Antitumor Effects of Newcastle Disease Virus Hemagglutinin-neuraminidase Used as a Molecular Adjuvant

Gene therapy is a potentially powerful tool used in cancer therapy. The strength of immune responses induced by some strategies is usually low, therefore, the development of agents capable of enhancing these responses is highlighted. The authors investigated the potential of an approach based on the hemagglutinin-neuraminidase(HN) of Newcastle disease virus(NDV) as a potential immune adjuvant. It was found that recombinant adenovirus(Ad) infected SGC7901 cells expressing HN exhibited both hemagglutinin(HA) and neuraminidase(NA) activities. It was demonstrated that administration of HN induced higher levels of the effector cytokines TNF-α, IFN-α and IFN-γ and increased natural killer(NK) cell activity. Based on the therapeutic tumor model, the results show that the administration of HN with Apoptin led to improved survival and tumor suppression. In conclusion, this study indicates that HN stimulates innate immune responses to make the activity of NK cells increased, which highlights the potential adjuvant activity of HN in cancer gene therapy.     

Anti-tumor Immune Response Mediated by Newcastle Disease Virus HN Gene

Hemagglutinin-neuramidinase(HN) is one of the most important surface structure proteins of the Newcastle disease virus(NDV). HN not only mediates receptor recognition but also possesses neuraminidase(NA) activity, which gives it the ability to cleave a component of those receptors, NAcneu. Previous studies have demonstrated that HN has interesting anti-neoplastic and immune-stimulating properties in mammalian species, including humans. To explore the application of the HN gene in cancer gene therapy, we con...     

Induction of Effective Antitumor Immune Response by Combined Administration of hIL-18 and NDV HN

To analyze the antitumor potential and mechanism of action of simultaneous Newcastle disease virus (NDV) hemagglutinin-neuraminidase(HN) and human interleukin 18(hIL-18) gene transfer in C57BL/6 mice with H22 hepatoma,the mouse model with H22 hepatoma was established in C57BL/6 mice, and the antitumor effects of the combined application of NDV HN and hIL-18 were evaluated in vivo. The results show that the growth of established tumors in mice immunized with adenovirus(Ad)-HN in conjunction with Ad-hIL-18 was significantly inhibited compared with that in mice immunized with Ad-HN, Ad-hIL-18 alone, or the empty vector(Ad-mock). Furthermore, the immunization of mice with Ad-HN in conjunction with Ad-hIL-18 elicited strong natural killer activity and H22 tumor-specific cytotoxic T lymphocyte(CTL) responses in vivo. In addition, T cells from the lymph nodes of mice immunized with Ad-hIL-18 or Ad-HN+Ad-hIL-18 secreted high levels of the Th1 cytokine IL-2 and interferon-γ (IFN-γ), indicating that the regression of tumor cells is related to a Th1-type dominant immune response. These results demonstrate that vaccination with NDV HN together with hIL-18 may be a novel and powerful strategy for cancer immunotherapy.     

新城疫病毒流式微球免疫检测新方法

本研究基于新城疫病毒多克隆抗体的制备及优化,以聚苯乙烯微球作为蛋白质载体建立免疫检测体系,建立了快速检测新城疫病毒的流式细胞术新方法。以藻红蛋白荧光染料对新城疫多克隆抗体荧光标记,取1μL荧光微球与100μL新配制的单克隆抗体溶液反应,依次加入一定量待测抗原和藻红蛋白标记的多克隆抗体,漩涡振荡10s,室温振荡反应充分,形成双抗体夹心复合物,用流式细胞仪进行检测。采用ELISA法对免疫试剂进行了匹配性筛选实验,优选了试剂用量,并与传统ELISA方法进行了对比分析。实验结果表明,单克隆抗体350mg/L、多克隆抗体300mg/L时可获得最佳分析效果,与传统的ELISA方法呈现出良好的相关性,不与鸡传染性支气管炎病毒、鸡痘病毒、鸡马立克氏病病毒等发生交叉反应。     

Oncolytic Newcastle Disease Virus Co-Delivered with Modified PLGA Nanoparticles Encapsulating Temozolomide against Glioblastoma Cells: Developing an Effective Treatment Strategy

Glioblastoma multiforme (GBM) is considered to be one of the most serious version of primary malignant tumors. Temozolomide (TMZ), an anti-cancer drug, is the most common chemotherapeutic agent used for patients suffering from GBM. However, due to its inherent instability, short biological half-life, and dose-limiting characteristics, alternatives to TMZ have been sought. In this study, the TMZ-loaded PLGA nanoparticles were prepared by employing the emulsion solvent evaporation technique. The prepared TMZ-PLGA-NPs were characterized using FT-IR, zeta potential analyses, XRD pattern, particle size estimation, TEM, and FE-SEM observations. The virotherapy, being safe, selective, and effective in combating cancer, was employed, and TMZ-PLGA-NPs and oncolytic Newcastle Disease Virus (NDV) were co-administered for the purpose. An AMHA1-attenuated strain of NDV was propagated in chicken embryos, and the virus was titrated in Vero-slammed cells to determine the infective dose. The in vitro cytotoxic effects of the TMZ, NDV, and the TMZ-PLGA-NPs against the human glioblastoma cancer cell line, AMGM5, and the normal cell line of rat embryo fibroblasts (REFs) were evaluated. The synergistic effects of the nano-formulation and viral strain combined therapy was observed on the cell lines in MTT viability assays, together with the Chou–Talalay tests. The outcomes of the in vitro investigation revealed that the drug combinations of NDV and TMZ, as well as NDV and TMZ-PLGA-NPs exerted the synergistic enhancements of the antitumor activity on the AMGM5 cell lines. The effectiveness of both the mono, and combined treatments on the capability of AMGM5 cells to form colonies were also examined with crystal violet dyeing tests. The morphological features, and apoptotic reactions of the treated cells were investigated by utilizing the phase-contrast inverted microscopic examinations, and acridine orange/propidium iodide double-staining tests. Based on the current findings, the potential for the use of TMZ and NDV as part of a combination treatment of GBM is significant, and may work for patients suffering from GBM.     

Potent antiviral effect of green synthesis silver nanoparticles on Newcastle disease virus

Newcastle disease virus (NDV) causes serious infectious diseases in birds, affecting poultry production. In addition to adverse side effects, almost all conventional drugs targeting viral proteins have drug resistance mutations. This study aimed to evaluate the antiviral activity of green silver nanoparticles using green tea leaf extract as a new strategy to control NDV in ovo. The Log embryo infective dose₅₀ (EID₅₀) virucidal reduction was used to measure the antiviral activity of silver nanoparticles against NDV. The treatment of Vero cells with the silver nanoparticles (AgNPs) at a noncytotoxic concentration significantly therapeutic value by inhibiting NDV entry and reduced viral replication, which led to a great reduction in the viral titer in ovo. In conclusion, silver nanoparticles are effective as a therapeutic antiviral agent against NDV and inhibit microbial resistance by making it difficult for the microbe to adapt.     

Detection of Newcastle disease virus with quantum dots-resonance light scattering system

A sensitive QDs-based RLS assay method for the detection of Newcastle disease virus (NDV) antibody has been developed. CdTe quantum dots (QDs) were conjugated with Newcastle disease virus and used as RLS-based probes to detect NDV antibody. The electrostatic interaction between CdTe QDs and NDV resulted in enhanced resonance light scattering (RLS) signal characterized at 555 nm. Upon the addition of NDV antibody, QDs-NDV formed dispersive immunocomplex that can decrease the RLS signal. The decreased RLS intensity at 555 nm (ΔI_RLS) was linearly proportional to the concentration of NDV antibody (C_anti-NDV) in the range of 0.5-50 ng/mL, with correlation coefficient of 0.974 and detection limit of 0.1 ng/mL under the optimization conditions. The proposed method was applied to the determination of NDV antibody in spiked samples with satisfactory results.     

Gold immunochromatographic strips for enhanced detection of Avian influenza and Newcastle disease viruses

A new technique that uses gold immunochromatographic strips enhances the detection sensitivity by inducing the clustering of additional gold nanoparticles (AuNPs) around the immunogold particles immobilized on nitrocellulose strips. The additional AuNPs provide an intense signal that can be detected by the naked eye. The AuNPs were synthesized and conjugated to monoclonal antibodies using self-assembly. Other antibodies were immobilized in a defined detection zone on the nitrocellulose membrane. The detection principle is based on a “sandwich” immunoreaction, where gold-labeled antibodies serve as signal vehicles. To improve the sensitivity of the strips, we use a mixture of 1% HAuCl₄ and 10 mmol L⁻¹ NH₂OH·HCl to “enlarge” the gold nanoparticles. The detecting limits of Avian influenza virus (AIV) and Newcastle disease virus (NDV) are significantly increased. Compared with commercial test strips, this method is 100-fold more sensitive. This method is easy to perform and can be carried out on-site in test laboratories.     

Vibrational Imaging of Glucose Uptake Activity in Live Cells and Tissues by Stimulated Raman Scattering

Glucose is a ubiquitous energy source for most living organisms. Its uptake activity closely reflects cellular metabolic demand in various physiopathological conditions. Extensive efforts have been made to specifically image glucose uptake, such as with positron emission tomography, magnetic resonance imaging, and fluorescence microscopy, but all have limitations. A new platform to visualize glucose uptake activity in live cells and tissues is presented that involves performing stimulated Raman scattering on a novel glucose analogue labeled with a small alkyne moiety. Cancer cells with differing metabolic activities can be distinguished. Heterogeneous uptake patterns are observed with clear cell–cell variations in tumor xenograft tissues, neuronal culture, and mouse brain tissues. By offering the distinct advantage of optical resolution but without the undesirable influence of fluorophores, this method will facilitate the study of energy demands of living systems with subcellular resolution.     

Anti-tumor Effects of a Recombinant Fowlpox Virus Expressing Apoptin on Human Cervical Carcinoma in Vivo and in Vitro

Apoptin is a chicken anemia virus-derived,p53-independent,bcl-2-insensitive apoptotic protein with the ability to specifically induce apoptosis of various human tumor cells,but not of normal diploid cells.To explore the application of apoptin in tumor gene therapy,we used a recombinant fowlpox virus expressing apoptin protein (vFV-Apoptin) to investigate the anti-tumor effectes of vFV-Apoptin on human cervical carcinoma(HeLa) cells in vivo and in vitro through 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetraz...     

Spleen SORT LNP Generated in situ CAR T Cells Extend Survival in a Mouse Model of Lymphoreplete B Cell Lymphoma

Chimeric Antigen Receptor (CAR) T cell immunotherapy is revolutionizing treatment for patients suffering from B-cell lymphoma (BL). However, the current method of CAR T cell production is complicated and expensive, requiring collection of patient blood to enrich the T cell population, ex vivo engineering/activation, and quality assessment before the patient can receive the treatment. Herein we leverage Spleen Selective ORgan Targeted (SORT) Lipid Nanoparticles (LNPs) to produce CAR T cells in situ and bypass the extensive and laborious process currently used. Optimized Spleen SORT LNPs containing 10 % 18 : 1 PA transfected CD3+, CD8+, and CD4+ T cells in wild-type mice. Spleen SORT LNPs delivered Cre recombinase mRNA and CAR encoding mRNA to T cells in reporter mice and in a lymphoreplete B cell lymphoma model (respectively) after intravenous injection without the need for active targeting ligands. Moreover, in situ CAR T cells increased the overall survival of mice with a less aggressive form of B cell lymphoma. In addition, in situ transfected CAR T cells reduced tumor metastasis to the liver by increasing tumor infiltrating lymphocytes. Overall, these results offer a promising alternative method for CAR T cell production with pre-clinical potential to treat hematological malignancies.     

硒化聚甘露糖醛酸的合成及抗阿尔茨海默症细胞氧化与凋亡

以聚甘露糖醛酸为原料, 采用先磺化、 再硒化的方法合成了硒化聚甘露糖醛酸, 产率为54%, 产物硒含量为437.25 μg/g. 在2.5 μmol/L硒浓度下, 硒化聚甘露糖醛酸促细胞生长能力达到最适范围, 能保护细胞免受过氧化氢损伤, 显著提高阿尔茨海默症(AD)模型细胞N2a-APP695-sw中的超氧化物歧化酶和谷胱甘肽过氧化物酶的活性, 降低细胞内活性氧自由基, 增加线粒体膜电位, 抑制细胞色素C的释放, 在促进Bcl-2表达的同时抑制Bax的表达, 从而具有抑制AD细胞凋亡的功能. 硒化聚甘露糖醛酸也能抑制AD病理相关蛋白BACE1和APP的表达. 结果表明, 硒化聚甘露糖醛酸在抗AD方面具有潜在的应用前景.     

催化材料对病毒的吸附和灭活作用及对哺乳动物细胞的毒性

 提出了以吸附和催化原理灭活病毒的设想，旨在开发出对病毒有过滤、吸附及灭活作用的高效非特异性催化材料，应用于各种防护设施，有效控制非典型肺炎（SARS）的传播．采用与SARS病毒相似的副流感病毒作为模拟对象，进行了吸附及灭活该病毒的催化材料研究，并考察了催化材料对哺乳动物细胞的毒性．结果表明，病毒气溶胶的阻留及吸附结果与基于DNA吸附的色谱分析结果相一致；部分材料可以强烈地吸附病毒（100％），甚至在强烈振荡下并洗脱至第3次，病毒也不能脱附；一些材料不仅可以吸附病毒，而且强烈振荡后的洗脱液虽然表现出一定的血凝效价，但接种鸡胚后，病毒并不增殖，说明材料具有明显的催化病毒灭活性能；对细胞毒性极低的材料可以用在与人体接触的防护材料和设施中．筛选出的性能优异的催化材料，拟进一步考察其对SARS病毒的灭活作用．     

混合价态钨硼稀土杂多蓝在MDCK细胞内抑制流感病毒的活性

合成了过渡金属取代型硼稀土杂多蓝Ln₂H₃[BW₉^ⅣW₂^ⅤCo(H₂O)O₃₉]·nH₂O(Ln=La,Ce,Pr,Nd,Sm,Eu,Gd),并对化合物进行了IR,UV-Vis,XPS和ESR等表征.采用CPE法测定了化合物在MDCK细胞内抗流感病毒(A/H1N1/Jingfang/1/91,A/H₃N₂/Jingfang/30/95,B/Hufang/1/87)的活性.经评定,化合物CeH₃[BW₁₁Co(H₂O)O₃₉]表现出优异的抗病毒活性,这些化合物在病毒吸收期间显示出很强的效力.比较了杂多蓝与母体杂多酸在细胞毒性及抗病毒活性方面的差异.结果表明,杂多蓝的抗病毒活性优于其母体杂多酸.对化合物的构效关系以及抗病毒机理方面的研究结果表明,杂多化合物可抑制病毒早期的复制过程,也可能干扰RNA转录酶的活性.     

基于分子识别的免疫层析技术用于新冠肺炎感染的快速诊断

目前由新型冠状病毒(SARS-CoV-2)引发的新冠肺炎疫情仍在全球蔓延. 快速筛查并隔离感染者(包括无症状感染者)是遏制疫情传播的重要手段之一. 免疫层析技术是一种相对成熟的快速检测技术, 由于其操作简单、 反应时间短且结果稳定, 在生物标志物检测领域具有广阔的应用前景. 本文总结了目前免疫层析检测技术在新冠肺炎感染筛查领域的研究进展, 涵盖病毒抗体、 蛋白、 核酸等检测靶标, 并对不同检测方法的优势、 局限性进行了简要评述, 最后简单介绍了目前用于新冠肺炎感染筛查的免疫层析试纸的实际应用情况.     

纳米Eu2O3对HEK-293T细胞的生物活性影响

稀土化合物的生物效应研究已经引起广泛关注. 利用激光共聚焦显微镜观察到纳米Eu₂O₃能进入活HEK-293T细胞中且位于细胞核周围. 在体外培养条件下, 利用流式细胞仪法检测了纳米Eu₂O₃对细胞的活力及对细胞周期的影响, 结果表明当纳米颗粒浓度小于200 μg•mL^－1时, 对细胞活性没有明显影响; 而当纳米颗粒浓度≥400 μg•mL^－1时, 对细胞活性明显抑制. 当Eu₂O₃纳米颗粒在低浓度范围(≤50 μg•mL^－1)时对细胞周期基本没有影响, 而高浓度(≥200 μg•mL^－1)时对细胞周期均有显著抑制, 停滞在DNA合成后期. 这些研究结果均表明纳米Eu₂O₃对HEK-293T细胞具有明显的生物效应.     

Chrysophanol Attenuates Manifestations of Immune Bowel Diseases by Regulation of Colorectal Cells and T Cells Activation In Vivo

Inflammatory bowel disease (IBD) is an immune disorder that develops due to chronic inflammation in several cells. It is known that colorectal and T cells are mainly involved in the pathogenesis of IBD. Chrysophanol is an anthraquinone family member that possesses several bioactivities, including anti-diabetic, anti-tumor, and inhibitory effects on T cell activation. However, it is unknown whether chrysophanol suppresses the activity of colorectal cells. In this study, we found that chrysophanol did not induce cytotoxicity in HT-29 colorectal cells. Pre-treatment with chrysophanol inhibited the mRNA levels of pro-inflammatory cytokines in tumor necrosis factor-α (TNF-α)-stimulated HT-29 cells. Western blot analysis revealed that pre-treatment with chrysophanol mitigates p65 translocation and the mitogen-activated protein kinase (MAPK) pathway in activated HT-29 cells. Results from the in vivo experiment confirmed that oral administration of chrysophanol protects mice from dextran sulfate sodium (DSS)-induced IBD. Chrysophanol administration attenuates the expression of pro-inflammatory cytokines in colon tissues of the DSS-induced IBD model. In addition, we found that oral administration of chrysophanol systemically decreased the expression of effector cytokines from mesenteric lymph nodes. Therefore, these data suggest that chrysophanol has a potent modulatory effect on colorectal cells as well as exhibiting a beneficial potential for curing IBD in vivo.     

基于"分段"-"完整"转化的G-四链体脱氧核糖核酸酶传感器用于多聚核苷酸激酶的检测

采用"分段"转化为"完整"G-四链体脱氧核糖核酸酶的策略,构建了检测多聚核苷激酶(T4 PNK)的传感器.将形成G-四链体的富G序列PS5.M序列拆分成5 '和3'端均为羟基的两条链:链S1OH和链S2OH,即分别具有12个碱基的"分段"的PS5.M.在ATP存在下,T4 PNK酶可以将链S2OH的5 '端的羟基磷酸化,转化为S2P;在S1OH、S2P以及Helper链S—H存在下,T4 DNA连接酶(T4 DNA Ligase)将链S1OH和链S2P连接成"完整"的PS5.M序列.在体系中再加入核酸外切酶Ⅲ(ExoⅢ),从S—H的3'端剪切S-H,释放出PS5.M.在K+存在下,PS5.M与氯化血红素(Hemin)作用,形成具有类过氧化物酶活性的复合物,催化H2O2氧化2,2'-联氮-二(3-乙基-苯并噻唑-6-磺酸)二铵盐(ABTS)的反应,通过检测氧化产物在418 nm处的吸收值变化,实现对T4 PNK活性的定量检测.线性检测范围为0.02 ～ 3.0 U/mL,检出限为0.014 U/mL(S/N=3).对Hela细胞和HEK293细胞实际样本的T4 PNK活性进行了检测,平均回收率为95.6％～105.7％.     

化学发光成像法测定人体血清中乙型肝炎病毒

基于对碘苯酚增强的luminol-H2O2-HRP化学发光反应,利用化学发光成像法检测乙肝病毒(HBV)。用该法对人体血清中的乙型肝炎表面抗原、表面抗体、e抗原、e抗体以及核心抗体进行测定,其结果与ELISA法所得结果一致,对表面抗原检测结果为阳性的病人血清测定9次,结果的相对标准偏差为4．2％。