Veterinary microbiology最新文献_第4页

Epidemiological investigation of equine rotavirus B outbreaks in horses in central Kentucky 肯塔基州中部马匹轮状病毒 B 爆发的流行病学调查。

IF 2.4 2区农林科学 Q3 MICROBIOLOGY

Veterinary microbiology

Pub Date : 2024-10-16 DOI: 10.1016/j.vetmic.2024.110278

Chithra C. Sreenivasan , Ahsan Naveed , Tirth Uprety, Shalini Soni, Olivia Jacob, Emma Adam, Dan Wang, Feng Li

Using metagenomic sequencing we identified equine rotavirus group B (ERVB) of ruminant origin in foal diarrhea outbreaks in the 2021 foaling season. To further investigate ERVB occurrence and determine its environmental stability, we collected mare and foal fecal samples from different farms in Central Kentucky during the 2022 foaling season. The RT-qPCR-based analyses showed that ERVB genome was detected in 16.67 % (42/252) of surveyed mare samples and 26.56 % (34/128) of foal samples. Furthermore, 94.12 % (16/17) of collected soil samples and 100 % (13/13) of water samples obtained from the ERVB-positive farm premises also tested weakly positive. In addition, ERVB genome fragments were detected in 58.33 % (7/12) of indoor samples collected from the equipment/barn/hospital wards during the outbreak period. Finally, the seroprevalence study showed 87 % (113/130) of surveyed horse serum samples were positive for ERVB antibodies. Despite unsuccessful attempts in ERVB cultivation, phylogenetic analyses showed that fecal ERVB strains representing 2022 and 2023 foal diarrhea outbreaks, like 2021 strains, were more closely related to ruminant rotavirus B than other viruses. Further sequence analyses revealed that none of the three viral capsid proteins, the primary targets of virus-neutralizing antibodies, exhibited notable mutations among ERVB strains circulated over the past three years. Our data demonstrated that ERVB was widespread in horses on affected farms with extreme stability in the farm environment. These findings continue to support the need for future surveillance of ERVB in horses and the surrounding environment, and the development of effective countermeasures to protect horses against this new viral disease.

通过元基因组测序，我们在 2021 年产驹季节爆发的马驹腹泻中发现了反刍动物来源的马轮状病毒 B 组（ERVB）。为了进一步调查 ERVB 的发生情况并确定其环境稳定性，我们在 2022 年的产驹季节收集了肯塔基州中部不同农场的母马和马驹粪便样本。基于 RT-qPCR 的分析表明，在调查的母马样本中有 16.67 %（42/252）和马驹样本中有 26.56 %（34/128）检测到 ERVB 基因组。此外，从ERVB阳性农场采集的94.12%（16/17）的土壤样本和100%（13/13）的水样本也检测出弱阳性。此外，在疫情爆发期间从设备/谷仓/医院病房采集的室内样本中，有 58.33 %（7/12）的样本检测到 ERVB 基因组片段。最后，血清流行率研究显示，87%（113/130）的马血清样本中ERVB抗体呈阳性。尽管ERVB的培养尝试并不成功，但系统进化分析表明，代表2022年和2023年马驹腹泻疫情的粪便ERVB毒株与2021年的毒株一样，与反刍轮状病毒B的亲缘关系比其他病毒更近。进一步的序列分析表明，在过去三年中流行的ERVB毒株中，病毒中和抗体的主要靶标--三种病毒外壳蛋白都没有出现明显的变异。我们的数据表明，ERVB 在受影响马场的马匹中普遍存在，且在马场环境中极为稳定。这些发现继续支持了今后对马匹和周围环境中的 ERVB 进行监测的必要性，以及开发有效对策保护马匹免受这种新型病毒性疾病侵害的必要性。

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引用次数: 0

A novel neuro-attenuated vaccine candidate with excellent safety and protective efficacy against highly virulent Feline Herpesvirus-1 一种新型神经减毒候选疫苗，对高致病性猫疱疹病毒-1 具有极佳的安全性和保护效力。

IF 2.4 2区农林科学 Q3 MICROBIOLOGY

Veterinary microbiology

Pub Date : 2024-10-15 DOI: 10.1016/j.vetmic.2024.110276

Mingyu Qi , Mengfang Yang , Ruxue Luo , Lingying Fang , Yixi Chen , Jianuo Gao , Zhe Jiao , Yuejun Shi , Guiqing Peng

Feline herpesvirus 1 (FHV-1) is a major pathogen responsible for respiratory, ocular and nervous system symptoms in felines. FHV-1 can remain latenct in ganglia and is difficult to eliminate completely with drug treatment. Currently, commercially FHV-1 vaccines are not sufficiently effective and provide only limited durations of protection. To enhance vaccine efficacy and reduce latent virus in tissues, two gene deletion mutants of FHV-1 conveyed excellent proliferation ability, genetic stability and attenuated FHV-1 virulence were constructed by CRISPR/Cas9-mediated homologous recombination, designated as FHV-△US3 and FHV-△UL50. Recombinant FHV-1 induce stronger cellular and humoral immune responses, as well as better protective effects than those of commercial vaccines. Notably, FHV-△US3 and FHV-△UL50 reveal neuro-attenuated, as viral residue in the trigeminal ganglia are significantly reduced. The knockout of the UL50 gene in FHV-1 has not been previously reported. In this study, we aimed to evaluate the safety and immunogenicity of FHV-△UL50, highlighting its potential as a novel neuroattenuated vaccine candidate.

猫疱疹病毒 1 (FHV-1) 是导致猫科动物出现呼吸道、眼部和神经系统症状的主要病原体。FHV-1 可潜伏在神经节中，很难通过药物治疗彻底清除。目前，市售的 FHV-1 疫苗效果不佳，只能提供有限的保护期。为了提高疫苗疗效并减少组织中的潜伏病毒，我们通过 CRISPR/Cas9 介导的同源重组技术构建了两个基因缺失的 FHV-1 突变体，这两个突变体分别被命名为 FHV-△US3 和 FHV-△UL50，它们具有优异的增殖能力、遗传稳定性和减毒能力。与商业疫苗相比，重组 FHV-1 能诱导更强的细胞和体液免疫反应，以及更好的保护效果。值得注意的是，FHV-△US3 和 FHV-△UL50 揭示了神经减毒作用，三叉神经节中的病毒残留明显减少。之前还没有关于 FHV-1 中 UL50 基因敲除的报道。在这项研究中，我们旨在评估FHV-△UL50的安全性和免疫原性，突出其作为新型神经减毒疫苗候选物的潜力。

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引用次数: 0

IFN-β contributes to astrocyte activation in the brain following coronavirus PHEV infection independent on peripheral immunity IFN-β有助于冠状病毒 PHEV 感染后大脑星形胶质细胞的活化，与外周免疫无关。

IF 2.4 2区农林科学 Q3 MICROBIOLOGY

Veterinary microbiology

Pub Date : 2024-10-15 DOI: 10.1016/j.vetmic.2024.110280

Junchao Shi , Qianhan Jia , Yanfang Yu , Jing Zhang , Zi Li , Wenqi He

Porcine hemagglutinating encephalomyelitis (PHE), caused by a betacoronavirus named porcine hemagglutinating encephalomyelitis virus (PHEV), is a highly fatal disease of pigs characterized by nonsuppurative encephalitis. Activation of astrocytes is a hallmark of viral encephalomyelitis; however, the mechanism of PHEV-induced astrocyte activation is currently unknown. Based on mouse model, we show that PHEV infection led to astrogliosis in mouse brain and brain slice cultures (BSCs), as indicated by increased expression of glial fibrillary acidic protein (GFAP). PHEV can neither infect nor activate primary astrocytes in vitro, indicating that activation of astrocytes maybe mediated by factors secreted from viral infected neurons but not by direct viral infection of astrocytes. PHEV infection results in increased interferon (IFN) response in later stage, we thereafter focused on whether IFN-β can activate astrocytes after PHEV infection similar to other neurotropic viruses. IFN-β treatment resulted in both the upregulation of GFAP and activation-associated cytokines/chemokines in mouse primary astrocytes. Furthermore, the addition of IFN-β neutralization antibody prevented PHEV-infected mouse brain tissue homogenate from activating astrocytes. Taken together, IFN-β triggers the activation of astrocytes in the central nervous system (CNS) following PHEV infection. Further understanding of the role of activated astrocytes during PHEV infection may provide new insights for treatment this disease.

猪血凝性脑脊髓炎（PHE）是由一种名为猪血凝性脑脊髓炎病毒（PHEV）的betacoronavirus引起的，是一种高度致命的猪疾病，以非化脓性脑炎为特征。星形胶质细胞活化是病毒性脑脊髓炎的特征之一；然而，PHEV 诱导星形胶质细胞活化的机制目前尚不清楚。基于小鼠模型，我们发现 PHEV 感染会导致小鼠大脑和脑片培养物（BSCs）中的星形胶质细胞增生，表现为胶质纤维酸性蛋白（GFAP）的表达增加。PHEV 既不能感染也不能激活体外原发性星形胶质细胞，这表明星形胶质细胞的激活可能是由病毒感染的神经元分泌的因子介导的，而不是由病毒直接感染星形胶质细胞介导的。PHEV 感染后期会导致干扰素（IFN）反应增强，因此我们关注 PHEV 感染后 IFN-β 是否能像其他神经毒性病毒一样激活星形胶质细胞。IFN-β 处理可导致小鼠原代星形胶质细胞的 GFAP 和活化相关细胞因子/趋化因子上调。此外，添加 IFN-β 中和抗体可防止 PHEV 感染的小鼠脑组织匀浆激活星形胶质细胞。综上所述，PHEV 感染后，IFN-β 会引发中枢神经系统（CNS）中星形胶质细胞的活化。进一步了解 PHEV 感染期间活化星形胶质细胞的作用可能会为治疗这种疾病提供新的见解。

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引用次数: 0

The prevalence and shedding of porcine epidemic diarrhea virus in intensive swine farms of China from 2022 to 2023 2022 至 2023 年中国集约化猪场猪流行性腹泻病毒的流行和脱落情况

IF 2.4 2区农林科学 Q3 MICROBIOLOGY

Veterinary microbiology

Pub Date : 2024-10-11 DOI: 10.1016/j.vetmic.2024.110273

Yuetao Xu , Meng Sun , Zhunxuan Wang , Xiaowen Li , Yijun Du , Ping Jiang

Porcine epidemic diarrhea has emerged as a significant threat to the global swine industry. The shedding and exposure status of porcine epidemic diarrhea virus (PEDV) in intensive farms is not completely understood. In this study, a total of 56,598 clinical samples collected from 256 intensive pig farms in 20 provinces in China from 2022 to 2023, were evaluated for PEDV using quantitative real-time PCR. The overall PEDV prevalence was 11.78 % and 28.45 % at the sample and farm levels, respectively, which are relatively high in Northern China and the fourth and first quarter of the year. The PEDV-positive rates and viral loads in suckling piglet herds were higher than those in growing-finishing pigs and multiparous sows. Meanwhile, 15.61 % of pig pens, 9.51 % of corridors, 9.4 % of office areas, 9.23 % of production personnel, and 8.33 % of pig cart driver samples were positive for PEDV, indicating potential biosafety gaps in intensive pig farms. In addition, 93.41 % of inguinal lymph node tissue samples contained viral nucleic acids, revealing a possible persistent infection of PEDV in pig herds. Our study presents the first report of the large-scale detection of PEDV in intensive pig farms, which constitutes indirect evidence of virus circulation in pig herds. This study provides valuable data for preventing and controlling PEDV infection in the future.

猪流行性腹泻已成为全球养猪业的一大威胁。猪流行性腹泻病毒（PEDV）在集约化养猪场的脱落和暴露状况尚不完全清楚。本研究采用实时定量 PCR 技术，对 2022 年至 2023 年期间从中国 20 个省的 256 个集约化猪场采集的 56598 份临床样本进行了 PEDV 评估。在样本和猪场层面，PEDV 的总体流行率分别为 11.78 % 和 28.45 %，在华北地区以及每年的第四季度和第一季度相对较高。乳仔猪群的 PEDV 阳性率和病毒载量均高于生长育成猪和多胎母猪。同时，15.61%的猪栏、9.51%的走廊、9.4%的办公区、9.23%的生产人员和 8.33%的猪车司机样本对 PEDV 呈阳性，这表明集约化猪场可能存在生物安全漏洞。此外，93.41% 的腹股沟淋巴结组织样本含有病毒核酸，揭示了 PEDV 可能在猪群中持续感染。我们的研究首次报告了在集约化养猪场大规模检测到 PEDV 的情况，这构成了猪群中病毒传播的间接证据。这项研究为今后预防和控制 PEDV 感染提供了宝贵的数据。

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引用次数: 0

Genetic Characteristics and Pathogenicity of Variant Porcine Parvovirus 1 in Northern China 华北地区变异猪细小病毒 1 型的遗传特征和致病性

IF 2.4 2区农林科学 Q3 MICROBIOLOGY

Veterinary microbiology

Pub Date : 2024-10-09 DOI: 10.1016/j.vetmic.2024.110274

Huiwei Deng, Mengting Wang, Guangyi Cong, Fang Fu, Li Feng

Porcine parvovirus type 1 (PPV1) can lead to reproductive disorders in pregnant sows, including stillbirth, mummification, embryonic death, and infertility (SMEDI syndrome). In this study, we isolated and identified 10 PPV1 strains in northern China, with genomes around 5 kb long and minor deletions in the 127-nt repeat region. The sequence analysis results showed that compared with strain NADL-2 (Reference strain), eight amino acid substitutions on the NS1 protein and fourteen amino acid substitutions on the VP2 protein were found in the ten isolates. Because the JX strain exhibited reduced neutralizing activity induced by commercially available vaccines in vitro, it was selected for challenge experiments in sows at 35 days of gestation. Mutant strain JX not only caused viremia, but also mild edema and mild inflammation in the trachea, lungs, lymph nodes, reproductive organs, and some intestines of the pregnant sows. Strain JX also caused fetal congestion and organ infection after penetrating the placental barrier. In conclusion, this study focused on the variation and evolution of PPV1 in northern China, screened a strain with reduced neutralizing activity, and determined that it has a certain degree of pathogenicity.

猪细小病毒 1 型（PPV1）可导致妊娠母猪繁殖障碍，包括死胎、木乃伊化、胚胎死亡和不孕（SMEDI 综合征）。本研究在中国北方分离并鉴定了10株PPV1病毒，其基因组长约5 kb，在127-nt重复区有轻微缺失。序列分析结果显示，与NADL-2株（参考株）相比，这10株分离株的NS1蛋白有8个氨基酸替换，VP2蛋白有14个氨基酸替换。由于 JX 株在体外接种市售疫苗后中和活性降低，因此被选中在妊娠 35 天的母猪身上进行挑战实验。变异株 JX 不仅会引起病毒血症，还会导致怀孕母猪的气管、肺部、淋巴结、生殖器官和部分肠道出现轻度水肿和轻度炎症。JX 株在穿透胎盘屏障后还会引起胎儿充血和器官感染。总之，本研究关注了 PPV1 在中国北方的变异和进化，筛选出一株中和活性降低的毒株，并确定其具有一定的致病性。

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引用次数: 0

Enhancement of protective efficacy of recombinant attenuated Salmonella typhimurium delivering H9N2 avian influenza virus hemagglutinins(HA) antigen vaccine candidate strains by C-C motif chemokine ligand 5 in chickens(chCCL5) C-C motif趋化因子配体5(chCCL5)增强重组减毒鼠伤寒沙门氏菌输送H9N2禽流感病毒血凝素(HA)抗原候选株疫苗对鸡的保护效力

IF 2.4 2区农林科学 Q3 MICROBIOLOGY

Veterinary microbiology

Pub Date : 2024-10-09 DOI: 10.1016/j.vetmic.2024.110264

Jingwen Ma , Shunshun Xu , Zewei Li , Yu-An Li , Shifeng Wang , Huoying Shi

The H9N2 inactivated avian influenza vaccine cannot induce cellular and mucosal immune responses, while the attenuated Salmonella vector as an intracellular bacterium can induce dominant cellular and mucosal immune responses. However, it provides low protection against the virus when delivering viral antigens and needs further optimization. Chicken C-C motif chemokine ligand 5 (chCCL5) is an important CC chemokine associated with immune cell chemotaxis, migration, and viral infection. This study connected the sequence of chCCL5 (CCL5) with the hemagglutinin sequence of the H9N2 avian influenza virus (yH9HA), utilizing the attenuated Salmonella typhimurium vector containing the delayed lysis system MazE/F regulated by arabinose as a carrier. A vaccine strain of recombinant attenuated Salmonella typhimurium and H9N2 avian influenza virus HA, rSC0130 (pS0017-yH9HA-CCL5), was successfully constructed. The experimental results indicate that yH9HA-CCL5 can be expressed in 293 T cells; compared to the strain without CCL5, rSC0130 (pS0017-yH9HA-CCL5) can induce significantly increased cellular immune responses and provide better protective effects in H9N2 virus challenge experiments. The above results indicate that chCCL5 can significantly enhance the protective effect of Salmonella delivering H9N2 avian influenza virus HA protein vaccine against H9N2 avian influenza virus infection, providing valuable theoretical support for further improving the protective efficiency of recombinant attenuated Salmonella vectors for delivering viral antigens.

H9N2 灭活禽流感疫苗不能诱导细胞和粘膜免疫反应，而作为细胞内细菌的减毒沙门氏菌载体可诱导细胞和粘膜免疫反应。然而，在传递病毒抗原时，它对病毒的保护能力较低，需要进一步优化。鸡 C-C motif 趋化因子配体 5（chCCL5）是一种重要的 CC 趋化因子，与免疫细胞趋化、迁移和病毒感染有关。本研究利用含有阿拉伯糖调控的延迟裂解系统 MazE/F 的鼠伤寒沙门氏菌减毒载体作为载体，将 chCCL5（CCL5）序列与 H9N2 禽流感病毒（yH9HA）的血凝素序列连接起来。成功构建了鼠伤寒沙门氏菌和 H9N2 禽流感病毒 HA 重组减毒疫苗株 rSC0130（pS0017-yH9HA-CCL5）。实验结果表明，yH9HA-CCL5可在293个T细胞中表达；与不含CCL5的毒株相比，rSC0130（pS0017-yH9HA-CCL5）可诱导细胞免疫反应显著增强，在H9N2病毒挑战实验中具有更好的保护作用。上述结果表明，chCCL5能显著增强沙门氏菌递送H9N2禽流感病毒HA蛋白疫苗对H9N2禽流感病毒感染的保护作用，为进一步提高重组减毒沙门氏菌载体递送病毒抗原的保护效率提供了有价值的理论支持。

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引用次数: 0

Characterization of the immunodominant regions of Senecavirus A-VP1 structural protein via ELISA epitope mapping 通过 ELISA 表位图谱确定塞内卡病毒 A-VP1 结构蛋白的免疫优势区。

IF 2.4 2区农林科学 Q3 MICROBIOLOGY

Veterinary microbiology

Pub Date : 2024-10-02 DOI: 10.1016/j.vetmic.2024.110266

Elizabeth Houston , Kepalee Saeng-chuto , Patumporn Jermsutjarit , Luis Giménez-Lirola , Avanti Sinha , Juan Carlos Mora-Díaz , Dachrit Nilubol , Nicolas F. Villarino , Pablo Piñeyro

Senecavirus A (SVA) is an RNA virus in the family Picornaviridae that has been detected in swine-production systems and is associated with vesicular disease and neonate mortality. The viral capsid is composed of four structural proteins: VP1–VP4. Although the VP1 protein has been reported to be the most immunogenic protein in vivo, no information on the immunodominant regions of the SVA polyprotein is available. The objective of this study was to identify the immunodominant regions of SVA polyprotein using an enzyme-linked immunosorbent assay (ELISA) epitope-mapping approach. The binding effect of SVA polyclonal antibody (SVA-pAb), SVA-VP1 monoclonal antibodies (SVA-mAb), and SVA-positive sera from clinically affected animals were characterized using a set of 18 overlapping SVA VP1-derived peptides by indirect and blocking ELISAs. All VP1 peptides yielded significant signal against SVA-pAb and SVA-VP1-mAb upon indirect ELISA. One peptide (aa 1–20) showed significantly high optical density on SVA recombinant VP1 protein (rVP1) and whole-virus-based indirect ELISAs. The blocking ELISA results demonstrated that peptides spanning aa 165–185 and 225–245 had a 50 % or greater inhibitory effect on SVA-pAb, while six groups of overlapping peptides spanning aa 1–35, 45–80, 90–140, 150–170, 195–230, and 240–264 and two groups of overlapping peptides spanning aa 1–50 and 60–264 showed a 50 % inhibitory effect or greater on swine VP1-mAb and SVA-seropositive swine serum, respectively, against SVA rVP1. Three-dimensional protein homology modeling showed that the peptides binding SVA-pAb are located on the outer surface of the viral capsid, while SVA mAbs and swine-positive sere can bind to epitopes located in both the inner and outer surfaces of the capsid. These linear epitopes showed differential binding and inhibitory activity on mAb and pAb; however, further studies will be necessary to evaluate whether they can act as decoy or neutralizing epitopes. Because mAb antibodies demonstrated a high binding affinity for this set of peptides, this information could lay the foundation for generating and screening specific antibodies for therapeutic potential.

猪流感病毒 A（SVA）是一种 RNA 病毒，属于 Picornaviridae 科，已在猪生产系统中检测到，与水泡病和新生仔猪死亡有关。病毒外壳由四种结构蛋白组成：VP1-VP4。尽管有报告称 VP1 蛋白是体内免疫原性最强的蛋白，但目前尚无关于 SVA 多聚蛋白免疫优势区的信息。本研究的目的是利用酶联免疫吸附试验（ELISA）表位映射法确定 SVA 多聚蛋白的免疫优势区。通过间接ELISA和阻断ELISA，使用一组18个重叠的SVA VP1衍生肽，鉴定了SVA多克隆抗体（SVA-pAb）、SVA-VP1单克隆抗体（SVA-mAb）和临床患病动物的SVA阳性血清的结合效应。在间接酶联免疫吸附试验中，所有 VP1 肽都能产生针对 SVA-pAb 和 SVA-VP1-mAb 的显著信号。其中一个多肽（aa 1-20）在 SVA 重组 VP1 蛋白（rVP1）和基于全病毒的间接 ELISA 中显示出明显的高光密度。阻断酶联免疫吸附试验结果表明，跨越 aa 165-185 和 225-245 的肽对 SVA-pAb 有 50% 或更大的抑制作用，而跨越 aa 1-35、45-80、90-140、150-170、195-230 和 240-264 的六组重叠肽对 SVA-pAb 有更大的抑制作用、195-230和240-264的六组重叠肽，以及跨越aa 1-50和60-264的两组重叠肽，对猪VP1-mAb和SVA-血清阳性猪血清对SVA rVP1的抑制效果分别为50%或更高。三维蛋白质同源建模显示，与 SVA-pAb 结合的肽位于病毒外壳的外表面，而 SVA mAbs 和猪阳性血清可与位于外壳内表面和外表面的表位结合。这些线性表位在 mAb 和 pAb 上显示出不同的结合和抑制活性；然而，要评估它们是否能作为诱饵或中和表位，还需要进一步的研究。由于 mAb 抗体对这组肽具有很高的结合亲和力，这些信息可为生成和筛选具有治疗潜力的特异性抗体奠定基础。

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引用次数: 0

NLRP12 inhibits PRRSV-2 replication by promoting GP2a degradation via MARCH8 NLRP12 通过 MARCH8 促进 GP2a 降解，从而抑制 PRRSV-2 的复制。

IF 2.4 2区农林科学 Q3 MICROBIOLOGY

Veterinary microbiology

Pub Date : 2024-09-30 DOI: 10.1016/j.vetmic.2024.110271

Huiyuan Jing , Yuzhen Song , Erzhen Duan , Jie Liu , Wenting Ke , Ran Tao , Yujin Lv , Pandeng Zhao , Wang Dong , Xianghui Li , Yongbin Guo , Huawei Li

NLRP12, a member of the NLR family, has been shown to exert a vital function in orchestrating immune responses. Here, using the immunosuppressive porcine reproductive and respiratory syndrome virus (PRRSV) as a model, the role of NLRP12 in virus infection was deciphered. We demonstrated that overexpression of NLRP12 significantly restrained PRRSV replication, while NLRP12 silencing resulted in increased viral titer. Mechanistically, NLRP12 interacts with glycoprotein 2a (GP2a) through its LRR domain and recruits the membrane-associated RING-CH E3 ubiquitin ligase 8 (MARCH8) via the PYD domain. NLRP12 facilitates the lysine-48 (K48)–linked polyubiquitination of GP2a at K128 and induces its lysosome degradation via the MARCH8-NDP52 (nuclear dot protein 52 kDa) pathway. To counteract this, PRRSV Nsp2 effectively prevented the polyubiquitination of GP2a induced by NLRP12 by its deubiquitinating activity. Meanwhile, the overexpression of Nsp4 decreased the mRNA of endogenous NLRP12 and cleaved NLRP12 in a 3C-like protease activity-dependent manner, which collaboratively counteracts the antiviral function of NLRP12. Collectively, this study revealed the mechanisms of the NLRP12-MARCH8-NDP52 axis in the host defense against PRRSV, which might be harnessed for the development of anti-PRRSV therapies.

NLRP12 是 NLR 家族的成员之一，已被证明在协调免疫反应中发挥着重要功能。在这里，我们以免疫抑制猪繁殖与呼吸综合征病毒（PRRSV）为模型，解读了 NLRP12 在病毒感染中的作用。我们证明，过表达 NLRP12 能显著抑制 PRRSV 的复制，而沉默 NLRP12 则会导致病毒滴度增加。从机理上讲，NLRP12通过其LRR结构域与糖蛋白2a（GP2a）相互作用，并通过PYD结构域招募膜相关RING-CH E3泛素连接酶8（MARCH8）。NLRP12 促进了 GP2a 在 K128 处的赖氨酸-48 (K48) 链接多泛素化，并通过 MARCH8-NDP52（核点蛋白 52 kDa）途径诱导其溶酶体降解。为了对抗这种情况，PRRSV Nsp2 通过其去泛素化活性有效地阻止了 NLRP12 诱导的 GP2a 的多泛素化。同时，Nsp4的过表达降低了内源性NLRP12的mRNA，并以3C样蛋白酶活性依赖的方式裂解NLRP12，共同抵消了NLRP12的抗病毒功能。总之，本研究揭示了NLRP12-MARCH8-NDP52轴在宿主防御PRRSV中的作用机制，可用于开发抗PRRSV疗法。

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引用次数: 0

Evaluation of Tembusu virus single-round infectious particle as vaccine vector in chickens 评估作为鸡疫苗载体的淡布苏病毒单轮感染颗粒。

IF 2.4 2区农林科学 Q3 MICROBIOLOGY

Veterinary microbiology

Pub Date : 2024-09-28 DOI: 10.1016/j.vetmic.2024.110270

Yunzhen Huang , Zhe Liu , Junqin Zhang , Jiawen Dong , Linlin Li , Yong Xiang , Ruihuan Kuang , Shimin Gao , Minhua Sun , Yongjie Liu

Orthoflaviviruses are single-stranded RNA viruses characterized by highly efficient self-amplification of RNA in host cells, which makes them attractive vehicles for vaccines. Numerous preclinical and clinical studies have demonstrated the efficacy and safety of orthoflavivirus replicon vectors for vaccine development. In this study, we constructed Tembusu virus (TMUV) replicon-based single-round infectious particles (SRIPs) as vaccine development platform. To evaluate the potential of TMUV SRIPs as vaccines, we generated SRIPs that express the heterologous Fowl adenovirus 4 (FAdV-4) fiber2 protein and fiber2 head domain, named TMUVRP-fiber2 and TMUVRP-fiber2H, respectively. To assess the immunogenicity of the TMUV SRIPs, SPF chicks were intramuscularly inoculated twice. Our results showed that the TMUVRP-fiber2 vaccines elicited high levels of neutralizing antibodies. Challenge experiments showed that TMUVRP-fiber2 provided full protection against virulent FAdV-4 and significantly reduced viral shedding. Moreover, the immunogenicity of TMUVRP-fiber2H was significantly lower than that of TMUVRP-fiber2, which was reflected in the neutralizing antibody titer, survival rate, and virus shedding after challenge. Therefore, our results suggested that TMUV SRIPs are a promising novel platform for the development of vaccines for existing and emerging poultry diseases.

正黄病毒是单链 RNA 病毒，其特点是 RNA 在宿主细胞中的高效自我扩增，这使它们成为极具吸引力的疫苗载体。大量临床前和临床研究都证明了正黄病毒复制载体用于疫苗开发的有效性和安全性。在本研究中，我们构建了基于坦布苏病毒（TMUV）复制子的单轮感染性颗粒（SRIPs）作为疫苗开发平台。为了评估TMUV SRIPs作为疫苗的潜力，我们生成了表达异源鸡腺病毒4（FAdV-4）纤维2蛋白和纤维2头部结构域的SRIPs，分别命名为TMUVRP-fiber2和TMUVRP-fiber2H。为了评估TMUV SRIPs的免疫原性，对SPF雏鸡进行了两次肌肉注射。结果表明，TMUVRP-纤维2疫苗可激发高水平的中和抗体。挑战实验表明，TMUVRP-纤维2对FAdV-4病毒提供了全面的保护，并显著减少了病毒的脱落。此外，TMUVRP-fiber2H 的免疫原性明显低于 TMUVRP-fiber2，这反映在挑战后的中和抗体滴度、存活率和病毒脱落上。因此，我们的研究结果表明，TMUV SRIPs 是一种很有前景的新型平台，可用于现有和新出现的家禽疾病疫苗的开发。

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引用次数: 0

Early antibody responses to lipid antigens in red deer infected with Mycobacterium bovis 红鹿感染牛分枝杆菌后对脂质抗原的早期抗体反应。

IF 2.4 2区农林科学 Q3 MICROBIOLOGY

Veterinary microbiology

Pub Date : 2024-09-27 DOI: 10.1016/j.vetmic.2024.110269

Paul S. Mason , Maria A. Risalde , Christian Gortázar , Joseba M. Garrido , Dafydd A. Thomas , Juma’a R. Al Dulayymi , Mark S. Baird

The problem: Early and rapid diagnosis of bovine tuberculosis remains an issue of great interest. Aim: The aim of this study was to evaluate the use of synthetic lipid antigens for diagnosis of tuberculosis in red deer (Cervus elaphus). The proposition: Synthetic mycolic acid derivatives, identical to components of mycobacterial cells, bind to antibodies to lipids produced in active human tuberculosis. Experimental infection studies in red deer (Cervus elaphus) allow the evaluation of such antigens for the serodiagnosis of bovine tuberculosis. Results: Antibody levels in plasma from deer experimentally infected with Mycobacterium bovis were evaluated in ELISA using synthetic antigens based on several classes of mycolic acid, using protein G as conjugate. All antigens gave significantly increased responses 60 days post-infection, when all animals had active disease. A significantly increased response was also observed with four antigens 15 days after infection. Conclusion: ELISA using synthetic lipid antigens not only detects antibodies in the plasma of deer experimentally infected with M. bovis, but a strong response occurs early in the infection. With a full analysis of responses with naturally infected animals, this may offer a useful supplement to current diagnostic methods.

问题：牛结核病的早期快速诊断仍然是一个备受关注的问题。研究目的：本研究旨在评估合成脂质抗原在赤鹿（Cervus elaphus）结核病诊断中的应用。命题：合成的霉酚酸衍生物与分枝杆菌细胞的成分相同，能与活动性人类结核病产生的脂质抗体结合。通过对红鹿（Cervus elaphus）的实验感染研究，可以对这种抗原进行评估，以用于牛结核病的血清诊断：结果：使用基于几类霉菌酸的合成抗原，以蛋白 G 为共轭物，通过酶联免疫吸附试验评估了实验性感染牛分枝杆菌的鹿血浆中的抗体水平。所有抗原在感染后 60 天（当时所有动物都有活动性疾病）产生的反应都明显增加。感染后 15 天，四种抗原的反应也明显增加：结论：使用合成脂质抗原的酶联免疫吸附试验不仅能在实验性感染牛海绵状芽孢杆菌的鹿血浆中检测到抗体，而且能在感染早期产生强烈的反应。在对自然感染动物的反应进行全面分析后，这可能会成为当前诊断方法的有益补充。

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引用次数: 0