Microbes and Infection最新文献_第10页

LPS-LBP complex induced endothelial cell pyroptosis in aortic dissection is associated with gut dysbiosis 主动脉夹层中 LPS-LBP 复合物诱导的内皮细胞脓毒症与肠道菌群失调有关

IF 2.6 4区医学 Q3 IMMUNOLOGY

Microbes and Infection

Pub Date : 2025-02-01 Epub Date: 2024-08-20 DOI: 10.1016/j.micinf.2024.105406

Gulinazi Yesitayi , Qi Wang , Mengmeng Wang , Mierxiati Ainiwan , Kaisaierjiang Kadier , Aliya Aizitiaili , Yitong Ma , Xiang Ma

Acute aortic dissection (AAD) is the most severe traumatic disease affecting the aorta. Pyroptosis-mediated vascular wall inflammation is a crucial trigger for AAD, and the exact mechanism requires further investigation. In this study, our proteomic analysis showed that Lipopolysaccharide (LPS)-binding protein (LBP) was significantly upregulated in the plasma and aortic tissue of patients with AAD. Further, 16S rRNA sequencing of stool samples suggested that patients with AAD exhibit gut dysbiosis, which may lead to an impaired intestinal barrier and LPS leakage. By comparing with control mice, we found that LBP, including Pyrin Domain Containing Protein3 (NLRP3), the CARD-containing adapter apoptosis-associated speck-like protein (ASC), and Cleaved caspase-1, were upregulated in the AAD aorta, whereas gut intestinal barrier-related proteins were downregulated. Moreover, treated with LBPK95A (an LBP inhibitor) attenuated the incidence of AAD, the expression levels of pyroptosis-related factors, and the extent of vascular pathological changes compared to those in AAD mice. In addition, LPS and LBP treatment of human umbilical vein endothelial cells (HUVECs) activated TLR4 signaling and intracellular reactive oxygen species (ROS) production, which stimulated NLRP3 inflammasome formation and mediated pyroptosis in endothelial cells. Our findings showed that gut dysbiosis mediates pyroptosis by the LPS-LBP complex, thus providing new insights into developing AAD.

急性主动脉夹层（AAD）是影响主动脉的最严重创伤性疾病。热蛋白沉积介导的血管壁炎症是诱发急性主动脉夹层的关键因素，其确切机制有待进一步研究。本研究的蛋白质组分析显示，AAD 患者血浆和主动脉组织中的脂多糖（LPS）结合蛋白（LBP）明显上调。此外，粪便样本的 16S rRNA 测序表明，AAD 患者的肠道菌群失调可能导致肠道屏障受损和 LPS 泄漏。通过与对照组小鼠进行比较，我们发现枸杞多糖（包括 Pyrin Domain Containing Protein3，NLRP3）、含 CARD 的适配器凋亡相关斑点样蛋白（apoptosis-associated speck-like protein，ASC）和裂解的 caspase-1 在 AAD 主动脉中上调，而肠道屏障相关蛋白下调。此外，与 AAD 小鼠相比，用 LBPK95A（一种 LBP 抑制剂）治疗可减轻 AAD 的发病率、热噬相关因子的表达水平以及血管病理变化的程度。此外，LPS和LBP处理人脐静脉内皮细胞（HUVECs）激活了TLR4信号传导和细胞内活性氧（ROS）的产生，从而刺激了NLRP3炎性体的形成并介导了内皮细胞的热蛋白沉积。我们的研究结果表明，肠道菌群失调通过 LPS-LBP 复合物介导了热蛋白沉积，从而为 AAD 的发展提供了新的见解。

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引用次数: 0

Murine C3 of the complement system affects infection by Leptospira interrogans 小鼠补体系统 C3 对钩端螺旋体感染的影响

IF 2.6 4区医学 Q3 IMMUNOLOGY

Microbes and Infection

Pub Date : 2025-02-01 Epub Date: 2024-09-14 DOI: 10.1016/j.micinf.2024.105413

Julia Avian Vassalakis , Denise Harumi Silva Yamashita , Leonardo Moura Midon , Bruno Cogliati , Marcos Bryan Heinemann , Thaís Akemi Amamura , Lourdes Isaac

Leptospirosis is an infectious neglected disease estimated to affect more than one million people worldwide each year. The Complement System plays a vital role in eliminating infectious agents. However, its precise role in leptospirosis remains to be fully understood. We investigated the importance of C3 in L. interrogans serovar Kennewicki strain Pomona Fromm (LPF) infection. Lack of C3 leads to decreased leukocyte number, impaired inflammatory response and failure to eliminate bacteria during the early stages of infection, which may cause interstitial nephritis later. These findings could be explained, at least in part, by the lower presence of local opsonins. Furthermore, antibody production against Leptospira was compromised in the absence of C3, highlighting the importance of CR2 in B lymphocyte proliferation and the adjuvant role of C3d in humoral immunity. Leptospires can be eliminated through the urine, and according to our study, the lack of C3 delays the elimination of LPF through urine during the early stages of the infection. These results strongly suggest the crucial role of C3 protein in orchestrating an appropriate inflammatory response against LPF infection and in effectively eliminating the bacteria from the body during the acute phase of leptospirosis.

钩端螺旋体病是一种被忽视的传染性疾病，据估计每年影响全球 100 多万人。补体系统在消除传染性病原体方面发挥着至关重要的作用。然而，它在钩端螺旋体病中的确切作用仍有待充分了解。我们研究了 C3 在 L. interrogans serovar Kennewicki strain Pomona Fromm（LPF）感染中的重要性。在感染的早期阶段，C3的缺乏会导致白细胞数量减少、炎症反应受损以及无法消灭细菌，这可能会在后期引起间质性肾炎。这些发现至少可以部分归因于当地存在较少的疏松素。此外，在缺乏 C3 的情况下，针对钩端螺旋体的抗体产生受到影响，这突出了 CR2 在 B 淋巴细胞增殖中的重要性以及 C3d 在体液免疫中的辅助作用。钩端螺旋体可通过尿液排出体外，而根据我们的研究，在感染的早期阶段，缺乏 C3 会延迟钩端螺旋体通过尿液排出体外。这些结果有力地表明，C3 蛋白在针对钩端螺旋体感染协调适当的炎症反应以及在钩端螺旋体病急性期有效地将细菌排出体外方面发挥着关键作用。

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引用次数: 0

Detection of various DNA and RNA viruses in bats in Yamaguchi Prefecture, Japan 日本山口县蝙蝠体内各种 DNA 和 RNA 病毒的检测。

IF 2.6 4区医学 Q3 IMMUNOLOGY

Microbes and Infection

Pub Date : 2025-02-01 Epub Date: 2024-09-24 DOI: 10.1016/j.micinf.2024.105425

Miyuka Nishizato , Urara Imai , Chisato Shigenaga , Miho Obata , Saki Mitsunaga , Marla Anggita , Samuel Nyampong , Shelly Wulandari , Weiyin Hu , Kazuki Kiuno , Lydia Mali Langata , Hiroyuki Imai , Masashi Sakurai , Tetsuya Yanagida , Ai Takano , Takashi Murakami , Chang-Gi Jeong , Jae-Ku Oem , Daisuke Hayasaka , Hiroshi Shimoda

Bats are important natural hosts of various zoonotic viruses, including Ebola virus, Lyssa virus, and severe acute respiratory syndrome coronavirus (SARS-CoV). Although investigation of bats is valuable for predicting emerging infectious diseases from these animals, few surveys of bat-derived viruses have been conducted in Japan. In the present study, samples were collected from a total of 132 bats of 4 different species from 4 different locations within Yamaguchi Prefecture; these sample were employed for comprehensive detection of bat-derived viruses by polymerase chain reaction (PCR) and reverse transcription (RT)-PCR using primers universal for each of 4 different viral classes. As a result of PCR and RT-PCR, various herpesviruses, astroviruses, coronaviruses, and adenoviruses were identified from a total of 80 bats. The detected herpesviruses belong to the Betaherpesvirinae or Gammaherpesvirinae subfamily, the detected adenoviruses to the genus Mastadenovirus, the detected astroviruses to the genus Mamastrovirus; and the detected coronaviruses belong to the genus Alphacoronavirus. The detected sequences of 12 strains of 4 families showed 100 % amino acid identity with viruses previously detected either in China or South Korea. These findings expand our understanding of viruses carried by bats, and provide insights into the nature of bat-derived viruses in Japan.

蝙蝠是各种人畜共患病病毒的重要天然宿主，包括埃博拉病毒、莱萨病毒和严重急性呼吸系统综合征冠状病毒（SARS-CoV）。虽然对蝙蝠的调查对于预测这些动物新出现的传染病很有价值，但日本很少对蝙蝠衍生病毒进行调查。本研究从山口县 4 个不同地点收集了 4 种不同种类的 132 只蝙蝠样本，并使用 4 种不同病毒类别的通用引物，通过聚合酶链式反应 (PCR) 和反转录 (RT)-PCR 对这些样本进行了蝙蝠衍生病毒的全面检测。通过聚合酶链式反应和反转录聚合酶链式反应，共从 80 只蝙蝠身上鉴定出了各种疱疹病毒、哮喘病毒、冠状病毒和腺病毒。检测到的疱疹病毒属于 Betaherpesvirinae 或 Gammaherpesvirinae 亚科，检测到的腺病毒属于 Mastadenovirus 属，检测到的星状病毒属于 Mamastrovirus 属，检测到的冠状病毒属于 Alphacoronavirus 属。检测到的 4 个科 12 个毒株的序列与之前在中国或韩国检测到的病毒的氨基酸同一性达到 100%。这些发现拓展了我们对蝙蝠所携带病毒的了解，并为了解日本蝙蝠衍生病毒的性质提供了线索。

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引用次数: 0

Bacterial and host factors involved in zoonotic Streptococcal meningitis 人畜共患链球菌脑膜炎涉及的细菌和宿主因素

IF 2.6 4区医学 Q3 IMMUNOLOGY

Microbes and Infection

Pub Date : 2025-01-01 Epub Date: 2024-04-04 DOI: 10.1016/j.micinf.2024.105335

Jiale Ma , Huizhen Wu , Zhe Ma , Zongfu Wu

Zoonotic streptococci cause several invasive diseases with high mortality rates, especially meningitis. Numerous studies elucidated the meningitis pathogenesis of zoonotic streptococci, some specific to certain bacterial species. In contrast, others are shared among different bacterial species, involving colonization and invasion of mucosal barriers, survival in the bloodstream, breaching the blood–brain and/or blood–cerebrospinal fluid barrier to access the central nervous system, and triggering inflammation of the meninges. This review focuses on the recent advancements in comprehending the molecular and cellular events of five major zoonotic streptococci responsible for causing meningitis in humans or animals, including Streptococcus agalactiae, Streptococcus equi subspecies zooepidemicus, Streptococcus suis, Streptococcus dysgalactiae, and Streptococcus iniae. The underlying mechanism was summarized into four themes, including 1) bacterial survival in blood, 2) brain microvascular endothelial cell adhesion and invasion, 3) penetration of the blood–brain barrier, and 4) activation of the immune system and inflammatory reaction within the brain. This review may contribute to developing therapeutics to prevent or mitigate injury of streptococcal meningitis and improve risk stratification.

人畜共患病链球菌可引起多种侵袭性疾病，死亡率很高，尤其是脑膜炎。许多研究阐明了人畜共患链球菌脑膜炎的发病机制，其中一些研究针对某些细菌种类。与此相反，另一些则是不同细菌种类共有的，包括定植和侵入粘膜屏障、在血液中存活、突破血脑和/或血脑脊液屏障进入中枢神经系统，以及引发脑膜炎症。本综述重点介绍了最近在理解导致人类或动物脑膜炎的五种主要人畜共患链球菌（包括、亚种、、和）的分子和细胞事件方面取得的进展。其基本机制归纳为四个主题，包括：1）细菌在血液中存活；2）脑微血管内皮细胞粘附和入侵；3）穿透血脑屏障；4）激活免疫系统和脑内炎症反应。本综述可能有助于开发预防或减轻链球菌脑膜炎损伤的疗法，并改善风险分层。

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引用次数: 0

Copyright page Elsevier 版权页面Elsevier

IF 2.6 4区医学 Q3 IMMUNOLOGY

Microbes and Infection

Pub Date : 2025-01-01 Epub Date: 2025-01-24 DOI: 10.1016/S1286-4579(25)00006-1

引用次数: 0

Comparative genome analysis of Streptococcus suis serotype 5 strains from humans and pigs revealed pathogenic potential of virulent, antimicrobial resistance, and genetic relationship 来自人和猪的猪链球菌血清型 5 菌株的基因组比较分析揭示了毒力、抗菌药耐药性和遗传关系的致病潜力

IF 2.6 4区医学 Q3 IMMUNOLOGY

Microbes and Infection

Pub Date : 2025-01-01 Epub Date: 2023-12-07 DOI: 10.1016/j.micinf.2023.105273

Anusak Kerdsin , Rujirat Hatrongjit , Thidathip Wongsurawat , Piroon Jenjaroenpun , Han Zheng , Peechanika Chopjitt , Parichart Boueroy , Nahuel Fittipaldi , Mariela Segura , Marcelo Gottschalk

Streptococcus suis is a causative agent of swine and human infections. Genomic analysis indicated that eight S. suis serotype 5 strains recovered from human patients and pigs carried many virulence-associated genes and markers defining pathogenic pathotypes. The strains were sequence types diverse and clustered within either minimum core genome group 3 (MCG-3) or MCG-7-3. Almost all the serotype 5 strains were non-susceptible to penicillin, ceftriaxone, erythromycin, and levofloxacin. Resistance to tetracycline and clindamycin was observed in all strains. The antimicrobial resistance genes tet(O), tet(O/W/32/O), tet(W), tet(44), erm(B), ant(6)-Ia, lsaE, and lnuB were found in these strains. Moderate-to-large numbers of substitutions were observed in three penicillin-binding proteins (PBP)—PBP1A, PBP2B, and PBP2X—in the penicillin-non-susceptible serotype 5 isolates that were involved in β-lactam-non-susceptibility. Comparative genomics between the serotype 5 and 2 strains revealed that only 15 genes absent from the serotype 2 strains were shared by all the serotype 5 strains. However, some additional genes were present only in some of the serotype 5 strains. This study highlighted the pathogenic potential of virulent serotype 5 strains in humans and pigs and the need for increased monitoring of penicillin-non-susceptibility in S. suis serotypes other than for serotype 2.

猪链球菌是猪和人类感染的致病菌。基因组分析表明，从人类患者和猪身上发现的八株猪链球菌血清型 5 菌株带有许多毒力相关基因和确定致病病型的标记。这些菌株的序列类型多种多样，并聚集在最小核心基因组群 3（MCG-3）或 MCG-7-3 中。几乎所有血清 5 型菌株都对青霉素、头孢曲松、红霉素和左氧氟沙星不敏感。所有菌株都对四环素和林可霉素有抗药性。在这些菌株中发现了抗菌药耐药基因 tet(O)、tet(O/W/32/O)、tet(W)、tet(44)、erm(B)、ant(6)-Ia、lsaE 和 lnuB。在青霉素不敏感的 5 号血清型分离株中，三种青霉素结合蛋白 (PBP)--PBP1A、PBP2B 和 PBP2X--出现了中度到大量的替换，这些蛋白与 β-内酰胺类药物不敏感性有关。5 号血清型和 2 号血清型菌株之间的基因组比较显示，只有 15 个 2 号血清型菌株不存在的基因为所有 5 号血清型菌株所共有。然而，还有一些基因只存在于部分血清 5 型菌株中。这项研究强调了毒力强大的血清 5 型菌株在人类和猪中的致病潜力，以及加强监测血清 2 型以外的猪链球菌血清型中青霉素不敏感性的必要性。

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引用次数: 0

Kinetic and proteomic studies in milk show distinct patterns among major Listeria monocytogenes clones 牛奶中的动力学和蛋白质组学研究显示，主要李斯特菌克隆之间存在不同的模式。

IF 2.6 4区医学 Q3 IMMUNOLOGY

Microbes and Infection

Pub Date : 2025-01-01 Epub Date: 2024-02-10 DOI: 10.1016/j.micinf.2024.105312

Alba Espí-Malillos , Carla Palacios-Gorba , Inmaculada López-Almela , Pilar Ruiz-García , María Carmen López-Mendoza , Francisco García-Del Portillo , M Graciela Pucciarelli , Juan J. Quereda

Listeria monocytogenes, a contaminant of raw milk, includes hypervirulent clonal complexes (CC) like CC1, CC4, and CC6, highly overrepresented in dairy products when compared to other food types. Whether their higher prevalence in dairy products is the consequence of a growth advantage in this food remains unknown. We examined growth kinetics of five L. monocytogenes isolates (CC1, CC4, CC6, CC9, and CC121) at 37 and 4 °C in ultra-high temperature (UHT) milk and raw milk. At 4 °C, hypovirulent CC9 and CC121 isolates exhibit better growth parameters in UHT milk compared to the hypervirulent CC1, CC4, and CC6 isolates. CC9 isolate in raw milk at 4 °C exhibited the fastest growth and the highest final concentrations. In contrast, hypervirulent isolates (CC1, CC4, and CC6) displayed better growth rates in UHT milk at 37 °C, the mammalian host temperature. Proteomic analysis of representative hyper- (CC1) and hypovirulent (CC9) isolates showed that they respond to milk cues differently with CC-specific traits. Proteins related to metabolism (such as LysA or different phosphotransferase systems), and stress response were upregulated in both isolates during growth in UHT milk. Our results show that there is a Listeria CC-specific and a Listeria CC-common response to the milk environment. These findings shed light on the overrepresentation of hypervirulent L. monocytogenes isolates in dairy products, suggesting that CC1 and CC4 overrepresentation in dairy products made of raw milk may arise from contamination during or after milking at the farm and discard an advantage of hypervirulent isolates in milk products when stored at refrigeration temperatures.

单核细胞增生李斯特菌是生牛奶中的一种污染物，其中包括 CC1、CC4 和 CC6 等超病毒克隆复合体（CC），与其他类型的食品相比，它们在乳制品中的比例很高。它们在乳制品中的高流行率是否是在这种食品中具有生长优势的结果，目前仍不得而知。我们研究了五种单核细胞增多性乳酸杆菌分离物（CC1、CC4、CC6、CC9 和 CC121）在超高温（UHT）牛奶和生奶中 37 ℃ 和 4 ℃ 的生长动力学。与高致病力的 CC1、CC4 和 CC6 分离物相比，低致病力的 CC9 和 CC121 分离物在 4 ℃ 超高温灭菌奶中的生长参数更好。CC9 分离物在 4 ℃ 的生乳中生长最快，最终浓度最高。相比之下，超病毒分离物（CC1、CC4 和 CC6）在 37 °C（哺乳动物宿主温度）超高温灭菌奶中的生长速度更快。对具有代表性的高病毒分离物（CC1）和低病毒分离物（CC9）进行的蛋白质组分析表明，它们对牛奶线索的反应不同，具有CC特异性。在超高温灭菌奶中生长期间，两种分离物中与新陈代谢（如 LysA 或不同的磷酸转移酶系统）和应激反应有关的蛋白质都出现了上调。我们的研究结果表明，李斯特菌 CC 对牛奶环境有特异性和共通性反应。这些发现揭示了乳制品中超病毒性单核细胞增生李斯特菌分离物比例过高的问题，表明生乳制成的乳制品中CC1和CC4比例过高可能是由于在牧场挤奶期间或挤奶后受到污染造成的，在冷藏温度下储存时，超病毒性分离物在乳制品中的优势就会消失。

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引用次数: 0

Cross-species transmission and animal infection model of hepatitis E virus 戊型肝炎病毒的跨物种传播和动物感染模型。

IF 2.6 4区医学 Q3 IMMUNOLOGY

Microbes and Infection

Pub Date : 2025-01-01 Epub Date: 2024-04-16 DOI: 10.1016/j.micinf.2024.105338

Ling-Dong Xu , Fei Zhang , Pinglong Xu , Yao-Wei Huang

Zoonotic hepatitis E virus (HEV) infection is an emerging global public health concern, and understanding the dynamics of HEV transmission between animals and humans is crucial for public health. Animal models are critical to advancing the understanding of HEV pathogenesis, drug screening, vaccine development, and other related areas. Here, we provide an overview of recent studies investigating the cross-species transmission of HEV, and also delve into the current research and application of animal HEV infection models including non-human primates, rodents, pigs, and chickens, offering a comprehensive assessment of the advantages and disadvantages of each model. This review highlights the findings related to viral replication, shedding patterns, and immune response in these animal models, and discusses the implications for our understanding of HEV transmission to humans. These advancements in the field enhance our understanding of the biological traits and pathogenic mechanisms of HEV, offering robust support for the development of highly effective and targeted prevention and treatment strategies.

人畜共患戊型肝炎病毒（HEV）感染是一个新兴的全球公共卫生问题，了解HEV在动物和人之间传播的动态对公共卫生至关重要。动物模型对于促进对HEV发病机制、药物筛选、疫苗开发和其他相关领域的理解至关重要。在此，我们概述了最近研究HEV跨物种传播的研究，并深入研究了目前动物HEV感染模型的研究和应用，包括非人类灵长类动物、啮齿动物、猪和鸡，并对每种模型的优缺点进行了全面评估。这篇综述强调了在这些动物模型中与病毒复制、脱落模式和免疫反应相关的发现，并讨论了我们对HEV向人类传播的理解的意义。该领域的这些进展增强了我们对HEV生物学特性和致病机制的理解，为制定高效和有针对性的预防和治疗策略提供了强有力的支持。

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引用次数: 0

The dual M protein systems have diverse biological characteristics, but both contribute to M18-type Group A Streptococcus pathogenicity 双M蛋白系统具有不同的生物学特性，但两者都有助于M18型A组链球菌的致病性。

IF 2.6 4区医学 Q3 IMMUNOLOGY

Microbes and Infection

Pub Date : 2025-01-01 Epub Date: 2023-08-17 DOI: 10.1016/j.micinf.2023.105209

Xiaorui Zhou , Haoshuai Song , Fei Pan , Chen Yuan , Lu Jia , Bing Wu , Hongjie Fan , Zhe Ma

M protein is a key surface virulence factor in Group A Streptococcus (GAS), Group C Streptococcus (GCS), and other streptococcal species. GAS encodes M protein using the emm gene, while GCS employs the szm (or sem) gene. In M18-type GAS, dual M protein systems exist, comprising both GAS and GCS M proteins (encoded separately by emm18 and spa18). The spa18 gene in M18-type GAS shares a conserved region highly similar to GCS's szm gene. Our study reveals that spa18 exhibits higher transcription levels than emm18 in M18-type GAS strains. The dual M protein systems defective mutant (Δemm18Δspa18) displays a smooth surface, whereas wild-type and single M protein gene mutants remain rough. M18 and SPA18 proteins possess distinct characteristics, showing varied binding properties and cytotoxicity effects on macrophages (THP-1) and keratinocytes (HaCaT). Both emm18 and spa18 genes contribute to the skin pathogenicity of M18-type GAS. Transcriptome analysis suggests the potential involvement of the mga gene in spa18 transcription regulation, while SpyM18_2047 appears to be specific to spa18 regulation. In summary, this research offers a crucial understanding of the biological characteristics of dual M protein systems in M18-type GAS, highlighting their contributions to virulence and transcriptional regulation.

M蛋白是a组链球菌（GAS）、C组链球菌（GCS）和其他链球菌物种的关键表面毒力因子。GAS使用emm基因编码M蛋白，而GCS使用szm（或sem）基因。在M18型GAS中，存在双M蛋白系统，包括GAS和GCS M蛋白（分别由emm18和spa18编码）。M18型GAS中的spa18基因与GCS的szm基因具有高度相似的保守区。我们的研究表明，在M18型GAS菌株中，spa18表现出比emm18更高的转录水平。双M蛋白系统缺陷突变体（Δemm18Δspa18）显示出光滑的表面，而野生型和单M蛋白基因突变体保持粗糙。M18和SPA18蛋白具有不同的特性，对巨噬细胞（THP-1）和角质形成细胞（HaCaT）表现出不同的结合特性和细胞毒性作用。emm18和spa18基因都参与了M18型GAS的皮肤致病性。转录组分析表明mga基因可能参与spa18转录调控，而SpyM18_2047似乎对spa18调控具有特异性。总之，这项研究对M18型GAS中双M蛋白系统的生物学特性有了至关重要的了解，突出了它们对毒力和转录调控的贡献。

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引用次数: 0

Rab27a via its effector JFC1 localizes to Anaplasma inclusions and promotes Anaplasma proliferation in leukocytes Rab27a 通过其效应物 JFC1 定位于阿那普拉斯原虫包涵体并促进阿那普拉斯原虫在白细胞中的增殖

IF 2.6 4区医学 Q3 IMMUNOLOGY

Microbes and Infection

Pub Date : 2025-01-01 Epub Date: 2023-12-16 DOI: 10.1016/j.micinf.2023.105278

Weiyan Huang, Mingqun Lin, Yasuko Rikihisa

Anaplasma phagocytophilum is an obligatory intracellular bacterium that causes tick-borne zoonosis called human granulocytic anaplasmosis. Mechanisms by which Anaplasma replicates inside of the membrane-bound compartment called “inclusion” in neutrophils are incompletely understood. A small GTPase Rab27a is found in the secretory granules and multivesicular endosomes. In this study we found Rab27a-containing granules were localized to Anaplasma inclusions in guanine nucleotide-dependent manner, and constitutively active Rab27a enhanced Anaplasma infection and dominant-negative Rab27a inhibited Anaplasma infection. Rab27a effector, JFC1 is known to mediate docking/fusion of Rab27a-bearing granules for exocytosis in leukocytes. shRNA stable knockdown of Rab27a or JFC1 inhibited Anaplasma infection in HL-60 cells. Similar to Rab27a, both endogenous and transfected JFC1 were localized to Anaplasma inclusions by immunostaining or live cell imaging. The JFC1 C2A domain that binds 3′-phosphoinositides, was sufficient and required for JFC1 and Rab27a localization to Anaplasma inclusions which were enriched with phosphatidylinositol 3-phosphate. Nexinhib20, the small molecule inhibitor specific to Rab27a and JFC1 binding, inhibited Anaplasma infection. Taken together, these results imply elevated phosphatidylinositol 3-phosphate in the inclusion membrane recruits JFC1 to mediate Rab27a-bearing granules/vesicles to dock/fuse with Anaplasma inclusions, the lumen of which is topologically equivalent to the exterior of the cell to benefit Anaplasma proliferation.

噬细胞无形体（Anaplasma phagocytophilum）是一种强制性细胞内细菌，可引起蜱传人畜共患疾病--人类粒细胞无形体病。目前还不完全清楚阿纳疟原虫在中性粒细胞内被称为 "包涵体 "的膜结合区内复制的机制。在分泌颗粒和多泡内体中发现了一种小 GTPase Rab27a。本研究发现，含 Rab27a 的颗粒以鸟嘌呤核苷酸依赖性方式定位于阿纳铂原虫包涵体，组成型活性 Rab27a 可增强阿纳铂原虫感染，显性阴性 Rab27a 可抑制阿纳铂原虫感染。已知 Rab27a 效应子 JFC1 在白细胞中介导带有 Rab27a 的颗粒的对接/融合以进行外渗。 shRNA 稳定敲除 Rab27a 或 JFC1 可抑制 HL-60 细胞中的阿纳疟原虫感染。与 Rab27a 相似，通过免疫染色和荧光探针转染活细胞，JFC1 也被定位到阿纳疟原虫包涵体中。JFC1 C2A结构域能结合3'-磷酸肌醇，是JFC1和Rab27a定位到阿纳普拉斯原虫内含物的充分条件和必要条件，阿纳普拉斯原虫内含物富含3-磷酸肌醇。对 Rab27a 和 JFC1 结合具有特异性的小分子抑制剂 Nexinhib20 可抑制阿纳普拉斯原虫感染。综上所述，这些结果表明包涵膜中磷脂酰肌醇-3-磷酸盐的升高招募了 JFC1，以介导含 Rab27a 的颗粒/囊泡与阿纳普拉斯原虫包涵体对接/融合，而阿纳普拉斯原虫包涵体的内腔在拓扑学上等同于细胞外部，有利于阿纳普拉斯原虫的增殖。

{"title":"Rab27a via its effector JFC1 localizes to Anaplasma inclusions and promotes Anaplasma proliferation in leukocytes","authors":"Weiyan Huang, Mingqun Lin, Yasuko Rikihisa","doi":"10.1016/j.micinf.2023.105278","DOIUrl":"10.1016/j.micinf.2023.105278","url":null,"abstract":"<div><div><em>Anaplasma phagocytophilum</em> is an obligatory intracellular bacterium that causes tick-borne zoonosis called human granulocytic anaplasmosis. Mechanisms by which <em>Anaplasma</em> replicates inside of the membrane-bound compartment called “inclusion” in neutrophils are incompletely understood. A small GTPase Rab27a is found in the secretory granules and multivesicular endosomes. In this study we found Rab27a-containing granules were localized to <em>Anaplasma</em> inclusions in guanine nucleotide-dependent manner, and constitutively active Rab27a enhanced <em>Anaplasma</em> infection and dominant-negative Rab27a inhibited <em>Anaplasma</em> infection. Rab27a effector, JFC1 is known to mediate docking/fusion of Rab27a-bearing granules for exocytosis in leukocytes. shRNA stable knockdown of Rab27a or JFC1 inhibited <em>Anaplasma</em> infection in HL-60 cells. Similar to Rab27a, both endogenous and transfected JFC1 were localized to <em>Anaplasma</em> inclusions by immunostaining or live cell imaging. The JFC1 C2A domain that binds 3′-phosphoinositides, was sufficient and required for JFC1 and Rab27a localization to <em>Anaplasma</em> inclusions which were enriched with phosphatidylinositol 3-phosphate. Nexinhib20, the small molecule inhibitor specific to Rab27a and JFC1 binding, inhibited <em>Anaplasma</em> infection. Taken together, these results imply elevated phosphatidylinositol 3-phosphate in the inclusion membrane recruits JFC1 to mediate Rab27a-bearing granules/vesicles to dock/fuse with <em>Anaplasma</em> inclusions, the lumen of which is topologically equivalent to the exterior of the cell to benefit <em>Anaplasma</em> proliferation.</div></div>","PeriodicalId":18497,"journal":{"name":"Microbes and Infection","volume":"27 1","pages":"Article 105278"},"PeriodicalIF":2.6,"publicationDate":"2025-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"138682491","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

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