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Spillover of Newcastle disease virus to Himalayan Griffon vulture: a possible food-based transmission. 新城疫病毒向喜马拉雅狮鹫的扩散:一种可能的食物传播。
IF 1.9 4区 医学 Q3 GENETICS & HEREDITY Pub Date : 2024-08-01 Epub Date: 2024-05-13 DOI: 10.1007/s11262-024-02072-9
Shinjini Bhattacharya, Pankaj Deka, Sangeeta Das, Samshul Ali, Bhaskar Choudhury, Parikshit Kakati, Sachin Kumar

The Newcastle disease virus (NDV) affects wild and domesticated bird species, including commercial poultry. Although the diversity of NDV in domestic chickens is well documented, limited information is available about Newcastle disease (ND) outbreaks in other bird species. We report an annotated sequence of NDV/Vulture/Borjuri/01/22, an avirulent strain of NDV reported from Borjuri, Northeast India, in Himalayan Griffon vulture. The complete genome is 15,186 bases long with a fusion protein (F) cleavage site 112GRQGR↓L117. The phylogenetic analysis based on the F protein gene and the whole genome sequence revealed that the isolate from the vulture belongs to genotype II, sharing significant homology with vaccine strain LaSota. The study highlights the possible spillover of the virus from domestic to wild species through the food chain.

新城疫病毒(NDV)会影响野生和驯养的鸟类物种,包括商品家禽。尽管 NDV 在家鸡中的多样性已被充分记录,但有关新城疫(ND)在其他鸟类中爆发的信息却很有限。我们报告了NDV/Vulture/Borjuri/01/22的注释序列,NDV/Vulture/Borjuri/01/22是NDV在喜马拉雅狮鹫中的一个无病毒毒株,报告来自印度东北部的Borjuri。完整的基因组长度为 15,186 个碱基,融合蛋白(F)裂解位点为 112GRQGR↓L117。基于 F 蛋白基因和全基因组序列的系统进化分析表明,该秃鹫分离株属于基因型 II,与疫苗株 LaSota 有显著的同源性。该研究强调了病毒可能通过食物链从家养物种向野生物种扩散。
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引用次数: 0
H6N2 reassortant avian influenza virus isolate in wild birds in Jiangxi Province, China. 中国江西省野鸟中的 H6N2 重型禽流感病毒分离株。
IF 1.6 4区 医学 Q3 GENETICS & HEREDITY Pub Date : 2024-06-01 Epub Date: 2024-05-09 DOI: 10.1007/s11262-024-02068-5
Yingjie Wang, Xiang Li, Xinru Lv, Yi Li, Qing An, Yang Xiu, Xiangtong Lv, Haozhuo Song, Changyu Zhou, Qiushi Xu, Dan Qiao, Siyuan Yang, Hongliang Chai, Xiangwei Zeng

H6 avian influenza virus is widely prevalent in wild birds and poultry and has caused human infection in 2013 in Taiwan, China. During our active influenza surveillance program in wild waterfowl at Poyang Lake, Jiangxi Province, an H6N2 AIV was isolated and named A/bean goose/JiangXi/452-4/2013(H6N2). The isolate was characterized as a typical low pathogenic avian influenza virus (LPAIV) due to the presence of the amino acid sequence PQIETR↓GLFGAI at the cleavage site of the hemagglutinin (HA) protein. The genetic evolution analysis revealed that the NA gene of the isolate originated from North America and exhibited the highest nucleotide identity (99.29%) with a virus recovered from wild bird samples in North America, specifically A/bufflehead/California/4935/2012(H11N2). Additionally, while the HA and PB1 genes belonged to the Eurasian lineage, they displayed frequent genetic interactions with the North American lineage. The remaining genes showed close genetic relationships with Eurasian viruses. The H6N2 isolate possessed a complex genome, indicating it is a multi-gene recombinant virus with genetic material from both Eurasian and North American lineages.

H6 禽流感病毒在野禽和家禽中广泛流行,2013 年曾在中国台湾造成人类感染。在对江西省鄱阳湖野生水禽进行流感监测期间,我们分离到了一种 H6N2 AIV,并将其命名为 A/江西/452-4/2013(H6N2)。由于该分离株的血凝素(HA)蛋白裂解位点存在氨基酸序列 PQIETR↓GLFGAI,因此被定性为典型的低致病性禽流感病毒(LPAIV)。遗传进化分析表明,该分离株的NA基因来自北美,与从北美野鸟样本中发现的病毒(特别是A/bufflehead/California/4935/2012(H11N2))具有最高的核苷酸同一性(99.29%)。此外,虽然 HA 和 PB1 基因属于欧亚系,但它们与北美系的基因相互作用频繁。其余基因与欧亚病毒的遗传关系密切。H6N2 病毒分离株拥有复杂的基因组,表明它是一种多基因重组病毒,其遗传物质既来自欧亚系,也来自北美系。
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引用次数: 0
Amsacta moorei entomopoxvirus encodes a protein kinase with dual activity and a broad substrate spectrum including two putative cellular substrates. Amsacta moorei entomopoxvirus编码的蛋白激酶具有双重活性和广泛的底物谱,包括两种假定的细胞底物。
IF 1.6 4区 医学 Q3 GENETICS & HEREDITY Pub Date : 2024-06-01 Epub Date: 2024-05-04 DOI: 10.1007/s11262-024-02069-4
Hacer Muratoğlu, Remziye Nalcacioglu, Basil M Arif, Zihni Demirbag

Amsacta moorei entomopoxvirus (AMEV) is a poxvirus that can only infect insects. This virus is an attractive research material because it is similar to smallpox virus. AMEV is one of many viruses that encode protein kinases that drive the host's cellular mechanisms, modifying immune responses to it, and regulating viral protein activity. We report here the functional characterization of a serine/threonine (Ser/Thr) protein kinase (PK) gene (ORF AMV197) of AMEV. Expression of the AMV197 gene in baculovirus expression system yielded a ~ 35.5 kDa protein. PK activity of expressed AMV197 was shown by standard PK assay. Substrate profiling of AMV197 protein by peptide microarray indicated that the expressed protein phosphorylated 81 of 624 substrates which belong to 28 families of PK substrates. While the hypothetical AMV197 protein phosphorylates Ser/Thr only, we demonstrated that the expressed PK also phosphorylates probes with tyrosine residues on the array which is a rare property among PKs. Pull-down assay of the AMV197 protein with the subcellular protein fractionations of Ld652 cells showed that it is using two cellular proteins (18 and 42 kDa) as novel putative substrates. Our results suggest that AMEV can regulate cellular mechanisms by phosphorylating cellular proteins through AMV197 PK. However, further experiments are needed to identify the exact role of this PK in the replication of AMEV.

Amsacta moorei entomopoxvirus(AMEV)是一种只能感染昆虫的痘病毒。这种病毒与天花病毒相似,因此是一种极具吸引力的研究材料。AMEV 是许多编码蛋白激酶的病毒之一,这些蛋白激酶能驱动宿主的细胞机制,改变对它的免疫反应,并调节病毒蛋白的活性。我们在此报告了 AMEV 的丝氨酸/苏氨酸(Ser/Thr)蛋白激酶(PK)基因(ORF AMV197)的功能特征。在杆状病毒表达系统中表达 AMV197 基因可获得约 35.5 kDa 的蛋白。表达的AMV197的PK活性由标准的PK测定法显示。用肽芯片对 AMV197 蛋白进行底物分析表明,表达的蛋白磷酸化了 624 种底物中的 81 种,这些底物属于 PK 底物的 28 个家族。虽然假定的 AMV197 蛋白只磷酸化 Ser/Thr,但我们证实表达的 PK 还磷酸化了肽阵列上带有酪氨酸残基的探针,这在 PK 中是罕见的。用 Ld652 细胞的亚细胞蛋白分馏物对 AMV197 蛋白进行的牵引试验表明,它将两种细胞蛋白(18 和 42 kDa)作为新的假定底物。我们的结果表明,AMEV 可通过 AMV197 PK 磷酸化细胞蛋白来调节细胞机制。然而,要确定这种 PK 在 AMEV 复制中的确切作用,还需要进一步的实验。
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引用次数: 0
Isolation and whole-genome sequence analysis of Balagodu virus in Japan. 日本 Balagodu 病毒的分离和全基因组序列分析。
IF 1.6 4区 医学 Q3 GENETICS & HEREDITY Pub Date : 2024-06-01 Epub Date: 2024-03-16 DOI: 10.1007/s11262-024-02060-z
Mayumi Honda, Hironori Setoyama, Ryosuke Nabekura, Katsunori Murota, Yuto Suda, Tohru Yanase

Whole-genome sequencing of a virus isolated from Culicoides biting midges in southern Japan in 2020 revealed that it is a strain of Balagodu virus (BLGV; genus Orthobunyavirus; family Peribunyaviridae; order Bunyavirales). A solitary instance of BLGV isolation occurred in India in 1963. All assembled segments comprise complete protein-coding sequences that are similar to those of other orthobunyaviruses. The consensus 3'- and 5'-terminal sequences of orthobunyaviruses' genomic RNAs are also conserved in the Japanese BLGV strain. Here, we update the geographic distribution of BLGV and provide its complete sequence, contributing to the clarification of orthobunyavirus phylogeny.

对 2020 年从日本南部咬蠓中分离出的一种病毒进行全基因组测序后发现,它是 Balagodu 病毒(BLGV;属 Orthobunyavirus;科 Peribunyaviridae;目 Bunyavirales)的一个毒株。1963年,印度曾分离出一株BLGV。所有组装的片段都包含完整的蛋白质编码序列,这些序列与其他正布尼亚病毒相似。在日本 BLGV 株系中,正圆病毒基因组 RNA 的 3'- 和 5'- 末端序列也是一致的。在此,我们更新了BLGV的地理分布,并提供了其完整序列,为澄清正圆病毒的系统发育做出了贡献。
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引用次数: 0
Donkey-like kirkovirus is associated with diarrhea in piglets. 驴样柯科病毒与仔猪腹泻有关。
IF 1.6 4区 医学 Q3 GENETICS & HEREDITY Pub Date : 2024-06-01 Epub Date: 2024-03-25 DOI: 10.1007/s11262-024-02066-7
Panpan Tong, Zunbao Wang, Yueyi Dang, Lei Zhang, Guangwei Song, Xiaozhen Song, Juanjuan Pan, Ling Kuang, Junhui Li, Gang Lu, Jinxin Xie

Kirkovirus (kirV), a seemingly novel virus family, has been found in horses and donkeys. The study's objectives are to investigate the presence of the virus in swine. In this study, donkey-like kirV was detected in rectal swabs of piglets with diarrhea, and the positive rate was found to be 100% (149/149). However, this virus was detected in only one of 261 clinically healthy piglets, which suggested a strong relationship between the kirV and the diarrheic disease. We obtained the whole-genome sequences of three kirVs (Cj-D5, Cj-D32, and Cj-D43), with a length of 3750 nucleotides (nt) and sharing 99.9% nt identity with donkey kirVs. Furthermore, the three viruses shared 88.5-100% and 23-51% of the Rep protein sequence, similar to available reference strains of Kirkoviridae and Circoviridae, respectively. Moreover, like horse and donkey kirVs, the RCR domain and P-loop NTPase domains of Rep protein and nonanucleotide motif (CAATATTAC) of the three viruses were similar to those of Circoviruses and Cycloviruses. Phylogenetic analysis showed that these viruses could be grouped with members in the proposed family Kirkoviridae. This is the first report to describe that kirV can circulate in piglets with diarrhea, and future studies are needed to determine the pathogenesis of this virus.

Kirkovirus (kirV)是一个看似新颖的病毒家族,曾在马和驴身上发现过。这项研究的目的是调查猪体内是否存在这种病毒。在这项研究中,从腹泻仔猪的直肠拭子中检测到了驴样 kirV,阳性率为 100%(149/149)。然而,在 261 头临床健康仔猪中,只有 1 头检测到该病毒,这表明 kirV 与腹泻病之间存在密切关系。我们获得了三种 kirV(Cj-D5、Cj-D32 和 Cj-D43)的全基因组序列,其长度为 3750 个核苷酸(nt),与驴 kirV 的 nt 一致度为 99.9%。此外,这三种病毒的 Rep 蛋白序列的共享率分别为 88.5%-100%和 23%-51%,与现有的柯科病毒科和圆环病毒科参考毒株相似。此外,与马和驴kirVs一样,这三种病毒的Rep蛋白的RCR结构域和P环NTP酶结构域以及非核苷酸基序(CAATATTAC)与圆环病毒和环状病毒相似。系统进化分析表明,这些病毒可与拟议的 Kirkoviridae 科的成员归为一类。这是第一份描述 kirV 可在腹泻仔猪中传播的报告,今后还需要对该病毒的致病机理进行研究。
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引用次数: 0
Ovine adenoviruses infecting sheep and goats in Türkiye: detection and molecular characterization of three different types. 感染土尔其绵羊和山羊的绵羊腺病毒:三种不同类型的检测和分子特征。
IF 1.6 4区 医学 Q3 GENETICS & HEREDITY Pub Date : 2024-06-01 Epub Date: 2024-03-15 DOI: 10.1007/s11262-024-02063-w
Ilke Karayel-Hacioglu, Buket Gul, Deniz Acun Yildiz, Feray Alkan

Adenoviruses (AdVs) have been detected in a wide variety of animals. To date, eight types of AdVs in sheep and two types in goats have been identified, which belong to two distinct genera, Mastadenovirus and Atadenovirus. Typically, the term pneumo-enteritis is used to describe adenovirus-induced disease in small ruminants, which has been associated with both enteric and respiratory symptoms of varying severity. The aim of this study was to detect and identify AdVs of small ruminants belonging to the genera Mastadenovirus and Atadenovirus. For this purpose, diagnostic samples (47 lung, 27 intestine, and two pooled tissue samples including intestine and lung) from 49 small ruminants (39 sheep and 10 goats) were used. Following the viral DNA extraction, PCR was carried out by using the primers targeting the hexon gene in order to detect both mast- and atadenoviruses. Sequencing the amplified fragments revealed the presence of three types of ovine adenovirus (OAdV): OAdV-3, OAdV-4, and OAdV-8. Specifically, OAdV-3 was detected in two sheep and a goat while OAdV-4 and OAdV-8 were found in only one sheep each. There is still limited data on the interaction between the viruses in different adenovirus genera and the detected disease, as well as the genetic diversity of adenoviruses, especially in small ruminants. In conclusion, the detection of AdVs in lung and intestinal tissues of small ruminants in this study suggests that these viruses may have contributed to the disease and/or predisposed to other agents.

腺病毒(AdVs)已在多种动物体内检测到。迄今为止,已在绵羊和山羊中分别发现了八种和两种腺病毒,它们分别属于两个不同的属,即马氏腺病毒属(Mastadenovirus)和阿特腺病毒属(Atadenovirus)。通常情况下,腺病毒引起的小反刍动物疾病被称为 "肺肠炎",这种疾病与严重程度不同的肠道和呼吸道症状有关。本研究的目的是检测和鉴定属于马氏腺病毒属(Mastadenovirus)和阿塔腺病毒属(Atadenovirus)的小反刍动物 AdVs。为此,研究人员使用了 49 只小反刍动物(39 只绵羊和 10 只山羊)的诊断样本(47 份肺样本、27 份肠道样本和两份包括肠道和肺的集合组织样本)。提取病毒 DNA 后,使用针对 hexon 基因的引物进行 PCR,以检测乳突病毒和阿塔登病毒。对扩增片段进行测序后发现了三种类型的绵羊腺病毒(OAdV):OAdV-3、OAdV-4 和 OAdV-8。具体来说,在两只绵羊和一只山羊身上检测到了 OAdV-3,而 OAdV-4 和 OAdV-8 只分别在一只绵羊身上发现。关于不同腺病毒属的病毒与所发现疾病之间的相互作用,以及腺病毒的遗传多样性(尤其是在小反刍动物中),目前的数据还很有限。总之,本研究在小反刍动物的肺部和肠道组织中检测到的 AdVs 表明,这些病毒可能导致了疾病和/或易感染其他病原体。
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引用次数: 0
Leucoma salicis nucleopolyhedrovirus (LesaNPV) genome sequence shed new light on the origin of the Alphabaculovirus orpseudotsugatae species 水蛭核多聚核酸病毒(LesaNPV)基因组序列揭示了阿尔法巴库尔病毒(Alphabaculovirus orpseudotsugatae)物种起源的新线索
IF 1.6 4区 医学 Q3 GENETICS & HEREDITY Pub Date : 2024-04-09 DOI: 10.1007/s11262-024-02062-x
Martyna Krejmer-Rabalska, Lukasz Rabalski, Maciej Kosinski, Iwona Skrzecz, Jadwiga Ziemnicka, Boguslaw Szewczyk

LesaNPV (Leucoma salicis nucleopolyhedrovirus) is an alphabaculovirus group Ib. Potentially, it can be an eco-friendly agent to control the white satin moth Leucoma salicis population. In this study, we have established the relationship between LesaNPV and other closely related alphabaculoviruses. Environmental samples of late instar of white satin moth collected in Poland infected with baculovirus have been homogenized, polyhedra were purified and subjected to scanning and transmission electron microscopy. Viral DNA was sequenced using the Illumina platform and the whole-genome sequence was established by de novo assembly of paired reads. Genome annotation and phylogenetic analyses were performed with the use of bioinformatics tools. The genome of LesaNPV is 132 549 bp long with 154 ORFs and 54.9% GC content. Whole-genome sequencing revealed deletion of dUTPase as well as ribonucleoside reductases small and large subunits region in LesaNPV genome compared to Dasychira pudibunda nucleopolyhedrovirus (DapuNPV) and Orgyia pseudotsugata multiple nucleopolyhedrovirus (OpMNPV) where this region is complete. Phylogenetic analysis of Baculoviridae family members showed that LesaNPV is less divergent from a common ancestor than closely related species DapuNPV and OpMNPV. This is interesting because their hosts do not occur in the same area. The baculoviruses described in this manuscript are probably isolates of one species and could be assigned to recently denominated species Alphabaculovirus orpseudotsugatae, historically originating from OpMNPV. This finding could have significant implications for the classification and understanding of the phylogeographical spread of baculoviruses.

LesaNPV(白缎蛾核多角体病毒)是一种Ⅰb组阿法巴库病毒。它有可能成为一种控制白缎纹夜蛾种群数量的环保药剂。在这项研究中,我们确定了 LesaNPV 与其他密切相关的阿尔法巴库洛病毒之间的关系。我们对在波兰收集到的感染了杆状病毒的白缎纹夜蛾晚鳞的环境样本进行了均质化处理,纯化了多面体,并对其进行了扫描和透射电子显微镜观察。利用 Illumina 平台对病毒 DNA 进行了测序,并通过对成对读数的从头组装建立了全基因组序列。利用生物信息学工具进行了基因组注释和系统进化分析。LesaNPV 的基因组长 132 549 bp,有 154 个 ORF,GC 含量为 54.9%。全基因组测序发现,与Dasychira pudibunda核多聚核酸病毒(DapuNPV)和Orgyia pseudotsugata多重核多聚核酸病毒(OpMNPV)相比,LesaNPV基因组中缺失了dUTP酶以及核糖核苷还原酶小亚基和大亚基区域,而Dasychira pudibunda核多聚核酸病毒(DapuNPV)和Orgyia pseudotsugata多重核多聚核酸病毒(OpMNPV)的这一区域是完整的。Baculoviridae 家族成员的系统进化分析表明,与密切相关的物种 DapuNPV 和 OpMNPV 相比,LesaNPV 与共同祖先的差异较小。这一点很有意思,因为它们的宿主并不在同一地区。本手稿中描述的杆状病毒可能是一个物种的分离物,可以归入最近命名的物种 Alphabaculovirus 或pseudotsugatae,历史上起源于 OpMNPV。这一发现可能会对巴库洛病毒的分类和系统地理学传播的理解产生重大影响。
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引用次数: 0
Characteristics and whole-genome analysis of a novel Pseudomonas syringae pv. tomato bacteriophage D6 isolated from a karst cave 从岩溶洞穴中分离出的新型噬菌体 D6(Pseudomonas syringae pv. tomato bacteriophage D6)的特征和全基因组分析
IF 1.6 4区 医学 Q3 GENETICS & HEREDITY Pub Date : 2024-04-09 DOI: 10.1007/s11262-024-02064-9
Qingshan Wu, Ni An, Zheng Fang, Shixia Li, Lan Xiang, Qiuping Liu, Leitao Tan, Qingbei Weng

Pseudomonas syringae is a gram-negative plant pathogen that infects plants such as tomato and poses a threat to global crop production. In this study, a novel lytic phage infecting P. syringae pv. tomato DC3000, named phage D6, was isolated and characterized from sediments in a karst cave. The latent period of phage D6 was found to be 60 min, with a burst size of 16 plaque-forming units per cell. Phage D6 was stable at temperatures between 4 and 40 °C but lost infectivity when heated to 70 °C. Its infectivity was unaffected at pH 6–10 but became inactivated at pH ≤ 5 or ≥ 12. The genome of phage D6 is a linear double-stranded DNA of 307,402 bp with a G + C content of 48.43%. There is a codon preference between phage D6 and its host, and the translation of phage D6 gene may not be entirely dependent on the tRNA library provided by the host. A total of 410 open reading frames (ORFs) and 14 tRNAs were predicted in its genome, with 92 ORFs encoding proteins with predicted functions. Phage D6 showed low genomic similarity to known phage genomes in the GenBank and Viral sequence databases. Genomic and phylogenetic analyses revealed that phage D6 is a novel phage. The tomato plants were first injected with phage D6, and subsequently with Pst DC3000, using the foliar spraying and root drenching inoculum approach. Results obtained after 14 days indicated that phage D6 inoculation decreased P. syringae-induced symptoms in tomato leaves and inhibited the pathogen’s growth in the leaves. The amount of Pst DC3000 was reduced by 150- and 263-fold, respectively. In conclusion, the lytic phage D6 identified in this study belongs to a novel phage within the Caudoviricetes class and has potential for use in biological control of plant diseases.

丁香假单胞菌(Pseudomonas syringae)是一种革兰氏阴性植物病原体,可感染番茄等植物,对全球作物生产构成威胁。本研究从岩溶洞穴的沉积物中分离并鉴定了一种感染 P. syringae pv. tomato DC3000 的新型裂殖噬菌体,命名为噬菌体 D6。研究发现,噬菌体 D6 的潜伏期为 60 分钟,每个细胞的爆发大小为 16 个斑块形成单位。噬菌体 D6 在 4 至 40 °C的温度下稳定,但加热到 70 °C时就失去了感染力。其感染力在 pH 值为 6-10 时不受影响,但在 pH 值≤ 5 或≥ 12 时会失活。噬菌体 D6 的基因组是 307,402 bp 的线性双链 DNA,G+C 含量为 48.43%。噬菌体 D6 与宿主之间存在密码子偏好,噬菌体 D6 基因的翻译可能并不完全依赖于宿主提供的 tRNA 文库。噬菌体 D6 基因组中共有 410 个开放阅读框(ORF)和 14 个 tRNA,其中 92 个 ORF 编码具有预测功能的蛋白质。噬菌体 D6 的基因组与 GenBank 和病毒序列数据库中的已知噬菌体基因组相似度较低。基因组和系统进化分析表明,噬菌体 D6 是一种新型噬菌体。首先用噬菌体 D6 对番茄植株进行注射,随后用 Pst DC3000 对植株进行注射,采用叶面喷洒和根部淋水接种的方法。14 天后的结果表明,接种噬菌体 D6 可减少西红柿叶片上由 P. syringae 引起的症状,并抑制病原体在叶片上的生长。Pst DC3000 的数量分别减少了 150 倍和 263 倍。总之,本研究发现的溶菌噬菌体 D6 属于 Caudoviricetes 类中的一种新型噬菌体,有望用于植物病害的生物防治。
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引用次数: 0
Analysis of SARS-CoV-2 omicron mutations that emerged during long-term replication in a lung cancer xenograft mouse model 分析肺癌异种移植小鼠模型长期复制过程中出现的 SARS-CoV-2 omicron 突变
IF 1.6 4区 医学 Q3 GENETICS & HEREDITY Pub Date : 2024-04-08 DOI: 10.1007/s11262-024-02067-6
Kyeongbin Baek, Dongbum Kim, Jinsoo Kim, Bo Min Kang, Heedo Park, Sangkyu Park, Ha-Eun Shin, Myeong-Heon Lee, Sony Maharjan, Minyoung Kim, Suyeon Kim, Man-Seong Park, Younghee Lee, Hyung-Joo Kwon

SARS-CoV-2 Omicron has the largest number of mutations among all the known SARS-CoV-2 variants. The presence of these mutations might explain why Omicron is more infectious and vaccines have lower efficacy to Omicron than other variants, despite lower virulence of Omicron. We recently established a long-term in vivo replication model by infecting Calu-3 xenograft tumors in immunodeficient mice with parental SARS-CoV-2 and found that various mutations occurred majorly in the spike protein during extended replication. To investigate whether there are differences in the spectrum and frequency of mutations between parental SARS-CoV-2 and Omicron, we here applied this model to Omicron. At 30 days after infection, we found that the virus was present at high titers in the tumor tissues and had developed several rare sporadic mutations, mainly in ORF1ab with additional minor spike protein mutations. Many of the mutant isolates had higher replicative activity in Calu-3 cells compared with the original SARS-CoV-2 Omicron virus, suggesting that the novel mutations contributed to increased viral replication. Serial propagation of SARS-CoV-2 Omicron in cultured Calu-3 cells resulted in several rare sporadic mutations in various viral proteins with no mutations in the spike protein. Therefore, the genome of SARS-CoV-2 Omicron seems largely stable compared with that of the parental SARS-CoV-2 during extended replication in Calu-3 cells and xenograft model. The sporadic mutations and modified growth properties observed in Omicron might explain the emergence of Omicron sublineages. However, we cannot exclude the possibility of some differences in natural infection.

在所有已知的 SARS-CoV-2 变体中,SARS-CoV-2 Omicron 的突变数量最多。这些突变的存在可能解释了为什么 Omicron 的传染性更强,尽管 Omicron 的毒力较低,但疫苗对 Omicron 的效力却低于其他变种。最近,我们用亲代 SARS-CoV-2 感染免疫缺陷小鼠的 Calu-3 异种移植瘤,建立了一个长期体内复制模型,发现在长期复制过程中,各种突变主要发生在尖峰蛋白中。为了研究亲代 SARS-CoV-2 和 Omicron 的变异谱和变异频率是否存在差异,我们在此将该模型应用于 Omicron。在感染后 30 天,我们发现病毒在肿瘤组织中的滴度很高,并出现了几种罕见的散发性突变,主要发生在 ORF1ab 中,另外还有一些小的尖峰蛋白突变。与原始的 SARS-CoV-2 Omicron 病毒相比,许多突变分离株在 Calu-3 细胞中的复制活性更高,这表明新型突变导致了病毒复制能力的增强。SARS-CoV-2 Omicron 在培养的 Calu-3 细胞中连续繁殖,导致各种病毒蛋白中出现几种罕见的零星突变,而尖峰蛋白中没有突变。因此,与亲本 SARS-CoV-2 相比,SARS-CoV-2 Omicron 的基因组在 Calu-3 细胞和异种移植模型的扩展复制过程中似乎基本稳定。在 Omicron 中观察到的零星突变和改变的生长特性可能是 Omicron 亚系出现的原因。不过,我们也不能排除自然感染中存在某些差异的可能性。
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引用次数: 0
Potentials as biomarker and therapeutic target of upregulated long non-coding RNA HLA-F antisense RNA 1 in hepatitis B virus-associated hepatocellular carcinoma 乙型肝炎病毒相关肝细胞癌中上调的长非编码 RNA HLA-F 反义 RNA 1 作为生物标记物和治疗靶点的潜力
IF 1.6 4区 医学 Q3 GENETICS & HEREDITY Pub Date : 2024-04-03 DOI: 10.1007/s11262-024-02065-8

Abstract

The tissue-specific characteristics have encouraged researchers to identify organ-specific lncRNAs as disease biomarkers. This study aimed to identify the clinical and functional roles of long non-coding RNA HLA-F antisense RNA 1 (HLA-F-AS1) in hepatitis B virus (HBV)-hepatocellular carcinoma (HCC). A total of 121 HBV-HCC, 81 chronic hepatitis B (CHB), and 85 normal liver tissues were evaluated in this study. Real-time quantitative PCR assay was used to evaluate the RNA expression levels. Performance in diagnosis was compared between alpha fetoprotein (AFP) and HLA-F-AS1 using Receiver Operating Characteristic (ROC) curves. Performance in post-hepatectomy prognosis with high or low HLA-F-AS1 was compared using Kaplan–Meier curves. Multi-variable analysis was used to determine the informative predictors. Downstream miRNAs for HLA-F-AS1 were predicted and miR-128-3p was confirmed by luciferase reporter assay and RNA pull-down assay. In vitro functional analysis was performed by MTS reagent for cell proliferation and transwell assay for cell migration. HLA-F-AS1 levels were significantly increased in the HBV-HCC compared to normal healthy tissue and CHB tissues. HLA-F-AS1 exhibited a well potential in making a distinction between HBV-HCC and health, as well as HBV-HCC and CHB. The survival analysis revealed that patients with high levels of HLA-F-AS1 tend to shorter overall survival times. The best prognostic performance was achieved by HLA-F-AS1 after multi-variable analysis (HR 2.290, 95% CI 1.191–4.403, p = 0.013). Functional analysis showed that HLA-F-AS1 promoted cell proliferation and migration via miR-128-3p. Up-regulation of HLA-F-AS1 could serve as a promising diagnostic and prognostic marker for HBV-HCC after surgery, maybe useful in the management of HBV-HCC patients. HLA-F-AS1 can promote the progression of HBV-HCC, may be useful in the targeting treatment of HBV-HCC patients.

摘要 组织特异性特征促使研究人员将器官特异性lncRNAs鉴定为疾病生物标志物。本研究旨在确定长非编码 RNA HLA-F 反义 RNA 1(HLA-F-AS1)在乙型肝炎病毒(HBV)-肝细胞癌(HCC)中的临床和功能作用。本研究共评估了 121 例 HBV-HCC、81 例慢性乙型肝炎(CHB)和 85 例正常肝组织。研究采用实时定量 PCR 法评估 RNA 表达水平。使用接收者操作特征曲线(ROC)比较了甲胎蛋白(AFP)和 HLA-F-AS1 的诊断效果。使用 Kaplan-Meier 曲线比较了高或低 HLA-F-AS1 对肝切除术后预后的影响。多变量分析用于确定有参考价值的预测因子。HLA-F-AS1的下游miRNA被预测出来,miR-128-3p通过荧光素酶报告实验和RNA牵引实验得到证实。体外功能分析采用 MTS 试剂检测细胞增殖,采用跨孔试验检测细胞迁移。与正常健康组织和 CHB 组织相比,HBV-HCC 中的 HLA-F-AS1 水平明显升高。HLA-F-AS1 在区分 HBV-HCC 和健康组织以及 HBV-HCC 和慢性阻塞性肺病方面具有很好的潜力。生存分析表明,HLA-F-AS1水平高的患者总生存时间往往较短。经过多变量分析,HLA-F-AS1 的预后效果最好(HR 2.290,95% CI 1.191-4.403,p = 0.013)。功能分析显示,HLA-F-AS1通过miR-128-3p促进细胞增殖和迁移。HLA-F-AS1的上调可作为术后HBV-HCC的诊断和预后标志物,或许有助于HBV-HCC患者的治疗。HLA-F-AS1能促进HBV-HCC的进展,可能有助于HBV-HCC患者的靶向治疗。
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Virus Genes
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