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Erratum for Choi et al., "Human saliva modifies growth, biofilm architecture, and competitive behaviors of oral streptococci". 对 Choi 等人 "人类唾液改变了口腔链球菌的生长、生物膜结构和竞争行为 "的勘误。
IF 3.7 2区 生物学 Q2 MICROBIOLOGY Pub Date : 2024-11-06 DOI: 10.1128/msphere.00868-24
Allen Choi, Kevin Dong, Emily Williams, Lindsey Pia, Jordan Batagower, Paige Bending, Iris Shin, Daniel I Peters, Justin R Kaspar
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引用次数: 0
Pneumococcal extracellular vesicles mediate horizontal gene transfer via the transformation machinery. 肺炎球菌细胞外囊泡通过转化机制介导横向基因转移。
IF 3.7 2区 生物学 Q2 MICROBIOLOGY Pub Date : 2024-11-06 DOI: 10.1128/msphere.00727-24
Sarah Werner Lass, Bailey E Smith, Shaw Camphire, Rory A Eutsey, Jojo A Prentice, Saigopalakrishna S Yerneni, Ashni Arun, Andrew A Bridges, Jason W Rosch, James F Conway, Phil Campbell, N Luisa Hiller

Bacterial cells secrete extracellular vesicles (EVs), the function of which is a matter of intense investigation. Here, we show that the EVs secreted by the human pathogen Streptococcus pneumoniae (pneumococcus) are associated with bacterial DNA on their surface and can deliver this DNA to the transformation machinery of competent cells. These findings suggest that EVs contribute to gene transfer in Gram-positive bacteria and, in doing so, may promote the spread of drug resistance genes in the population.IMPORTANCEThis work extends our understanding of horizontal gene transfer and the roles of extracellular vesicles in pneumococcus. This bacterium serves as the model for transformation, a process by which bacteria can take up naked DNA from the environment. Here, we show that extracellular vesicles secreted by the pneumococcus have DNA on their surface and that this DNA can be imported by the transformation machinery, facilitating gene transfer. Understanding EV-mediated gene transfer may provide new avenues to manage the spread of antibiotic drug resistance.

细菌细胞会分泌胞外囊泡(EVs),其功能一直是人们研究的热点。在这里,我们发现人类病原体肺炎链球菌(肺炎球菌)分泌的细胞外囊泡表面与细菌 DNA 相关联,并能将 DNA 运送到合格细胞的转化机制中。这些研究结果表明,细胞外囊泡有助于革兰氏阳性细菌的基因转移,从而可能促进耐药基因在群体中的传播。这种细菌是转化的典范,通过转化,细菌可以从环境中吸收裸 DNA。在这里,我们发现肺炎球菌分泌的胞外囊泡表面有 DNA,这种 DNA 可以被转化机器导入,从而促进基因转移。了解由细胞外小泡介导的基因转移可能会为控制抗生素耐药性的传播提供新的途径。
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引用次数: 0
mSphere of Influence: Venturing outside the biology canon with sex and gender. mSphere of Influence:跳出生物学对性和性别的束缚。
IF 3.7 2区 生物学 Q2 MICROBIOLOGY Pub Date : 2024-11-06 DOI: 10.1128/msphere.00594-24
Victoria Prieto-Echagüe

Victoria Prieto-Echagüe works in the field of signaling by primary cilia, adipogenesis, and obesity. In this mShpere of Influence article, she reflects on how gender studies, feminism, and societal movements such as #metoo may inform all areas of biomedical and health research. She describes how they inspired her to incorporate sex as a biological variable (SABV) principle to her research exploring sex-specific mechanisms in obesity and metabolic diseases and argues that incorporating SABV is crucial for advancing precision medicine and addressing healthcare inequities.

Victoria Prieto-Echagüe从事初级纤毛信号、脂肪生成和肥胖领域的研究。在这篇《影响力》(mShpere of Influence)文章中,她探讨了性别研究、女权主义和#metoo等社会运动如何为生物医学和健康研究的各个领域提供信息。她描述了这些运动如何启发她将性别作为生物变量(SABV)的原则纳入她探索肥胖和代谢性疾病中性别特异性机制的研究中,并认为纳入 SABV 对于推进精准医学和解决医疗保健不平等问题至关重要。
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引用次数: 0
The promiscuous biotin ligase TurboID reveals the proxisome of the T3SS chaperone IpgC in Shigella flexneri. 杂合生物素连接酶 TurboID 揭示了 flexneri 志贺氏菌中 T3SS 合酶 IpgC 的近端体。
IF 3.7 2区 生物学 Q2 MICROBIOLOGY Pub Date : 2024-10-31 DOI: 10.1128/msphere.00553-24
Nathaline Haidar-Ahmad, Kyle Tomaro, Mathieu Lavallée-Adam, François-Xavier Campbell-Valois

Promiscuous biotin ligases derived from the bacterial enzyme BirA are used to identify proteins vicinal to a bait protein, thereby defining its proxisome. Despite the popularity of this approach, surprisingly little is known about its use in prokaryotes. Here, we compared the activity of four widely used promiscuous biotin ligases in the cytoplasm of Shigella flexneri, a pathogenic subgroup of Escherichia coli. Our data indicate that the kinetics of TurboID's biotinylating activity is the highest of those tested. In addition, TurboID showed reduced interaction with the natural BirA binding partners, BccP and the biotin operator, when compared to its ancestor BioID. We therefore evaluated the ability of TurboID to probe the proxisome of the type III secretion system (T3SS) chaperone IpgC and the transcriptional activator MxiE. When the T3SS is inactive (off-state), these proteins are inhibited by forming complexes with the T3SS substrates OspD1 and IpaBC, respectively. In contrast, when the T3SS is active (on-state), OspD1 and IpaBC are secreted allowing MxiE and IpgC to interact together and activate their target genes. The results obtained with the IpgC and TurboID fusions capture a good fraction of these known interactions. It also suggests that the availability of IpgC increases in the on-state, resulting in a greater number of proteins detected in its vicinity. Among these is the T3SS ATPase SpaL (also known as Spa47 or SctN), further supporting the notion that chaperones escort their substrate to the T3SS. Interestingly, a specific subset of proteins conserved in E. coli completes the IpgC proxisome in the on-state.IMPORTANCEPromiscuous biotin ligases are widely used to study protein function in eukaryotes. Strikingly, their use in prokaryotes has been rare. Indeed, the small volume and the cytoplasmic location of the biotin ligase's natural binding partners in these organisms pose unique challenges that can interfere with the study of the proxisome of proteins of interest. Here, we evaluated four of the most common promiscuous biotin ligases and found TurboID was best suited for use in the cytoplasm of Shigella flexneri. Using this method, we extended the proxisome of IpgC beyond its known direct binding partners involved in the regulation of the type III secretion system (T3SS) signaling cascade. Of particular interest for further study are transcription factors and housekeeping proteins that are enriched around IpgC when the T3SS is active. We propose a model in which the increased availability of IpgC in the on-state may allow cross-talk of the T3SS with other cellular processes.

源于细菌酶 BirA 的杂合生物素连接酶可用于识别诱饵蛋白附近的蛋白质,从而确定其近端体。尽管这种方法很受欢迎,但令人惊讶的是,人们对它在原核生物中的应用知之甚少。在这里,我们比较了四种广泛使用的杂合生物素连接酶在柔性志贺氏菌(大肠杆菌的一个致病亚群)细胞质中的活性。我们的数据表明,TurboID 的生物素连接活性的动力学是所测试的生物素连接活性中最高的。此外,与其祖先 BioID 相比,TurboID 与天然 BirA 结合伙伴 BccP 和生物素算子的相互作用减少了。因此,我们评估了 TurboID 探测 III 型分泌系统(T3SS)伴侣 IpgC 和转录激活剂 MxiE 的近端体的能力。当 T3SS 处于非活性(关闭状态)时,这些蛋白分别与 T3SS 底物 OspD1 和 IpaBC 形成复合物而受到抑制。相反,当 T3SS 处于活动状态(开启状态)时,OspD1 和 IpaBC 被分泌出来,使 MxiE 和 IpgC 能够相互作用并激活它们的目标基因。利用 IpgC 和 TurboID 融合体获得的结果捕捉到了这些已知相互作用的大部分。它还表明,IpgC 在开启状态下的可用性增加,导致在其附近检测到更多的蛋白质。其中包括 T3SS ATP 酶 SpaL(又称 Spa47 或 SctN),这进一步支持了伴侣蛋白护送底物到 T3SS 的观点。有趣的是,在大肠杆菌中保守的特定蛋白质亚群在导通状态下完成了 IpgC 近端体。令人吃惊的是,它们在原核生物中的应用却很少。事实上,生物素连接酶在这些生物体内的天然结合伙伴体积小且位于细胞质中,这给研究相关蛋白质的近端体带来了独特的挑战。在这里,我们评估了四种最常见的杂合生物素连接酶,发现 TurboID 最适合用于柔性志贺氏杆菌的细胞质。利用这种方法,我们扩展了 IpgC 的近端体,使其超越了参与调节 III 型分泌系统(T3SS)信号级联的已知直接结合伙伴。当 T3SS 活跃时,IpgC 周围富集的转录因子和管家蛋白尤其值得进一步研究。我们提出了一个模型,在该模型中,IpgC 在开启状态下的可用性增加可能会使 T3SS 与其他细胞过程发生交叉对话。
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引用次数: 0
Adaptation to zinc restriction in Streptococcus agalactiae: role of the ribosomal protein and zinc-importers regulated by AdcR. 无乳链球菌对锌限制的适应:受 AdcR 调节的核糖体蛋白和锌输入器的作用。
IF 3.7 2区 生物学 Q2 MICROBIOLOGY Pub Date : 2024-10-31 DOI: 10.1128/msphere.00614-24
M Melet, S Blanchet, P Barbarin, E A Maunders, S L Neville, V Rong, L Mereghetti, C A McDevitt, A Hiron

Zinc (Zn) is an essential cofactor for numerous bacterial proteins and altering Zn availability is an important component of host innate immunity. During infection, adaptation to both Zn deprivation and excess is critical for pathogenic bacteria development. To understand the adaptive responses to Zn availability of Streptococcus agalactiae, a pathogen causing invasive infections of neonates, global transcriptional profiling was conducted. Results highlight that in response to Zn limitation, genes belonging to the AdcR regulon, the master regulator of Zn homeostasis in streptococci, were overexpressed. Through a combination of in silico analysis and experimental validation, new AdcR-regulated targets were identified. Among them, we identified a duplicated ribosomal protein, RpsNb, and an ABC transporter, and examined the role of these genes in bacterial growth under Zn-restricted conditions. Our results indicated that, during Zn restriction, both the RpsNb protein and a potential secondary Zn transporter are important for S. agalactiae adaptation to Zn deficiency.

Importance: Streptococcus agalactiae is a bacterial human pathobiont causing invasive diseases in neonates. Upon infection, S. agalactiae is presented with Zn limitation and excess but the genetic systems that allow bacterial adaptation to these conditions remain largely undefined. A comprehensive analysis of S. agalactiae global transcriptional response to Zn availability shows that this pathogen manages Zn limitation mainly through upregulation of the AdcR regulon. We demonstrate that several AdcR-regulated genes are important for bacterial growth during Zn deficiency, including human biological fluids. Taken together, these findings reveal new mechanisms of S. agalactiae adaptation under conditions of metal deprivation.

锌(Zn)是许多细菌蛋白质的重要辅助因子,改变锌的供应是宿主先天免疫的重要组成部分。在感染过程中,对锌缺乏和过剩的适应对病原菌的发展至关重要。为了了解引起新生儿侵袭性感染的病原体--无乳链球菌对锌供应的适应性反应,我们进行了全局转录谱分析。结果表明,为了应对锌限制,属于链球菌锌平衡主调节器 AdcR 调节子的基因被过度表达。通过硅学分析和实验验证,我们发现了新的 AdcR 调控靶标。其中,我们发现了一个重复的核糖体蛋白 RpsNb 和一个 ABC 转运体,并研究了这些基因在锌限制条件下细菌生长中的作用。我们的研究结果表明,在锌限制条件下,RpsNb 蛋白和一个潜在的次级锌转运体对于无乳链球菌适应锌缺乏非常重要:重要意义:无乳链球菌是一种人类致病细菌,可导致新生儿侵袭性疾病。感染后,S. agalactiae 会出现锌限制和过量,但使细菌适应这些条件的遗传系统在很大程度上仍未确定。对 S. agalactiae 对锌可用性的全局转录反应的全面分析表明,这种病原体主要通过 AdcR 调节子的上调来管理锌限制。我们证明,在锌缺乏期间,包括在人体生物液体中,几个由 AdcR 调节的基因对细菌的生长非常重要。总之,这些发现揭示了 S. agalactiae 在金属匮乏条件下的新适应机制。
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引用次数: 0
Analytical measuring interval, linearity, and precision of serology assays for detection of SARS-CoV-2 antibodies according to CLSI guidelines. 根据 CLSI 指南检测 SARS-CoV-2 抗体的血清学测定的分析测量间隔、线性度和精确度。
IF 3.7 2区 生物学 Q2 MICROBIOLOGY Pub Date : 2024-10-31 DOI: 10.1128/msphere.00393-24
Katarzyna Haynesworth, Troy J Kemp, Sarah A Loftus, Jordan Metz, Nicholas C Castro, Jimmie Bullock, David Fetterer, Ligia A Pinto
<p><p>Serology testing is commonly used to evaluate the immunogenicity of COVID-19 vaccines and measure antibodies as a marker of previous infection with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). In this study, four laboratory-developed serology enzyme-linked immunosorbent assays (SARS-CoV-2 anti-Spike and anti-Nucleocapsid immunoglobin G [IgG] and immunoglobin M [IgM]) calibrated to the WHO International Standard 20/136 were validated via analytical measuring interval (limit of blank [LOB], limit of detection [LOD], and limit of quantification [LOQ]), linearity, and precision according to the Clinical and Laboratory Standards Institute (CLSI) guidelines EP17-A2, EP06 2nd Edition, and EP05-A3. For Spike IgG, LOB was 3.0 binding antibody units per milliliter (BAU/mL), LOD was 4.1 BAU/mL, and LOQ was 27.1 BAU/mL. For Nucleocapsid IgG, LOB was 1.9 BAU/mL, LOD was 3.2 BAU/mL, and LOQ was 24.6 BAU/mL. For Spike IgM, LOB was 57.1 BAU/mL, LOD was 69.0 BAU/mL, and LOQ was 113.5 BAU/mL. For Nucleocapsid IgM, LOD was 242.2 BAU/mL, LOD was 289.9 BAU/mL, and LOQ was 572.4 BAU/mL. Each assay displayed good linearity (max % deviation from linearity (≥LOQ) = 10.7%). The result of within-run repeatability evaluation for medium positive samples was 7.7% for Spike IgG, 4.6% for Nucleocapsid IgG, 7.5% for Spike IgM, and 10.1% for Nucleocapsid IgM. The total precision, including medium positive sample variability across 20 days, three reagent kits, and two operators, was 13.5% for Spike IgG, 14.5% for Nucleocapsid IgG, 17.6% for Spike IgM, and 16.2% for Nucleocapsid IgM. The assays were successfully validated following the applicable CLSI guidelines. All assays met the ±20% deviation from linearity and the ±20% coefficient of variation specification for precision and repeatability.</p><p><strong>Importance: </strong>Reliable and validated serology assays are of increasing importance as the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) virus continues to evolve and cause outbreaks. Validation of serology assays along with calibration to the International and National Standards (such as anti-SARS-CoV-2 Immunoglobulin WHO International Standard 20/136 or Frederick National Laboratory for Cancer Research's National Serology Standard COVID-NS01097) is critical to ensuring that results from clinical studies are reliable and comparable among various assays and laboratories. We describe the design and execution of a comprehensive study that established the analytical measuring intervals, linearity, precision, and repeatability of four in-house developed serology enzyme-linked immunosorbent assays (SARS-CoV-2 anti-Spike immunoglobin G [IgG] and immunoglobin M [IgM] and anti-Nucleocapsid IgG and IgM) following applicable Clinical and Laboratory Standards Institute (CLSI) guidelines. Overall, this study provides practical guidance on experimental design strategies and data analysis techniques, pertaining to the validation of COVID-19 serology
血清学检测通常用于评估 COVID-19 疫苗的免疫原性,并测量作为严重急性呼吸系统综合症冠状病毒 2(SARS-CoV-2)既往感染标志物的抗体。在这项研究中,根据世界卫生组织国际标准 20/136 校准的四种实验室开发的血清学酶联免疫吸附测定(SARS-CoV-2 抗梭状病毒和抗核头壳免疫球蛋白 G [IgG] 和免疫球蛋白 M [IgM])通过分析测量间距(空白限 [LOB]、检出限 [LOD]、分析限 [LOB]、分析限 [LOD])进行了验证、检测限[LOD]和定量限[LOQ])、线性度和精确度进行了验证,符合临床和实验室标准协会(CLSI)指南 EP17-A2、EP06 第 2 版和 EP05-A3。Spike IgG 的 LOB 为 3.0 结合抗体单位/毫升(BAU/mL),LOD 为 4.1 BAU/mL,LOQ 为 27.1 BAU/mL。核壳 IgG 的 LOB 为 1.9 BAU/mL,LOD 为 3.2 BAU/mL,LOQ 为 24.6 BAU/mL。对于 Spike IgM,LOB 为 57.1 BAU/mL,LOD 为 69.0 BAU/mL,LOQ 为 113.5 BAU/mL。对于核壳 IgM,LOD 为 242.2 BAU/mL,LOD 为 289.9 BAU/mL,LOQ 为 572.4 BAU/mL。每种检测方法都显示出良好的线性(线性偏差的最大% (≥LOQ) = 10.7%)。中等阳性样品的重复性评估结果为:斯派克 IgG 7.7%,核头壳 IgG 4.6%,斯派克 IgM 7.5%,核头壳 IgM 10.1%。总精密度(包括 20 天、3 种试剂盒和 2 名操作员的中等阳性样本变异性)为:尖峰 IgG 13.5%,核头状病毒 IgG 14.5%,尖峰 IgM 17.6%,核头状病毒 IgM 16.2%。根据适用的 CLSI 指南,这些检测方法成功通过了验证。所有测定的精确度和重复性均符合线性偏差±20%和变异系数±20%的规范:随着严重急性呼吸系统综合症冠状病毒 2(SARS-CoV-2)病毒的不断演变和爆发,可靠且经过验证的血清学测定变得越来越重要。血清学测定的验证以及根据国际和国家标准(如抗 SARS-CoV-2 免疫球蛋白的世界卫生组织国际标准 20/136 或弗雷德里克国家癌症研究实验室的国家血清学标准 COVID-NS01097)进行的校准对于确保临床研究结果的可靠性以及不同测定和实验室之间的可比性至关重要。我们介绍了一项综合研究的设计和执行情况,该研究按照适用的临床和实验室标准协会 (CLSI) 指南,确定了四种内部开发的血清学酶联免疫吸附测定(SARS-CoV-2 抗梭状病毒免疫球蛋白 G [IgG] 和免疫球蛋白 M [IgM],以及抗核头壳 IgG 和 IgM)的分析测量范围、线性度、精确度和可重复性。总之,本研究为临床研究中根据 CLSI 指南验证 COVID-19 血清学检测方法的实验设计策略和数据分析技术提供了实用指导。
{"title":"Analytical measuring interval, linearity, and precision of serology assays for detection of SARS-CoV-2 antibodies according to CLSI guidelines.","authors":"Katarzyna Haynesworth, Troy J Kemp, Sarah A Loftus, Jordan Metz, Nicholas C Castro, Jimmie Bullock, David Fetterer, Ligia A Pinto","doi":"10.1128/msphere.00393-24","DOIUrl":"10.1128/msphere.00393-24","url":null,"abstract":"&lt;p&gt;&lt;p&gt;Serology testing is commonly used to evaluate the immunogenicity of COVID-19 vaccines and measure antibodies as a marker of previous infection with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). In this study, four laboratory-developed serology enzyme-linked immunosorbent assays (SARS-CoV-2 anti-Spike and anti-Nucleocapsid immunoglobin G [IgG] and immunoglobin M [IgM]) calibrated to the WHO International Standard 20/136 were validated via analytical measuring interval (limit of blank [LOB], limit of detection [LOD], and limit of quantification [LOQ]), linearity, and precision according to the Clinical and Laboratory Standards Institute (CLSI) guidelines EP17-A2, EP06 2nd Edition, and EP05-A3. For Spike IgG, LOB was 3.0 binding antibody units per milliliter (BAU/mL), LOD was 4.1 BAU/mL, and LOQ was 27.1 BAU/mL. For Nucleocapsid IgG, LOB was 1.9 BAU/mL, LOD was 3.2 BAU/mL, and LOQ was 24.6 BAU/mL. For Spike IgM, LOB was 57.1 BAU/mL, LOD was 69.0 BAU/mL, and LOQ was 113.5 BAU/mL. For Nucleocapsid IgM, LOD was 242.2 BAU/mL, LOD was 289.9 BAU/mL, and LOQ was 572.4 BAU/mL. Each assay displayed good linearity (max % deviation from linearity (≥LOQ) = 10.7%). The result of within-run repeatability evaluation for medium positive samples was 7.7% for Spike IgG, 4.6% for Nucleocapsid IgG, 7.5% for Spike IgM, and 10.1% for Nucleocapsid IgM. The total precision, including medium positive sample variability across 20 days, three reagent kits, and two operators, was 13.5% for Spike IgG, 14.5% for Nucleocapsid IgG, 17.6% for Spike IgM, and 16.2% for Nucleocapsid IgM. The assays were successfully validated following the applicable CLSI guidelines. All assays met the ±20% deviation from linearity and the ±20% coefficient of variation specification for precision and repeatability.&lt;/p&gt;&lt;p&gt;&lt;strong&gt;Importance: &lt;/strong&gt;Reliable and validated serology assays are of increasing importance as the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) virus continues to evolve and cause outbreaks. Validation of serology assays along with calibration to the International and National Standards (such as anti-SARS-CoV-2 Immunoglobulin WHO International Standard 20/136 or Frederick National Laboratory for Cancer Research's National Serology Standard COVID-NS01097) is critical to ensuring that results from clinical studies are reliable and comparable among various assays and laboratories. We describe the design and execution of a comprehensive study that established the analytical measuring intervals, linearity, precision, and repeatability of four in-house developed serology enzyme-linked immunosorbent assays (SARS-CoV-2 anti-Spike immunoglobin G [IgG] and immunoglobin M [IgM] and anti-Nucleocapsid IgG and IgM) following applicable Clinical and Laboratory Standards Institute (CLSI) guidelines. Overall, this study provides practical guidance on experimental design strategies and data analysis techniques, pertaining to the validation of COVID-19 serology","PeriodicalId":19052,"journal":{"name":"mSphere","volume":null,"pages":null},"PeriodicalIF":3.7,"publicationDate":"2024-10-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142546502","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
In vivo RNA sequencing reveals a crucial role of Fus3-Kss1 MAPK pathway in Candida glabrata pathogenicity. 体内 RNA 测序揭示了 Fus3-Kss1 MAPK 通路在念珠菌致病性中的关键作用。
IF 3.7 2区 生物学 Q2 MICROBIOLOGY Pub Date : 2024-10-30 DOI: 10.1128/msphere.00715-24
Xinreng Mo, Xiangtai Yu, Hao Cui, Kang Xiong, Shan Yang, Chang Su, Yang Lu

Candida glabrata is an important and increasingly common pathogen of humans, particularly in immunocompromised hosts. Despite this, little is known about how this fungus causes disease. Here, we applied RNA sequencing and an in vivo invasive infection model to identify the attributes that allow this organism to infect hosts. Fungal transcriptomes show a dramatic increase in the expression of Fus3 and Kss1, two mitogen-activated protein kinases (MAPKs), during invasive infection. We further demonstrate that they are both highly induced under a combination of serum and high CO2 conditions. Deletion of both FUS3 and KSS1, but neither gene alone, results in a reduced fungal burden in organs, as well as in the gastrointestinal tract in the DSS (Dextran Sulfate Sodium)-induced colitis model. Similarly, the defect in persistence in macrophages and attenuated adhesion to epithelial cells are observed when FUS3 and KSS1 are both disrupted. The fus3 kss1 double mutant also displays defects in the induction of virulence attributes such as genes required for iron acquisition and adhesion and in the anti-fungal drug tolerance. The putative downstream transcription factors Ste12 (1), Ste12 (2), Tec1, and Tec2 are found to be involved in the regulation of these virulence attributes. Collectively, our study indicates that an evolutionary conserved MAPK pathway, which regulates mating and filamentous growth in Saccharomyces cerevisiae, is critical for C. glabrata pathogenicity.

Importance: The MAPK signaling pathway, mediated by closely related kinases Fus3 and Kss1, is crucial for controlling mating and filamentous growth in Saccharomyces cerevisiae, but this pathway does not significantly impact hyphal development and pathogenicity in Candida albicans, a commensal-pathogenic fungus of humans. Furthermore, deletion of Cpk1, the ortholog of Fus3 in pathogenic fungus Cryptococcus neoformans, has no effect on virulence. Here, we demonstrate that the MAPK pathway is crucial for the pathogenicity of Candida glabrata, a fungus that causes approximately one-third of cases of hematogenously disseminated candidiasis in the United States. This pathway regulates multiple virulence attributes including the induction of iron acquisition genes and adhesins, as well as persistence in macrophages and organs. Our work provides insights into C. glabrata pathogenesis and highlights an example in which regulatory rewiring of a conserved pathway confers a virulent phenotype in a pathogen.

光滑念珠菌是一种重要的人类病原体,而且越来越常见,尤其是在免疫力低下的宿主中。尽管如此,人们对这种真菌如何致病却知之甚少。在这里,我们应用 RNA 测序和体内侵袭感染模型来确定这种真菌感染宿主的属性。真菌转录组显示,在入侵感染过程中,两种丝裂原活化蛋白激酶(MAPK)Fus3 和 Kss1 的表达量急剧增加。我们进一步证明,在血清和高二氧化碳条件下,这两种酶都会被高度诱导。在右旋糖酐硫酸钠(DSS)诱导的结肠炎模型中,同时缺失 FUS3 和 KSS1(而非单独缺失这两个基因)会导致器官以及胃肠道中的真菌负担减轻。同样,当 FUS3 和 KSS1 同时被破坏时,巨噬细胞中的持久性缺陷和对上皮细胞的粘附力也会减弱。fus3 kss1 双突变体在诱导毒力属性(如铁获取和粘附所需的基因)和抗真菌药物耐受性方面也表现出缺陷。推测的下游转录因子 Ste12 (1)、Ste12 (2)、Tec1 和 Tec2 参与了这些毒力属性的调控。总之,我们的研究表明,进化保守的 MAPK 通路(调节酿酒酵母的交配和丝状生长)对 C. glabrata 的致病性至关重要:由密切相关的激酶 Fus3 和 Kss1 介导的 MAPK 信号通路对控制酿酒酵母的交配和丝状生长至关重要,但这一通路对白色念珠菌(一种人类共生致病真菌)的菌丝发育和致病性并无显著影响。此外,致病真菌新生隐球菌中 Fus3 的直向同源物 Cpk1 的缺失对致病性也没有影响。在这里,我们证明了 MAPK 通路对于光滑念珠菌的致病性至关重要,这种真菌导致了美国约三分之一的血源性播散念珠菌病病例。该途径调节多种致病性属性,包括诱导铁获取基因和粘附蛋白,以及在巨噬细胞和器官中的持久性。我们的研究深入了解了玻璃样念珠菌的致病机理,并突出展示了一个实例,即保守通路的调控重构赋予病原体一种毒性表型。
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引用次数: 0
A novel antibody treatment reduces deformed wing virus loads in the western honey bee (Apis mellifera). 一种新型抗体疗法可减少西方蜜蜂(Apis mellifera)的畸形翅病毒载量。
IF 3.7 2区 生物学 Q2 MICROBIOLOGY Pub Date : 2024-10-30 DOI: 10.1128/msphere.00497-24
N J J MacMillan, B M Hause, T Nordseth, A Felden, J W Baty, J L Pitman, P J Lester

The deformed wing virus (Iflavirus aladeformis) (DWV) is a key driver of colony loss in the western honey bee (Apis mellifera). Here, we demonstrate that orally delivered anti-DWV antibodies can act systemically to reduce DWV loads in naturally infected honey bees. Immunoglobulin Y (IgY) was produced in adult chickens against two DWV proteins, harvested from their eggs, and fed to bees in a sucrose solution. An enzyme-linked immunosorbent assay demonstrated that orally delivered anti-DWV IgY migrated to the hemolymph. We next assessed the ability of orally delivered anti-DWV IgY to reduce DWV viral loads in naturally infected bees using qPCR. An antibody treatment resulted in a significant eightfold viral load reduction in DWV-infected bees. Our findings demonstrate the potential for antibody treatments to help mitigate the losses attributed to DWV in A. mellifera.

Importance: Deformed wing virus (DWV) is considered to be a key component of declining honey bee health which threatens global food production. The virus can result in significantly shortened lifespan, deformities in developing bees, and impaired cognition. There is currently no method to directly control the virus. The virus can be indirectly controlled with acaricidal treatments that target a key vector, the parasitic varroa mite (Varroa destructor). But acaricide resistance and a lack of effective alternatives for the control of both Varroa and DWV are major threats to beekeeping and the wider agricultural industry. Our research presents a significant development in the ability to reduce DWV burden in honey bees using IgY antibodies. Moreover, immunoglobulin Y has the potential to be more broadly established as a new treatment modality to combat other pathogens and parasites in A. mellifera.

畸形翅病毒(Iflavirus aladeformis)(DWV)是造成西方蜜蜂(Apis mellifera)蜂群损失的主要原因。在这里,我们证明了口服抗 DWV 抗体可在自然感染的蜜蜂中发挥系统性作用,减少 DWV 的负荷。我们在成年鸡体内产生了抗两种DWV蛋白的免疫球蛋白Y(IgY),从它们的卵中提取,并用蔗糖溶液喂给蜜蜂。酶联免疫吸附试验表明,口服抗DWV IgY可迁移到血淋巴中。接下来,我们使用 qPCR 评估了口服抗 DWV IgY 降低自然感染蜜蜂体内 DWV 病毒载量的能力。经抗体处理后,受 DWV 感染的蜜蜂体内的病毒载量明显降低了 8 倍。我们的研究结果表明,抗体疗法有可能帮助减轻畸形翼病毒给 A. mellifera 造成的损失:畸形翅病毒(DWV)被认为是蜜蜂健康状况下降的一个关键因素,它威胁着全球的粮食生产。该病毒可导致蜜蜂寿命明显缩短、发育中的蜜蜂畸形以及认知能力受损。目前还没有直接控制这种病毒的方法。病毒可以通过杀螨剂来间接控制,杀螨剂针对的是一种关键的媒介--寄生变节螨(Varroa destructor)。但是,杀螨剂的抗药性以及缺乏有效的替代品来控制变螨和 DWV,是养蜂业和更广泛的农业产业面临的主要威胁。我们的研究在利用 IgY 抗体减少蜜蜂 DWV 负担方面取得了重大进展。此外,免疫球蛋白 Y 有可能被更广泛地确立为一种新的治疗方式,用于防治蜜蜂体内的其他病原体和寄生虫。
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引用次数: 0
Detection of Hepatovirus A (HAV) in wastewater indicates widespread national distribution and association with socioeconomic indicators of vulnerability. 在废水中检测到甲型肝炎病毒(HAV)表明该病毒在全国广泛分布,并与易感性的社会经济指标有关。
IF 3.7 2区 生物学 Q2 MICROBIOLOGY Pub Date : 2024-10-30 DOI: 10.1128/msphere.00645-24
Alessandro Zulli, Elana M G Chan, Alexandria B Boehm

Wastewater-based epidemiology, which seeks to assess disease occurrence in communities through measurements of infectious disease biomarkers in wastewater, may represent a valuable tool for understanding the occurrence of hepatitis A infections in communities. In this study, we measured concentrations of Hepatovirus A (HAV) RNA, in samples from 191 wastewater treatment plants spanning 40 US states and the District of Columbia from September 2023 to June 2024 and compared the measurements with traditional measures of disease occurrence. Nationally, 13.76% of the 21,079 wastewater samples were positive for HAV RNA, and both concentrations and positivity rates were associated with NNDSS hepatitis A case data nationally (Kendall rank correlation coefficient = 0.20, concentrations; and 0.33, positivity rate; both P < 0.05). We further demonstrated that higher rates of wastewater HAV detection were positively associated with socioeconomic indicators of vulnerability including homelessness and drug overdose deaths (both P < 0.0001). Areas with above average levels of homelessness were 48% more likely to have HAV wastewater detections, while areas with above average levels of drug overdose deaths were 14% more likely to have HAV wastewater detections. Using more granular case data, we present a case study in the state of Maine that reinforces these results and suggests a potential lead time for wastewater over clinical case detection and exposure events. The ability to detect HAV RNA in wastewater before clinical cases emerge could allow public health officials to implement targeted interventions like vaccination campaigns.IMPORTANCEDespite the existence of a highly effective vaccine for hepatitis A, outbreaks in vulnerable populations remain common. The disease can be asymptomatic or subclinical, and disproportionately impacts populations with inadequate access to healthcare, leading to a severe underestimation of the occurrence of this viral infection. This study investigates the potential for wastewater measurements of biomarkers of the causative agent of hepatitis A (HAV RNA) to provide insights into disease occurrence. Results highlight the potential for wastewater-based epidemiology to be a complementary tool to traditional surveillance for monitoring and controlling HAV transmission.

基于废水的流行病学旨在通过测量废水中的传染病生物标志物来评估社区中的疾病发生情况,它可能是了解社区中甲型肝炎感染发生情况的重要工具。在这项研究中,我们测量了 2023 年 9 月至 2024 年 6 月期间来自美国 40 个州和哥伦比亚特区的 191 家污水处理厂样本中甲型肝炎病毒 (HAV) RNA 的浓度,并将测量结果与传统的疾病发生率测量方法进行了比较。从全国范围来看,21,079 份废水样本中有 13.76% 呈 HAV RNA 阳性,浓度和阳性率均与全国 NNDSS 甲型肝炎病例数据相关(浓度方面的 Kendall 秩相关系数 = 0.20;阳性率方面的 Kendall 秩相关系数 = 0.33;P 均小于 0.05)。我们进一步证明,较高的废水甲型肝炎病毒检测率与无家可归者和吸毒过量死亡等社会经济弱势指标呈正相关(均 P < 0.0001)。无家可归人数高于平均水平的地区检测到 HAV 废水的可能性要高出 48%,而吸毒过量死亡人数高于平均水平的地区检测到 HAV 废水的可能性要高出 14%。通过使用更精细的病例数据,我们介绍了缅因州的一个案例研究,该研究证实了这些结果,并表明废水可能比临床病例检测和暴露事件更早出现。在临床病例出现之前检测废水中甲型肝炎病毒 RNA 的能力可以让公共卫生官员实施有针对性的干预措施,如开展疫苗接种活动。这种疾病可能没有症状或处于亚临床状态,对无法获得充分医疗保健服务的人群的影响尤为严重,导致对这种病毒感染发生率的严重低估。本研究调查了废水中甲型肝炎致病因子(HAV RNA)生物标志物的测量潜力,以便深入了解疾病的发生情况。研究结果突出表明,基于废水的流行病学有可能成为传统监测的补充工具,用于监测和控制甲型肝炎病毒的传播。
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引用次数: 0
Cell aggregation mediated by ACE2 deletion in Candida auris modulates fungal colonization and host immune responses in the skin. 白色念珠菌中 ACE2 缺失介导的细胞聚集调节真菌在皮肤中的定植和宿主免疫反应。
IF 3.7 2区 生物学 Q2 MICROBIOLOGY Pub Date : 2024-10-30 DOI: 10.1128/msphere.00734-24
Abishek Balakumar, Abigail Cox, Shankar Thangamani

Candida auris is an emerging multi-drug-resistant fungal pathogen that colonizes the skin and causes invasive infections in hospitalized patients. Multi-cellular aggregative phenotype is widely reported in the C. auris isolates, but its role in skin colonization and host immune response is not yet known. In this study, we generated aggregative phenotype by deleting the ACE2 gene in C. auris and determined the fungal colonization and host immune response using an intradermal mouse model of C. auris skin infection. Our results indicate that mice infected with ace2Δ strain had significantly lower fungal load after 3 and 14 days post-infections compared to the non-aggregative wild-type and the ACE2 reintegrated strain. The colonization of ace2Δ is associated with increased recruitment of CD11b+ Ly6G+ neutrophils and decreased accumulation of CD11b+ Ly6 Chi inflammatory monocytes and CD11b+ MHCII+ CD64+ macrophages. Furthermore, Th17 cells and type 3 innate lymphoid cells (ILCs) were significantly increased in the skin tissue of ace2Δ infected mice. Our findings suggest that aggregative phenotype mediated by ACE2 deletion in C. auris induces potent neutrophil and IL-17-mediated immune response and reduces fungal colonization in the skin.IMPORTANCEC. auris is a rapidly emerging fungal pathogen that can colonize hospitalized patients, especially in skin tissue, and cause invasive infections. C. auris isolates exhibit morphological heterogeneity, and the multicellular aggregative phenotype of C. auris is reported frequently in clinical settings. Understanding the role of fungal morphotypes in colonization, persistence, and immune response in the skin microenvironment will have potential applications in clinical diagnosis and novel preventive and therapeutic measures. Here, we utilized the murine model of intradermal infection and determined that the aggregative phenotype of C. auris as the result of ACE2 gene deletion elicits potential innate and adaptive immune responses in mice. These observations will help explain the differences in the skin colonization and immune responses of the aggregative morphotype of C. auris and open the door to developing novel antifungal therapeutics.

白色念珠菌(Candida auris)是一种新出现的多重耐药真菌病原体,可在皮肤上定植并导致住院病人的侵袭性感染。多细胞聚集表型在 C. auris 分离物中被广泛报道,但其在皮肤定植和宿主免疫反应中的作用尚不清楚。在本研究中,我们通过删除 C. auris 中的 ACE2 基因产生了聚集表型,并使用 C. auris 皮肤感染小鼠皮内模型测定了真菌定植和宿主免疫反应。我们的结果表明,与非聚集野生型和 ACE2 重整合株相比,感染 ace2Δ 株的小鼠在感染后 3 天和 14 天的真菌负荷量明显较低。ace2Δ的定植与CD11b+ Ly6G+中性粒细胞的招募增加以及CD11b+ Ly6 Chi炎性单核细胞和CD11b+ MHCII+ CD64+巨噬细胞的聚集减少有关。此外,ace2Δ感染小鼠皮肤组织中的Th17细胞和3型先天性淋巴细胞(ILCs)显著增加。我们的研究结果表明,ACE2缺失介导的C. auris聚集表型可诱导有效的中性粒细胞和IL-17介导的免疫反应,并减少真菌在皮肤中的定植。C. auris 分离物表现出形态异质性,临床上经常报告 C. auris 的多细胞聚集表型。了解真菌形态在皮肤微环境中的定植、持续存在和免疫反应中的作用将可能应用于临床诊断和新型预防与治疗措施。在这里,我们利用小鼠皮内感染模型,确定了由于 ACE2 基因缺失导致的 C. auris 的聚集表型会引起小鼠潜在的先天性和适应性免疫反应。这些观察结果将有助于解释C. auris聚集表型在皮肤定植和免疫反应方面的差异,并为开发新型抗真菌疗法打开大门。
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