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Cigarette smoke compromises macrophage innate sensing in response to pneumococcal infection. 香烟烟雾会损害巨噬细胞对肺炎球菌感染的先天感应。
IF 4.5 2区 医学 Q2 IMMUNOLOGY Pub Date : 2024-10-28 DOI: 10.1016/j.jmii.2024.10.001
Wei-Chih Liao, Chia-Huei Chou, Mao-Wang Ho, Jo-Tsen Chen, Shu-Ling Chou, Yu-Tsen Huang, Ngoc-Niem Bui, Hui-Yu Wu, Chi-Fan Lee, Wei-Chien Huang, Chih-Ho Lai

Background: Cigarette smoking remains a leading cause of mortality worldwide. Streptococcus pneumoniae, also known as pneumococcus, is one of the most common pathogens that colonizes the human respiratory tract, causing life-threatening infections. Several studies have reported that cigarette smoke (CS) exposure promotes pneumococcal infectivity; however, the underlying mechanisms remain to be illustrated.

Methods: In this study, we prepared cigarette smoke extract (CSE) from tobacco containing nicotine (0.8 mg/cigarette) and tar (10 mg/cigarette) to investigate the effects of CSE on innate immune response using murine macrophage models.

Results: The results from the cytokine array showed that the production of C-C Motif Chemokine Ligand 2 (CCL2), CCL4, CCL3, C-X-C Motif Chemokine Ligand 2 (CXCL2), and CXCL-10, in pneumococcus-infected cells was reduced upon 5 % CSE treatment. Our results further demonstrated that 5 % CSE exposure, followed by pneumococcal challenge, significantly decreased CCL2 and type I interferon (IFN) production in macrophages by inhibiting nuclear factor (NF)-κB and IFN regulatory factor 3 (IRF3) signaling pathways. Moreover, CSE disrupts macrophage polarization and impedes innate immune signaling to suppress pneumococcal phagocytosis by macrophages.

Conclusion: Our results provide evidence that CS manipulates the signaling molecules to subvert macrophage functions, thereby hindering the innate response against pneumococcal infection.

背景:吸烟仍然是导致全球死亡的主要原因。肺炎链球菌又称肺炎球菌,是人类呼吸道中最常见的病原体之一,可引起危及生命的感染。有几项研究报告称,接触香烟烟雾(CS)会促进肺炎球菌的感染性;然而,其根本机制仍有待说明:在这项研究中,我们从含有尼古丁(0.8 毫克/支)和焦油(10 毫克/支)的烟草中制备了香烟烟雾提取物(CSE),并利用小鼠巨噬细胞模型研究了 CSE 对先天性免疫反应的影响:细胞因子阵列的结果表明,5 % CSE 处理后,肺炎球菌感染细胞中 C-C Motif Chemokine Ligand 2 (CCL2)、CCL4、CCL3、C-X-C Motif Chemokine Ligand 2 (CXCL2) 和 CXCL-10 的产生量减少。我们的研究结果进一步表明,暴露于 5 % CSE 后再接受肺炎球菌挑战,可通过抑制核因子 (NF)-κB 和 IFN 调节因子 3 (IRF3) 信号通路,显著减少巨噬细胞中的 CCL2 和 I 型干扰素 (IFN) 的产生。此外,CSE 还能破坏巨噬细胞的极化,阻碍先天性免疫信号传导,从而抑制巨噬细胞对肺炎球菌的吞噬作用:结论:我们的研究结果提供了 CS 操纵信号分子以颠覆巨噬细胞功能的证据,从而阻碍了针对肺炎球菌感染的先天性免疫反应。
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引用次数: 0
Impacts of Pta-AckA pathway on CPS biosynthesis and type 3 fimbriae expression in Klebsiella pneumoniae. 肺炎克雷伯氏菌中 Pta-AckA 通路对 CPS 生物合成和 3 型纤毛膜表达的影响。
IF 4.5 2区 医学 Q2 IMMUNOLOGY Pub Date : 2024-10-24 DOI: 10.1016/j.jmii.2024.10.002
Tien-Huang Lin, Chen-Yu Wang, Chien-Chen Wu, Ching-Ting Lin

Background: Klebsiella pneumoniae is a Gram-negative bacterium that can cause infections, especially in individuals with diabetes. Recently, more hypervirulent strains have emerged, posing a threat even to healthy individuals. Understanding how K. pneumoniae regulates its virulence factors is crucial. Acetyl-phosphate (AcP) is essential for bacterial metabolism and can affect virulence factor expression. However, the role of the Pta-AckA pathway, which regulates AcP levels, in K. pneumoniae pathogenesis remains unclear.

Methods: Deletion mutants lacking the pta and ackA, involved in AcP production and hydrolysis, were generated in K. pneumoniae CG43S3. Their effects on AcP levels, the patterns of global acetylated protein, capsular polysaccharide (CPS) amount, serum resistance, type 3 fimbriae expression, biofilm formation, and virulence in G. mellonella larva were assessed.

Results: Deletion of ackA in K. pneumoniae CG43S3 led to AcP accumulation, while pta deletion abolished AcP synthesis when grown in TB7+1 % glucose. This pathway influenced global protein acetylation, with pta deletion decreasing acetylation and ackA deletion increasing it. Additionally, pta deletion decreased the CPS amount, serum resistance, and type 3 fimbriae expression, while ackA deletion increased these factors. Furthermore, deleting pta and ackA attenuated the infected larva's virulence and death rate.

Conclusion: Our findings highlight the critical role of the Pta-AckA pathway in K. pneumoniae pathogenesis. This pathway regulates AcP levels, global protein acetylation, CPS production, serum resistance, and type 3 fimbriae expression, ultimately impacting virulence. The information provides insights into potential therapeutic targets for combating K. pneumoniae infection.

背景:肺炎克雷伯菌是一种革兰氏阴性菌,可引起感染,尤其是糖尿病患者。最近,出现了更多的高毒力菌株,甚至对健康人也构成了威胁。了解肺炎克氏菌如何调节其毒力因子至关重要。乙酰磷酸(AcP)对细菌的新陈代谢至关重要,并可影响毒力因子的表达。然而,调节 AcP 水平的 Pta-AckA 通路在肺炎双球菌致病过程中的作用仍不清楚:方法:在肺炎双球菌 CG43S3 中产生了缺失参与 AcP 生成和水解的 pta 和 ackA 的缺失突变体。方法:在肺炎双球菌 CG43S3 中产生了缺乏 pta 和 ackA 的缺失突变体,它们参与 AcP 的产生和水解,评估了它们对 AcP 水平、全局乙酰化蛋白模式、荚膜多糖(CPS)量、血清抗性、3 型缘毛表达、生物膜形成和对 G. mellonella 幼虫的毒力的影响:结果:当肺炎克氏菌 CG43S3 在 TB7+1 % 葡萄糖中生长时,ackA 的缺失会导致 AcP 的积累,而 pta 的缺失则会阻碍 AcP 的合成。这一途径影响了全局蛋白质乙酰化,pta 缺失会降低乙酰化,ackA 缺失则会增加乙酰化。此外,pta 缺失会减少 CPS 量、血清抗性和 3 型叶状体的表达,而 ackA 缺失则会增加这些因素。此外,缺失 pta 和 ackA 会降低受感染幼虫的毒力和死亡率:我们的研究结果凸显了 Pta-AckA 通路在肺炎双球菌致病过程中的关键作用。该途径调控 AcP 水平、全局蛋白乙酰化、CPS 生成、血清抗性和 3 型缘膜表达,最终影响毒力。这些信息提供了抗击肺炎克氏菌感染的潜在治疗靶点。
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引用次数: 0
Serum Mpox-specific IgG titers before and after breakthrough Mpox infection in an HIV-infected individual with viral suppression and prior 2-dose Mpox vaccination. 一名病毒抑制并接种过 2 剂 Mpox 疫苗的 HIV 感染者在突破性 Mpox 感染前后的血清 Mpox 特异性 IgG 滴度。
IF 4.5 2区 医学 Q2 IMMUNOLOGY Pub Date : 2024-10-22 DOI: 10.1016/j.jmii.2024.10.003
Wang-Da Liu, Tai-Ling Chao, Sui-Yuan Chang, Chien-Ching Hung
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引用次数: 0
Alternations of the gut microbiota and the Firmicutes/Bacteroidetes ratio after biologic treatment in inflammatory bowel disease. 炎症性肠病患者接受生物治疗后肠道微生物群和固缩菌/类杆菌比例的变化。
IF 4.5 2区 医学 Q2 IMMUNOLOGY Pub Date : 2024-10-01 DOI: 10.1016/j.jmii.2024.09.006
Yu-Chieh Tsai, Wei-Chen Tai, Chih-Ming Liang, Cheng-Kun Wu, Ming-Chao Tsai, Wan-Hsiang Hu, Pao-Yuan Huang, Chien-Hung Chen, Yuan-Hung Kuo, Chih-Chien Yao, Seng-Kee Chuah

Background: The inflammatory bowel disease (IBD), comprising Crohn's disease (CD) and ulcerative colitis (UC) is a complex disease with multifactorial etiology. The intestinal dysbiosis have been investigated to play an important role in IBD pathogenesis and disease activity. The aim of our study was to analyze the intestinal microbiota composition in IBD across different severity levels and the impact of biologic therapy on microbiota modulation.

Methods: In this study, 27 IBD patients were recruited, including 14 patients undergoing biologic therapy for moderate to severe disease activity and 13 controls with inactive disease. The gut microbial composition was determined by 16 S ribosomal RNA gene sequencing of stool samples.

Results: Biologic therapy led to significant clinical improvement in IBD disease activity after 48 weeks. About species richness, community alpha diversity was significant lower in active CD patients and enriched gradually after biologic therapy. The beta-diversity regard to the difference of bacterial community composition showed significant difference between patients in biologic and control group. A decrease in Firmicutes and increase in Bacteroidetes abundance were observed in patients with active disease, both in CD and UC. Biologic treatment induced shifts in gut microbiota, with increased Firmicutes and decreased Bacteroidetes, as well as improved F/B ratio gradually after treatment, correlating with disease activity.

Conclusions: Our study suggested that gut microbiota differences changed after biologic therapies among IBD with different disease activity, and a rising Firmicutes/Bacteroidetes ratio could be a potential predictor for disease activity and treatment response monitoring.

背景:炎症性肠病(IBD),包括克罗恩病(CD)和溃疡性结肠炎(UC),是一种病因复杂的多因素疾病。据研究,肠道菌群失调在 IBD 发病机制和疾病活动中发挥着重要作用。我们的研究旨在分析不同严重程度的IBD患者的肠道微生物群组成,以及生物治疗对微生物群调节的影响:本研究共招募了 27 名 IBD 患者,包括 14 名因中重度疾病活动而接受生物治疗的患者和 13 名疾病不活跃的对照组患者。通过对粪便样本进行16 S核糖体RNA基因测序,确定肠道微生物组成:结果:48周后,生物疗法明显改善了IBD疾病的临床活动。在物种丰富度方面,活动性 CD 患者的群落α多样性明显较低,生物治疗后逐渐丰富。关于细菌群落组成差异的贝塔多样性显示,生物治疗组和对照组患者之间存在显著差异。在活动性疾病患者中,无论是 CD 还是 UC,都观察到了固醇菌的减少和类杆菌的增加。生物制剂治疗诱导肠道微生物群发生变化,固缩菌增加,类杆菌减少,治疗后F/B比值逐渐改善,这与疾病活动相关:我们的研究表明,不同疾病活动性的 IBD 患者在接受生物制剂治疗后肠道微生物群的差异发生了变化,而固缩菌/类杆菌比值的升高可作为疾病活动性和治疗反应监测的潜在预测指标。
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引用次数: 0
Unveiling unique effector function-related bulk antibody profiles in long-term hemodialysis patients following COVID-19 mRNA booster vaccination. 揭示长期血液透析患者接种 COVID-19 mRNA 强化疫苗后独特的效应功能相关批量抗体概况。
IF 4.5 2区 医学 Q2 IMMUNOLOGY Pub Date : 2024-10-01 DOI: 10.1016/j.jmii.2024.09.007
Chia-Yi Chou, Chung-Yi Cheng, Chih-Hsin Lee, Makoto Kuro-O, Tso-Hsiao Chen, San-Yuan Wang, Yung-Kun Chuang, Yun-Jung Yang, Yun-Hsuan Lin, I-Lin Tsai

Background: Hemodialysis patients exhibit a reduced response to vaccination and have different vaccine dose regimens. Vaccines induce antibodies and affect the inflammatory balance through antibody glycosylation and effector functions. Therefore, we aimed to analyze the antibody glycosylation profiles in hemodialysis patients who were vaccinated against severe acute respiratory syndrome coronavirus 2, infected with the virus, or both, and compare them with those of dialysis patients in a control group.

Methods: Plasma samples from 112 hemodialysis patients were assigned to four groups: control, infected, vaccinated, and post-vaccine-infected. Paired plasma samples from 47 people with vaccination (vaccinees) were analyzed before and after the booster dose. The same analytical approach was applied to the four groups for a cross-sectional comparison.

Results: Our study found that both vaccination and infection groups showed decreased fucosylation of IgG1, which is associated with a proinflammatory biosignature. However, vaccination also leads to increased galactosylation and bisection of IgG antibodies, which are associated with anti-inflammatory effects and the additional regulation of immune responses. In contrast, infection led to an additional decrease in the fucosylation of IgG2 and IgA, demonstrating a more intense proinflammatory biosignature than vaccination.

Conclusions: Our findings emphasize the proinflammatory biosignature of afucosylation in both vaccination and infection groups. Additionally, we uncovered further regulated profiles related to galactosylation in vaccinees. These findings suggest that antibody investigation for vaccination or infection should not solely focus on neutralization but should also consider effector function-related glycosylation profiling. This comprehensive information can be valuable for fine-tuning vaccine development in the future.

背景:血液透析患者对疫苗接种的反应减弱,疫苗剂量方案也不同。疫苗会诱导抗体,并通过抗体糖基化和效应器功能影响炎症平衡。因此,我们旨在分析接种过严重急性呼吸道综合征冠状病毒 2 疫苗、感染过该病毒或两者兼有的血液透析患者的抗体糖基化谱,并与对照组透析患者的抗体糖基化谱进行比较:将 112 名血液透析患者的血浆样本分为四组:对照组、感染组、接种组和接种后感染组。对接种疫苗的 47 人(接种者)在加强剂量前后的配对血浆样本进行了分析。对四个组别采用相同的分析方法进行横向比较:我们的研究发现,接种组和感染组的 IgG1 肌糖基化都有所下降,而这与促炎生物标志有关。然而,接种疫苗也会导致 IgG 抗体的半乳糖基化和双链化增加,这与抗炎作用和免疫反应的额外调节有关。相比之下,感染导致 IgG2 和 IgA 的岩藻糖基化进一步降低,显示出比接种疫苗更强烈的促炎生物特征:我们的研究结果表明,疫苗接种组和感染组中的岩藻糖基化都具有促炎症生物特征。此外,我们还发现了疫苗接种者中与半乳糖基化相关的进一步调控特征。这些发现表明,针对疫苗接种或感染的抗体调查不应只关注中和作用,还应考虑与效应器功能相关的糖基化谱分析。这些全面的信息对未来疫苗开发的微调很有价值。
{"title":"Unveiling unique effector function-related bulk antibody profiles in long-term hemodialysis patients following COVID-19 mRNA booster vaccination.","authors":"Chia-Yi Chou, Chung-Yi Cheng, Chih-Hsin Lee, Makoto Kuro-O, Tso-Hsiao Chen, San-Yuan Wang, Yung-Kun Chuang, Yun-Jung Yang, Yun-Hsuan Lin, I-Lin Tsai","doi":"10.1016/j.jmii.2024.09.007","DOIUrl":"https://doi.org/10.1016/j.jmii.2024.09.007","url":null,"abstract":"<p><strong>Background: </strong>Hemodialysis patients exhibit a reduced response to vaccination and have different vaccine dose regimens. Vaccines induce antibodies and affect the inflammatory balance through antibody glycosylation and effector functions. Therefore, we aimed to analyze the antibody glycosylation profiles in hemodialysis patients who were vaccinated against severe acute respiratory syndrome coronavirus 2, infected with the virus, or both, and compare them with those of dialysis patients in a control group.</p><p><strong>Methods: </strong>Plasma samples from 112 hemodialysis patients were assigned to four groups: control, infected, vaccinated, and post-vaccine-infected. Paired plasma samples from 47 people with vaccination (vaccinees) were analyzed before and after the booster dose. The same analytical approach was applied to the four groups for a cross-sectional comparison.</p><p><strong>Results: </strong>Our study found that both vaccination and infection groups showed decreased fucosylation of IgG1, which is associated with a proinflammatory biosignature. However, vaccination also leads to increased galactosylation and bisection of IgG antibodies, which are associated with anti-inflammatory effects and the additional regulation of immune responses. In contrast, infection led to an additional decrease in the fucosylation of IgG2 and IgA, demonstrating a more intense proinflammatory biosignature than vaccination.</p><p><strong>Conclusions: </strong>Our findings emphasize the proinflammatory biosignature of afucosylation in both vaccination and infection groups. Additionally, we uncovered further regulated profiles related to galactosylation in vaccinees. These findings suggest that antibody investigation for vaccination or infection should not solely focus on neutralization but should also consider effector function-related glycosylation profiling. This comprehensive information can be valuable for fine-tuning vaccine development in the future.</p>","PeriodicalId":56117,"journal":{"name":"Journal of Microbiology Immunology and Infection","volume":null,"pages":null},"PeriodicalIF":4.5,"publicationDate":"2024-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142481864","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Development of the novel gene chip and restriction fragment length polymorphism (RFLP) methods for rapid detection of Mycobacterium tuberculosis complex in broth culture. 开发新型基因芯片和限制性片段长度多态性(RFLP)方法,用于肉汤培养液中结核分枝杆菌复合体的快速检测。
IF 4.5 2区 医学 Q2 IMMUNOLOGY Pub Date : 2024-09-21 DOI: 10.1016/j.jmii.2024.09.003
Wen-Hung Wang, Chun-Yu Lin, Shu-Huei Jain, Po-Liang Lu, Yen-Hsu Chen

Background: Tuberculosis (TB) is a major global public health issue. Prompt and accurate TB diagnosis is crucial for starting appropriate treatments and preventing the disease's spread. Current diagnostic techniques are either slow or expensive. This study aimed to create and evaluate a new, fast, highly reliable, and cost-effective TB detection method using a gene chip and Restriction Fragment Length Polymorphism (RFLP) analysis on Mycobacteria Growth Indicator Tubes (MGIT) specimens.

Methods: We assessed the effectiveness of a novel gene chip and RFLP methods targeting the 16S rRNA gene of Mycobacterium tuberculosis in 2000 MGIT culture-positive specimens. RFLP analysis identified the AfeI restriction site within the M. tuberculosis complex (MTBC) genome. Discrepancies were investigated through extensive sequencing and Cobas TaqMan PCR analysis, along with reviewing patient profiles.

Results: Both methods showed high efficacy in detecting MTBC in broth cultures, with the gene chip method achieving a sensitivity of 99.27 %, specificity of 98.35 %, and the RFLP method showing a sensitivity of 98.18 %, specificity of 99.31 %. False negatives in two isolates were due to a mutation in the AfeI site. Additionally, five cases showed MTBC presence when nontuberculous Mycobacterium species grew in cultures.

Conclusion: Our novel gene chip and RFLP methods are effective for rapid highly-reliable and cost-effective M. tuberculosis detection in MGIT specimens. Both gene chip and RFLP methods are suitable for resource-limited settings, offering an economical advantage. These methods have significant potential to improve clinical TB diagnosis.

背景:结核病(TB)是一个重大的全球公共卫生问题。及时准确的结核病诊断对于开始适当的治疗和防止疾病蔓延至关重要。目前的诊断技术要么缓慢,要么昂贵。本研究旨在利用基因芯片和对分枝杆菌生长指示管(MGIT)标本的限制性片段长度多态性(RFLP)分析,创建并评估一种新型、快速、高度可靠且经济高效的结核病检测方法:我们在 2000 份 MGIT 培养阳性标本中评估了针对结核分枝杆菌 16S rRNA 基因的新型基因芯片和 RFLP 方法的有效性。RFLP 分析确定了结核分枝杆菌复合体(MTBC)基因组中的 AfeI 限制位点。通过广泛的测序和 Cobas TaqMan PCR 分析,并查阅患者资料,对差异进行了调查:基因芯片法的灵敏度为 99.27%,特异性为 98.35%;RFLP 法的灵敏度为 98.18%,特异性为 99.31%。两个分离物的假阴性是由于 AfeI 位点发生了突变。此外,有五个病例在培养物中生长出非结核分枝杆菌时显示出 MTBC 的存在:结论:我们的新型基因芯片和 RFLP 方法可在 MGIT 标本中快速、高可靠、低成本地检测结核杆菌。基因芯片和 RFLP 方法都适用于资源有限的环境,具有经济优势。这些方法在改善临床结核病诊断方面具有巨大潜力。
{"title":"Development of the novel gene chip and restriction fragment length polymorphism (RFLP) methods for rapid detection of Mycobacterium tuberculosis complex in broth culture.","authors":"Wen-Hung Wang, Chun-Yu Lin, Shu-Huei Jain, Po-Liang Lu, Yen-Hsu Chen","doi":"10.1016/j.jmii.2024.09.003","DOIUrl":"https://doi.org/10.1016/j.jmii.2024.09.003","url":null,"abstract":"<p><strong>Background: </strong>Tuberculosis (TB) is a major global public health issue. Prompt and accurate TB diagnosis is crucial for starting appropriate treatments and preventing the disease's spread. Current diagnostic techniques are either slow or expensive. This study aimed to create and evaluate a new, fast, highly reliable, and cost-effective TB detection method using a gene chip and Restriction Fragment Length Polymorphism (RFLP) analysis on Mycobacteria Growth Indicator Tubes (MGIT) specimens.</p><p><strong>Methods: </strong>We assessed the effectiveness of a novel gene chip and RFLP methods targeting the 16S rRNA gene of Mycobacterium tuberculosis in 2000 MGIT culture-positive specimens. RFLP analysis identified the AfeI restriction site within the M. tuberculosis complex (MTBC) genome. Discrepancies were investigated through extensive sequencing and Cobas TaqMan PCR analysis, along with reviewing patient profiles.</p><p><strong>Results: </strong>Both methods showed high efficacy in detecting MTBC in broth cultures, with the gene chip method achieving a sensitivity of 99.27 %, specificity of 98.35 %, and the RFLP method showing a sensitivity of 98.18 %, specificity of 99.31 %. False negatives in two isolates were due to a mutation in the AfeI site. Additionally, five cases showed MTBC presence when nontuberculous Mycobacterium species grew in cultures.</p><p><strong>Conclusion: </strong>Our novel gene chip and RFLP methods are effective for rapid highly-reliable and cost-effective M. tuberculosis detection in MGIT specimens. Both gene chip and RFLP methods are suitable for resource-limited settings, offering an economical advantage. These methods have significant potential to improve clinical TB diagnosis.</p>","PeriodicalId":56117,"journal":{"name":"Journal of Microbiology Immunology and Infection","volume":null,"pages":null},"PeriodicalIF":4.5,"publicationDate":"2024-09-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142333172","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
High hemolytic activity in Staphylococcus aureus t1081/ST45 due to increased hla protein production and potential RNAIII-independent regulation. 金黄色葡萄球菌 t1081/ST45 的高溶血活性是由于 hla 蛋白生成增加和潜在的 RNAIII 非依赖性调控所致。
IF 4.5 2区 医学 Q2 IMMUNOLOGY Pub Date : 2024-09-21 DOI: 10.1016/j.jmii.2024.09.005
Yu-Tzu Lin, Ngoc-Niem Bui, Yu-Syuan Cheng, Cheng-Wen Lin, Chun-Li Lee, Tai-Fen Lee, Po-Ren Hsueh

Background: α-Hemolysin, encoded by hla, is a major virulence factor of Staphylococcus aureus. Sequence type (ST) 45 is a globally spread clone with increasing clinical prevalence in Taiwan. Our previous study showed that among the CC45 isolates, the spa type t1081 isolates presented greater hemolytic activity.

Materials and methods: The hemolytic activity of 67 CC45 isolates (44 t1081 and 23 non-t1081) from clinical blood cultures was assessed using rabbit red blood cells. The sequences of hla and its upstream regulatory regions and RNAIII were compared between the two groups. The expression of hla and its regulators RNAIII, mgrA, and saeR was analyzed via qRT‒PCR, while Hla protein levels were measured via Western blotting.

Results: Compared with non-t1081 isolates, t1081 isolates presented increased hemolytic activity. No significant differences in hla sequences, upstream regulatory regions, or gene expression levels were detected between the two groups. The expression of the transcriptional regulators mgrA and saeR was also similar between the two groups. Western blotting revealed increased Hla protein in the t1081 isolates. However, neither the sequence or expression of RNAIII, a regulator of hla at both the transcriptional and posttranscriptional levels, differed between the groups.

Conclusion: Our study revealed that, compared with other CC45 isolates, the t1081/ST45 isolates presented greater hemolytic activity. This heightened activity was due mainly to increased Hla protein levels. Moreover, the higher translation levels may be independent of the known regulator RNAIII, indicating a potential RNAIII-independent mechanism for Hla regulation.

背景:由 hla 编码的 α 溶血素是金黄色葡萄球菌的主要毒力因子。序列类型(ST)45 是一种全球传播的克隆,在台湾的临床流行率越来越高。我们之前的研究表明,在 CC45 分离物中,t1081 型 spa 分离物具有更强的溶血活性:用兔红细胞评估了临床血液培养物中 67 个 CC45 分离物(44 个 t1081 型和 23 个非 t1081 型)的溶血活性。比较了两组细胞中 hla 及其上游调控区和 RNAIII 的序列。通过 qRT-PCR 分析了 hla 及其调控因子 RNAIII、mgrA 和 saeR 的表达,并通过 Western 印迹检测了 Hla 蛋白水平:结果:与非 t1081 分离物相比,t1081 分离物的溶血活性增强。两组分离物的 hla 序列、上游调控区和基因表达水平均无明显差异。两组之间转录调控因子 mgrA 和 saeR 的表达也相似。Western 印迹显示,t1081 分离物中的 Hla 蛋白增加了。然而,RNAIII(hla 在转录和转录后水平的调控因子)的序列和表达在两组间均无差异:我们的研究表明,与其他 CC45 分离物相比,t1081/ST45 分离物具有更强的溶血活性。这种活性的增强主要是由于 Hla 蛋白水平的提高。此外,较高的翻译水平可能与已知的调节因子 RNAIII 无关,这表明 Hla 的调节机制可能与 RNAIII 无关。
{"title":"High hemolytic activity in Staphylococcus aureus t1081/ST45 due to increased hla protein production and potential RNAIII-independent regulation.","authors":"Yu-Tzu Lin, Ngoc-Niem Bui, Yu-Syuan Cheng, Cheng-Wen Lin, Chun-Li Lee, Tai-Fen Lee, Po-Ren Hsueh","doi":"10.1016/j.jmii.2024.09.005","DOIUrl":"https://doi.org/10.1016/j.jmii.2024.09.005","url":null,"abstract":"<p><strong>Background: </strong>α-Hemolysin, encoded by hla, is a major virulence factor of Staphylococcus aureus. Sequence type (ST) 45 is a globally spread clone with increasing clinical prevalence in Taiwan. Our previous study showed that among the CC45 isolates, the spa type t1081 isolates presented greater hemolytic activity.</p><p><strong>Materials and methods: </strong>The hemolytic activity of 67 CC45 isolates (44 t1081 and 23 non-t1081) from clinical blood cultures was assessed using rabbit red blood cells. The sequences of hla and its upstream regulatory regions and RNAIII were compared between the two groups. The expression of hla and its regulators RNAIII, mgrA, and saeR was analyzed via qRT‒PCR, while Hla protein levels were measured via Western blotting.</p><p><strong>Results: </strong>Compared with non-t1081 isolates, t1081 isolates presented increased hemolytic activity. No significant differences in hla sequences, upstream regulatory regions, or gene expression levels were detected between the two groups. The expression of the transcriptional regulators mgrA and saeR was also similar between the two groups. Western blotting revealed increased Hla protein in the t1081 isolates. However, neither the sequence or expression of RNAIII, a regulator of hla at both the transcriptional and posttranscriptional levels, differed between the groups.</p><p><strong>Conclusion: </strong>Our study revealed that, compared with other CC45 isolates, the t1081/ST45 isolates presented greater hemolytic activity. This heightened activity was due mainly to increased Hla protein levels. Moreover, the higher translation levels may be independent of the known regulator RNAIII, indicating a potential RNAIII-independent mechanism for Hla regulation.</p>","PeriodicalId":56117,"journal":{"name":"Journal of Microbiology Immunology and Infection","volume":null,"pages":null},"PeriodicalIF":4.5,"publicationDate":"2024-09-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142333173","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Streptococcus canis infection in Taiwan. 台湾的犬链球菌感染。
IF 4.5 2区 医学 Q2 IMMUNOLOGY Pub Date : 2024-09-21 DOI: 10.1016/j.jmii.2024.09.004
Pei-Yun Tsai, Wan-Hsuan Hsu, Yu-Hsin Chiu, Chih-Cheng Lai, Hung-Jen Tang
{"title":"Streptococcus canis infection in Taiwan.","authors":"Pei-Yun Tsai, Wan-Hsuan Hsu, Yu-Hsin Chiu, Chih-Cheng Lai, Hung-Jen Tang","doi":"10.1016/j.jmii.2024.09.004","DOIUrl":"https://doi.org/10.1016/j.jmii.2024.09.004","url":null,"abstract":"","PeriodicalId":56117,"journal":{"name":"Journal of Microbiology Immunology and Infection","volume":null,"pages":null},"PeriodicalIF":4.5,"publicationDate":"2024-09-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142367665","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Comparison of global transcriptomes for nontyphoidal Salmonella clinical isolates from pediatric patients with and without bacteremia after their interaction with human intestinal epithelial cells in vitro. 患有和未患有菌血症的儿科临床分离株的非伤寒沙门氏菌在体外与人类肠上皮细胞相互作用后的全局转录组比较。
IF 4.5 2区 医学 Q2 IMMUNOLOGY Pub Date : 2024-09-19 DOI: 10.1016/j.jmii.2024.09.002
Buyandelger Batsaikhan, Pei-Chun Lin, Katsumi Shigemura, Yu-Wei Wu, Reo Onishi, Pei-Ru Chang, Hung-Yen Cheng, Shiuh-Bin Fang

Background: Nontyphoidal Salmonella (NTS) outbreaks of invasive diseases are increasing. Whether the genetic diversity of invasive NTS correlates with the clinical characteristics and bacteremia development in NTS infections remains unclear. In this study, we compared the global transcriptomes between bacteremic and nonbacteremic NTS strains after their interaction with human intestinal epithelial cells in vitro.

Methods: We selected clinical isolates obtained from stool and blood samples of patients with or without bacteremia and patients with high and low C-reactive protein (CRP) levels. The bacterial RNA samples were isolated after coculturing with Caco-2 cells for RNA sequencing and subsequent analyses.

Results: CRP is an unreliable predictive maker for NTS bacteremia with a median CRP level of 1.6 mg/dL. Certain Salmonella Pathogenicity Island (SPI)-1 genes (sipC, sipA, sicA, sipD, and sipB), SPI-2 genes (ssaP, ssrA, and ssaS), and six SPI-4 genes (siiA, siiB, siiC, siiD, siiE, and siiF) remained upregulated in the bacteremic blood-derived strains but significantly downregulated in the nonbacteremic strains after their interaction with Caco-2 cells. The Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways analysis identified that arginine biosynthesis, ascorbate and aldarate metabolism, and phosphotransferase system pathways were activated in bacteremic NTS strains after Caco-2 cell priming.

Conclusion: CRP levels were not correlated with bacteremia development. Significant regulation of certain SPI genes in bacteremic NTS strains after Caco-2 cell priming; bacteremia development might be influenced by the host immune response and the extent to which specific metabolism pathways in NTS strains can be prevented from invading the bloodstream.

背景:非伤寒沙门氏菌(NTS)引起的侵袭性疾病爆发日益增多。侵袭性 NTS 的遗传多样性是否与 NTS 感染的临床特征和菌血症发展相关仍不清楚。在本研究中,我们比较了菌血症和非菌血症NTS菌株在体外与人类肠上皮细胞相互作用后的全局转录组:我们选择了从菌血症或非菌血症患者以及C反应蛋白(CRP)水平高和低的患者的粪便和血液样本中获得的临床分离株。细菌 RNA 样本在与 Caco-2 细胞共培养后分离出来,进行 RNA 测序和后续分析:结果:CRP 是预测 NTS 菌血症的不可靠指标,CRP 水平中位数为 1.6 mg/dL。某些沙门氏菌致病性岛(SPI)-1基因(sipC、sipA、sicA、sipD和sipB)、SPI-2基因(ssaP、ssrA和ssaS)以及6个SPI-4基因(siiA、siiB、siiC、siiD、siiE和siiF)在菌血症血源性菌株中保持上调,但在与Caco-2细胞相互作用后,在非菌血症菌株中则显著下调。京都基因和基因组百科全书(KEGG)通路分析表明,Caco-2细胞引物作用后,菌血症NTS菌株的精氨酸生物合成、抗坏血酸和醛酸代谢以及磷酸转移酶系统通路被激活:结论:CRP水平与菌血症的发生无关。Caco-2细胞诱导后,菌血症NTS菌株中的某些SPI基因受到显著调控;菌血症的发生可能受到宿主免疫反应的影响,以及NTS菌株中特定代谢途径可阻止其入侵血液的程度。
{"title":"Comparison of global transcriptomes for nontyphoidal Salmonella clinical isolates from pediatric patients with and without bacteremia after their interaction with human intestinal epithelial cells in vitro.","authors":"Buyandelger Batsaikhan, Pei-Chun Lin, Katsumi Shigemura, Yu-Wei Wu, Reo Onishi, Pei-Ru Chang, Hung-Yen Cheng, Shiuh-Bin Fang","doi":"10.1016/j.jmii.2024.09.002","DOIUrl":"https://doi.org/10.1016/j.jmii.2024.09.002","url":null,"abstract":"<p><strong>Background: </strong>Nontyphoidal Salmonella (NTS) outbreaks of invasive diseases are increasing. Whether the genetic diversity of invasive NTS correlates with the clinical characteristics and bacteremia development in NTS infections remains unclear. In this study, we compared the global transcriptomes between bacteremic and nonbacteremic NTS strains after their interaction with human intestinal epithelial cells in vitro.</p><p><strong>Methods: </strong>We selected clinical isolates obtained from stool and blood samples of patients with or without bacteremia and patients with high and low C-reactive protein (CRP) levels. The bacterial RNA samples were isolated after coculturing with Caco-2 cells for RNA sequencing and subsequent analyses.</p><p><strong>Results: </strong>CRP is an unreliable predictive maker for NTS bacteremia with a median CRP level of 1.6 mg/dL. Certain Salmonella Pathogenicity Island (SPI)-1 genes (sipC, sipA, sicA, sipD, and sipB), SPI-2 genes (ssaP, ssrA, and ssaS), and six SPI-4 genes (siiA, siiB, siiC, siiD, siiE, and siiF) remained upregulated in the bacteremic blood-derived strains but significantly downregulated in the nonbacteremic strains after their interaction with Caco-2 cells. The Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways analysis identified that arginine biosynthesis, ascorbate and aldarate metabolism, and phosphotransferase system pathways were activated in bacteremic NTS strains after Caco-2 cell priming.</p><p><strong>Conclusion: </strong>CRP levels were not correlated with bacteremia development. Significant regulation of certain SPI genes in bacteremic NTS strains after Caco-2 cell priming; bacteremia development might be influenced by the host immune response and the extent to which specific metabolism pathways in NTS strains can be prevented from invading the bloodstream.</p>","PeriodicalId":56117,"journal":{"name":"Journal of Microbiology Immunology and Infection","volume":null,"pages":null},"PeriodicalIF":4.5,"publicationDate":"2024-09-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142333171","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Sequential use of capsular typing and whole-genome sequencing-based analysis for transmission of carbapenem-resistant Acinetobacter baumannii in a tertiary medical center. 在一家三级医疗中心,依次使用胶囊分型和全基因组测序分析耐碳青霉烯类鲍曼不动杆菌的传播。
IF 4.5 2区 医学 Q2 IMMUNOLOGY Pub Date : 2024-09-12 DOI: 10.1016/j.jmii.2024.08.014
Yi-An Way, Chong-Wei Huang, Wei-Chao Liao, Shiao-Wen Li, Ruei-Lin Chiang, En-Wei Hsing, Yi-Jiun Pan, Shian-Sen Shie, Yu-Chia Hsieh

Background: During the COVID-19 pandemic, there has been an increasing trend in healthcare-associated infections (HAIs) caused by carbapenem-resistant Acinetobacter baumannii (CRAB), posting a global public health concern. The heightened sensitivity of whole-genome sequencing (WGS) renders it an optimal and potent tool for monitoring outbreaks and tracing the transmission routes of nosocomial pathogens.

Method: We collected CRAB isolates from March 1, 2023, to April 6, 2023 in Chang Gung Memorial Hospital Lin Kou branch, a tertiary medical center in northern Taiwan. Any two or more isolates with the same identifiable capsular K-locus (KL) types were selected, and analyzed via WGS to identify putative transmission clusters, combined with epidemiologic and retrospective analysis on medical records to confirm risk factors and hidden transmission chains.

Result: A total of 48 non-redundant CRAB isolates were collected, belonging to ST2 of Pasteur MLST scheme and identifiable KL types of KL2, KL3, KL9, KL10, KL22, KL52. Excluding the KL types that was only found in 1 case, KL2 (n = 9, 22.5 %), KL3 (n = 24, 60 %), KL9 (n = 3, 7.5 %), and KL10 (n = 4, 10 %) were selected for further WGS analysis. Four distinct transmission clusters comprised of 2, 3, 10, and 23 cases were identified on a basis of phylogenetic status. 12 probable transmission chains were revealed, and 2 hidden transmission routes can be speculated.

Conclusion: This study referred to some hidden transmission chains that may be missed from traditional surveillance measures. Despite its low prevalence and high cost currently, implementing WGS could be a efficient, prompt, and unequivocal option for future MDRO infection control.

背景:在 COVID-19 大流行期间,耐碳青霉烯类鲍曼不动杆菌(CRAB)引起的医疗相关感染(HAIs)呈上升趋势,引起了全球公共卫生的关注。全基因组测序(WGS)的高灵敏度使其成为监测疫情和追踪鼻腔病原体传播途径的最佳有力工具:我们从 2023 年 3 月 1 日至 2023 年 4 月 6 日在台湾北部的三级医疗中心长庚纪念医院林口分院收集了 CRAB 分离物。我们选择了两个或两个以上具有相同可识别囊膜 K-病灶(KL)类型的分离株,并通过 WGS 进行分析,以确定潜在的传播集群,同时结合流行病学和病历回顾性分析,以确认风险因素和隐藏的传播链:结果:共收集到48个非冗余的CRAB分离株,属于巴斯德MLST方案的ST2,可识别的KL类型为KL2、KL3、KL9、KL10、KL22和KL52。除去仅在 1 个病例中发现的 KL 类型,KL2(n = 9,22.5%)、KL3(n = 24,60%)、KL9(n = 3,7.5%)和 KL10(n = 4,10%)被选中用于进一步的 WGS 分析。根据系统发育状况,确定了由 2、3、10 和 23 个病例组成的四个不同的传播集群。发现了 12 条可能的传播链,并推测出 2 条隐性传播途径:本研究发现了一些隐藏的传播链,这些传播链可能会被传统的监测措施所遗漏。尽管目前 WGS 的流行率较低且成本较高,但在未来的 MDRO 感染控制中,实施 WGS 可能是一种高效、迅速且明确的选择。
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Journal of Microbiology Immunology and Infection
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