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Viral Biology

Research Lines

Content with Investigacion Genética Bacteriana .

Bacterial Genetics

Our group has been studying for more than 30 years the mechanisms of antibiotic resistance in Streptococcus pneumoniae (Spn). Our objectives are to understand the molecular basis of antimicrobial action, to search for new targets of action and new compounds. Seconeolitsine (SCN) is one of these new compounds targeting topoisomerase I (Topo I). As for the search for new targets, our research has focused in recent years on the factors that organize the topology of the chromosome, allowing optimal compaction (about 1000-fold) to harmonize its replication, chromosome segregation and gene expression. This compaction is mediated both by the level of DNA supercoiling (Sc) and by association with nucleoid-binding proteins (NAPs). The level of Sc depends mainly on the enzymatic activities of their DNA topoisomerases, reaching a homeostatic equilibrium by the opposite activities of the topoisomerases that relax DNA (Topo I and Topo IV), and of gyrase, which introduces negative Sc. Our group has characterized the three Spn topoisomerases and two NAPs: HU and SatR. In addition, the availability of antimicrobials that inhibit each of the Spn topoisomerases has allowed us to analyze their transcriptome under conditions of local or global change of the Sc level and to define gene domains of coordinated transcription and similar functions. Fluoroquinolones, which inhibit Topo IV and gyrase, produce local changes in Sc that induce alterations in 6% of the transcriptome, altering metabolic pathways that originate an increase in reactive oxygen species (ROS) that contribute to lethality, in accordance with the general mechanism of bactericidal antibiotics. On the other hand, the induction of global changes in Sc by novobiocin (NOV, gyrase inhibitor), or by SCN (Topo I inhibitor), has allowed us to define topological domains. Global changes in Sc include the regulation of topoisomerase genes: its decrease activates the transcription of gyrase genes (gyrA, gyrB) and inhibits those of Topo IV (parEC) and Topo I (topA); the increase in Sc regulates the expression of topA. Decreased Sc affects 37% of the genome, with >68% of genes clustered in 15 domains. Increased Sc affects 10% of the genome, with 25% of the genes clustered in 12 domains. The AT content in the genome correlates with the domains, being higher in UP domains than in DOWN domains. The genes in the different domains have common functional characteristics, indicating that they have been subjected to topological selective pressure to determine the location of genes involved in metabolism, virulence and competition. 

The current objectives of the group are:
1.    Identification of factors that stabilize chromosome topology: NAPs, ncRNAs, intra-chromosomal interactions.
2.    Regulation of transcription in response to topological stress: in vivo localization of DNA topoisomerases, RNA polymerase and NAPs.
3.    Topo I as a new antimicrobial target and action of SCN. 
4.    Design of antisense RNAs and use of the CRISPR system as new antibacterial agents.

Research projects

Content with Investigacion Genética Bacteriana .

1) Project Title: Interaction Between DNA Supercoiling and Transcription in the Human Pathogen  Streptococcus pneumoniae

Principal Investigator:   Adela González de la Campa  
Funding Entity:   Ministry of Science and Innovation, State Research Agency (Call for "R&D&I Projects" 2020 – "Research Challenges" and "Knowledge Generation" Modalities).  
Reference:   PID2021-124738OB-100.  
Duration:   2022-2025.  
Funding Amount:   €108,900.
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2) Project Title:   Study of the Factors Organizing the Chromosome of  Streptococcus pneumoniae: New Antibiotic Targets and Resistance Mechanisms.

Principal Investigator:   Adela González de la Campa  
Funding Entity:   Ministry of Economy, Industry, and Competitiveness. State Research Agency.  
Reference:   BIO2017-82951-R.  
Duration:   2018-2020.  
Funding Amount:   €169,400.  

3) Project Title:   Role of DNA Topoisomerases and Nucleoid-Associated Proteins in the Chromosome Organization of  Streptococcus pneumoniae: Response to Antibiotics and Virulence.  

Principal Investigator:   Adela González de la Campa  
Funding Entity:   Ministry of Economy and Competitiveness. Secretariat of State for Research, Development, and Innovation.  
Reference:   BIO2014-55462.  
Duration:   2015-2017.  
Funding Amount:   €193,600.  

4) Project Title:   The Control of Supercoiling Level in  Streptococcus pneumoniae  as an Antimicrobial Target.  

Principal Investigator:   Adela González de la Campa  
Funding Entity:   Ministry of Economy and Competitiveness. Secretariat of State for Research, Development, and Innovation.  
Reference:   BIO2011-25343.  
Duration:   2012-2015.  
Funding Amount:   €209,000.  

5) Project Title:   Role of Small Non-Coding RNAs in the Pathogenicity of  Streptococcus pneumoniae.   

Principal Investigator:   Mónica Amblar Esteban  
Funding Entity:   Ministry of Economy and Competitiveness. Strategic Health Action (AES).  
Reference:   PI11/00656.  
Duration:   2012-2015.  
Funding Amount:   €198,714.
 

Publications

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Epidemiology and predictive factors for early and late mortality in Candida bloodstream infections: a population-based surveillance in Spain

11: Puig-Asensio M, Padilla B, Garnacho-Montero J, Zaragoza O, Aguado JM, Zaragoza R, Montejo M, Muñoz P, Ruiz-Camps I, Cuenca-Estrella M, Almirante B; CANDIPOP Project; GEIH-GEMICOMED (SEIMC); REIPI. Epidemiology and predictive factors for early and late mortality in Candida bloodstream infections: a population-based surveillance in Spain. Clin Microbiol Infect. 2014 Apr;20(4):O245-54.

PUBMED DOI

B Pérez de Val, B Romero, MT Tórtola, L Herrera-León, P Pozo, I Mercader, JL Sáez, M Domingo, E Vidal. Poly-resistant Mycobacterium bovis infection in a human and sympatric sheep, Spain, 2017-2018

B Pérez de Val, B Romero, MT Tórtola, L Herrera-León, P Pozo, I Mercader, JL Sáez, M Domingo, E Vidal. Poly-resistant Mycobacterium bovis infection in a human and sympatric sheep, Spain, 2017-2018. Emerg Infect Dis. 2021 Apr;27(4):1241-1243. doi: 10.3201/eid2704.204467. PMID: 33755008.

DOI

L Bernal-Martínez; L Herrera-Leon; C Valero; P de la Cruz; L Ghimpu; AC Mesa-Arango; G Santoni; L Goterris; R Millán; MJ Buitrago. Differential Diagnosis of Fungal Pneumonias vs.Tuberculosis in AIDS Patients by Using Two New Molecular Methods.

L Bernal-Martínez; L Herrera-Leon; C Valero; P de la Cruz; L Ghimpu; AC Mesa-Arango; G Santoni; L Goterris; R Millán; MJ Buitrago. Differential Diagnosis of Fungal Pneumonias vs.Tuberculosis in AIDS Patients by Using Two New Molecular Methods. J. Fungi 2021, 7, 336. doi.org: 10.3390/jof7050336. PMID: 33925404.

DOI

E Tagliani, RAnthony, TA Kohl, A de Neeling, V Nikolayevskyy, C Ködmön, FP Maurer, S Niemann, D van Soolingen, MJ van der Werf, D Cirillo, ECDC molecular surveillance project participants. Use of a whole genome sequencing-based approach for Mycobacterium tuberculosis surveillance in Europe in 2017-2019: an ECDC pilot study

E Tagliani, RAnthony, TA Kohl, A de Neeling, V Nikolayevskyy, C Ködmön, FP Maurer, S Niemann, D van Soolingen, MJ van der Werf, D Cirillo, ECDC molecular surveillance project participants. Use of a whole genome sequencing-based approach for Mycobacterium tuberculosis surveillance in Europe in 2017-2019: an ECDC pilot study. Eur Respir J. 2021 Jan 5;57(1):2002272. doi: 10.1183/13993003.02272-2020. Print 2021 Jan. PMID: 32732329.

DOI

MJ Iglesias, D Ibarz, A Cebollada, J Comín, MS Jiménez, MC Vázquez, S Samper, Spanish Working Group on MDRTB. The value of the continuous genotyping of multidrug resistant tuberculosis over 20 years in Spain.

MJ Iglesias, D Ibarz, A Cebollada, J Comín, MS Jiménez, MC Vázquez, S Samper, Spanish Working Group on MDRTB. The value of the continuous genotyping of multidrug resistant tuberculosis over 20 years in Spain. Sci Rep. 2020 Nov 24;10(1):20433. doi: 10.1038/s41598-020-77249-x. PMID: 33235225.

DOI

S Campos-Gutierrez, MJ Ramos-Real, R Abreu, MS Jimenez, M Lecuona. Pseudo-ourbreak of Mycobacterium fortuitum, in a hospital bronchoscopy unit.

S Campos-Gutierrez, MJ Ramos-Real, R Abreu, MS Jimenez, M Lecuona. Pseudo-ourbreak of Mycobacterium fortuitum, in a hospital bronchoscopy unit. Am J Infect Control. 2020 Jul;48(7):765-769. doi: 10.1016/j.ajic.2019.11.019. Epub 2019 Dec 24. PMID: 31882175.

DOI

Gascha , Y Meijeb, M Espasac, B Fonta, MS Jiménez, N Fernández-Hidalgo. Disseminated Infection Due to Mycobacterium chimaera After Aortic Valve Replacement.

Gascha , Y Meijeb, M Espasac, B Fonta, MS Jiménez, N Fernández-Hidalgo. Disseminated Infection Due to Mycobacterium chimaera After Aortic Valve Replacement. Revista Española de cardiología. 2019. Vol 72 (6):502-503. DOI: 10.1016/j.rec.2018.06.026. PMID: 30029979

DOI

PBMCs gene expression signature of advanced cirrhosis with high risk for clinically significant portal hypertension in HIV/HCV coinfected patients: A cross-control study

2. Salgüero S, Brochado-Kith O, Virseda Verdices A, Berenguer J, González-García J, Martínez I, Díez C, Hontañón V, Pérez-Latorre L, Fernández-Rodríguez A (‡), Jiménez-Sousa MA (‡,*), and Resino S (‡, *). PBMCs gene expression signature of advanced cirrhosis with high risk for clinically significant portal hypertension in HIV/HCV coinfected patients: A cross-control study. Biomed Pharmacother 2023, 159:114220. (A; FI= 7.42; D1, Pharmacology & Pharmacy; JCR 2021). PMID: 36628818. DOI: 10.1016/j.biopha.2023.114220.

PUBMED

List of staff

Additional Information

The research activity of the Viral Biology group since its beginnings in the 1980s has focused on respiratory viruses, especially on the study of the mechanisms of virus entry into the cell, evolutionary aspects, antigenic properties and vaccine development.

Currently, the group's objectives are focused on the characterisation of the immune response and the development of vaccines against human pneumoviruses: human respiratory syncytial virus (hRSV) and human metapneumovirus (hMPV).

Both viruses are considered to be important respiratory pathogens of high clinical relevance, especially in the paediatric population.

Safe and effective vaccines against these viruses are currently not available.  Soluble protein subunits based on the fusion protein (F-protein) of hRSV and hMPV are being developed in the laboratory by protein engineering for use as vaccines against human pneumoviruses.

On the other hand, and thanks to the characterisation of the type of humoral response induced by the F proteins of these viruses, the laboratory is also involved in the isolation of monoclonal antibodies and nanoantibodies for use as treatments against these viruses.

The research activity of the Viral Biology group since its beginnings in the 1980s has focused on respiratory viruses, especially on the study of the mechanisms of virus entry into the cell, evolutionary aspects, antigenic properties and vaccine development.

Currently, the group's objectives are focused on the characterisation of the immune response and the development of vaccines against human pneumoviruses: human respiratory syncytial virus (hRSV) and human metapneumovirus (hMPV).

Both viruses are considered to be important respiratory pathogens of high clinical relevance, especially in the paediatric population.

Safe and effective vaccines against these viruses are currently not available.  Soluble protein subunits based on the fusion protein (F-protein) of hRSV and hMPV are being developed in the laboratory by protein engineering for use as vaccines against human pneumoviruses.

On the other hand, and thanks to the characterisation of the type of humoral response induced by the F proteins of these viruses, the laboratory is also involved in the isolation of monoclonal antibodies and nanoantibodies for use as treatments against these viruses.

The current director of CNM is Dr. José Miguel Rubio Muñoz.

Dr. José Miguel Rubio has a degree in Biological Sciences from the Universidad Autónoma de Madrid (1986) and a PhD in Biological Sciences from the same university (1992). He carried out his doctoral thesis at the Department of Genetics of the Universidad Autónoma de Madrid, as Associate Professor (1988-1989), and at the School of Biology of the University of East Anglia in Norwich, UK, as Senior Research Assistant (1989-1992).

During his postdoctoral period he obtained a grant from the European Commission within the Human Capital and Mobility Program to be carried out at the University of “La Sapienza” in Rome, Italy and the Institute of Molecular Biology and Biotechnology in Crete, Greece (1993-1994). Subsequently, he made a further stay funded by the WHO and the university itself at the Department of Entomology, Wageningen University, The Netherlands (1994-1996).

Since 1997 he has been a member of the Instituto de Salud Carlos III (ISCIII), where he joined the Department of Parasitology of the National Center of Microbiology, as an EU-INCO postdoctoral fellow and later with a grant from the Autonomous Community of Madrid (CAM). She was part of the founding group of the National Center for Tropical Medicine (2003-2006) and of the 24/7 Alerts and Emergencies Unit (2006-2018) and is currently Head of the Malaria and Emerging Parasitosis Unit of the National Microbiology Center and is part, as research staff, of the Center for Biomedical Research Network on Infectious Diseases (CIBERINFEC/ISCIII).

During his scientific career he has been Visiting Scientist at the Leonidas e Marie Dean Center (FIOCRUZ-AMAZONAS, Manaus, Brazil) and is an External Consultant of the Parasitology Departments of Cairo University (Egypt) and the Medical Research Center (MRC) of Kuala Lumpur (Malaysia).  He also belongs or has belonged to different national and international committees:  Member of the expert group for malaria control of the European Centre for Disease Control (ECDC) since 2011; Expert-Evaluator for health programs of the European Commission since 2004; Spanish Representative (commissioned by ISCIII and MSC) in the Technical Scientific Committee of the TDR (WHO) 2007-2008; Spanish Deputy Focal Point for microbiology at the European Centre for Disease Control (ECDC) from 2012 to 2020; and, member of the Research Ethics Committee of ISCIII until 2019.

In this period he has published more than 100 articles in international indexed journals, 10 book chapters and has been co-editor of two books in the area of malaria, tropical medicine and neglected diseases. He has participated in 58 competitively funded research projects, 20 of them international, having been the principal investigator in 8 national and 11 international projects as PI of the project or WP leader. In addition, he has led five agreements with companies. Currently he has been awarded four sexenios of research, being presented this year 2025 to the fifth. In the teaching field, he participates in different postgraduate programs in the areas of microbiology and parasitology, having directed seven doctoral theses and more than 20 Master's or Degree final projects, both nationally and internationally. ​​​​​

El laboratorio de Referencia e Investigación en Resistencia a Antibióticos ofrece una amplia cartera de servicios al Sistema Nacional de Salud, las cuales pueden solicitarse en cnm-laboratorios.isciii.es. Jefe del Laboratorio: Jesús Oteo Iglesias (Punto focal Nacional de Resistencia antibiótica).

Dispone de dos programas de Vigilancia oficiales y gratuitos que engloban los ensayos ofertados ya sea como aislamientos individuales o mediante estudio de brotes. El Laboratorio utiliza asimismo técnicas de PCR en tiempo real para la detección de genes de resistencia, estas técnicas se han adaptado a un formato multiplex que permite detectar varios genes en la misma reacción. En los últimos años se han incluido metodologías basadas en la secuenciación de genomas completos para el análisis de bacterias multiresistentes (WGS).

Programa de vigilancia de Haemophilus influenzae. Responsables: María Pérez Vázquez (Punto focal Nacional de Haemophilus influenzae) y Belén Aracil. Laboratorio encargado de la identificación, estudio de sensibilidad y análisis genotípico de aislados de Haemophilus influenzae, centrándose esencialmente en la patología invasiva debida este patógeno. 

Programa de vigilancia de Resistencia a Antibióticos. Responsables: María Pérez Vázquez  y Belén Aracil (Punto focal Nacional de Resistencia antibiótica). Laboratorio encargado de la identificación, el estudio de sensibilidad antibiótica, y el diagnóstico fenotípico y genotípico de los diferentes mecanismos de resistencia a antibióticos fundamentalmente en enterobacterias y gram-negativos no fermentadores y Enterococcus spp.

Estudio de brotes. Responsables: Belén Aracil y María Pérez Vázquez. El programa incluye la caracterización de brotes nosocomiales y clones emergentes de alto riesgo mediante diferentes técnicas moleculares (tabla resumen). Éstas, nos permiten realizar estudios filogenéticos con el fin de obtener una información detallada acerca la relación entre los diferentes aislados y su trazabilidad. El objetivo final es generar datos que se transfieren a los hospitales como ayuda para la prevención o control de la propagación del brote.

Acreditación y Calidad. Responsable: Belén Aracil. El laboratorio Referencia e Investigación en Resistencia a Antibióticos ha sido de los primeros en el ISCIII en la utilización de técnicas acreditadas por la Entidad Nacional de Acreditaciones (ENAC). Este laboratorio consiguió la primera acreditación homologada de técnicas diagnósticas en 2012, programa que ha sido ampliado, de manera que en la actualidad más de la mitad de las técnicas ofrecidas al Sistema Nacional de Salud están debidamente acreditadas por ENAC.

Técnicos responsables de las técnicas realizadas en el Laboratorio: Noelia Lara Fuella y Verónica Bautista Sánchez.

En la siguiente imagen se resumen las técnicas ofrecidas al Sistema Nacional de Salud.

PROGRAMAS NOMBRE CARTERA SERVICIO PATÓGENO DETERMINACIÓN, DETECCIÓN, ANÁLISIS MÉTODOS

Programa de vigilancia de Haemophilus

Programa de vigilancia de resistencia a antibióticos.

Identificación bacteriana

Haemophilus sp.

Enterobacterias, gram-negativos no fermentadores, Enterococcus spp

Identificación bacteriana

Bioquímicos

MALDI TOF

Secuenciación de RNAr

Identificación capsular

Haemophilus influenzae

 

Identificación capsular fenotípica y genotípica

Aglutinación serológica en latex

PCR ind/multiplex

Determinación de Sensibilidad

Haemophilus sp.

Enterobacterias, gram-negativos no fermentadores, Enterococcus

 

Determinación de Sensibilidad

Microdilución                

Tiras epsilon               

Kirby Bauer

Métodos fenotípicos de detección de mecanismos de resistencia

Enterobacterias, gram-negativos no fermentadores,

 

Métodos fenotípicos de detección de mecanismos de resistencia

Discos y tabletas combinados con inhibidores                

Tiras combinadas     

Test de Hodge modificado

CabaNP                               

Inmunocromatografía CBP

Métodos genotípicos de detección de mecanismos de resistencia

Haemophilus sp.

Enterobacterias, gram-negativos no fermentadores, Enterococcus

 

ADN, PCR y secuenciación

PCR ind/multiplex

Análisis comparativo de las secuencias

Tipificación molecular/análisis filogenéticos

Haemophilus sp.

Enterobacterias, gram-negativos no fermentadores, Enterococcus

 

Corte enzimas de restricción, electroforesis

ADN, PCR y secuenciación

Preparación de librerías y secuenciación y análisis de genomas completos

 

PFGE

 

MLST

 

WGS