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Research Lines

Content with Investigacion Inmunobiología .

The Immunobiology group has been working for years on the following lines of research:
1) The mechanisms of haematopoietic cell generation throughout ontogeny and the influence that the first haematopoietic cells exert on the innate and adaptive immune system present in the adults. We have identified and characterised a new population of B lymphocytes called B1-Rel (B220lo), which produce high levels of natural IgG/IgA antibodies. We sought to understand their role in the immune response in animal models of infection, analysing their impact on immune cell populations and on the production of soluble mediators (cytokines and immunoglobulins). In this regard, we have evaluated the generation of embryonic megakaryocytes (and their differentiation niches), their functionality and that of platelets, and their influence on haematopoietic development. For lymphoid populations, we have carried out extensive characterisation by flow cytometry and single cell RNA sequencing (scRNAseq) methodology. To carry out these cellomic studies, we have designed complex panels for use in multiparametric phenotypic analysis, and single cell cytometry and RNAseq omics technologies on purified cell populations.


In parallel, we are interested in understanding local immune responses in respiratory infections at times of particular susceptibility due to the fragility of the immune system (childhood and old age), both in mouse animal models, which allow their manipulation, and in humans. 

2) Mouse models studied during neonatal life, in which we evaluated the effect of antibiotic (AB) treatment and addressed the role of TLR receptors in innate, pseudo-innate and adaptive immune cell populations. In these models, we observed that AB administration was able to modulate B-lymphoid populations, as well as their ability to secrete proinflammatory cytokines in culture and their differentiation into plasma cells, with differentiated immunoglobulin repertoires. Furthermore. These effects were mediated through the Toll-like receptor-2 (TLR2).

3) Mouse models with accelerated senescence (SAMP8) and senescent animals (over 20 months of age) to map lymphoid populations and soluble mediators of the immune response (immunoglobulins and cytokines). In these models, the B lymphoid populations (B1Rel and marginal zone B lymphocytes) are observed to be altered, accompanied by an increase in IgG1 with great restriction of their VDJ repertoires.


4) Role of the B1Rel population in animal models of local or systemic infection. We analysed the response to Streptoccoccus pneumoniae (SPN) locally in the lung and systemically in the spleen, as well as the role of TLR4 in these responses.

5) In humans, we are studying immune responses in children with respiratory syncytial virus (RSV) viral primo-infection. In this case we studied the immune response that occurs locally in the nasal mucosa (by analysis of nasal washings, NW) in a cohort of infected children versus healthy controls, stratified by age. We found that lymphomyeloid cells accumulate in these nasal washings in patients with diverse lymphocyte populations, as well as cytokines and immunoglobulins.

6) Analysis and characterisation of extracellular vesicles produced during respiratory infection both in lung supernatants from models of SPN infection and in LN in the case of children with RSV infection.

7) In parallel, we carry out studies of the genetic rearrangements of immunoglobulins and their use in the generation of chimeric receptors for possible use in immunotherapy.

Research projects

Content with Investigacion Inmunobiología .

-Project “Induction, differentiation and modulation of resident B lymphocytes in the lung in response to pneumococcus (NEUBLUNG)”. Ministry of Science and Innovation, PID2022-141754OB-I00 Call 2022 "Knowledge Generation Projects". 09/01/2023-08/31/2026. Financed by MICIU/AEI /10.13039/501100011033 and by ERDF, EU. PI: Belén by Andrés Muguruza. CoPI: María Luisa Gaspar Alonso-Vega.


 

-Project." Immune response of the nasal mucosa in childhood bronchiolitis” Instituto de Salud Carlos III-AESI. AESI-PI22CIII/00030 PI: Belén by Andrés Muguruza. CoPI Maria Luisa Gaspar Alonso-Vega. 01/01/2023-12/31/2025..

-Project. BenBedPhar. CA20121, European Union. Antonio Cuadrado. (CNM-ISCIII).10/19/2021-10/18/2025.

-Spanish Association Against Cancer Project “Novel comprehensive immunotherapy to specifically target the malignant clone in Sézary syndrome, an ultra-rare cancer of mature T lymphocytes”, number PROYE20084REGU. PI: José Ramón Regueiro, PI group Maria Luisa Gaspar. 01/01/2021-12/31/2023.

Project “The pulmonary immune system in homeostasis and infection: characterization and function of immature and pseudoinnate lymphoid populations.” MINECO-RETOS RTI2018-099114-B-100. PI: Maria Luisa Gaspar, CoPI: Belén de Andrés 01/01/2019-12/31/2022. Financed by MICIU/AEI /10.13039/501100011033/ and by FEDER A way of making Europe.


 

-Project “New B lymphoid populations: B1-rel pseudoinnate cells, homeostatic maintenance and their response under infection conditions.” MINECO-RETOS SAF2015-70880-R. PI: Maria Luisa Gaspar. 01/01/2016-12/31/2019.


 

-Project “Role of CD19+CD45R lymphocytes- in perinatal immune responses. Implications related to respiratory diseases in neonates. AESI PI14CIII/00049; PI Belén de Andrés. 2015-2018.

-Project “Study of the pseudo-innate population of CD19+CD45R- B lymphocytes in TLR-dependent infection models”. AESI PI11/01733FIS. PI Belén de Andrés. 2012-2015.

-Project." Cellular interactions in the establishment of B lymphoid differentiation niches: role of megakaryocytes and their implications in pathology. MINECO; SAF2012-33916. Maria Luisa Gaspar. 01/01/2013-12/31/2015.

-ISCIII Platforms Project to support R&D&I in Biomedicine and Health Sciences. PT23CIII/00006. 2023. Participating researcher: Isabel Cortegano.

-Research contracts between the Carlos III Health Institute and Inmunotek S.L. for the development of the Bactek-mv130 and Uromune-MV140 study in protection against S. pneumoniae infections. Immunotek. IP: Belen de Andrés 2019-2021.

-Research contract between the Carlos III Health Institute and Inmunotek S.L. “MV130 as a vaccine model based on trained immunity against respiratory infections due to pneumococcus and respiratory syncytial virus”, CAM Call. Industrial Doctorates. IND2023/BMD-27071. PI: Belén by Andrés Muguruza. 12/01/2023-11/30/2026.

Publications

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Alastruey-Izquierdo A, Alcazar-Fuoli L, Rivero-Menéndez O, Ayats J, Castro C, García-Rodríguez J, Goterris-Bonet L, Ibáñez-Martínez E, Linares-Sicilia MJ, Martin-Gomez MT, Martín-Mazuelos E, Pelaez T, Peman J, Rezusta A, Rojo S, Tejero R, Anza DV, Viñuelas J, Zapico MS, Cuenca-Estrella M; the FILPOP2 Project from GEMICOMED (SEIMC) and REIPI. Molecular Identification and Susceptibility Testing of Molds Isolated in a Prospective Surveillance of Triazole Resistance in Spain (FILPOP2 Study). Antimicrob Agents Chemother. 2018 Aug 27;62(9):e00358-18. doi: 10.1128/AAC.00358-18. PMID: 29941643; PMCID: PMC6125503.

Alastruey-Izquierdo A, Alcazar-Fuoli L, Rivero-Menéndez O, Ayats J, Castro C, García-Rodríguez J, Goterris-Bonet L, Ibáñez-Martínez E, Linares-Sicilia MJ, Martin-Gomez MT, Martín-Mazuelos E, Pelaez T, Peman J, Rezusta A, Rojo S, Tejero R, Anza DV, Viñuelas J, Zapico MS, Cuenca-Estrella M; the FILPOP2 Project from GEMICOMED (SEIMC) and REIPI. Molecular Identification and Susceptibility Testing of Molds Isolated in a Prospective Surveillance of Triazole Resistance in Spain (FILPOP2 Study). Antimicrob Agents Chemother. 2018 Aug 27;62(9):e00358-18. doi: 10.1128/AAC.00358-18. PMID: 29941643; PMCID: PMC6125503.

PUBMED DOI

Gonçalves SM, Lagrou K, Rodrigues CS, Campos CF, Bernal-Martínez L, Rodrigues F, Silvestre R, Alcazar-Fuoli L, Maertens JA, Cunha C, Carvalho A. Evaluation of Bronchoalveolar Lavage Fluid Cytokines as Biomarkers for Invasive Pulmonary Aspergillosis in At-Risk Patients. Front Microbiol. 2017 Nov 29;8:2362. doi:10.3389/fmicb.2017.02362. PMID: 29238334; PMCID: PMC5712575.

Gonçalves SM, Lagrou K, Rodrigues CS, Campos CF, Bernal-Martínez L, Rodrigues F, Silvestre R, Alcazar-Fuoli L, Maertens JA, Cunha C, Carvalho A. Evaluation of Bronchoalveolar Lavage Fluid Cytokines as Biomarkers for Invasive Pulmonary Aspergillosis in At-Risk Patients. Front Microbiol. 2017 Nov 29;8:2362. doi:10.3389/fmicb.2017.02362. PMID: 29238334; PMCID: PMC5712575.

PUBMED DOI

Alcazar-Fuoli L, Buitrago M, Gomez-Lopez A, Mellado E. An alternative host model of a mixed fungal infection by azole susceptible and resistant Aspergillus spp strains. Virulence. 2015;6(4):376-84. doi: 10.1080/21505594.2015.1025192. PMID: 26065322; PMCID: PMC4601236.

Alcazar-Fuoli L, Buitrago M, Gomez-Lopez A, Mellado E. An alternative host model of a mixed fungal infection by azole susceptible and resistant Aspergillus spp strains. Virulence. 2015;6(4):376-84. doi: 10.1080/21505594.2015.1025192. PMID: 26065322; PMCID: PMC4601236.

PUBMED DOI

Alcazar-Fuoli L, Cairns T, Lopez JF, Zonja B, Pérez S, Barceló D, Igarashi Y, Bowyer P, Bignell E. A modified recombineering protocol for the genetic manipulation of gene clusters in Aspergillus fumigatus. PLoS One. 2014 Nov 5;9(11):e111875. doi: 10.1371/journal.pone.0111875. PMID: 25372385; PMCID:PMC4221250.

Alcazar-Fuoli L, Cairns T, Lopez JF, Zonja B, Pérez S, Barceló D, Igarashi Y, Bowyer P, Bignell E. A modified recombineering protocol for the genetic manipulation of gene clusters in Aspergillus fumigatus. PLoS One. 2014 Nov 5;9(11):e111875. doi: 10.1371/journal.pone.0111875. PMID: 25372385; PMCID:PMC4221250.

PUBMED DOI

Bertuzzi M, Schrettl M, Alcazar-Fuoli L, Cairns TC, Muñoz A, Walker LA, Herbst S, Safari M, Cheverton AM, Chen D, Liu H, Saijo S, Fedorova ND, Armstrong-James D, Munro CA, Read ND, Filler SG, Espeso EA, Nierman WC, Haas H, Bignell EM. The pH-responsive PacC transcription factor of Aspergillus fumigatus governs epithelial entry and tissue invasion during pulmonary aspergillosis. PLoS Pathog. 2014 Oct 16;10(10):e1004413. doi: 10.1371/journal.ppat.1004413.

Bertuzzi M, Schrettl M, Alcazar-Fuoli L, Cairns TC, Muñoz A, Walker LA, Herbst S, Safari M, Cheverton AM, Chen D, Liu H, Saijo S, Fedorova ND, Armstrong-James D, Munro CA, Read ND, Filler SG, Espeso EA, Nierman WC, Haas H, Bignell EM. The pH-responsive PacC transcription factor of Aspergillus fumigatus governs epithelial entry and tissue invasion during pulmonary aspergillosis. PLoS Pathog. 2014 Oct 16;10(10):e1004413. doi: 10.1371/journal.ppat.1004413.

DOI

Yasmin S, Alcazar-Fuoli L, Gründlinger M, Puempel T, Cairns T, Blatzer M, Lopez JF, Grimalt JO, Bignell E, Haas H. Mevalonate governs interdependency of ergosterol and siderophore biosyntheses in the fungal pathogen Aspergillus fumigatus. Proc Natl Acad Sci U S A. 2012 Feb 21;109(8):E497-504. doi: 10.1073/pnas.1106399108. Epub 2011 Nov 21. PMID: 22106303; PMCID: PMC3286978.

Yasmin S, Alcazar-Fuoli L, Gründlinger M, Puempel T, Cairns T, Blatzer M, Lopez JF, Grimalt JO, Bignell E, Haas H. Mevalonate governs interdependency of ergosterol and siderophore biosyntheses in the fungal pathogen Aspergillus fumigatus. Proc Natl Acad Sci U S A. 2012 Feb 21;109(8):E497-504. doi: 10.1073/pnas.1106399108. Epub 2011 Nov 21. PMID: 22106303; PMCID: PMC3286978.

PUBMED DOI

Impaired Cytotoxic Response in PBMCs From Patients With COVID-19 Admitted to the ICU: Biomarkers to Predict Disease Severity.

Impaired Cytotoxic Response in PBMCs From Patients With COVID-19 Admitted to the ICU: Biomarkers to Predict Disease Severity. Vigón L, Fuertes D, García-Pérez J, Torres M, Rodríguez-Mora S, Mateos E, Corona M, Saez-Marín AJ, Malo R, Navarro C, Murciano-Antón MA, Cervero M, Alcamí J, García-Gutiérrez V, Planelles V, López-Huertas MR, Coiras M (AC). Front Immunol. 2021 May 26;12:665329. doi: 10.3389/fimmu.2021.665329. eCollection 2021. PMID: 34122423.

PUBMED DOI

Identification of Immunological Parameters as Predictive Biomarkers of Relapse in Patients with Chronic Myeloid Leukemia on Treatment-Free Remission

Identification of Immunological Parameters as Predictive Biomarkers of Relapse in Patients with Chronic Myeloid Leukemia on Treatment-Free Remission. Vigón L, Luna A, Galán M, Rodríguez-Mora S, Fuertes D, Mateos E, Piris-Villaespesa M, Bautista G, San José E, Rivera-Torres J, Steegmann JL, de Ory F, Pérez-Olmeda M, Alcamí J, Planelles V, López-Huertas MR, García-Gutiérrez V, Coiras M (AC). J Clin Med. 2020 Dec 25;10(1):42. doi: 10.3390/jcm10010042. PMID: 33375572.

PUBMED DOI

Cytotoxic cell populations developed during treatment with tyrosine kinase inhibitors protect autologous CD4+ T cells from HIV-1 infection

Cytotoxic cell populations developed during treatment with tyrosine kinase inhibitors protect autologous CD4+ T cells from HIV-1 infection. Vigón L, Rodríguez-Mora S, Luna A, Sandonís V, Mateos E, Bautista G, Steegmann JL, Climent N, Plana M, Pérez-Romero P, de Ory F, Alcamí J, García-Gutierrez V, Planelles V, López-Huertas MR, Coiras M (AC). Biochem Pharmacol. 2020 Dec;182:114203. doi: 10.1016/j.bcp.2020.114203. PMID: 32828803.

PUBMED DOI

Tyrosine Kinase Inhibition: a New Perspective in the Fight against HIV

Tyrosine Kinase Inhibition: a New Perspective in the Fight against HIV. Rodríguez-Mora S, Spivak AM, Szaniawski MA, López-Huertas MR, Alcamí J, Planelles V, Coiras M (AC). Curr HIV/AIDS Rep. 2019 Oct;16(5):414-422. doi: 10.1007/s11904-019-00462-5. PMID: 31506864. Review.

PUBMED DOI

Dasatinib protects humanized mice from acute HIV-1 infection

Dasatinib protects humanized mice from acute HIV-1 infection. Salgado M, Martinez-Picado J, Gálvez C, Rodríguez-Mora S, Rivaya B, Urrea V, Mateos E, Alcamí J, Coiras M (AC). Biochem Pharmacol. 2020 Apr;174:113625. doi: 10.1016/j.bcp.2019.113625. PMID: 31476293.

PUBMED DOI

Evaluation of resistance to HIV-1 infection ex vivo of PBMCs isolated from patients with chronic myeloid leukemia treated with different tyrosine kinase inhibitors.

Evaluation of resistance to HIV-1 infection ex vivo of PBMCs isolated from patients with chronic myeloid leukemia treated with different tyrosine kinase inhibitors. Bermejo M, Ambrosioni J, Bautista G, Climent N, Mateos E, Rovira C, Rodríguez-Mora S, López-Huertas MR, García-Gutiérrez V, Steegmann JL, Duarte R, Cervantes F, Plana M, Miró JM, Alcamí J, Coiras M (AC). Biochem Pharmacol. 2018 Oct;156:248-264. doi: 10.1016/j.bcp.2018.08.031. PMID: 30142322.

PUBMED DOI

Staphylococcus aureus Nasal Colonization in Spanish Children. The COSACO Nationwide Surveillance Study.

Staphylococcus aureus Nasal Colonization in Spanish Children. The COSACO Nationwide Surveillance Study. Del Rosal T, Méndez-Echevarría A, Garcia-Vera C, Escosa-Garcia L, Agud M, Chaves F, Román F, Gutierrez-Fernandez J, Ruiz de Gopegui E, Ruiz-Carrascoso G, Ruiz-Gallego MDC, Bernet A, Quevedo SM, Fernández-Verdugo AM, Díez-Sebastian J, Calvo C; COSACO Study Group.

PUBMED

Antimicrobial Resistance and Distribution of Staphylococcus spp. Pulsotypes Isolated from Goat and Sheep Bulk Tank Milk in Southern Spain

Antimicrobial Resistance and Distribution of Staphylococcus spp. Pulsotypes Isolated from Goat and Sheep Bulk Tank Milk in Southern Spain. Barrero-Domínguez B, Luque I, Galán-Relaño Á, Vega-Pla JL, Huerta B, Román F, Astorga RJ. Foodborne Pathog Dis. 2019 Oct;16(10):723-730. doi: 10.1089/fpd.2018.2593. Epub 2019 Jun 3.Foodborne Pathog Dis. 2019. PMID: 31157980

PUBMED

Prevalence of pSCFS7-like vectors among cfr-positive staphylococcal population in Spain

Prevalence of pSCFS7-like vectors among cfr-positive staphylococcal population in Spain. Nguyen LTT*, Román F*, Morikawa K, Trincado P, Marcos C, Rojo-Martín MD, Cafini F. Int J Antimicrob Agents. 2018 Aug;52(2):305-306.

PUBMED

Methodology for the Study of Horizontal Gene Transfer in Staphylococcus aureus.

Methodology for the Study of Horizontal Gene Transfer in Staphylococcus aureus. Cafini F, Thi Le Thuy N, Román F, Prieto J, Dubrac S, Msadek T, Morikawa K. J Vis Exp. 2017 Mar 10;(121).

PUBMED

Emergence of cfr-Mediated Linezolid Resistance in a Methicillin-Resistant Staphylococcus aureus Epidemic Clone Isolated from Patients with Cystic Fibrosis.

Emergence of cfr-Mediated Linezolid Resistance in a Methicillin-Resistant Staphylococcus aureus Epidemic Clone Isolated from Patients with Cystic Fibrosis. de Dios Caballero J, Pastor MD, Vindel A, Máiz L, Yagüe G, Salvador C, Cobo M, Morosini MI, del Campo R, Cantón R; GEIFQ Study Group. Antimicrob Agents Chemother. 2015 Dec 14;60(3):1878-82.

PUBMED

Molecular epidemiology of community-associated methicillin-resistant Staphylococcus aureus in Spain: 2004-12.

Molecular epidemiology of community-associated methicillin-resistant Staphylococcus aureus in Spain: 2004-12. Vindel A, Trincado P, Cuevas O, Ballesteros C, Bouza E, Cercenado E. J Antimicrob Chemother. 2014 Nov;69(11):2913-9.

PUBMED

Draft Genome Sequence of Strain SA_ST125_MupR of Methicillin-Resistant Staphylococcus aureus ST125, a Major Clone in Spain.

Draft Genome Sequence of Strain SA_ST125_MupR of Methicillin-Resistant Staphylococcus aureus ST125, a Major Clone in Spain. Barrado L, Viedma E, Vindel A, Otero JR, Chaves F. Genome Announc. 2013 Aug 8;1(4).

PUBMED

Detection of linezolid-resistant Staphylococcus aureus with 23S rRNA and novel L4 riboprotein mutations in a cystic fibrosis patient in Spain.

Detection of linezolid-resistant Staphylococcus aureus with 23S rRNA and novel L4 riboprotein mutations in a cystic fibrosis patient in Spain. Román F, Roldán C, Trincado P, Ballesteros C, Carazo C, Vindel A. Antimicrob Agents Chemother. 2013 May;57(5):2428-9.

PUBMED

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List of staff

Additional Information

The research objectives focus on the areas of Bacterial Taxonomy and Microbiology in Public Health:

- Identification of pathogenic bacteria that are difficult to assign taxonomically
- Description of new species
- Study of bacterial population biology through typing
- Epidemiology of antimicrobial resistance and virulence factors for certain bacterial pathogens in different nosocomial/community settings.

These objectives are addressed from the perspective of polyphasic taxonomy (phenotypic and genotypic taxonomy) through the study of targets for genus/species assignment and phylogenetic analyses. The analysis of the virulome and resistome of certain species is approached from the perspective of complete genome sequencing.

Research on invasive infection by Streptococcus pyogenes and other beta-hemolytic streptococci has been carried out since 1994, through the Surveillance Program of the National Center for Microbiology. The typing of invasive S. pyogenes strains circulating in Spain includes determination of: serotype (emm-type); the genetic profile of streptococcal pyrogenic exotoxins; the antimicrobial resistance phenotype; and the genotypes of epidemiologically related strains and/or outbreaks.

The research objectives focus on the areas of Bacterial Taxonomy and Microbiology in Public Health:

- Identification of pathogenic bacteria that are difficult to assign taxonomically
- Description of new species
- Study of bacterial population biology through typing
- Epidemiology of antimicrobial resistance and virulence factors for certain bacterial pathogens in different nosocomial/community settings.

These objectives are addressed from the perspective of polyphasic taxonomy (phenotypic and genotypic taxonomy) through the study of targets for genus/species assignment and phylogenetic analyses. The analysis of the virulome and resistome of certain species is approached from the perspective of complete genome sequencing.

Research on invasive infection by Streptococcus pyogenes and other beta-hemolytic streptococci has been carried out since 1994, through the Surveillance Program of the National Center for Microbiology. The typing of invasive S. pyogenes strains circulating in Spain includes determination of: serotype (emm-type); the genetic profile of streptococcal pyrogenic exotoxins; the antimicrobial resistance phenotype; and the genotypes of epidemiologically related strains and/or outbreaks.

Content with Investigacion Inmunobiología .