Xingyan Ma, Dendi K. Nugraha, Yukihiro Hiramatsu, Yasuhiko Horiguchi
The Gram-negative pathogenic bacterium Bordetella bronchiseptica is a respiratory pathogen closely related to Bordetella pertussis, the causative agent of whooping cough. Despite sharing homologous virulence factors, B. bronchiseptica infects a broad range of mammalian hosts, including some experimental animals, whereas B. pertussis is strictly adapted to humans. Therefore, B. bronchiseptica is often used as a representative model to explore the pathogenicity of Bordetella in infection experiments with laboratory animals. Although Bordetella virulence factors, including toxins and adhesins have been studied well, our recent study implied that unknown virulence factors are involved in tracheal colonization and infection. Here, we investigated bacterial genes contributing to tracheal colonization by high-throughput transposon sequencing (Tn-seq). After the screening, we picked up 151 candidate genes of various functions and found that a rpoN-deficient mutant strain was defective in tracheal colonization when co-inoculated with the wild-type strain. rpoN encodes σ54, a sigma factor that regulates the transcription of various genes, implying its contribution to various bacterial activities. In fact, we found RpoN of B. bronchiseptica is involved in bacterial motility and initial biofilm formation. From these results, we propose that RpoN supports bacterial colonization by regulating various bacteriological functions.
{"title":"RpoN (sigma factor 54) contributes to bacterial fitness during tracheal colonization of Bordetella bronchiseptica","authors":"Xingyan Ma, Dendi K. Nugraha, Yukihiro Hiramatsu, Yasuhiko Horiguchi","doi":"10.1111/1348-0421.13109","DOIUrl":"10.1111/1348-0421.13109","url":null,"abstract":"<p>The Gram-negative pathogenic bacterium <i>Bordetella bronchiseptica</i> is a respiratory pathogen closely related to <i>Bordetella pertussis</i>, the causative agent of whooping cough. Despite sharing homologous virulence factors, <i>B. bronchiseptica</i> infects a broad range of mammalian hosts, including some experimental animals, whereas <i>B. pertussis</i> is strictly adapted to humans. Therefore, <i>B. bronchiseptica</i> is often used as a representative model to explore the pathogenicity of <i>Bordetella</i> in infection experiments with laboratory animals. Although <i>Bordetella</i> virulence factors, including toxins and adhesins have been studied well, our recent study implied that unknown virulence factors are involved in tracheal colonization and infection. Here, we investigated bacterial genes contributing to tracheal colonization by high-throughput transposon sequencing (Tn-seq). After the screening, we picked up 151 candidate genes of various functions and found that a <i>rpoN</i>-deficient mutant strain was defective in tracheal colonization when co-inoculated with the wild-type strain. <i>rpoN</i> encodes σ<sup>54</sup>, a sigma factor that regulates the transcription of various genes, implying its contribution to various bacterial activities. In fact, we found RpoN of <i>B. bronchiseptica</i> is involved in bacterial motility and initial biofilm formation. From these results, we propose that RpoN supports bacterial colonization by regulating various bacteriological functions.</p>","PeriodicalId":18679,"journal":{"name":"Microbiology and Immunology","volume":"68 2","pages":"36-46"},"PeriodicalIF":2.6,"publicationDate":"2023-12-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/1348-0421.13109","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"138714334","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}
Mako Yamaguchi, Yohana S. Mtali, Hitomi Sonokawa, Ken Takashima, Yoshimi Fukushima, Takahisa Kouwaki, Hiroyuki Oshiumi
Cervical cancer is caused mostly by human papillomavirus (HPV), and several HPV vaccines have been developed to prevent its onset. Vaccines include antigens as well as adjuvants, with adjuvants playing an important role in activating the innate immune responses necessary for inducing adaptive immunological responses. Recent research has shown the presence of trained immunity inside the innate immune system. However, trained immunity conferred by HPV vaccinations is not well understood. In this work, we explored the innate immune responses and trained immunity caused by two HPV vaccines, Cervarix and Gardasil. Cervarix includes monophosphoryl lipid A and an aluminum adjuvant, and it significantly increased the expression of IL-6 and IFN-β mRNAs in RAW264.7 cells. On the contrary, Gardasil, which only includes an aluminum adjuvant, exhibited little cytokine expression but increased the expression of TLRs. Furthermore, Cervarix significantly increased IL-1β secretion from mouse macrophages, while Gardasil only mildly induced IL-1β secretion. Interestingly, initial stimulation with Gardasil enhanced the expression of IL-6 and TNF-α mRNAs upon secondary stimulation with TLR ligands, indicating that Gardasil induced trained immunity in macrophages. Moreover, Gardasil injection into mice resulted in enhanced TNF-α production in sera following secondary TLR stimulation. Our findings suggest that HPV vaccinations have the ability to induce trained immunity that modulate TLR ligand responses.
{"title":"HPV vaccines induce trained immunity and modulate pro-inflammatory cytokine expression in response to secondary Toll-like receptor stimulations","authors":"Mako Yamaguchi, Yohana S. Mtali, Hitomi Sonokawa, Ken Takashima, Yoshimi Fukushima, Takahisa Kouwaki, Hiroyuki Oshiumi","doi":"10.1111/1348-0421.13108","DOIUrl":"10.1111/1348-0421.13108","url":null,"abstract":"<p>Cervical cancer is caused mostly by human papillomavirus (HPV), and several HPV vaccines have been developed to prevent its onset. Vaccines include antigens as well as adjuvants, with adjuvants playing an important role in activating the innate immune responses necessary for inducing adaptive immunological responses. Recent research has shown the presence of trained immunity inside the innate immune system. However, trained immunity conferred by HPV vaccinations is not well understood. In this work, we explored the innate immune responses and trained immunity caused by two HPV vaccines, Cervarix and Gardasil. Cervarix includes monophosphoryl lipid A and an aluminum adjuvant, and it significantly increased the expression of IL-6 and IFN-β mRNAs in RAW264.7 cells. On the contrary, Gardasil, which only includes an aluminum adjuvant, exhibited little cytokine expression but increased the expression of TLRs. Furthermore, Cervarix significantly increased IL-1β secretion from mouse macrophages, while Gardasil only mildly induced IL-1β secretion. Interestingly, initial stimulation with Gardasil enhanced the expression of IL-6 and TNF-α mRNAs upon secondary stimulation with TLR ligands, indicating that Gardasil induced trained immunity in macrophages. Moreover, Gardasil injection into mice resulted in enhanced TNF-α production in sera following secondary TLR stimulation. Our findings suggest that HPV vaccinations have the ability to induce trained immunity that modulate TLR ligand responses.</p>","PeriodicalId":18679,"journal":{"name":"Microbiology and Immunology","volume":"68 2","pages":"65-74"},"PeriodicalIF":2.6,"publicationDate":"2023-12-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"138714561","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Yoshiki Kawamura, Satoshi Komoto, Saori Fukuda, Masanori Kugita, Shuang Tang, Amita Patel, Julianna R. Pieknik, Shizuko Nagao, Koki Taniguchi, Philip R. Krause, Tetsushi Yoshikawa
Vaccine development for herpes simplex virus 2 (HSV-2) has been attempted, but no vaccines are yet available. A plasmid-based reverse genetics system for Rotavirus (RV), which can cause gastroenteritis, allows the generation of recombinant RV containing foreign genes. In this study, we sought to develop simian RV (SA11) as a vector to express HSV-2 glycoprotein D (gD2) and evaluated its immunogenicity in mice. We generated the recombinant SA11-gD2 virus (rSA11-gD2) and confirmed its ability to express gD2 in vitro. The virus was orally inoculated into suckling BALB/c mice and into 8-week-old mice. Serum IgG and IgA titers against RV and gD2 were measured by ELISA. In the 8-week-old mice inoculated with rSA11-gD2, significant increases in not only antibodies against RV but also IgG against gD2 were demonstrated. In the suckling mice, antibodies against RV were induced, but gD2 antibody was not detected. Diarrhea observed after the first inoculation of rSA11-gD2 in suckling mice was similar to that induced by the parent virus. A gD2 expressing simian RV recombinant, which was orally inoculated, induced IgG against gD2. This strategy holds possibility for genital herpes vaccine development.
{"title":"Development of recombinant rotavirus carrying herpes simplex virus 2 glycoprotein D gene based on reverse genetics technology","authors":"Yoshiki Kawamura, Satoshi Komoto, Saori Fukuda, Masanori Kugita, Shuang Tang, Amita Patel, Julianna R. Pieknik, Shizuko Nagao, Koki Taniguchi, Philip R. Krause, Tetsushi Yoshikawa","doi":"10.1111/1348-0421.13107","DOIUrl":"10.1111/1348-0421.13107","url":null,"abstract":"<p>Vaccine development for herpes simplex virus 2 (HSV-2) has been attempted, but no vaccines are yet available. A plasmid-based reverse genetics system for Rotavirus (RV), which can cause gastroenteritis, allows the generation of recombinant RV containing foreign genes. In this study, we sought to develop simian RV (SA11) as a vector to express HSV-2 glycoprotein D (gD2) and evaluated its immunogenicity in mice. We generated the recombinant SA11-gD2 virus (rSA11-gD2) and confirmed its ability to express gD2 in vitro. The virus was orally inoculated into suckling BALB/c mice and into 8-week-old mice. Serum IgG and IgA titers against RV and gD2 were measured by ELISA. In the 8-week-old mice inoculated with rSA11-gD2, significant increases in not only antibodies against RV but also IgG against gD2 were demonstrated. In the suckling mice, antibodies against RV were induced, but gD2 antibody was not detected. Diarrhea observed after the first inoculation of rSA11-gD2 in suckling mice was similar to that induced by the parent virus. A gD2 expressing simian RV recombinant, which was orally inoculated, induced IgG against gD2. This strategy holds possibility for genital herpes vaccine development.</p>","PeriodicalId":18679,"journal":{"name":"Microbiology and Immunology","volume":"68 2","pages":"56-64"},"PeriodicalIF":2.6,"publicationDate":"2023-12-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"138692892","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
A.-G. Leroy, J. Caillon, A. Broquet, V. Lemabecque, S. Delanou, N. Caroff, K. Asehnoune, A. Roquilly, L. Crémet
Pseudomonas aeruginosa (PA) remains one of the leading causes of nosocomial acute pneumonia. The array of virulence factors expressed by PA and the intense immune response associated with PA pneumonia play a major role in the severity of these infections. New therapeutic approaches are needed to overcome the high resistance of PA to antibiotics and to reduce the direct damage to host tissues. Through its immunomodulatory and anti-virulence effects, azithromycin (AZM) has demonstrated clinical benefits in patients with chronic PA respiratory infections. However, there is relatively little evidence in PA acute pneumonia. We investigated the effects of AZM, as an adjunctive therapy combined with ceftazidime (CAZ), in a murine model of PA acute pneumonia. We observed that the combined therapy (i) reduces the weight loss of mice 24 h post-infection (hpi), (ii) decreases neutrophil influx into the lungs at 6 and 24 hpi, while this effect is absent in a LPS-induced pneumonia or when PA is pretreated with antibiotics and mice do not receive any antibiotics, and that (iii) AZM, alone or with CAZ, modulates the expression of PA quorum sensing regulators and virulence factors (LasI, LasA, PqsE, PhzM, ExoS). Our findings support beneficial effects of AZM with CAZ on PA acute pneumonia by both bacterial virulence and immune response modulations. Further investigations are needed to clarify the exact underlying mechanisms responsible for the reduction of the neutrophils influx and to better discriminate between direct immunomodulatory properties of AZM, and indirect effects on neutrophilia resulting from bacterial virulence modulation.
铜绿假单胞菌(Pseudomonas aeruginosa,PA)仍然是引起医院内急性肺炎的主要原因之一。铜绿假单胞菌表达的一系列毒力因子以及与铜绿假单胞菌肺炎相关的强烈免疫反应对这些感染的严重程度起着重要作用。需要新的治疗方法来克服 PA 对抗生素的高度耐药性,并减少对宿主组织的直接损害。通过免疫调节和抗病毒作用,阿奇霉素(AZM)已在慢性 PA 呼吸道感染患者中显示出临床疗效。然而,有关 PA 急性肺炎的证据相对较少。我们在 PA 急性肺炎小鼠模型中研究了 AZM 与头孢他啶(CAZ)联合辅助治疗的效果。我们观察到,联合疗法(i)减少了小鼠感染后 24 小时(hpi)的体重下降,(ii)减少了中性粒细胞在感染后 6 小时和 24 小时涌入肺部、(iii)AZM单独或与CAZ一起可调节PA法定量感应调节因子和毒力因子(LasI、LasA、PqsE、PhzM、ExoS)的表达。我们的研究结果表明,AZM 联合 CAZ 可通过调节细菌毒力和免疫反应对 PA 急性肺炎产生有益影响。还需要进一步研究,以明确减少中性粒细胞流入的确切机制,并更好地区分 AZM 的直接免疫调节特性和细菌毒力调节对中性粒细胞增多的间接影响。
{"title":"Azithromycin regulates bacterial virulence and immune response in a murine model of ceftazidime-treated Pseudomonas aeruginosa acute pneumonia","authors":"A.-G. Leroy, J. Caillon, A. Broquet, V. Lemabecque, S. Delanou, N. Caroff, K. Asehnoune, A. Roquilly, L. Crémet","doi":"10.1111/1348-0421.13106","DOIUrl":"10.1111/1348-0421.13106","url":null,"abstract":"<p><i>Pseudomonas aeruginosa</i> (PA) remains one of the leading causes of nosocomial acute pneumonia. The array of virulence factors expressed by PA and the intense immune response associated with PA pneumonia play a major role in the severity of these infections. New therapeutic approaches are needed to overcome the high resistance of PA to antibiotics and to reduce the direct damage to host tissues. Through its immunomodulatory and anti-virulence effects, azithromycin (AZM) has demonstrated clinical benefits in patients with chronic PA respiratory infections. However, there is relatively little evidence in PA acute pneumonia. We investigated the effects of AZM, as an adjunctive therapy combined with ceftazidime (CAZ), in a murine model of PA acute pneumonia. We observed that the combined therapy (i) reduces the weight loss of mice 24 h post-infection (hpi), (ii) decreases neutrophil influx into the lungs at 6 and 24 hpi, while this effect is absent in a LPS-induced pneumonia or when PA is pretreated with antibiotics and mice do not receive any antibiotics, and that (iii) AZM, alone or with CAZ, modulates the expression of PA <i>quorum sensing</i> regulators and virulence factors (LasI, LasA, PqsE, PhzM, ExoS). Our findings support beneficial effects of AZM with CAZ on PA acute pneumonia by both bacterial virulence and immune response modulations. Further investigations are needed to clarify the exact underlying mechanisms responsible for the reduction of the neutrophils influx and to better discriminate between direct immunomodulatory properties of AZM, and indirect effects on neutrophilia resulting from bacterial virulence modulation.</p>","PeriodicalId":18679,"journal":{"name":"Microbiology and Immunology","volume":"68 2","pages":"27-35"},"PeriodicalIF":2.6,"publicationDate":"2023-12-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"138567237","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Cover photograph: illustrates the analysis of the known 2VSM.PDB complex, which consists of the G protein and Ephrin B2 receptor. The stability of the complex was evaluated using the covariance of the residue index. Microbiol Immunol: 67:501-513. Article link here