白细胞介素1B和1RN基因多态性与骨关节炎易感性的关系

目的在中国青岛地区膝骨性关节炎患者中,探讨白细胞介素-1B(-31C/T、-511C/T、＋3954C/T)及其拮抗基因IL-1RN基因多态性与骨关节炎易感性的关系。 方法采用病例对照研究,对就诊青岛市胶州中心医院经临床诊断为膝关节骨关节炎的216例患者和233例年龄、性别匹配的健康对照者,排除既往有膝关节外伤史、类风湿性关节炎及合并其他内科疾病的患者,以聚合酶链反应和限制性片段长度多态性(PCR-RFLP)方法检测IL-1B的－31C/T、－511C/T、＋3954 C/T位点及IL-1RN的多态性,采用logistic回归分析方法比较不同基因型与骨关节炎发病风险的关系。 结果骨关节炎组中IL-1B-511CT、IL-1B＋3954CT基因型频率明显高于对照组,与对照组差异有统计学意义(IL-1B-511CT χ²＝2.034,P＝0.026;IL-1B＋3954CT χ²＝5.526,P＝0.017),IL-1B＋3954C/T位点未发现TT基因型,通过logistic回归分析,结果显示与野生型的纯合子相比,杂合子-511CT、＋3954CT基因型与骨性关节炎高风险发病率具有相关性[IL-1B-511CT,OR＝1.89,95%CI(0.56,1.39),P＝0.026;IL-1B＋3954CT,OR＝2.51,95%CI(1.18,5.35),P＝0.017]。骨关节炎组和对照组在IL-1B-31C/T位点及IL-1RN基因型分布未见明显差异。 结论IL-1B-511TC、＋3954CT基因型人群可能增加对膝骨关节炎的易感性,而IL-1B-31C/T, IL-1RN*2基因多态性可能与膝骨关节炎遗传易感性不具相关性。     

The impact of donor and recipient diabetes on renal transplant outcomes

The use of diabetic kidneys is increasing worldwide with better outcome than being on waitlist and possible reversal of diabetic changes in transplanted kidneys. But particular caution is warranted in diabetic donor-recipient combination. Total 1223 deceased donor kidney transplants were performed at our center between 2008 and 2018. 689 from non-diabetic donor (NDD) to non-diabetic recipient, 400 from non-diabetic donor to diabetic recipient, 97 from diabetic to non-diabetic recipient, and 32 from diabetic donor (DD) to diabetic recipient. The DD was older than NDDs (median age 48 vs 39 years, P < 0.0001). DD had higher BMI (35.6 vs 26.9, P < 0.0001), higher KDPI (74% vs 37%, P < 0.0001), and higher terminal creatinine (1.10 mg/dl vs 0.95 mg/dl, p 0.0046) than the NDD. Diabetes recipients were comparatively older (57 vs 54, P < 0.001). DD recipients had higher serum creatinine at 6 months (1.70 vs 1.50 mg/dl, p 0.00304) and 2 years post-transplant (1.70 vs 1.50 mg/dl P < 0.0002). DD recipients had more favorable end CPRA than NDD recipients (77.5% at 0% vs 67.4% at 0, P = 0.0074). Ten-year patient and graft survival was best in NDD-recipient pair and worse in DD-recipient pair. Diabetic donor kidneys to diabetic recipients have lower 1-, 3-, and 5-year graft survival.     

The impact of donor and recipient weight incompatibility on renal transplant outcomes

International Urology and Nephrology - Donor/recipient size mismatching and correlation to allograft outcome remains poorly defined. This study assessed the impact of donor body weight (DBW) to...     

Administration of interleukin-1 receptor antagonist ameliorates renal ischemia-reperfusion injury.

Interleukin (IL)-1 is a major contributor to inflammation and apoptosis during ischemia/reperfusion (I/R) injury. Its deleterious effects are primarily mediated by the activation of nuclear factor-kappaB (NF-kappaB). Receptor-binding and signaling of IL-1 can be blocked by the IL-1 receptor antagonist (IL-1ra). The aim of our study was to characterize effects and mechanisms of IL-1ra administration on inflammation, apoptosis, and infiltration in renal I/R injury. Renal ischemia was induced in Lewis rats by clamping of the left renal artery for 45 min. Kidneys were removed for histological and molecular analysis 24 h or 5 days after reperfusion. IL-1ra ameliorated I/R induced renal injury and inflammation. Furthermore, the number of apoptotic tubular cells was lower in IL-1ra-treated animals 24 h after ischemia, which was paralleled by a Bax/Bcl-2 mRNA ratio towards anti-apoptotic effects. IL-1ra reduced the expression of monocyte chemoattractant protein-1 (MCP-1) mRNA at 24 h and 5 days and that of intracellular adhesion molecule-1 (ICAM-1) expression at 24 h in the ischemic reperfused kidneys. Our results indicate that IL-1ra treatment ameliorates renal I/R injury and this protective effect might be mediated by reduced induction of NF-kappaB mediated MCP-1, ICAM-1, and a decreased ratio between Bax and Bcl-2 mRNA expression.     

Association of interleukin-1beta gene and receptor antagonist polymorphisms with calcium oxalate urolithiasis

BACKGROUND AND PURPOSE: Genetic polymorphisms of the interleukin-1beta (IL-1beta) promoter region (-511) and exon 5 +3954 and a variable number of tandem repeats in the IL receptor antagonist (IL-1RA) gene have been proposed as markers for calcium oxalate urolithiasis. Because the prevalence of these polymorphisms could be influenced by racial variation/ethnicity, we explored the association of IL-1 gene-cluster polymorphisms with stone formation in a north India population. PATIENTS AND METHODS: The case-control study involved 150 stone-free control subjects (mean age 46.5 +/- 10.5 years) and 130 patients (mean age 40.0 +/- 11.5 years) with calcium oxalate urolithiasis. Biallelic polymorphisms of two loci, IL-1beta (-511) and IL-1beta (+3954), as well as the penta-allelic variable number of tandem repeats of IL-1RA, were genotyped by polymerase chain reaction-based restriction analysis. Haplotypes were constructed for the IL-1 gene cluster using SNP Analyzer software. RESULTS: We observed a significant association between stone disease and IL-1beta -511 and IL-1RA polymorphisms (P < 0.001 and P = 0.039, respectively), whereas no association was observed for IL-1beta +3954 (P = 0.408). The frequency of the TT (-511) and I/II (410/240; IL-1RA) genotypes was higher in patients than in control subjects (50/130 v 16/150 and 55/130 v 38/150, respectively), whereas the frequencies of the haplotypes were similar (P = 0.485). Significant linkage disequilibrium showed that three genes were strongly linked (P < 0.0001). Patients with a combination of high IL-1beta (-511 and +3954) and low IL-1RA genotypes were at significantly higher risk for urolithiasis (P < 0.001; odds ratio = 5.448, 0.013, and 2.560, respectively). CONCLUSION: Our study demonstrated a strong association of IL-1RA and IL-1beta-511 and suggested that differences in the IL-1 gene cluster could be linked to the risk of urolithiasis. A combination of IL-1beta and IL-1RA associations exhibiting gene-gene interaction further substantiates the finding of risk.     

Genetic association of interleukin-1beta and receptor antagonist (IL-1Ra) gene polymorphism with allograft function in renal transplant patients

Cytokines are known to be important mediators during renal graft outcome. The present study was therefore, conducted to determine the impact of IL-1beta and its receptor antagonist polymorphism on allograft outcome. We evaluated single nucleotide polymorphism (SNPs) in interleukin-1 gene cluster, IL-1beta (promoter region -511 and exon-5 +3954) and IL-1Ra (86-bp VNTR) in 136 renal transplant recipients and 150 normal healthy controls by polymerase chain restriction based (PCR-RFLP) analysis. Recipients were HLA matched and clinically characterized including delayed graft function (DGF), rejection episode (RE) and stable graft function (SGF). Haplotypes and linkage disequilibrium (LD) were determined using SNPAnalyzer software. Significant difference was observed for the frequency distribution of the three sites of IL-1 gene among patients and controls (p<0.001, 0.022 and <0.001 respectively). When RE and DGF were compared to SGF, only IL-1Ra showed significant differences among RE and SGF (p=0.014) and DGF and SGF (p=0.020). The presence of 1/2 genotype showed 18 folds risk in RE and 10 folds in DGF (OR=18.000 and OR=10.667 respectively). The majority of recipients with SGF had 1-4 HLA mismatch whereas RE had 5-8 mismatches. Risk for rejection increased >6 folds (OR=6.571; p<0.01) for 5-8 mismatches. Haplotypes constructed with the combination of three polymorphisms in IL-1 gene cluster showed significant difference between RE and SGF group. LD value for IL-1beta (promoter region) and IL-1Ra and IL-1beta promoter and exon-5 gene in the control group indicated strong association among the variants (D'=0.37, p<0.0001 and D'=0.29, p=0.002). Our study demonstrate that genetically determined low production of IL-1Ra may be a risk factor for RE and DGF and that IL-1beta/IL-1Ra haplotype influences the impact of allograft outcome. These findings may significantly abet in better perception of the survival of the graft.     

Demographics of 353 laparoscopic renal donor and recipient pairs at the Johns Hopkins Medical Institutions

BACKGROUND AND PURPOSE: The demographics of laparoscopic donor nephrectomy (LDN) have not been characterized in detail. The aim of this study was to review our LDN experience with respect to donor and recipient demographic characteristics and trends. PATIENTS AND METHODS: Over a 6-year period, 353 patients underwent LDN. A retrospective chart review was performed to identify the donor and recipient demographic characteristics and trends associated with the procedure. RESULTS: Among the donors, the mean age was 41 years, with a predominance of females (59.2%), whites (76.2%), and blood relations (72%). Siblings were the most common related-donor-to-recipient relationship, and spouses were the most common unrelated relationships. Among the recipients, the mean age was 43 years, with a predominance of males (58.4%), whites (73.7%), and dialysis-dependent patients (55%). Diabetes mellitus and hypertension were the most common causes of end-stage renal failure. With the introduction of laparoscopy, there was a nearly twofold increase in the total number of live renal donations, and there was a significant expansion in the unrelated-donor pool. CONCLUSIONS: Laparoscopic harvest of donated kidneys is associated with new trends that may help alleviate the current organ shortage.     

The impact of donor and recipient common clinical and genetic variation on estimated glomerular filtration rate in a European renal transplant population

Genetic variation across the human leukocyte antigen loci is known to influence renal‐transplant outcome. However, the impact of genetic variation beyond the human leukocyte antigen loci is less clear. We tested the association of common genetic variation and clinical characteristics, from both the donor and recipient, with posttransplant eGFR at different time‐points, out to 5 years posttransplantation. We conducted GWAS meta‐analyses across 10 844 donors and recipients from five European ancestry cohorts. We also analyzed the impact of polygenic risk scores (PRS), calculated using genetic variants associated with nontransplant eGFR, on posttransplant eGFR. PRS calculated using the recipient genotype alone, as well as combined donor and recipient genotypes were significantly associated with eGFR at 1‐year posttransplant. Thirty‐two percent of the variability in eGFR at 1‐year posttransplant was explained by our model containing clinical covariates (including weights for death/graft‐failure), principal components and combined donor‐recipient PRS, with 0.3% contributed by the PRS. No individual genetic variant was significantly associated with eGFR posttransplant in the GWAS. This is the first study to examine PRS, composed of variants that impact kidney function in the general population, in a posttransplant context. Despite PRS being a significant predictor of eGFR posttransplant, the effect size of common genetic factors is limited compared to clinical variables.     

In vitro production of interleukin-1 receptor antagonist in chronic renal failure, CAPD and HD.

Dialysis-related symptoms are believed to be mediated, at least in part, by monocyte/macrophage-derived pro-inflammatory cytokines including interleukin-1 (IL-1) and tumor necrosis factor (TNF). Measuring the production of interleukin-1 receptor antagonist (IL-Ra), a naturally occurring inhibitor of IL-1, opens avenues to study the balance between these two cytokines in patients. We studied the cell content and production of IL-1 beta and IL-Ra by unstimulated and endotoxin- or IgG-stimulated peripheral blood mononuclear cells (PBMC) in undialyzed patients with chronic renal failure (CRF), patients on continuous ambulatory peritoneal dialysis (CAPD) and patients on chronic hemodialysis with reuse cuprophan membranes (HD), and compared them to healthy controls. IL-1 beta and IL-Ra were measured by specific radioimmunoassay. IL-1 beta was undetectable in freshly harvested PBMC from healthy controls, CRF, CAPD or HD. In contrast, the content of IL-Ra in HD patients (2828 +/- 466 pg/ml) was significantly higher than that in healthy controls (643 +/- 53 pg/ml, P < 0.01), CRF (1097 +/- 320 pg/ml, P < 0.01) or CAPD (1398 +/- 390 pg/ml, P < 0.05). In endotoxin-stimulated PBMC, IL-1 beta production by HD patients (9375 +/- 1687 pg/ml) was not significantly different from healthy controls (8429 +/- 1621 pg/ml). However, endotoxin-stimulated IL-Ra production by HD patients (32,350 +/- 8276 pg/ml) was greater than that from healthy controls (11,284 +/- 1250 pg/ml, P < 0.001), CRF (12,263 +/- 2680 pg/ml, P < 0.01) or CAPD patients (11,822 +/- 1797 pg/ml, P < 0.01).(ABSTRACT TRUNCATED AT 250 WORDS)     

Induction of interleukin-1 and interleukin-1 receptor antagonist during contaminated in-vitro dialysis with whole blood

BACKGROUND: Previous studies on the permeability of cellulosic and syntheticdialysers for bacterial-derived cytokine-inducing substancesgave conflicting results. We tried to study this issue as closeto the in-vivo situation as possible. METHODS: An in-vitro dialysis circuit with whole human blood presentin the blood compartment of cuprophane (Cup), polysulphone (PS),and polyamide (PA) dialysers was employed; sterile filtratesderived from Pseudomonas aeruginosa cultures were added to thedialysate. We studied the induction of interleukin-1ß(IL-1ß) by plasma samples taken from the blood compartmentas well as the induction of IL-1ß and interleukin-1receptor antagonist (IL-1Ra) in mononuclear cells separatedfrom whole blood after circulation by radioimmunoassay and polymerasechain reaction. RESULTS: Plasma samples from the blood side of all dialysers inducedIL-1ß from non-circulated mononuclear cells afteraddition of pseudomonas filtrates to the dialysate; the maximalamount of IL-1ß induced by samples from the bloodcompartment was 4.8±1.2 ng/ml for Cup, 1.9±0.5ng/ml for PS, and 2.0±0.6 ng/ml for PA. Mononuclear cellsseparated after contaminated dialysis with all types of dialysersexpressed increased mRNA levels for IL-1ß and IL-1Ra.Production of IL-1Ra by cells separated after contaminated dialysiswas determined after Cup and PS dialysis; there was increasedproduction of IL-1Ra by these cells (Cup, 10.3±4.2; PS,7.3±2.5 ng/ml) compared to cells separated after steriledialysis (Cup, 5.6±2.1, P<0.05; PS, 4.5±1.1ng/ml, n.s.) or from non-circulated blood (Cup experiments,4.7±1.5, P<0.05; PS experiments, 4.1±1.2 ng/ml,n.s.). CONCLUSIONS: These data suggest penetration of cytokine-inducing substancesthrough both cellulosic and synthetic dialysers. Differencesbetween dialysers may exist regarding extent and time courseof penetration. The detection of cytokine mRNA as well as themeasurement of IL-1Ra synthesis is more sensitive substancesthrough dialyser membranes than the measurement of IL-1ßprotein synthesis.     

Tacrolimus dose requirement in relation to donor and recipient ABCB1 and CYP3A5 gene polymorphisms in Chinese liver transplant patients.

The aim of this study was to investigate whether the heterogeneity in tacrolimus dose requirement is associated with ABCB1 and CYP3A5 gene polymorphisms in Chinese liver transplant patients during the first month after transplantation. ABCB1 and CYP3A5 genotyping was performed using the polymerase chain reaction restriction sites polymorphism-based procedure in Chinese liver transplant recipients (n = 50) and their corresponding donors (n = 50). Tacrolimus whole-blood trough concentrations were measured by immunoassays on the IMx analyzers (Abbott Diagnostics Laboratories, Abbott-Park, IL). Doses required to achieve target blood concentrations and dose-adjusted trough concentrations (concentration/dose [C/D] ratios) were compared among patients according to allelic status of ABCB1 and CYP3A5. The ABCB1 3435CC was observed in 23 subjects (23%), whereas 64 (64%) carried 3435CT and 13 (13%) carried 3435TT. The CYP3A5*1/*1 was observed in 13 subjects (13%), 50 (50%) carried *1/*3, and 37 (37%) carried*3/*3. The tacrolimus C/D ratios were obviously lower in recipients carrying ABCB1 3435CC genotype. For CYP3A5, recipients who received organs from CYP3A5*3/*3 donors had higher C/D ratios. But the donors' ABCB1 and recipients' CYP3A5 genotype did not affect the recipients' pharmacokinetics. Analysis of the combination of recipients' ABCB1 and donors' CYP3A5 genotypes revealed that the tacrolimus C/D ratios were significantly lower in the ABCB1 3435CC-carrying recipients, regardless of donors' CYP3A5 genotype. In conclusion, our finding suggests that the recipients' ABCB1 and donors' CYP3A5 genotype affect the tacrolimus dose requirements. ABCB1 C3435T polymorphism is a major determinant of tacrolimus trough concentration in Chinese liver transplant recipients, and recipients with 3435CC genotype will require higher dose of tacrolimus.     

Influence of genetic polymorphisms in GSTM1, GSTM3, GSTT1 and GSTP1 on allograft outcome in renal transplant recipients

Abstract: Introduction: Glutathione S‐transferases (GSTs) are important in protection against xenobiotic compounds and toxicity caused by immunosuppressants in renal transplant recipients. In the present study we hypothesize that genetic variability in GSTM1, GSTM3, GSTP1 and GSTT1 genes may be associated with allograft outcome. Methods: The study included 223 controls and 273 transplant recipients categorized into 184 stable graft function (SGF), 57 rejection episodes (RE) and 32 delayed graft function (DGF). The polymorphism was studied using multiplex PCR and PCR‐RFLP. Results: GSTM1 null genotype showed a 3.35‐fold higher risk for rejection in SGF vs. RE category [95% confidence interval (CI) 1.27–8.84, p = 0.014]. Mutant (G) allele of GSTP1 was associated with a 5.52‐fold risk for DGF (95% CI 1.37–22.17, p = 0.016). Kaplan–Meier analysis revealed significantly lower mean time to first RE in null genotype as compared with GSTM1 present patients (Log p = 0.002). The dose adjusted C₂ levels in null genotype was higher as compared with GSTM1 present patients at one (p = 0.007) and three months (p = 0.027) post transplantation. Conclusion: Patients with variant genotype of GSTM1 and GSTP1 were at higher risk for rejection and delayed functioning of the allograft, respectively, supporting the hypothesis for involvement of GST isoform variants in allograft outcome in renal transplant recipients.     

Suppression of early experimental osteoarthritis by gene transfer of interleukin-1 receptor antagonist and interleukin-10.

Gene therapy offers a radically different approach to the treatment of arthritis. We demonstrated that cDNA coding for human interleukin-1 receptor-antagonist protein (IL-1Ra) and cDNA coding for human interleukin-10 (IL-10) can be delivered, by ex vivo techniques, to the synovial lining of joints, intra-articular expression of gene significantly reduced cartilage matrix degradation and cartilage breakdown. To achieve this, lapine synoviocytes were first transduced in culture by retroviral infection. The genetically modified synoviocytes were then transplanted by intra-articular injection into the knee joints of OA rabbits, assay of joint lavages confirmed that the gene expression was not lost 14 days after transfer. Knees receiving the IL-1Ra gene had significantly reduced cartilage breakdown. Delivery of the IL-10 gene was less effective, having only a moderate effect on cartilage breakdown. When both genes were injected together, there was a greater inhibition of cartilage breakdown, suggesting that simultaneous gene delivery may be necessary to treat OA by targeting the activities of multiple inflammatory effectors.