1例血浆置换对抗中性粒细胞胞浆抗体患者相关性血管炎肺损害的护理

正抗中性粒细胞胞浆抗体(antineutrophil cytoplasmic antibodies,ANCA)相关性血管炎(ANCA-associated vasculitis,AAV)是一种严重累及小和中等血管引起严重全身多系统器官功能衰竭,其病理特点是小血管壁的炎症和纤维素样坏死为病理基础,主要指韦格纳肉芽肿、显微镜下多血管炎、过敏性肉芽肿性血管炎。AAV可广泛累及全身多个脏器,但肾脏、肺脏受     

抗中性粒细胞胞浆抗体相关性血管炎研究进展

抗中性粒细胞胞浆抗体(ANCA)相关性血管炎(AAV)是一类以小血管壁炎症和纤维素样坏死为特征,累及全身多系统的自身免疫性疾病。亚洲地区AAV以显微镜下多血管炎(MPA)为主,最常累及肾和肺。近几年发现补体,例如C5a,通过替代途径在AAV的发病机制中发挥着重要作用,所以了解AAV补体与凝血过程之间的联系有利于从发病机制出发找到治疗疾病的靶点。本综述总结AAV发病机制中凝血及纤溶系统变化及与补体系统相互作用的研究进展。     

原发性小血管炎5例临床分析

原发性小血管炎主要指韦格纳肉芽肿病(wegener’s granulo-matosis)，显微镜多血管炎(microscopic polyangiitis)和局灶节段坏死性肾小球肾炎。此病为系统性自身免疫性疾病，可累及全身多个脏器。其中肾脏受累明显并进展迅速，肺部受累病情亦凶险，及时诊断和治疗非常重要，现将我院1998     

儿童抗中性粒细胞胞浆抗体相关性血管炎3例

正抗中性粒细胞胞浆抗体(antineutrophil cytoplasmic antibody,ANCA)相关性血管炎(ANCA associated vasculitis,AAV)是一种多系统累及的小血管炎,按照2012Chanpel Hill标准,可分为显微镜下多血管炎(microscopic polyangiitis,MPA)、肉芽肿性血管炎(Granulomatosis with polyangiitis,GPA)和嗜酸性肉芽肿血管炎     

CRRT联合血浆置换对AAV肾损害的护理1例

抗中性粒细胞胞浆抗体(ANCA)相关性血管炎(AAV)是一种累及多系统的小血管炎症性疾病,包括三个亚组,即肉芽肿性多血管炎、显微镜下多血管炎和嗜酸性肉芽肿性多血管炎 [1].其特征是产生丝氨酸蛋白酶-3(PR3-ANCA)或髓过氧化物酶(MPO-ANCA)自身抗体.由于肾脏血流丰富,因此是最常见的受损器官.当肾、肺、心...     

显微镜下多血管炎肺损害发病机制及治疗进展

显微镜下多血管炎(microscopic polyangiitis,MPA)是一种以累及小血管为主的系统性血管炎,可造成全身多个重要组织、脏器的损害。肺是其常见的受累器官,主要可表现为肺间质纤维化、弥漫性肺泡出血(diffuse alveolar hemorrhage,DAH)等。近年来随着对MPA肺损害发病机制认识的不断深入,对MPA肺受累的治疗也取得了新的进展。     

ANCA相关性原发性小血管炎

目的：探讨原发性小血管炎的临床特点。方法：总结18例原发性小血管炎的临床资料。结果：原发性小血管炎的临床表现复杂，全身各个系统均可受累，肾脏是最易受累器官之一，抗中性粒细胞胞浆抗体对原发性小血管炎的诊断具有非常重要的意义。结论：原发性小血管炎临床表现复杂，累及全身多个器官或系统，早期治疗效果较好。     

结节性多动脉炎2例并文献分析

结节性多动脉炎是一种主要累及中小肌性动脉的坏死性血管炎.全身各组织器官的中小动脉均可受累、临床表现复杂,病情轻重差异大,本科收住结节性多动脉炎患者2例,现报告如下.     

1例ANCA相关性血管炎合并急性肾损伤病人行血液净化的护理

正抗中性粒细胞胞浆抗体(antineutrophil cytoplasmic antibody,ANCA)相关性血管炎是一类起病隐匿的自身免疫性疾病,可累及全身各处小血管~([1])。肾脏和肺脏是ANCA相关性血管炎损害的最主要靶器官。文献报道,肾累及率达70%~80%~([2-3]),肺脏受累达61%~75%~([4-7]),且多进展迅速,治疗不及时会导致呼     

韦格纳肉芽肿病误诊1例分析

韦格纳肉芽肿病(WG)是一种特殊类型的以中小动脉、静脉及毛细血管受累为主的肉芽肿性或坏死性血管炎,可累及全身多系统,以血管壁坏死性炎症为特征。我科收治1例WG患者,病初误诊为鼻窦炎,现报告如下。     

Transendocardial delivery of AAV6 results in highly efficient and global cardiac gene transfer in rhesus macaques

Heart disease is the leading cause of morbidity and mortality, and cardiac gene transfer has potential as a novel therapeutic approach. We previously demonstrated safe and efficient gene transfer to the canine heart using a percutaneous transendocardial injection procedure to deliver self-complementary (sc) adeno-associated virus 6 (AAV6) vector. In the present study, we proceed with our vertical translation study to evaluate cardiac gene transfer in nonhuman primates (NHPs). We screened approximately 30 adult male rhesus macaques for the presence of neutralizing antibodies against AAV6, AAV8, and AAV9, and then selected seven monkeys whose antibody titers against these three serotypes were lower than 1/5. The animals were then randomized to receive either scAAV6 (n=3), scAAV8 (n=1), or scAAV9 (n=3) vector expressing the enhanced green fluorescent protein (EGFP) reporter gene at a dose of 5.4×10(12) genome copies/kg, which was administered according to a modified version of our previously developed transendocardial injection procedure. One animal treated with scAAV6 died secondary to esophageal intubation. The remaining animals were euthanized 7 days after gene transfer, at which time tissue was collected for analysis of EGFP expression, histopathology, and biodistribution of the vector genome. We found that (i) transendocardial delivery of AAV is safe in the NHP, (ii) AAV6 and AAV8 provide efficient cardiac gene transfer at similar levels and are superior to AAV9, and (iii) AAV6 is more cardiac-specific than AAV8 and AAV9. The results of this NHP study may help guide the development AAV vectors for the treatment of cardiovascular disease in humans.     

Adeno-associated virus (AAV) serotype 9 provides global cardiac gene transfer superior to AAV1, AAV6, AAV7, and AAV8 in the mouse and rat

Heart disease is the leading cause of morbidity and mortality. Cardiac gene transfer may serve as a novel therapeutic approach. This investigation was undertaken to compare cardiac tropisms of adeno-associated virus (AAV) serotypes 1, 6, 7, 8, and 9. Neonatal mice were injected with 2.5 x 10(11) genome copies (GC) of AAV serotype 1, 6, 7, 8, or 9 expressing LacZ under the control of the constitutive chicken beta-actin promoter with cytomegalovirus enhancer promoter via intrapericardial injection and monitored for up to 1 year. Adult rats were injected with 5 x 10(11) GC of the AAV vectors via direct cardiac injection and monitored for 1 month. Cardiac distribution of LacZ expression was assessed by X-Gal histochemistry, and beta-galactosidase activity was quantified in a chemiluminescence assay. Cardiac functional data and biodistribution data were also collected in the rat. AAV9 provided global cardiac gene transfer stable for up to 1 year that was superior to other serotypes. LacZ expression was relatively cardiac specific, and cardiac function was unaffected by gene transfer. AAV9 provides high-level, stable expression in the mouse and rat heart and may provide a simple alternative to the creation of cardiac-specific transgenic mice. AAV9 should be used in rodent cardiac studies and may be the vector of choice for clinical trials of cardiac gene transfer.     

Immunosuppression decreases inflammation and increases AAV6-hSERCA2a-mediated SERCA2a expression

The calcium pump SERCA2a (sarcoplasmic reticulum calcium ATPase 2a), which plays a central role in cardiac contraction, shows decreased expression in heart failure (HF). Increasing SERCA2a expression in HF models improves cardiac function. We used direct cardiac delivery of adeno-associated virus encoding human SERCA2a (AAV6-hSERCA2a) in HF and normal canine models to study safety, efficacy, and the effects of immunosuppression. Tachycardic-paced dogs received left ventricle (LV) wall injection of AAV6-hSERCA2a or solvent. Pacing continued postinjection for 2 or 6 weeks, until euthanasia. Tissue/serum samples were analyzed for hSERCA2a expression (Western blot) and immune responses (histology and AAV6-neutralizing antibodies). Nonpaced dogs received AAV6-hSERCA2a and were analyzed at 12 weeks; a parallel cohort received AAV-hSERCA2a and immunosuppression. AAV-mediated cardiac expression of hSERCA2a peaked at 2 weeks and then declined (to ~50%; p<0.03, 6 vs. 2 weeks). LV end diastolic and end systolic diameters decreased in 6-week dogs treated with AAV6-hSERCA2a (p<0.05) whereas LV diameters increased in control dogs. Dogs receiving AAV6-hSERCA2a developed neutralizing antibodies (titer ≥1:120) and cardiac cellular infiltration. Immunosuppression dramatically reduced immune responses (reduced inflammation and neutralizing antibody titers <1:20), and maintained hSERCA2a expression. Thus cardiac injection of AAV6-hSERCA2a promotes local hSERCA2a expression and improves cardiac function. However, the hSERCA2a protein level is reduced by host immune responses. Immunosuppression alleviates immune responses and sustains transgene expression, and may be an important adjuvant for clinical gene therapy trials.     

Cardiac gene transfer of short hairpin RNA directed against phospholamban effectively knocks down gene expression but causes cellular toxicity in canines

Derangements in calcium cycling have been described in failing hearts, and preclinical studies have suggested that therapies aimed at correcting this defect can lead to improvements in cardiac function and survival. One strategy to improve calcium cycling would be to inhibit phospholamban (PLB), the negative regulator of SERCA2a that is upregulated in failing hearts. The goal of this study was to evaluate the safety and efficacy of using adeno-associated virus (AAV)-mediated cardiac gene transfer of short hairpin RNA (shRNA) to knock down expression of PLB. Six dogs were treated with self-complementary AAV serotype 6 (scAAV6) expressing shRNA against PLB. Three control dogs were treated with empty AAV6 capsid, and two control dogs were treated with scAAV6 expressing dominant negative PLB. Vector was delivered via a percutaneously inserted cardiac injection catheter. PLB mRNA and protein expression were analyzed in three of six shRNA dogs between days 16 and 26. The other three shRNA dogs and five control dogs were monitored long-term to assess cardiac safety. PLB mRNA was reduced 16-fold, and PLB protein was reduced 5-fold, with treatment. Serum troponin elevation and depressed cardiac function were observed in the shRNA group only at 4 weeks. An enzyme-linked immunospot assay failed to detect any T cells reactive to AAV6 capsid in peripheral blood mononuclear cells, heart, or spleen. Microarray analysis revealed alterations in cardiac expression of several microRNAs with shRNA treatment. AAV6-mediated cardiac gene transfer of shRNA effectively knocks down PLB expression but is associated with severe cardiac toxicity. Toxicity may result from dysregulation of endogenous microRNA pathways.     

Widespread and early myocardial gene expression by adeno-associated virus vector type 6 with a beta-actin hybrid promoter.

Gene therapy for acute cardiac events such as myocardial infarction requires early gene expression over an entire region of myocardium, which has not been possible using adeno-associated virus (AAV) vectors to date. Here we demonstrate marked improvement in the distribution and rapidity of gene expression in myocardium using the AAV pseudotype 6 (AAV6) vector, compared to the standard serotype 2 (AAV2) vector. An alkaline phosphatase (AP) reporter construct driven by the chicken beta-actin promoter was packaged in either AAV6 or AAV2 capsids and delivered to rat hearts in vivo by direct injection. AP expression was evident in both AAV6 and AAV2 vector-treated hearts as early as 1 day after injection, but increased rapidly in AAV6 vector-treated hearts during the first 7 days. The amplitude of AP activity produced by the AAV6 vector was 5-fold greater than that produced by the equivalent AAV2 vector at both 3 and 7 days postinjection. Additionally, the AAV6 vector transduced a myocardial volume that was 10-fold larger than the AAV2 vector. These results indicate the significant potential of AAV6 serotype vectors for early gene expression and widespread regional transduction of myocardium, both auspicious results for in vivo applications in acute cardiac disease.     

Topoisomerase inhibition accelerates gene expression after adeno-associated virus-mediated gene transfer to the mammalian heart.

Utility of adeno-associated virus 2 (AAV2) vectors for cardiac gene therapy is limited by the prolonged lag phase before maximal gene expression. Topoisomerase inhibition can induce AAV2-mediated gene expression in vivo, but with variable success in different tissues. In this study, we demonstrate that topoisomerase inhibition can accelerate AAV2-mediated gene expression in the mouse heart. We used an AAV2 vector expressing firefly luciferase and monitored expression kinetics using non-invasive bioluminescence imaging. In the group receiving vector alone, cardiac luciferase activity was evident from week 2 onward and increased progressively to reach a steady plateau by 9 weeks postinjection. In the group receiving vector and camptothecine (CPT), luciferase expression was evident from days 2 to 4 onward and increased rapidly to reach a steady plateau by 3-4 weeks postinjection, nearly three times faster than in the absence of CPT (P<0.05). Southern blot analysis of AAV2 genomes in cardiac tissue showed rapid conversion of the AAV2 genome from its single-stranded to double-stranded form in CPT-treated mice. Non-invasive determinations of luciferase expression correlated well with in vitro luciferase assays. Direct injection of the AAV2 vector and long-term luciferase gene expression had no detectable effects on normal cardiac function as assessed by magnetic resonance imaging.     

Engineering Liver-detargeted AAV9 Vectors for Cardiac and Musculoskeletal Gene Transfer

We report the generation of a new class of adeno-associated virus serotype 9 (AAV9)-derived vectors displaying selective loss of liver tropism and demonstrating potential for cardiac and musculoskeletal gene transfer applications. Random mutagenesis of residues within a surface-exposed region of the major AAV9 capsid protein yielded a capsid library with mutations clustered at the icosahedral threefold symmetry axis. Using a combination of sequence analysis, structural models, and in vivo screening, we identified several functionally diverse AAV9 variants. The latter were classified into three functional subgroups, with respect to parental AAV9 displaying: (i) decreased transduction efficiency across multiple tissues; (ii) a selective decrease in liver transduction, or (iii) a similar transduction profile. Notably, variants 9.45 and 9.61 (subgroup II) displayed 10- to 25-fold lower gene transfer efficiency in liver, while transducing cardiac and skeletal muscle as efficiently as AAV9. These results were further corroborated by quantitation of vector genome copies and histological analysis of reporter (tdTomato) gene expression. The study highlights the feasibility of generating AAV vectors with selectively ablated tissue tropism, which when combined with other targeting strategies could allow sharply segregated gene expression. Liver-detargeted AAV9 variants described herein are excellent candidates for preclinical evaluation in animal models of cardiac and musculoskeletal disease.     

AAV serotype-1 mediates early onset of gene expression in mouse hearts and results in better therapeutic effect

Adeno-associated viral vectors (AAV) are attractive tool for gene therapy for coronary artery disease. However, gene expression in myocardium mediated by AAV serotype 2 (AAV2) does not peak until 4-6 weeks after gene transfer. This delayed gene expression may reduce its therapeutic potential for acute cardiac infarction. To determine whether earlier gene expression and better therapeutic effect could be achieved using a different serotype, CMV promoter driving the EPO gene (AAV-EPO) was packaged into AAV serotypes 1-5 capsids and injected into mouse myocardium. EPO expression was studied by measuring the hematocrits and EPO mRNA. After we found that AAV1 mediates the highest gene expression after 4 days of gene transduction, AAV-LacZ (CMV promoter driving LacZ gene expression) and MLCVEGF (hypoxia-inducible and cardiac-specific VEGF expression) were packaged into AAV1 and 2 capsids. LacZ expression was detected in AAV1-LacZ but not in AAV2-LacZ-injected hearts 1 day after vector injection. Compared to AAV2-MLCVEGF that mediated no significant VEGF expression, AAV1-MLCVEGF mediated 13.7-fold induction of VEGF expression in ischemic hearts 4 days after gene transduction and resulted in more neovasculatures, better cardiac function and less myocardial fibrosis. Thus, AAV1 mediates earlier and higher transgene expression in myocardium and better therapeutic effects.