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Pneumococcus Reference Laboratory (LRN)

Specialized Group:

Pneumococcus Unit. Reference Laboratory and Research in Bacterial Diseases Preventable by Vaccines
National Center of Microbiology. Carlos III Health Institute

Jose Yuste, Fernando Gonzalez-Camacho, and Mirian Domenech

 

Laboratory members (from left to right): José Yuste, Beatriz López,
Mirella Llamosí, Idoia del Río, Fernando González, Julio Sempere, Mirian Domenech.

 

The Pneumococcus Reference Laboratory (LRN) began its care activity supporting the National Health System in 1979, dedicating itself since then to the microbiological surveillance of Streptococcus pneumoniae, analyzing the serotypes, genotypes and antibiotic resistance of pneumococcal clinical isolates from hospitals in the different autonomous communities. Since then, nearly 100,000 invasive clinical isolates have been identified and characterized and are available in our strain collection. Every year we report cases of invasive pneumococcal disease to Europe through the ECDC, being the third European country that reports the most cases. The LRN is coordinated by Dr. José Yuste and is subdivided into two units; on the one hand, the diagnostic and epidemiological part formed by two research assistants (Idoia del Río and Beatriz López) and the research part, formed by Dr. Fernando González Camacho, Dr. Mirian Domenech, Julio Sempere and Mirella Llamosí, who are doing their doctoral theses. The main lines of research of the laboratory are set out below.

  • Research in epidemiology and public health. Pneumococcus is the main bacterium responsible for community-acquired pneumonia and one of the main etiological agents of sepsis and bacterial meningitis, mainly affecting the pediatric population and adults over 65 years of age. Our laboratory participates in the characterization of circulating strains, providing very valuable information to the National Health System on the epidemiology of pneumococcus, which is essential for the early detection of emerging clones and serotypes, which can be very useful in evaluating the impact of current and future vaccines available. Among the techniques used, the characterization of pneumococci by dot blot with antisera, MLST, PCR-sequencing of capsular genes and real-time PCR for negative culture samples stands out.
  • Resistance mechanisms. For several years, our laboratory has been focused on the study of the molecular mechanisms related to the different stages of the infectious process, among which the colonization of the upper respiratory tract, the establishment of pneumonia and its dissemination, producing invasive disease, stand out. To achieve these goals, our laboratory uses mutants in various virulence factors, epithelial cell lines, phagocytic cells (macrophages and neutrophils), as well as mice deficient in important receptors of the host immune system. In order to address these studies, flow cytometry and confocal microscopy are used as the main methodological techniques.
  • Search for vaccine antigens based on conserved proteins. The currently available vaccines are based on capsular polysaccharides that may or may not be conjugated to a carrier protein. Due to the high variability of pneumococcus (100 serotypes), the limitation in the number of serotypes that can be included in a polysaccharide vaccine and the phenomenon of replacement of serotypes, alternative vaccines to the current ones are being studied. In our group we are characterizing the protective role of some highly conserved surface proteins.
  • Importance of pneumococcus biofilms in pathogenesis and lethality. One of the lines in which our laboratory is interested is based on the study of pneumococcal biofilms from a public health perspective, analyzing their impact on the virulence of emerging serotypes. With the incorporation of Dr. Domenech, an expert in bacterial biofilms, we are analyzing the relationship between lethality due to serotypes and their ability to form biofilms, since the formation of biofilm allows the bacteria to evade the immune system, as well as to develop high levels of antibiotic resistance.

AFFILIATION

National Center for Microbiology
Carlos III Health Institute
Madrid

CONTRIBUTIONS AND SELECTED PUBLICATIONS

  1. de Miguel S, Domenech M, González-Camacho F, Sempere J, Vicioso D, Sanz JC, García Comas L, Ardanuy C, Fenoll A, Yuste J. Nationwide trends of invasive pneumococcal disease in Spain (2009-2019) in children and adults during the pneumococcal conjugate vaccine era. Clin Infect Dis. 2020:ciaa1483.
  2. Sempere J, by 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; 11: 309.
  3. Tarancon R, Dominguez-Andrews J, Uranga S, Ferreira AV, Groh LA, Domenech M, Gonzalez-Camacho F, Riksen NP, Aguilo N, Yuste J, Martin C, Netea MG. New live attenuated tuberculosis vaccine MTBVAC induces trained immunity and confers protection against experimental lethal pneumonia. PLoS Pathog. 2020; 16(4):e1008404.
  4. Domenech M, Sempere J, de Miguel S, Yuste J. Domenech M, Sempere J, de Miguel S, Yuste J.
  5. Corsini B , Diez-Martinez R , Aguinagalde L , Gonzalez- Camacho F , Garcia-Fernandez E , Letrado P , Garcia P , Yuste J . Chemotherapy with Phage Lysins Reduces Pneumococcal Colonization of the Respiratory Tract. Antimicrob Agents Chemother. 2018 25;62 (6).
  6. Domenech M, Sempere J, de Miguel S, Yuste J. Immunization with LytB protein of Streptococcus pneumoniae activates complement-mediated phagocytosis and induces protection against pneumonia and sepsis. Vaccine. 2016; 34 (50):6148-6157.
  7. 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. 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.
  8. 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. Emergence of amoxicillin-resistant variants of Spain9V-ST156 pneumococci expressing serotype 11A correlates with their ability to evade the host immune response. PLoS One. 2015;10 (9): e0137565.
  9. Aguinagalde L, Diez-Martinez R, Yuste J, Royo I, Gil C, Lasa I, Martin-Fontecha M, Marin-Ramos NI, Ardanuy C, Liñares J, Garcia P, Garcia E, Sanchez-Puelles JM. Auranofin efficacy against MDR Streptococcus pneumoniae and Staphylococcus aureus infections. J Antimicrob Chemother. 2015; 70(9): 2608-17
  10. , Díez-Martínez R, Yuste J, Royo I, Gil C, Lasa Í, Martín-Fontecha M, Marín-Ramos NI, Ardanuy C, Liñares J, García P, García E, Sánchez-Puelles JM. Pleiotropic effects of cell wall amidase LytA on Streptococcus pneumoniae sensitivity to the host immune response. Infect Immun. 2015; 83 (2): 591-603.
  11. , Díez-Martínez R, Yuste J, Royo I, Gil C, Lasa Í, Martín-Fontecha M, Marín-Ramos NI, Ardanuy C, Liñares J, García P, García E, Sánchez-Puelles JM. Emerging non-PCV13 serotypes 11A and 35B of Streptococcus pneumoniae show high potential for biofilm formation in vitro. PLoS One. 2015; 10(4): e0125636.
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