Human adenovirus replication and persistence in hypertrophic adenoids and palatine tonsils in children

The role of human adenovirus (HAdV) infection in different acute diseases, such as febrile exudative tonsillitis, conjunctivitis, and pharyngoconjunctival fever is well established. However, the relationships, if any, of HAdV persistence and reactivation in the development of the chronic adenotonsillar disease is not fully understood. The present paper reports a 3-year cross-sectional hospital-based study aimed at detecting and quantifying HAdV DNA and mRNA of the HAdV hexon gene in adenoid and palatine tonsil tissues and nasopharyngeal secretions (NPS) from patients with adenotonsillar hypertrophy or recurrent adenotonsillitis. HAdV C, B, and E were detectable in nearly 50% of the patients, with no association with the severity of airway obstruction, nor with the presence of recurrent tonsillitis, sleep apnea or otitis media with effusion (OME). Despite the higher rates of respiratory viral coinfections in patients with HAdV, the presence of other viruses, including DNA and RNA viruses, had no association with HAdV replication or shedding in secretions. Higher HAdV loads in adenoids showed a significant positive correlation with the presence of sleep apnea and the absence of OME. Although this study indicates that a significant proportion (~85%) of individuals with chronic adenotonsillar diseases have persistent nonproductive HAdV infection, including those by HAdV C, B, and E, epithelial and subepithelial cells in tonsils seem to be critical for HAdV C production and shedding in NPS in some patients, since viral antigen was detected in these regions by immunohistochemistry in four patients, all of which were also positive for HAdV mRNA detection.     

Epidemiology of enteric viruses in children with gastroenteritis in Ogun State,Nigeria

Acute gastroenteritis (AGE) remains a global public health concern and Nigeria is one of the two countries accounting for 42% of global under-5 deaths attributable to gastroenteritis. This study aimed to determine the prevalence, seasonality, and risk factors of enteric viruses (EVs) in children with AGE in Ogun State, Nigeria. Stool samples collected from children under-5 from three different hospitals between February 2015 and April 2017 were analyzed using molecular methods for the presence of four EVs (group A rotavirus [RVA], norovirus [NoV], human astrovirus [HAstV], and human adenovirus [HAdV]). Among the 175 samples analyzed, 63 (36%) were positive for at least one EV. The most prevalent was HAstV (19.4%), followed by RVA (16.6%), NoV (5.1%), and HAdV (5.1%). Mixed infections were found in 17 cases. No significant association was observed with age, sex, and risk factors. Though not significant, EV prevalence was higher in the dry season. Positive cases (asides HAdV) had no correlation with temperature and/or humidity. This study provides information on the prevalence and seasonal fluctuations of EVs, which will be of value in the effective management of patients and control strategies for viral gastroenteritis in the country.     

Infection and direct injury in human hepatocyte explants and a hepatoblastoma cell line due to hepatiticomimetic (non-hepatitis) viruses

Hepatitis is caused by hepatitis viruses, but hepatitis or hepatocellular enzyme abnormalities is sometimes associated with infection by the hepatiticomimetic viruses. The direct and indirect effects of infection with hepatiticomimetic viruses were examined in two human hepatocyte systems. Poliovirus, adenovirus, and herpes simplex virus (HSV) induced cytopathology in Hep G2 cells. Measles virus caused no change in hepatocytes. Poliovirus infection did not affect cellular protein synthesis, and the peak of hepatocellular enzyme release coincided with the peak of virus release. The increase in adenovirus protein synthesis correlated with the decrease of transferrin synthesis, and enzyme release was not prominent. HSV induced viral protein synthesis with enhanced processing and inhibition of synthesis of alpha1-antitrypsin. The peak of enzyme release was later than the peak of virus release. In primary hepatocytes, poliovirus, adenovirus, and induced extensive cytopathology and enzyme release, and VZV caused cytopathology and significant but minute enzyme release. The ratio of lactate dehydrogenase to aspartate aminotransferase release was larger in poliovirus infection in both hepatocytes than in HSV or VZV infection. Although poliovirus and adenovirus are released by cytolysis and HSV and VZV are secreted by exocytosis of cytoplasmic vacuoles, enzyme release was independent of the type of virus release. Adenovirus showed strong cytotoxicity but did not modify the membrane nor cause enzyme release. Enzyme release was associated with modification of the surface membrane due to apoptosis with poliovirus and necrosis with HSV. Consequently hepatocellular injury by viral infection did not reflect the amount or pattern of hepatocellular enzyme release.     

TACE联合重组人p53腺病毒治疗对原发性肝癌患者免疫应答及预后的影响

目的:探讨TACE联合重组人p53腺病毒治疗对原发性肝癌患者免疫应答及预后的影响。方法:选取我院于2016年01月至2017年12月收治的60例中晚期原发性肝癌患者作为研究对象,分为对照组和研究组,每组30例。对照组采取经导管肝动脉化疗栓塞术（TACE)治疗,研究组在对照组基础上联合重组人p53腺病毒注射液治疗。比较两组患者近期临床疗效、两组患者治疗后1年存活率、生活质量情况,比较两组患者术前1天、术后1周、术后2周、术后4周的外周血免疫细胞水平以及甲胎蛋白（AFP）水平,同时观测两组患者治疗期间不良反应发生情况。结果:研究组近期临床治疗总有效率60.00%(18/30)显著高于对照组33.33%（10/30）;两组患者治疗后1周各免疫细胞水平与治疗前相比无明显差异（P＞0.05）,但治疗2周和4周后两组患者CD8＋水平均显著下降,CD3＋、CD4＋水平以及CD4＋/CD8＋比值显著上升,且研究组改善程度显著优于对照组（P＜0.05）;研究组患者甲胎蛋白（AFP）水平相比对照组下降更明显（P＜0.05）;治疗期间研究组不良反应发生率20.00%与对照组16.67%相比,差异无统计学意义（P＞0.05）;对两组患者1年生存随访,结果显示研究组死亡3例,存活27例,对照组死亡10例,存活20例,两组生存期差异存在显著意义(log-rank P= 0.030);且研究组半年后和1年后生活质量改善程度显著优于对照组（P＜0.05）。结论:TACE基础上联合重组人p53腺病毒可以显著改善原发性肝癌患者免疫细胞水平和机体免疫应答功能,提高1年生存率以及改善患者预后。     

Gene Therapy Corrects Brain and Behavioral Pathologies in CLN6-Batten Disease

1. Download : Download high-res image (205KB)
2. Download : Download full-size image

     

提高腺病毒载体转染造血细胞效率的研究进展

重组腺病毒作为基因转移的载体已经广泛应用于基因治疗的基础研究和临床试验，但造血细胞由于缺乏腺病毒特异性受体柯萨奇-腺病毒受体(coxsacke virus and adenovirus receptor，CAR)导致转染效率较低，从而限制了腺病毒载体在造血系统的应用。多种策略可以提高腺病毒转染造血细胞的效率，这些策略包括腺病毒本身的改造、造血细胞CAR表达的提高等。本文就这一领域的主要研究进展作一综述。     

存活蛋白重组腺病毒载体的构建及其在树突状细胞的表达

本研究旨在构建含有人存活蛋白(survivin)基因的重组腺病毒载体,并探讨其在转染树突状细胞中的表达。以质粒pcDNA3.0-survivin为模板,通过PCR扩增获得survivin基因全长序列。PCR产物回收后经酶切,定向插入腺病毒穿梭质粒,获得重组质粒pShuttle-CMV-survivin。通过双酶切、PCR及插入片段测序鉴定后,将正确重组体pShuttle-CMV-survivin转化E.coliBJ5183菌(含腺病毒骨架质粒)并进行同源重组,然后筛选阳性克隆,提取质粒,将此重组腺病毒质粒分别进行酶切、线性化、纯化,用脂质体LipofectamineTM2000介导转染293细胞。制备病毒上清并测定其滴度,将病毒上清转染树突状细胞,应用Westernblot方法分析survivin的表达。结果表明成功构建了含有人survivin基因的重组腺病毒,病毒滴度为2.65×109pfu/ml。Westernblot鉴定显示,经重组腺病毒转染的DC可有效表达survivin。结论含survivin基因的重组腺病毒载体的构建成功,为下一步开展抗白血病免疫研究奠定实验基础。     

Adenovirus based HPV L2 vaccine induces broad cross-reactive humoral immune responses

Oncogenic high-risk human papillomavirus (HPV) infections cause a substantial number of genital and non-genital cancers worldwide. Approximately 70% of all cervical cancers are caused by the high-risk HPV16 and 18 types. The remaining 30% can be attributed to twelve other high-risk HPV-types. Highly efficacious 2-valent, 4-valent and 9-valent L1 protein based prophylactic HPV vaccines are available however with limited cross-protection. To further increase the coverage, development of a multivalent cross-protective HPV vaccine is currently focused on the conserved N-terminus of HPV’s L2 protein. We have developed a vaccine candidate based on the rare human adenovirus type 35 (HAdV35) vector that displays a concatemer of L2 protein epitopes from four different HPV-types via protein IX (pIX). A mix of two heterologous HAdV35 pIX-L2 display vectors present highly conserved linear epitopes of nine HPV-types. Each HAdV35 pIX-L2 display vector exhibits a good manufacturability profile. HAdV35 pIX-L2 display vaccine vectors were immunogenic and induced neutralizing antibodies against HPV-types included in the vaccine and cross-neutralizing antibodies against distant a HPV-type not included in the vaccine in mice. The HAdV35 pIX-L2 display vectors offer an opportunity for a multivalent HAdV-based prophylactic HPV vaccine.