Preemptive plasma therapy prevents atypical hemolytic uremic syndrome relapse in kidney transplant recipients

BackgroundAtypical hemolytic uremic syndrome (aHUS) frequently leads to renal failure, and kidney transplantation bears a high risk of disease recurrence and graft loss.MethodsPatients who received a kidney graft in our center were retrospectively identified using our Vienna Thrombotic Microangiopathy Cohort. Since 2005, the majority of aHUS patients received perioperative plasma exchange (PE) followed by plasma infusions (PI). Patients were switched to eculizumab in case of plasma intolerance or failure. Those with no preemptive therapy served as controls. We used proportional Cox regression and logistic regression to examine predictors of graft survival.Results19 aHUS patients received 32 grafts and had a follow-up > 1 year. Eight patients received preventive plasma therapy for eight transplants and 13 patients (including 2 patients who received plasma therapy for their last transplant) had no preventive therapy for 24 grafts. The median graft survival was 2.372 days in patients, that received preemptive therapy and 411 days in patients, that did not receive preemptive treatment (hazard ratio: 0.11; p= 0.03). Four patients were switched to eculizumab because of plasma intolerance or failure. Additionally, one patient, that was not transplanted according to the above-mentioned protocol, received eculizumab for aHUS relapse. Additionally, relapse of aHUS (p = 0.01) and year of transplantation (p<0.01) had an effect on graft failure.ConclusionsThis study shows that preemptive plasma therapy and eculizumab rescue in selected cases improve graft survival among kidney transplant recipients with aHUS.     

乙型肝炎病毒基因型、BCP及PC区变异与拉米夫定治疗后HBV DNA反弹的关系

目的:探讨HBV基因型、C区基本核心启动子(BcP)及前C(PC)区变异与拉米夫定抗病毒治疗后HBV DNA反弹的关系.方法:应用多引物对巢式PCR法,PCR-序列分析法,检测拉米夫定治疗27例乙型肝炎患者(治疗组),以及19例从未用过抗病毒治疗患者(对照组)的HBV基因型PC区,BCP的突变位点.结果:27例HBV DNA反弹的患者9例检出G1896A变异率高于对照组(33.33% vs 5.26%,P<0.05),4例检出C1856T变异(14.81%).治疗组4份治疗前标本未检出G1896A、C1856T和BCP变异.与对照组比较,治疗组PC(G1896A)及BCP(A1762T G1764A)双变异的患者中B基因型的构成比增高,分别为75%和50%,C基因型的构成比下降,分别为25%和50%.其中在BCP(A1762T G1764A)变异患者中B、C基因型构成比与对照组比较有显著性差异(P<0.05).4例HBV DNA反弹患者治疗前未检出有基因变异,治疗后有2例检出变异,BCP变异1例,BCP PC变异1例.27例HBV DNA反弹患者BCP变异4例,PC变异2例,BCP PC变异8例.结论:BCP(T1762/A1764)变异、PC区(G1896A)变异可能与拉米夫定治疗后HBV DNA反弹有关.病毒变异导致的HBV DNA反弹可以是单基因变异引起,也可以是多个基因联合变异引起,拉米夫定治疗后B基因型患者更易发生A1762T G1764A变异.     

Identification of HBV DNA sequences that are predictive of response to lamivudine therapy

Numerous studies have shown that resistance to long-term lamivudine therapy occurs in as many as (2/3) of hepatitis B virus (HBV) chronic carriers. Additional studies have shown that reversion of HBV mutations in the precore/core promoter region conferring an HBeAg-negative phenotype/genotype can occur in up to 30% of lamivudine-treated patients. In this study, sequences of the HBV polymerase and precore/core coding regions in 26 HBV-infected patients (24 with HBeAg-negative virus infection, 25 genotype D, 1 genotype A) treated for 27 to 53 months with lamivudine were analyzed to determine the relationship between pretreatment HBV DNA sequence patterns and long-term treatment response, and the effect of therapy on the status of HBV precore mutations. Reversions of precore mutations A1762T/G1764A and G1896A were observed in 29% and 25% of patients, respectively, but none became HBeAg-positive. These data are consistent with previously published reversion frequencies for 2 other groups of lamivudine-treated patients. Two naturally-occurring DNA polymorphisms at aa91 and aa256 of the HBV polymerase were observed in the pretreatment serum samples, which correlated with extended treatment failure. In conclusion, reversion of mutations conferring an HBeAg-negative phenotype occur relatively frequently under lamivudine therapy. Furthermore, at least in HBeAg-negative patients infected predominantly with HBV genotype D, specific viral DNA sequences which are present before therapy appear to be useful as predictors of long-term response to lamivudine treatment.     

Preemptive therapy is not adequate for prevention of cytomegalovirus disease in pancreas–kidney transplant recipients

Background. Cytomegalovirus (CMV) remains the most common viral infection after pancreas–kidney transplantation (PKT). Comparative studies about CMV prophylaxis in PKT have not been developed.
Methods. We analyzed CMV disease in a cohort of 84 PKT recipients. All received intravenous ganciclovir during treatment with anti-thymocyte globulin and later one of the following options for pre-transplant CMV-seropositive recipients: (a) no prophylaxis ( n =10 patients), (b) preemptive therapy (PT) ( n =13), or (c) continuous prophylaxis (CP) for 12 weeks ( n =29). Pre-transplant CMV-seronegative recipients received CP ( n =21).
Results. Eleven patients were excluded because of organ explantation in the first 15 days. Incidence of CMV disease in seropositive recipients was 30% under no prophylaxis, 23% under PT, and 6.9% under CP. Incidence of CMV disease under CP was 33.3% in seronegative recipients. Six of 9 episodes of CMV disease under CP occurred after finishing prophylaxis. Under CP, the incidence of CMV disease was significantly higher in seronegative than in seropositive recipients ( P <0.05).
Conclusion. According to the results of our study, for CMV-seropositive PKT recipients, CP is a better strategy than PT. For CMV-seronegative recipients, 3 months of CP is an inadequate strategy.     

拉米夫定治疗慢性乙型肝炎病毒感染的近期疗效

目的评价拉米夫定治疗不同临床类型慢性乙型肝炎病毒（HBV）感染的近期疗效。方法口服拉米夫定150mg,每日1次,连服6个月,治疗慢性乙型肝炎病人40例,肝炎肝硬化18例,慢性重型肝炎10例。观察其临床、生物化学、血清学和病毒学改变。结果（1）慢性乙型肝炎病情缓解。对照组病毒血症持续,27．5％病人于随访期内肝炎复发（P＜0．001）。同时观察拉米夫定联用干扰素治疗病人20例,未见提高疗效。（2）肝炎肝硬化病情渐趋稳定,肝功能好转,Child—Pugh积分下降。（3）慢性重型肝炎除2例服药不足3个月死亡外,余8例病情缓解,随着肝功能改善,生活质量显著好转。结论拉米夫定适用于治疗慢性乙型肝炎,对处于HBV复制状态的肝硬化和重型肝炎也有效。     

拉米夫定治疗相关的乙型肝炎病毒变异研究进展

拉米夫定治疗相关HBV变异多发生于活性最强、包含第546－575位氨基酸的C区段酪氨酸－蛋氨酸－天门冬氨酸－天门冬氨酸(YMDD)位点,亦涉及第501－545位氨基酸的B区段。YMDD位点是HBV逆转录酶的活性部分,属高度保守序列。在HBV 的逆转录过程中,     

Efficiency and safety of lamivudine therapy in patients with chronic HBV infection, dialysis or after kidney transplantation

AIM: To analyze the effectiveness and safety of lamivudine treatment in patients with chronic HBV infection undergoing hemodialysis or after kidney transplantation, and to study the frequency of tyrosine - methionine - aspartate - aspartate (YMDD) mutation occurrence after lamivudine treatment. METHODS: We analyzed 91 patients with chronic hepatitis B, among whom, 16 patients underwent hemodialysis, 7 patients had kidney transplantation and 68 patients had normal function of kidney. The hemodialysis patients were treated by lamivudine 300 mg/wk. patients after kidney transplantation and patiente with normal function of kidney were treated with lamivudine 100 mg/d. Therapy lasted for 12 mo. HBV-DNA, HBsAg, HBeAg and anti-HBe, and anti-HCV antibodies were assessed in sera of patients. The analysis was performed before and 6 mo after the end of lamivudine treatment. Before, during and after the lamivudine therapy, the number of erythrocytes, leukocytes, platelets and hemoglobin concentration, ALT and AST activity, as well as bilirubin, urea and creatinine concentrations were analyzed in sera from patients. RESULTS: After the 12-mo lamivudine treatment, elimination of HBV - DNA was observed in 56% patients undergoing hemodialysis and in 53% patients with normal kidney function. Only 1 from 7 (14%) kidney-transplanted patients eliminated HBV-DNA. Furthermore, HBeAg elimination was observed in 36% hemodialysis patients, in 51% patients with normal function of kidneys and in 43% kidney transplanted patients. Among the patients undergoing dialysis, no YMDD mutation was found after 12 mo of therapy, while it was detected in 9 patients (13%) with normal function of kidney and in 2 kidney-transplanted patients (29%, P<0.006). We did not observe significant side effecte of lamivudine treatment in studied patiente. CONCLUSION: Effectiveness of lamivudine therapy in dialysis patients is comparable with that in patiente with normal function of kidney. Lamivudine treatment is well tolerated and safe in patiente with renal insufficiency undergoing hemodialysis and kidney-transplantation. However, in the latter group, high incidence of YMDD mutation after lamivudine treatment was observed.     

Detection of HBV DNA in HBsAg-positive sera after amplification using the polymerase chain reaction

The presence of hepatitis B virus (HBV) DNA in serum as detected by molecular hybridization is considered the most reliable marker for the presence of complete virions and, therefore, infectivity. This technique, however, has a lower limit of detection of 0.1 pg HBV DNA. Using the polymerase chain reaction (PCR), a technique by which DNA sequences can be amplified selectively, we investigated sera from 30 HBsAg carriers, 6 also positive for HBeAg and 24 negative for HBeAg. After PCR followed by Southern blot, 27 sera were found to be positive for HBV DNA, whereas only 7 sera were positive for HBV DNA in the conventional dot blot. PCR followed by Southern blot analysis lowered the limit of detection to 0.5 fg HBV DNA. Amplified HBV DNA fragments from some samples were directly sequenced without previous cloning. We conclude that PCR is a suitable method to amplify parts of viral genomes present in human sera, and that PCR with subsequent Southern blot analysis allows the detection of hepatitis B virions in the majority of HBsAg-positive sera.     

HBsAg阳性非霍奇金淋巴瘤患者抗HBV治疗期间HBV再激活的危险因素分析

目的:探讨HBsAg阳性非霍奇金淋巴瘤患者抗病毒治疗期间HBV再激活的危险因素,从而早期识别高危人群,指导临床治疗。方法:采用临床回顾性研究的方法,选择2011年10月至2018年3月期间在武汉大学中南医院住院治疗的90例HBsAg阳性非霍奇金淋巴瘤患者为研究对象,根据是否接受抗病毒治疗分为治疗组58例(抗病毒治疗)及对照组32例(无抗病毒治疗),观察两组患者的一般情况及实验室检查结果,比较两组患者化疗后HBV再激活、化疗延迟及肝功能损害的情况,分析HBsAg阳性非霍奇金淋巴瘤患者抗病毒治疗期间HBV再激活的危险因素。结果:两组患者平均年龄分别为(55.2±10.7)岁和(47.4±12.8)岁,差异有统计学意义(P<0.05)。治疗组患者发生HBV再激活10例(17.2%),较对照组HBV再激活12例(37.5%)显著减少,差异有统计学意义(P<0.05);两组患者化疗延迟率分别为24.1%、46.9%,差异有统计学意义(P<0.05);肝功能损害发生率分别为56.9%、40.6%,差异无统计学意义(P>0.05)。危险因素单因素分析结果显示年龄、化疗前血清HBeAg阳性、抗病毒药物种类、停用抗病毒药物及化疗周期方面,差异具有统计学意义(P<0.05)。多因素分析结果显示化疗前HBeAg阳性[OR=1.6,95%可信区间(CI):1.5~3.5],停用抗病毒药物[OR=15.5,95%可信区间(CI):1.5~171.9],化疗周期≥5[OR=3.0,95%可信区间(CI):1.2~7.7]。结论:预防性应用核苷(酸)类似物抗病毒药物能显著降低HBsAg阳性非霍奇金淋巴瘤患者化疗后HBV再激活率及化疗延迟率。化疗前血清HBeAg阳性、停用抗病毒药物及化疗周期≥5是HBsAg阳性非霍奇金淋巴瘤患者抗病毒治疗期间发生HBV再激活的独立危险因素。     

Effect of lamivudine in HBeAg-positive chronic hepatitis B： Discordant effect on HBeAg and HBV DNA according to pretreatment ALT level

AIM: To clarify differences in antiviral effect of the drug in patients with different ALT levels, we examined the changes in HBV markers in patients with high or low ALT levels with or without lamivudine treatment. METHODS: Thirty-seven HBeAg-positive patients were studied. Ten patients with ALT levels higher than 200 IU/L (group 1) and 8 patients with ALT below 200 IU/L (group 2) were treated orally with 100 mg/d of lamivudine. As untreated control, 9 patients with ALT above 200 IU/L (group 3) and 10 patients with ALT below 200 IU/L (group 4) were examined. ALT level, HBeAg/HBeAb status, and HBV DNA level were examined monthly for 11.9+/-0.4 mo. RESULTS: The ALT level normalized in all 10 patients of group 1, 7/8 of group 2, 4/9 of group 3, and 1/10 of group 4 within 6 mo (groups 1 vs 2, P = NS; groups 1 vs 3, P = 0.002; groups 1 vs 4, P<0.0001). HBV DNA fell below the detection limit in all 10 patients of group 1, 7/8 of group 2, 0/9 of group 3, and 0/10 of group 4 within 6 mo (groups 1 vs 2, P = NS). HBeAg became seronegative in 7/10 patients of group 1, 1/8 of group 2, 3/9 of group 3, and 0/10 of group 4 within 12 mo (groups 1 vs 2, P = 0.02; groups 1 vs 3, P = NS). CONCLUSION: Our data suggest that HBeAg-positive patients with higher ALT levels can be considered good candidates for lamivudine therapy, probably because lamivudine accelerates the natural seroconversion of HBeAg, accompanied by HBV DNA loss, in these patients.     

拉米夫定所致YMDD变异的个体化治疗对策

随着拉米夫定应用时间的延长,病毒变异的问题日渐突出,不同肝病患者出现病毒变异后情况不同,因此应给予不同的治疗对策。 1.资料与方法:3例患者均为在我科住院或门诊治疗的乙型肝炎病例。肝功能使用全自动生化分析仪检测,乙型肝炎病毒(HBV)标志物采用微粒子酶免荧光分析法(MEIA)检测,试剂使用美国雅培公司提供的Abbott试剂盒,HBV DNA采用荧光聚合酶链反应(PCR)定量检测,YMDD变异采用限制性内切酶长度多态性(IFLP)分析法检测。病例1,男,41岁。发现HBV感染12年,肝功能异常2年,HBV复制指标阳性。诊断:慢性乙     

Baseline HBV DNA level is the most important factor associated with virologic breakthrough in chronic hepatitis B treated with lamivudine

AIM： To identify the factors associated with virologic breakthrough and to select a subgroup of patients who respond well to lamivudine without developing virologic breakthrough （VBT）.
METHODS： Of 79 patients who had received lamivudine therapy for 9-57 mo, 34 were HBeAg-positive and 45 were HBeAg-negative, 24 developed virologic breakthrough and 55 did not. Clinical and virologic factors were compared between the two groups.
RESULTS： The median duration of therapy was 25 （9-57） mo. Virologic breakthrough was defined as a 〉 1 log HBV DNA increase following initial suppression. When several factors, including gender, duration of infection, baseline HBV DNA, and baseline ALT in HBeAg-positive chronic hepatitis patients were analyzed by logistic regression, the most important predictor of virologic breakthrough was the baseline HBV DNA （r^2 = 0.12, P 〈 0.05）. When HBeAg-postitive chronic hepatitis patients were divided into two groups by a point of 6.6 log HBV DNA, the incidence of virologic breakthough between two groups was significantly different.
CONCLUSION： Lamivudine may remain an effective first line therapy for those HBeAg-positive patients with a baseline HBV DNA 〈 6.6 log10 copies/mL.     

Long-term therapy with lamivudine in renal transplant recipients with chronic hepatitis B

BACKGROUND: Lamivudine is a potent inhibitor of hepatitis B virus (HBV) replication. As other available HBV therapies, its efficacy is hampered by relapse after discontinuation and by the risk of viral breakthrough. A recent study suggests that pre-emptive lamivudine therapy improves survival in HBV renal transplants, but few data are available regarding its long-term use in this population. The clinical features, course and viral mutations associated with the emergence of viral resistance in this population have not been well studied. METHODS: We followed 14 consecutive renal transplant patients treated with lamivudine for chronic hepatitis B. Breakthrough was defined as the reappearance of HBV DNA by hybridization. In patients with breakthrough, lamivudine was always continued and patients were followed up monthly. Mutations associated with viral resistance were determined by sequencing the polymerase encoding gene at the beginning of treatment and at the time of breakthrough. RESULTS: The median duration of treatment was 64.5 months. Resistance to lamivudine appeared in eight patients (57%) after a median duration of treatment of 15 (9-24) months. During a 51 month follow-up after breakthrough, only three of eight patients had a flare-up with alanine aminotransferase levels more than 5 ULN, and no hepatic decompensation was observed. Analysis of HBV sequencing after breakthrough revealed specific resistance mutations in both the B and C domains of the polymerase (rtL180M/M204V, n = 5; rtM204I, n = 2). CONCLUSION: Lamivudine is a safe and effective treatment of active hepatitis B in renal transplant patients. Resistance to treatment is frequent but seems to have little clinical impact over the considered period. In our experience, the YMDD mutation accounts for most cases of virological escape in patients with good compliance.     

乙型肝炎相关肝移植受体供肝植入前后乙型肝炎病毒标志物的变化

目的 了解乙型肝炎相关受体在拉米夫定或拉米夫定 乙型肝炎免疫球蛋白(HBIG)预防下供肝植入前后移植物乙型肝炎病毒(HBV)标志物的变化，探讨移植物HBV再感染的可能机制，为预防复发寻找切入点。方法用酶联放射免疫法、HBVDNA荧光定量法、免疫组织化学LSAB法定期检测78例受体手术前后血清及90例供肝活检组织，重点观察供肝植入前后HBV标志物在肝组织及血清中的动态变化。结果移植时无论受体为HBV活跃复制状态或非活跃复制状态，供肝植入后2h内活检肝组织既无HBVDNA又无HBsAg、HBcAg阳性证据。结论在现有预防措施下，无论受体术前HBV复制状态如何，健康供肝无术中HBV颗粒直接感染的证据，其机制及在远期HBV再感染中的意义尚待进一步研究。