Nanoscale
·
2023
·Vol. 15
·期 7
·3475-3481
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IF 6.7
Rui Gong,Chenyang Yang,Ghulam Abbas,Dianbing Wang,Xianen Zhang,Guohui Nie,Hui Ding
The detection of the antibody of Epstein–Barr virus (EBV) is critical for the diagnosis of nasopharyngeal carcinoma (NPC). An accurate and scalable point-of-care detection method would support the screening, diagnosis, and monitoring of NPC patients. In this study, firstly, we made an antibody enrichment element, antigen-MNPs, which can screen out specific antibodies in a complex sample. Secondly, signal-amplifying elements were synthesized by labelling inorganic quantum dots (QDs) and anti-antibodies on the surface of flop-ferritin. A sandwich structure is formed among antigen-MNPs, target-antibodies, and anti-antibodies-flop-ferritin@QDs. The antibodies are quantified by fluorescence intensity with a limit of detection (LOD) as low as 10−11 g mL−1. Moreover, the method can detect different types of antibodies and was employed to examine 10 sera from NPC patients and 10 sera from healthy individuals. The result indicates that the simultaneous detection of anti-EBNA-IgG and anti-EBNA-IgA provides an efficient route for early diagnosis of NPC.
研究领域:
医学
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免疫学
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DOI
PubMed
高影响因子
Journal of Analytical Methods in Chemistry
·
2022
·Vol. 2022
·期 1
·5711384
·
IF 2.594
Miao Yu,Ming Liu,Yuan Li
This work describes a label-free electrochemical immunosensor for the sensing of Epstein-Barr virus (EBV) with high sensitivity. First, a monolayer of 1,6-hexanedithiol (HDT) was fabricated on the screen-printed electrode surface by the interaction between sulfur atoms and SPE. AuNPs can be modified on the electrode by the Au-S bond formed between the HDT-free group and Au atom in AuNPs. Protein A is then adsorbed onto AuNPs. Several parameters were optimized. The optimum concentration of protein A is 0.6 mg/mL. The optimum immobilization time for protein A is 90 min. The optimum concentration of antibody is 80 μ g/mL. The optimum immobilization time for antibody is 90 min. Directional immobilization of EBV antibody is achieved by high affinity binding of protein A to the Fc segment of antibody. When antigen specifically binds to antibody, the formation of immune complexes blocks electron transfer of [Fe(CN)6] 4-/3- and is reflected in the detection of cyclic voltammetry/electrochemical impedance spectroscopy. The detection range is 1 pg/mL–l00 ng/mL with a LOD of 0.1 pg/mL. In addition, the proposed sensor exhibited an excellent antiinterference property.
研究领域:
医学检测
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生物传感
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DOI
高影响因子
Advanced Healthcare Materials
·
2025
·
IF 9.6
Yanping Xie,Yunfei Zhu,Ya Wen,Siqi Hu,Ju Jiao,Yue Wu,Lijun Jiang,Bruno Viana,Ka-Leung Wong,Jing Wang,Rui Zou
Epstein–Barr virus (EBV) is a well-recognized oncogenic virus that promotes several lymphoid and epithelial cancers. The Epstein–Barr nuclear antigen 1 (EBNA1), which is known to be expressed in all EBV-positive cancers, plays a vital role in viral genome replication and maintenance and is therefore emerged as an attractive target for clinical intervention. Several EBNA1 inhibitors have shown potency in the growth inhibition of EBV-positive cancers, yet low bioavailability and in vivo unmonitored nature hamper their further implementation. Here a novel EBNA1 nano-inhibitor based on EBNA1-specific peptide inhibitor (P4) functionalized ZnGa 2 O 4 :Cr 3+ ultra-small near-infrared persistent luminescent (NIR-PL) nanoparticles (ZGOC-P4) is developed. Owing to the specific binding to EBNA1, ZGOC-P4 nano-inhibitor can quickly achieve nuclear internalization in EBV-positive nasopharyngeal carcinoma (NPC) cells (C666-1) and selectively inhibit their growth. In sharp contrast, ZGOC-P4 nano-inhibitor shows no inhibition effect on EBV-negative NPC cells (HK-1). Moreover, the results indicate that the well-designed nano-inhibitor enables efficient tumor-targeting accumulation in NPC xenograft model under the monitoring of autofluorescence interference-free NIR-PL imaging in vivo and suppresses EBV-associated tumor growth with an inhibition rate of 61.6%. This work highlights the potency of ZGOC-P4 on NPC treatment and may provide new sight into future research on EBV-associated diseases.
研究领域:
生物医学工程
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肿瘤学
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病毒学
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产品关联
DOI
PubMed
高影响因子
EBioMedicine
·
2022
·
IF 11.205
Ruo-Wen Xiao,Fang Wang,Tong-Min Wang,Jiang-Bo Zhang,Zi-Yi Wu,Chang-Mi Deng,Ying Liao,Ting Zhou,Da-Wei Yang,Si-Qi Dong,Wen-Qiong Xue,Yong-Qiao He,Xiao-Hui Zheng,Xi-Zhao Li,Pei-Fen Zhang,Shao-Dan Zhang,Ye-Zhu Hu,Yu-Ying Liu,Yun-Fei Xia,Song Gao,Jian-Bing Mu,Lin Feng,Wei-Hua Jia
Summary Background Nasopharyngeal carcinoma (NPC) exhibits significant familial aggregation; however, its susceptibility genes are largely unknown. Thus, this study aimed to identify germline mutations that might contribute to the risk of familial NPC, and explore their biological functions. Methods Whole-exome sequencing was performed in 13 NPC pedigrees with multiple cases. Mutations co-segregated with disease status were further validated in a cohort composed of 563 probands from independent families, 2,953 sporadic cases, and 3,175 healthy controls. Experimental studies were used to explore the functions of susceptibility genes and their disease-related mutations. Findings The three rare missense mutations in POLN (DNA polymerase nu) gene, P577L, R303Q, and F545C, were associated with familial NPC risk (5/576 [0·87%] in cases vs. 2/3374 [0·059%] in healthy controls with an adjusted OR of 44·84 [95% CI:3·91-514·34, p = 2·25 × 10 −3 ]). POLN was involved in Epstein-Barr virus (EBV) lytic replication in NPC cells in vitro. POLN promoted viral DNA replication, immediate-early and late lytic gene expression, and progeny viral particle production, ultimately affecting the proliferation of host cells. The three mutations were located in two pivotal functional domains and were predicted to alter the protein stability of POLN in silico . Further assays demonstrated that POLN carrying any of the three mutations displayed reduced protein stability and decreased expression levels, thereby impairing its ability to promote complete EBV lytic replication and facilitate cell survival. Interpretation We identified a susceptibility gene POLN for familial NPC and elucidated its function. Funding This study was funded by the National Key Research and Development Program of China (2021YFC2500400); the National Key Research and Development Program of China (2020YFC1316902); the Basic and Applied Basic Research Foundation of Guangdong Province, China (2021B1515420007); the National Natural Science Foundation of China (81973131); the National Natural Science Foundation of China (82003520); the National Natural Science Foundation of China (81903395).
研究领域:
肿瘤学
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分子生物学
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遗传学
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产品关联
DOI
PubMed
高影响因子
Cancer Management and Research
·
2020
·
IF 2.886
Mei Zeng,Yuhua Chen,Xintao Jia,Yan Liu
Background Epstein-Barr virus (EBV) has been indicated in the development of some tumors, including lymphoma. However, the potential role of latent membrane protein 1 (LMP1) encoded by EBV in the tumorigenesis of lymphoma remains debated. Herein, we examined the function of LMP1 in lymphoma. Methods The expression of LMP1 was downregulated or upregulated in EBV negative cell line SNT-8 and positive cell line KHYG-1, respectively. Subsequently, the cell viability, apoptosis, as well as the expression patterns of p53, mouse double minute 2 (MDM2), B-cell CLL/lymphoma 2 (Bcl-2) and NF-κB were evaluated. Next, the binding relationship between MDM2 and p53 along with p53 ubiquitination in cells was tested by Western blot and co-immunoprecipitation. Finally, the effects of LMP1 on lymphoma cell growth through p53, Bcl-2 and NF-κB pathways were verified by functional rescue experiments. Results Overexpression of LMP1 promoted KHYG-1 cell growth and inhibited cell apoptosis. Moreover, LMP1 upregulation significantly enhanced the activation of NF-κB pathway, thus increasing MDM2 binding to p53, leading to p53 ubiquitination and degradation as well as Bcl-2 expression enhancement. Further inhibition of the NF-κB pathway or Bcl-2 expression significantly weakened the promotive role of LMP1 in the growth of KHYG-1 cells. Conclusion EBV-LMP1 promoted the p53 ubiquitination and degradation by activating NF-κB signaling pathway and the following binding of MDM2 and p53 in cells to enhance Bcl-2 expression, thus promoting the growth of lymphoma cells and inhibiting cell apoptosis.
研究领域:
肿瘤学
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分子生物学
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细胞生物学
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病毒学
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产品关联
DOI
PubMed
高影响因子
Nano Research
·
2023
·
IF 9.9
Wang Daji,Wang Jie,Liu Dan,He Jiuyang,Wang Meiying,Huang Haibing,Nie Guohui,Ding Hui,Yan Xiyun
The correlation between Epstein-Barr virus (EBV) infection and nasopharyngeal carcinoma (NPC) risk has been extensively researched. The continual monitoring of EBV-IgAs provides a promising approach of NPC screening in its early stage. In this study, we successfully synthesized a single-atom nanozyme (SANzyme) through the application of iron-porphyrin based metal organic framework (MOF-FeP). The MOF-FeP possesses precisely-defined electronic and geometric structures that accurately mimic highly-evolved catalytic site of natural peroxidase. The peroxidase-like activity of MOF-FeP enables it to catalyze the chemiluminescence of luminol substrate. By integrating MOF-FeP into a traditional strip, we created a rapid and highly-sensitive evaluation tool for detecting EBV-IgAs. Importantly, the MOF-FeP strip enables the simultaneous detection of three EBV-IgAs, greatly improving the accuracy of EBV-associated NPC screening. The sensitivities of the MOF-FeP strip (75.56%–93.30%) surpass those of current enzyme-linked immunosorbent assay (ELISA) methods (64.44%–82.22%). This test takes only 16 min to perform as opposed to the customary 1–2 h required for standard ELISA. Additionally, the MOF-FeP strip is suitable for whole blood samples, thereby significantly simplifying the sample preparation and detection process. In conclusion, the MOF-FeP strip combines the simplicity of traditional strip with the high catalytic activity of SANzyme. Our innovative MOF-FeP strip offers a new point-of-care strategy for EBV-IgAs detection, which is expected to markedly facilitate early screening for EBV-associated diseases.
研究领域:
癌症生物学
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分子诊断
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纳米技术
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生物医学工程
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产品关联
DOI
高影响因子
Scientific Reports
·
2024
·Vol. 14
·期 1
·1-15
·
IF 3.8
Chen Linli,Guo Xiaojing,Lin Wen,Huang Yingying,Zhuang Suling,Li Qianfeng,Xu Jianhua,Ye Shengnan
Lytic induction therapy was devised to selectively combat malignancies associated with Epstein–Barr virus (EBV) by triggering viral reactivation from latency. At present, the major challenges of lytic induction therapy are to maximize reactivating efficiencies and meanwhile minimize infectious virion production. C210, a novel curcumin derivative with potent Hsp90 inhibitory activity, was explored for EBV-reactivating and virion-producing effects in EBV-positive nasopharyngeal carcinoma (NPC) and gastric carcinoma (GC) cell lines. And the molecular mechanisms underlying these effects were determined. Follow C210 treatment, EBV lytic RNAs and proteins were upregulated, but infectious virions were not produced. Knockdown of heat shock protein 90 (Hsp90) induced expression of lytic RNAs and proteins, and diminished C210-driven EBV lytic induction. Pretreatment with an X box binding protein 1 (XBP1) inhibitor reduced C210-induced EBV lytic RNA. Furthermore, we demonstrated that C210 inhibited the binding of Hsp90 with its clients, signal transducer and activator of transcription 3 (STAT3) and xeroderma pigmentosum group B-complementing protein (XPB), which subsequently promoted their proteasomal degradation. Degradation of STAT3 by C210 enhanced the EBV-reactivating and anticancer capacity of suberoylanilide hydroxamic acid (SAHA). Depletion of XPB blocked SAHA-induced expression of late viral genes and production of infectious virions. These results elucidate a novel Hsp90 inhibitor targeting EBV lytic phase and extend the research on lytic induction strategy, which may offer reference value in the treatment of EBV-positive malignancies.
研究领域:
肿瘤学
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病毒学
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分子生物学
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药理学
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产品关联
DOI
PubMed
高影响因子
Frontiers in Immunology
·
2023
·
IF 7.3
Xi Zhang,Tiaoxia Wang,Xiaona Zhu,Yong Lu,Mingpeng Li,Zhihong Huang,Deping Han,Longzhen Zhang,Yang Wu,Liantao Li,Frank Klawonn,Renata Stripecke
Introduction Epstein-Barr virus (EBV) is a widely spread pathogen associated with lymphoproliferative diseases, B/ T/ NK cell lymphomas, nasopharyngeal carcinoma (NPC) and gastric carcinoma (GC). EBV lytic reactivations contribute to the genomic instability, inflammation and tumorigenesis of NPC, promoting cancer progression. Patients with NPC refractory to standard therapies show dismal survival. EBV gp350 is an envelope protein detectable in NPC specimens intracellularly and on the cell membrane of malignant cells, and is a potential viral antigen for T cell-directed immunotherapies. The potency of T cells engineered with a chimeric antigen receptor (CAR) targeting gp350 against EBV + lymphoproliferative disease was previously shown. Methods Here, we advanced towards preclinical and non-clinical developments of this virus-specific CAR-T cell immunotherapy against NPC. Different gp350CAR designs were inserted into a lentiviral vector (LV) backbone. Results A construct expressing the scFv 7A1-anti−gp350 incorporating the CD8 transmembrane and CD28.CD3ζ signaling domain (ZT002) was selected. High titer ZT002 (~1x10 8 TU/ml) was manufactured in HEK 293T/17 suspension cells in serum free media as large-scale production under good manufacturing practices (GMP). A LV multiplicity of infection (MOI) of 1 resulted in high frequencies of functional gp350CAR + T cells (>70%) at a low (<2) vector copy numbers in the genome. ZT002 was therefore used to establish gp350CAR-T batch run production methods. GMP upscaling and validation of T cell transduction and expansion in several runs resulted in average 3x10 9 gp350CAR-T cells per batch. >80% CD3 + gp350CAR-T cells bound to purified gp350 protein. In vitro cytotoxicity and cytokine secretion assays (IFN-γ and TNF-α) confirmed the specificity of gp350CAR-T cells against gp350 + NPC, GC and lymphoma cell targets. Immunocompromised B-NDG mice (NOD.CB17- PrkdcscidIl2rgtm1 /Bcgen) were challenged s.c. with a EBV + NPC C666.1 cell line expressing gp350 and then treated with escalating doses of gp350CAR-T cells or with non-transduced T cells. gp350CAR-T cells promoted antitumor responses, bio-distributed in several tissues, infiltrated in tumors and rejected gp350 + tumor cells. Discussion These results support the use of gp350CAR-T cells generated with ZT002 as an Innovative New Drug to treat patients with solid and liquid EBV-associated malignancies.
研究领域:
免疫治疗
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肿瘤学
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分子生物学
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病毒学
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临床前研究
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产品关联
DOI
PubMed
高影响因子
Journal of Advanced Research
·
2024
·
IF 11.4
Songwei Tan,Caiyan Yuan,Yuhe Zhu,Shuangyan Chang,Qianru Li,Jiahui Ding,Xueqin Gao,Rui Tian,Zhiqiang Han,Zheng Hu
Introduction CRISPR/Cas9 gene editing technology has significantly advanced gene therapy, with gene vectors being one of the key factors for its success. Poly (beta-amino ester) (PBAE), a distinguished non-viral cationic gene vector, is known to elevate intracellular reactive oxygen species (ROS) levels, which may cause cytotoxicity and, consequently, impact gene transfection efficacy (T.E.). Objectives To develop a simple but efficient strategy to improve the gene delivery ability and biosafety of PBAE both in vivo and in vitro . Methods We used glutathione (GSH), a clinically utilized drug with capability to modulating intracellular ROS level, to prepare a hybrid system with PBAE-plasmid nanoparticles (NPs). This system was characterized by flow cytometry, RNA-seq, Polymerase Chain Reaction (PCR) and Sanger sequencing in vitro , and its safety and efficacy in vivo was evaluated by imaging, PCR, Sanger sequencing and histology analysis. Results The particle size of GSH-PBAE-plasmid NPs were 168.31 nm with a ζ-potential of 15.21 mV. An enhancement in T.E. and gene editing efficiency, ranging from 10 % to 100 %, was observed compared to GSH-free PBAE-plasmid NPs in various cell lines. In vitro results proved that GSH-PBAE-plasmid NPs reduced intracellular ROS levels by 25 %–40 %, decreased the total number of upregulated/downregulated genes from 4,952 to 789, and significantly avoided the disturbance in gene expression related to cellular oxidative stress-response and cell growth regulation signaling pathway compared to PBAE-plasmid NPs. They also demonstrated lower impact on the cell cycle, slighter hemolysis, and higher cell viability after gene transfection. Furthermore, GSH hybrid PBAE-plasmid NPs exhibited superior safety and improved tumor suppression ability in an Epstein–Barr virus (EBV)-infected murine tumor model, via targeting cleavage the EBV related oncogene by delivering CRISPR/Cas9 gene editing system and down-regulating the expression levels. This simple but effective strategy is expected to promote clinical applications of non-viral vector gene delivery.
研究领域:
基因治疗
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纳米技术
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分子生物学
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药学
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癌症研究
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产品关联
DOI
PubMed
高影响因子
危险品化学品经营许可证（带存储）