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Bacterial Genetics

Research Lines

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Research

The Molecular Virology group focuses its research on the study of HIV-1 genetic variation and viral evolution using both in vitro and ex vivo approaches, structured around the following research lines:

- Non-progressor patients. These patients maintain control of the disease in the absence of antiretroviral therapy and have therefore been proposed as a model of functional cure. Our objective is to study the contribution of viral factors to disease control through biological characterization and analysis of viral evolution in individuals with undetectable viral loads (elite controllers, EC), compared with individuals showing other patterns of viral control.

- Viral envelope. This viral protein is key in determining viral fitness. Therefore, its functionality significantly affects infection progression. In collaboration with Dr. Blanco and Dr. Valenzuela, we study which specific events (CD4 binding, fusogenicity, etc.) are associated with envelope functionality. To this end, we have analyzed envelopes from individuals with different patterns of disease progression. Some of these have been contributed to the AIDS Research Network envelope biobank for broader use.

- Dual infection. Infection with more than one viral variant (either through co-infection or superinfection) may have consequences for infection pathogenesis. Within our group, different aspects of DI have been analyzed, including its detection in non-progressor patients, its prevalence and incidence in Spain, and its influence on the neutralizing antibody response.

- Molecular Epidemiology. The group has analyzed viral evolution throughout the epidemic in Spain and in other countries (the Netherlands, Italy, Germany, Uruguay, Panama, Brazil, etc.).

- Role of amino acid residues in reverse transcriptase. We study the role of specific amino acid residues in HIV-1 reverse transcriptase in enzymatic function and replication capacity using an infectious molecular clone previously obtained by the group.

- “In vitro” variability. Serial passage studies have been used to detect the mechanisms responsible for the gain or loss of viral fitness.

- Antiviral studies. We have analyzed the selection of resistance mutations in vitro against different antivirals, as well as the effect of these mutations on viral fitness, and the activity of new antivirals such as ATR inhibitors.

 

Virological Diagnosis and Reference in HIV and HTLV Infections

The research group provides diagnostic and reference activities through the service portfolio of the National Center for Microbiology to the entire Spanish National Health System.

These services include:

  • Diagnosis and reference of HIV infection (types 1 and 2) through detection of specific antibodies and detection of proviral DNA by PCR.

  • Diagnosis and reference of HTLV-I/II infection through detection of specific antibodies and detection of proviral DNA by PCR. Quantification of HTLV-1 proviral load by real-time PCR.

European Union Reference Laboratory (EURL) in the field of in vitro diagnostic medical devices for microbiological diagnosis (IVD) of HIV and HTLV (Regulation 2023/2713 of December 5th, 2023). Our role is to confirm the reliability and effectiveness of devices for detecting these pathogens and to ensure their specific performance requirements through laboratory testing before they can be marketed within the European Union.

Research projects

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- Towards a functional cure: Implications of early antiretroviral therapy and hormonal changes on the HIV reservoir in perinatally infected adolescents. Health Research Fund (FIS) – Carlos III Health Institute (01/01/2026 – 31/12/2028). €72,000. PI: María Pernas, Concepción Casado.

- Determination of factors associated with protection against Human Immunodeficiency Virus type 1 reinfection: Identification of correlates of protection. 9th Gilead Fellowship Program for Biomedical Research, Gilead Sciences, S.L. (01/07/2023 – 30/06/2025). €16,330. PI: María Pernas.

- Impact of the envelope on HIV viral replication: New avenues for vaccine development. Health Research Fund (FIS) – Carlos III Health Institute (01/01/2020 – 31/12/2023). €53,000. PI: María Pernas, Concepción Casado.

- Study of HIV-1 virulence in recently infected patients and its contribution, together with clinical and epidemiological factors, to disease progression. Ministry of Economy and Competitiveness. State Program for Scientific and Technical Research and Innovation (30/12/2016 – 30/06/2021). €145,000. PI: Concepción Casado, Cecilio López-Galíndez.

-Contribution of HIV-1 dual infection to virological and clinical evolution in homo/bisexual men. Health Research Fund (FIS) – Carlos III Health Institute (01/01/2014 – 31/01/2016). €74,410. PI: Cecilio López-Galíndez.

- Characterization of non-pathogenic HIV variants obtained “ex vivo” and “in vitro” for the study of disease pathogenesis. Ministry of Science and Innovation (01/01/2011 – 31/01/2014). €169,400. PI: Cecilio López-Galíndez.

- Spanish AIDS Research Network (RIS-RETIC). Carlos III Health Institute (02/01/2017 – 02/01/2022). €195,212. PI: Cecilio López-Galíndez, Concepción Casado.

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Publications

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Leon KE, Schubert RD, Casas-Alba D, Hawes IA, Ramachandran PS, Ramesh A, Pak JE, Wu W, Cheung CK, Crawford ED, Khan LM, Launes C, Sample HA, Zorn KC, Cabrerizo M, Valero-Rello A, Langelier C, Muñoz-Almagro C, DeRisi JL, Wilson MR. Genomic and serologic characterization of enterovirus A71 brainstem encephalitis. Neurol Neuroimmunol Neuroinflamm. 2020

Leon KE, Schubert RD, Casas-Alba D, Hawes IA, Ramachandran PS, Ramesh A, Pak JE, Wu W, Cheung CK, Crawford ED, Khan LM, Launes C, Sample HA, Zorn KC, Cabrerizo M, Valero-Rello A, Langelier C, Muñoz-Almagro C, DeRisi JL, Wilson MR. Genomic and serologic characterization of enterovirus A71 brainstem encephalitis. Neurol Neuroimmunol Neuroinflamm. 2020 Mar 5;7(3):e703. doi: 10.1212/NXI.0000000000000703. PMID: 32139440; PMCID: PMC7136061.

González-Sanz R, Casas-Alba D, Launes C, Muñoz-Almagro C, Ruiz-García MM, Alonso M, González-Abad MJ, Megías G, Rabella N, Del Cuerpo M, Gozalo-Margüello M, González-Praetorius A, Martínez-Sapiña A, Goyanes-Galán MJ, Romero MP, Calvo C, Antón A, Imaz M, Aranzamendi M, Hernández-Rodríguez Á, Moreno-Docón A, Rey- Cao S, Navascués A, Otero A, Cabrerizo M. Molecular epidemiology of an enterovirus A71 outbreak associated with severe neurological disease, Spain, 2016. Euro Surveill. 2019

González-Sanz R, Casas-Alba D, Launes C, Muñoz-Almagro C, Ruiz-García MM, Alonso M, González-Abad MJ, Megías G, Rabella N, Del Cuerpo M, Gozalo-Margüello M, González-Praetorius A, Martínez-Sapiña A, Goyanes-Galán MJ, Romero MP, Calvo C, Antón A, Imaz M, Aranzamendi M, Hernández-Rodríguez Á, Moreno-Docón A, Rey- Cao S, Navascués A, Otero A, Cabrerizo M. Molecular epidemiology of an enterovirus A71 outbreak associated with severe neurological disease, Spain, 2016. Euro Surveill. 2019 Feb;24(7):1800089. doi: 10.2807/1560-7917.ES.2019.24.7.1800089. PMID: 30782267; PMCID: PMC6381658.

Spanish Afp Surveillance Working Group. Acute flaccid paralysis (AFP) surveillance: challenges and opportunities from 18 years' experience, Spain, 1998 to 2015. Euro Surveill.

Spanish Afp Surveillance Working Group. Acute flaccid paralysis (AFP) surveillance: challenges and opportunities from 18 years' experience, Spain, 1998 to 2015. Euro Surveill. 2018 Nov;23(47):1700423. doi: 10.2807/1560-7917.ES.2018.23.47.1700423. PMID: 30482263; PMCID: PMC6341937.

Molecular Epidemiology of Human Parechoviruses in Children With Acute Respiratory Infection in Spain

M Cabrerizo*, C Calvo, G Trallero, ML García-García, M Arroyas, V Sánchez, F Pozo, I Casas. Molecular epidemiology of human parechoviruses children with acute respiratory infection in Spain. Pediatric Infect Dis J 32:802-3 (2013).

PUBMED DOI

Identification of novel Betaherpesviruses in iberian bats reveals parallel evolution

Pozo F, Juste J, Vázquez-Morón S., Aznar-López C, Ibáñez C, Garin I, Aihartza J, Casa I, Tenorio A, Echevarría JE. Identification of novel Betaherpesviruses in iberian bats reveals parallel evolution. PLoS ONE. 2016. 11(12): e0169153. doi:10.1371/journal.pone.0169153

PUBMED DOI

Detection of Rhabdovirus viral RNA in oropharyngeal swabs and ectoparasites of Spanish bats

Aznar C, Vazquez-Moron S, Martson D, Juste J, Ibáñez C, Berciano JM, Salsamendi E, Aihartza J, Banyard AC, McElhinney L, Fooks AR, Echevarria JE. Detection of Rhabdovirus viral RNA in oropharyngeal swabs and ectoparasites of Spanish bats. Journal of General Virology. 2013. 94: 69-75.

PUBMED DOI

Genomic non-coding regions reveal hidden patterns of mumps virus circulation in Spain, 2005 to 2015

Gavilán AM, Fernández-García A*, Rueda A, Castellanos A, Masa J, López-Perea N, Torres de Mier MV, de Ory F, Echevarría JE. Non-coding sequences reveal hidden patterns of mumps virus circulation in Spain, 2005 to 2015. Eurosurveillance,2018, 23(15): 1-8. *Corresponding author.

PUBMED DOI

First cases of European Bat Lyssavirus type 1 in Iberian serotine bats: implications for the molecular epidemiology of bat rabies in Europe.

Mingo-Casas P, Sandonís V, Obón E, Berciano JM, Vázquez-Morón S, Juste J, Echevarría JE. First cases of European Bat Lyssavirus type 1 in Iberian serotine bats: implications for the molecular epidemiology of bat rabies in Europe. Plos Neglected Tropical Diseases, 2018: 12(4): e0006290.

PUBMED DOI

Last cases of rubella and congenital rubella syndrome in Spain, 1997–2016: The success of a vaccination program

Seppälä EM, López-Perea N, Torres de Mier MV, Echevarría JE, Fernández García A, Masa-Calles J. Last cases of rubella and congenital rubella syndrome in Spain, 1997–2016: The success of a vaccination program. Vaccine, 2019, 37(1):169-175.

PUBMED DOI

Combination of Cefditoren and N-acetyl-l-Cysteine Shows a Synergistic Effect against Multidrug-Resistant Streptococcus pneumoniae Biofilms

Llamosí M, Sempere J, Coronel P, Gimeno M, Yuste J, Domenech M. Combination of Cefditoren and N-acetyl-l-Cysteine Shows a Synergistic Effect against Multidrug-Resistant Streptococcus pneumoniae Biofilms. Microbiol Spectr. 2022 Dec 21;10(6):e0341522

PUBMED DOI

Clearance of mixed biofilms of Streptococcus pneumoniae and methicillin-susceptible/resistant Staphylococcus aureus by antioxidants N-acetyl-L-cysteine and cysteamine

Sempere J, Llamosí M, Román F, Lago D, González-Camacho F, Pérez-García C, Yuste J, Domenech M. Clearance of mixed biofilms of Streptococcus pneumoniae and methicillin-susceptible/resistant Staphylococcus aureus by antioxidants N-acetyl-L-cysteine and cysteamine. Sci Rep. 2022 Apr 23;12(1):6668

PUBMED DOI

Clinical Relevance and Molecular Pathogenesis of the Emerging Serotypes 22F and 33F of Streptococcus pneumoniae in Spain

Sempere J, de Miguel S, González-Camacho F, Yuste J, Domenech M. Clinical Relevance and Molecular Pathogenesis of the Emerging Serotypes 22F and 33F of Streptococcus pneumoniae in Spain. Front Microbiol. 2020 Feb 27;11:309.

PUBMED DOI

Combination of Antibodies and Antibiotics as a Promising Strategy Against Multidrug-Resistant Pathogens of the Respiratory Tract

Domenech M, Sempere J, de Miguel S, Yuste J. Combination of Antibodies and Antibiotics as a Promising Strategy Against Multidrug-Resistant Pathogens of the Respiratory Tract. Front Immunol. 2018 Nov 20;9:2700. doi: 10.3389/fimmu.2018.02700. PMID: 30515172; PMCID: PMC6256034.

DOI

Chemotherapy with Phage Lysins Reduces Pneumococcal Colonization of the Respiratory Tract

Corsini B, Díez-Martínez R, Aguinagalde L, González-Camacho F, García-Fernández E, Letrado P, García P, Yuste J. Chemotherapy with Phage Lysins Reduces Pneumococcal Colonization of the Respiratory Tract. Antimicrob Agents Chemother. 2018 May 25;62(6):e02212-17. doi: 10.1128/AAC.02212-17. PMID: 29581113; PMCID: PMC5971604.

DOI

Impact of Biological Therapies on the Immune Response after Pneumococcal Vaccination in Patients with Autoimmune Inflammatory Diseases

Richi P, Yuste J, Navío T, González-Hombrado L, Salido M, Thuissard-Vasallo I, Jiménez-Díaz A, Llorente J, Cebrián L, Lojo L, Steiner M, Cobo T, Martín MD, García-Castro M, Castro P, Muñoz-Fernández S. Impact of Biological Therapies on the Immune Response after Pneumococcal Vaccination in Patients with Autoimmune Inflammatory Diseases. Vaccines. 2021 Feb 28;9(3):203. doi: 10.3390/vaccines9030203. PMID: 33671007; PMCID: PMC7997274.

DOI

Pleiotropic Effects of Cell Wall Amidase LytA on Streptococcus pneumoniae Sensitivity to the Host Immune Response

Ramos-Sevillano E, Urzainqui A, Campuzano S, Moscoso M, González-Camacho F, Domenech M, Rodríguez de Córdoba S, Sánchez-Madrid F, Brown JS, García E, Yuste J. Pleiotropic effects of cell wall amidase LytA on Streptococcus pneumoniae sensitivity to the host immune response. Infect Immun. 2015 Feb;83(2):591-603. doi: 10.1128/IAI.02811-14. PMID: 25404032; PMCID: PMC4294232.

DOI

PSGL-1 on Leukocytes is a Critical Component of the Host Immune Response against Invasive Pneumococcal Disease

Ramos-Sevillano E, Urzainqui A, de Andrés B, González-Tajuelo R, Domenech M, González-Camacho F, Sánchez-Madrid F, Brown JS, García E, Yuste J. PSGL-1 on Leukocytes is a Critical Component of the Host Immune Response against Invasive Pneumococcal Disease. PLoS Pathog. 2016 Mar 14;12(3):e1005500. doi: 10.1371/journal.ppat.1005500. PMID: 26975045; PMCID: PMC4790886.

DOI

Comparison of methods and characterization of small RNAs from plasma extracellular vesicles of HIV/HCV coinfected patients

Martínez-González E; Brochado-Kith O; Gómez-Sanz A; et al; Fernández-Rodríguez A (AC). (9/9). 2020. Small RNA sequencing from plasma extracellular vesicles of HIV/HCV coinfected patients: a protocol comparison SCIENTIFIC REPORTS. 9. ISSN 2045-2322.

DOI

Relative telomere length impact on mortality of COVID-19: Sex differences

Virseda-Berdices A; Concostrina-Martinez L; Martínez-González O;et al; Fernández-Rodríguez A (AC). (14/14). 2022. Relative telomere length impact on mortality of COVID-19: Sex differences.Journal of medical virology. 95, pp.e28368. ISSN 0146-6615.

DOI

Hepatitis C Virus Influences HIV-1 Viral Splicing in Coinfected Patients

Martínez-Román P; López-Huertas MR; Crespo-Bermejo C; et al; Briz V (AC). (16/15). 2020. Hepatitis C virus influences HIV-1 viral splicing in coinfected patients JOURNAL OF CLINICAL MEDICINE. MDPI. ISSN 2077-0383.

DOI

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Additional Information

Streptococcus pneumoniae is a human pathogen that, despite the development of vaccines, continues to be an important cause of mortality and morbidity. We investigate the mechanisms of antibiotic resistance in this bacterium. On the one hand by identifying new therapeutic targets and on the other hand by investigating the molecular basis of the action of antibiotics already used in clinical practice (the fluoroquinolones levofloxacin and moxifloxacin) or not yet used (seconeolitsine). For this purpose, we used a multidisciplinary analysis involving genomics, transcriptomics and proteomics to understand the organization of the S. pneumoniae chromosome and the identification of the factors that stabilize this organization, including ncRNAs. Changes in the level of global supercoiling, either by inhibition of gyrase (decrease) or by inhibition of topoisomerase I (increase) alter the transcriptome. The modulated genes are located in domains, whose genes show specific functional characteristics. The aim is to identify new factors essential for S. pneumoniae physiology and to characterize transcriptional regulation in response to topological stress. In addition, RNA interference technology and CRISPR systems will be used as novel antibacterials. These studies will establish the bases for translational research aimed at the development of new therapeutic targets for the treatment of pneumococcal diseases.

Streptococcus pneumoniae is a human pathogen that, despite the development of vaccines, continues to be an important cause of mortality and morbidity. We investigate the mechanisms of antibiotic resistance in this bacterium. On the one hand by identifying new therapeutic targets and on the other hand by investigating the molecular basis of the action of antibiotics already used in clinical practice (the fluoroquinolones levofloxacin and moxifloxacin) or not yet used (seconeolitsine). For this purpose, we used a multidisciplinary analysis involving genomics, transcriptomics and proteomics to understand the organization of the S. pneumoniae chromosome and the identification of the factors that stabilize this organization, including ncRNAs. Changes in the level of global supercoiling, either by inhibition of gyrase (decrease) or by inhibition of topoisomerase I (increase) alter the transcriptome. The modulated genes are located in domains, whose genes show specific functional characteristics. The aim is to identify new factors essential for S. pneumoniae physiology and to characterize transcriptional regulation in response to topological stress. In addition, RNA interference technology and CRISPR systems will be used as novel antibacterials. These studies will establish the bases for translational research aimed at the development of new therapeutic targets for the treatment of pneumococcal diseases.

Content with Investigacion Virología Molecular .