polymorphisms, previously confirmed as type 1 diabetes susceptibility loci. PTPN22 and UBASH3A predicted both IA and diabetes in regression models controlling for family history of type 1 diabetes and presence of HLA-DR3/4-DQB1*0302 genotype. In addition, PTPN2 predicted IA whereas INS predicted type 1 diabetes. The final multivariate regression models for both IA and type 1 diabetes included PTPN22, UBASH3A, and INS, in addition to family history of type 1 diabetes and HLA-DR3/4. In general population children, the most frequent combinations including these five significant predictors conferred hazard ratio of up to 13 for IA and >40 for type 1 diabetes. Non-HLA susceptibility alleles may help estimate risk for development of type 1 diabetes in the general population. These findings require replication in different populations.     

TNF-alpha, IL-6, IFN-gamma, and IL-10 gene expression polymorphisms and the IL-4 receptor alpha-chain variant Q576R: effects on renal allograft outcome.

BACKGROUND: There has been much interest recently in the effects of various cytokine gene expression polymorphisms on graft outcome. However, the results of these investigations reveal the outcomes to be organ-specific and center-specific. We sought to confirm and add to some of the earlier findings by studying the impact of tumor necrosis factor-alpha (TNF-alpha), interleukin-10 (IL-10), interferon-gamma (IFN-gamma), and interleukin-6 (IL-6) polymorphisms and the interleukin-4 (IL-4) receptor alpha-chain variant on posttransplant renal allograft outcome. METHOD: TNF-alpha, IL-6, IFN-gamma, and IL-10 gene promoter region polymorphisms were assayed genotypically by PCR-SSP on 120 patients transplanted at the Albany Medical Center. These patients were also typed for the IL-4 receptor alpha-chain variant Q576R. RESULTS: Producers of high levels of the proinflammatory cytokine TNF-alpha were found to be at increased risk for acute rejection episodes if the allograft was mismatched for the molecular products of the class II region of the human major histocompatibility complex (HLA-DR). Expression level polymorphisms of the IL-6, IFN-gamma, and IL-10 genes were not associated with acute rejection episodes, nor was the IL-4 receptor alpha-chain variant Q576R. CONCLUSIONS: These data would suggest that the production of high levels of the cytokine TNF-alpha is especially detrimental to graft survival when the recipient's T-helper lymphocytes are being activated by mismatched donor HLA-class II antigens. Typing all potential kidney recipients for TNF-alpha, and providing well-matched organs for high producers of this cytokines, may be expected to increase rejection-free graft survival in these patients.     

Non-HLA immunogenetic polymorphisms and the risk of complications after allogeneic hemopoietic stem-cell transplantation

BACKGROUND: Existing data indicate that non-human leukocyte antigen (HLA) immunogenetic polymorphisms influence the risk of complications after allogeneic hemopoietic stem-cell transplantation. However, prior studies have been limited by small sample size and limited genotyping. METHODS: We examined 22 polymorphisms in 11 immunoregulatory genes including cytokines, mediators of apoptosis, and host-defense molecules by polymerase chain reaction using sequence-specific primers in 160 related myeloablative transplants. Associations were confirmed in two independent cohorts. RESULTS: An intronic polymorphism in the tumor necrosis factor gene (TNF 488A) was associated with the risk of acute graft-versus-host disease (GVHD) (odds ratio [OR] 16.9), grades II to IV acute GVHD (OR 3.3), chronic GVHD (OR 12.5), and early death posttransplant (OR 3.4). Recipient Fas -670G and donor interleukin (IL)-6 -174G were independent risk factors for acute GVHD. Recipient IL-10 ATA and Fas -670 genotype were independent risk factors for chronic GVHD. Recipient IL-1beta +3953T was associated with hepatic acute GVHD, and Fas -670G was associated with major infection. CONCLUSIONS: These results highlight the potential importance of cytokine and apoptosis gene polymorphisms in stem-cell transplantation, and indicate that non-HLA genotyping may be useful to identify individuals at the highest risk of complications and new targets for therapeutic intervention.     

Donor genomics influence graft events: the effect of donor polymorphisms on acute rejection and chronic allograft nephropathy

BACKGROUND: Organs procured from deceased donors emanate from individuals with diverse genetic backgrounds. Donor organs, therefore, may vary in their response to injury and immune stimuli in a genetically determined manner. We assessed polymorphisms from 244 renal allograft donors to better understand the impact of donor polymorphisms on selected transplant outcomes. METHODS: Donor genomic DNA restriction fragment length polymorphisms were assayed for evidence of common cytokine [interleukin (IL)-2, IL-6, IL-10, tumor necrosis factor (TNF)-alpha, TGF-beta, interferon (IFN)-gamma] and chemokine (CCR2, CCR5) polymorphisms. Associations between donor polymorphisms and graft events were determined using chi-square, linear regression, and Kaplan-Meier analyses. RESULTS: Several genotypic polymorphisms demonstrated a modest association with acute rejection, including the transforming growth factor (TGF)-beta T/C codon 10 (P= 0.027) and the CCR5 G/A 59029 (P= 0.039) genes by chi-square analysis. Notably, the presence of the T allele in the IFN-gamma gene (+874) demonstrated a highly significant association with biopsy-proven chronic allograft nephropathy (P < 0.008). This association remained highly significant in a multiple linear regression model that incorporated biopsy-proven acute rejection as a covariate. CONCLUSION: These data suggest that many of the donor polymorphisms studied in this analysis may influence a recipient's immune response to a renal allograft. However, their greatest impact may be demonstrated in long-term outcomes.     

Molecular analysis of adrenergic receptor genes and interleukin-4/interleukin-4 receptor genes in patients with interstitial cystitis

PURPOSE: Interstitial cystitis is a chronic and sterile inflammatory disease of the bladder. Clinically many patients with interstitial cystitis also have allergic or autoimmune diseases. Recent studies have suggested that allelic variations in the adrenergic receptor beta2 gene, the interleukin (IL)-4 gene and its receptor gene are closely associated with immune diseases. Therefore, we studied adrenergic receptor genes alpha1d (ADRA1d), beta2 (ADRB2) and beta3 (ADRB3), and IL-4/IL-4 receptor genes in patients with interstitial cystitis. MATERIALS AND METHODS: We studied 55 patients with interstitial cystitis and 228 controls. Genomic DNA was extracted from a whole blood sample collected from each subject. We analyzed 5 gene polymorphisms (G404781T, Arg16Gly, Trp64Arg, C589T and Ile50Val) per gene (ADRA1d, ADRB2, ADRB3, IL-4 and IL-4 receptor, respectively) and their relationship to interstitial cystitis. RESULTS: The frequency of the Arg/Arg genotype of the ADRB2 gene and the TT genotype of the IL-4 gene were significantly higher in patients with interstitial cystitis than in controls. A significant difference of ADRA1d genotype prevalence was also observed. However, there were no remarkable differences of IL-4 receptor or ADRB3 genotype prevalences. CONCLUSIONS: Variants of the ADRB2, ADRA1d and IL-4 genes may be related to a predisposition to interstitial cystitis.     

Influence of cytokine genes polymorphisms on long-term outcome in renal transplantation

BACKGROUND: Recently, polymorphisms of cytokine genes have been associated with modified gene expression and increased cytokine production. We evaluated the influence of interleukin-10 (IL-10) gene G-1082A, tumour necrosis factor alpha (TNFalpha) gene G-308A and IL-6 gene G-174C polymorphisms on the rejection rate, renal function and long-term outcome in renal transplantation. PATIENTS AND METHODS: We studied n = 224 consecutive patients, who underwent renal transplantation at our centre from 1998 to 2001 (cadaveric: n = 175, living related: n = 49) followed up for 4.9 +/- 2.0 yr and n = 100 healthy volunteers. IL-10 gene G-1082A, TNFalpha gene G-308A and IL-6 gene G-174C polymorphisms were determined by polymerase chain reaction (PCR) amplification. RESULTS: The genotype distribution of the investigated polymorphisms was similar in patients and controls (ns). The age of donor and the recipient, the number of HLA mismatches and cold and warm ischemic time did not differ among patients with different genotypes (ns). No association between cytokine polymorphisms and the incidence of acute rejection episodes was detected (ns). The cytokine genotypes did not correlate with serum creatinine or creatinine clearance at any time during follow up (ns). Furthermore, there was no significant difference in the genotype frequencies among patients experiencing graft failure (ns). Patients with different cytokine gene polymorphisms showed similar outcomes in the Kaplan-Meier analysis of graft survival (ns). Finally, cytokine polymorphisms had no influence on the acute rejection rate or graft outcome also in the subgroup of HLA-DR mismatched grafts (ns). CONCLUSION: Our results suggest that IL-10 gene G-1082A, TNFalpha gene G-308A and IL-6 gene G-174C polymorphisms are no major risk factors in renal transplantation.     

Interleukin-1β gene and receptor antagonist gene polymorphisms in patients with calcium oxalate stones

Interleukin-1 (IL-1) might play a role in the process of bone loss and hypercalciuria and is therefore considered to be involved in the formation of urinary stones. The aim of this study is to test whether the IL-1beta promoter region, exon 5 region and IL-1 receptor antagonist gene intron 2 polymorphisms could be genetic markers for the susceptibility to the formation of urinary stones. A control group of 152 healthy people and a group of 105 patients with recurrent calcium oxalate stone were examined in this study. Polymerase chain reaction (PCR) analyzed the variable number tandem repeats at intron 2 of the IL-1Ra gene for the polymorphisms. PCR-based restriction analysis was done for the IL-1beta gene polymorphisms of the promoter region and exon 5 by the endonucleases Ava I and Taq I, respectively. The polymorphisms studied in the IL-1beta genes did not reveal a strong association with calcium oxalate stone disease when compared with the control group (promoter region by chi-square test, P=0.627: exon 5 region by Fisher's exact test, P = 0.403). Only two frequent alleles of the IL-1Ra gene corresponding to one and two copies of an 86-bp sequence repeat were identified by PCR. The result revealed significant differences between control individuals and stone patients (P < 0.01. Fisher's exact test). In addition, the frequency of the type I allele in the stone group (99.0%) was higher than in the control group (94.0%). The odds ratio for the type I allele of the IL-1Ra gene in calcium oxalate stone disease is 6.041 (95% CI: 1.683 approximately 21.687). There is an association between urolithiasis and polymorphism in the IL-1Ra gene. No significant difference was found when dividing the stone patients into groups with normocalciuria and hypercalciuria in relation to these genetic polymorphisms. Further studies of the type I allele of the IL-IRa gene are worthwhile because of its correlation with stone disease. In our study, neither the IL-1beta promoter region nor the exon 5 polymorphisms were significantly different when comparing control subjects and calcium oxalate stone patients.     

供者细胞因子基因多态性对肾移植受者急性排斥反应发生的影响

目的 研究供者的细胞因子和细胞因子受体基因多态性对肾移植受者急性排斥反应发生的影响.方法 (1)将126例肾移植受者分成急性排斥组和无排斥组,比较可能影响发生急性排斥反应的因素在两组中的分布情况;比较两组受者中供者的13种细胞因子及受体22个位点的基因型及部分细胞因子表达型的分布情况.(2)根据HLA-DR配型分成0～1个HLA-DR位点错配、HLA-DR完全错配两种情况,分别比较阳性基因多态性在急性排斥组和无排斥组中的分布情况.结果 (1)急性排斥组的HLA-DR错配数明显高于无排斥组;无排斥组供者的白细胞介素(IL)-1α889 C/C、IL-1Ra raspI 11100 T/T、IL-4Rd+1902 A/A、转化生长因子(TGF)-β1密码子10 C/C、IL-10-1082A/A、IL-10低表达型的频率明显较高,IL-12～1 188 A/A、IL-2-330 G/G、IL-10 GCC/ATA频率明显较低;(2)HLA-DR 0～1个位点错配时,两组供者中IL-1Rα msp I 11100 T/T、IL-4Rα+1902 A/A、TGF-β1密码子10 C/C、IL-2-330 G/G、IL-10低表达型频率明显不同,而在HLA-DR完全错配时,只有IL-12-1188 A/A表达频率明显不同.结论 供者的IL-1α-889 C/C、IL-1Rα msp I 11100 T/T、IL-4Rα+1902 A/A、TGF-β1,密码子10 C/C、IL-10-1082 A/A和IL-10低表达型是不发生肾移植急性排斥反应的遗传学指标,而IL-12-1188 A/A、IL-2-330 G/G则是发生急性排斥反应的遗传学危险因素.HLA-DR错配状况可干扰供者细胞因子基因多态性对急性排斥反应发生的影响.     

Influence of cytokine gene polymorphisms on graft rejection in Turkish patients with renal transplants from living related donors

Background

Certain cytokine gene polymorphisms (CGPs) have been shown to be associated with renal transplant rejection episodes or graft outcomes. We sought to evaluate the relationships between gene polymorphisms and acute rejection episodes (RG, n = 19) versus stable graft function (NRG, n = 71) in transplant recipients compared with healthy control subjects (HCG, n = 150). The follow-up time period was 18 months. Using polymerase chain reaction sequence-specific primers with the Heidelberg kit we genotyped 22 single nucleotide polymorphisms distributed across 13 cytokine and cytokine receptor genes.

Results

Interleukin (IL)-2 TT/GT haplotype was found in 36.8% of RG patients and 6.7% of HCG but not among the NRG (P < .0001; .0007). The IL-2 GG/TT haplotype was observed among 13 NRG and nine HCG patients (P = .007); the IL-2 GG/GG haplotype, 18.7% HCG and 4.2% NRG patients (P = .0033); and the IL-2 TT/TT haplotype, five NRG and eight HCG patients, but none of the RG cohort (P > .05). The transforming-growth factor-beta 1 CG/CC haplotype was noted in 15 NRG (21.1%) and four HCG but no RG patients (P < .0001). The IL-2 +166 GT genotype was detected in 36.8% of RG, 8.5% of NRG, and 14.7% of HCG patients (P = .005, .0244). The IL-2 −330 GG genotype was demonstrated in 32 healthy controls and three nonrejection transplant patients (P = .0007). Significant differences were concluded between NRG and HCG for IL-6 565 AG, IL-1beta −511 TT and +3962 CC/CT/TT genotypes.

Discussion

We observed significant differences among the frequencies of IL-2 gene polymorphisms among RG and NRG subjects, which agreed with previous clinical, but not in vitro studies.