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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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Rabbit trypanosome detection in Phlebotomus perniciosus sand flies from the leishmaniasis outbreak in Madrid,

González E, Molina R, Jiménez M. Rabbit trypanosome detection in Phlebotomus perniciosus sand flies from the leishmaniasis outbreak in Madrid, Spain. Acta Trop 2018, 187:201-206.

PUBMED DOI

Phlebotomine sand fly survey in the focus of leishmaniasis of Madrid, Spain (2012–2014): seasonal dynamics, Leishmania infantum infection rates and blood meal preferences.

González E, Jiménez M, Hernández S, Martín-Martín I, Molina R. Phlebotomine sand fly survey in the focus of leishmaniasis of Madrid, Spain (2012–2014): seasonal dynamics, Leishmania infantum infection rates and blood meal preferences. Parasit Vectors 2017, 10:368.

PUBMED DOI

Methods in Sand Fly Research

Molina R, Jiménez M, Alvar J, González E, Hernández-Taberna S, Martín-Martín Inés. 2017. Methods in Sand Fly Research (R. Molina, M. Jiménez & J. Alvar, edits.). Servicio de Publicaciones Universidad de Alcalá de Henares. ISBN: 978-84-16978-28-1

Kinetics of anti-Phlebotomus perniciosus saliva antibodies in experimentally bitten mice and rabbits.

Martín-Martín I, Molina R, Jiménez M. Kinetics of anti-Phlebotomus perniciosus saliva antibodies in experimentally bitten mice and rabbits. PLoS ONE, November 16, 2015; 10(11):

PUBMED DOI

Identifying salivary antigens of Phlebotomus argentipes by a 2DE approach.

Martín-Martín I, Molina R, Jiménez M. Identifying salivary antigens of Phlebotomus argentipes by a 2DE approach. Acta Trop 2013, 126: 229–239.

PUBMED DOI

Factors associated with Leishmania asymptomatic infection: results from a cross-sectional survey in highland northern Ethiopia

Custodio E, Gadisa E, Sordo L, Cruz I, Moreno J, Nieto J, Chicharro C, Aseffa A, Abraham Z, Hailu T, Cañavate C. Factors associated with Leishmania asymptomatic infection: results from a cross-sectional survey in highland northern Ethiopia. PLoS Negl Trop Dis. 2012;6(9):e1813.

PUBMED DOI

Cytokine Release Assays as Tests for Exposure to Leishmania, and for Confirming Cure from Leishmaniasis, in Solid Organ Transplant Recipients.

Carrillo E, Carrasco-Antón N, López-Medrano F, Salto E, Fernández L, San Martín JV, Alvar J, Aguado JM, Moreno J. Cytokine Release Assays as Tests for Exposure to Leishmania, and for Confirming Cure from Leishmaniasis, in Solid Organ Transplant Recipients. PLoS Negl Trop Dis. 2015 Oct 23;9(10):e0004179.

PUBMED DOI

Chemotactic Protein 1 in Plasma from Soluble Leishmania Antigen-Stimulated Whole Blood as a Potential Biomarker of the Cellular Immune Response to Leishmania infantum

Ibarra-Meneses AV, Sanchez C, Alvar J, Moreno J, Carrillo E. Monocyte Chemotactic Protein 1 in Plasma from Soluble Leishmania Antigen-Stimulated Whole Blood as a Potential Biomarker of the Cellular Immune Response to Leishmania infantum. Front Immunol. 2017 Sep 29;8:1208.

PUBMED DOI

Cytokines and chemokines measured in dried SLA-stimulated whole blood spots for asymptomatic Leishmania infantum and Leishmania donovani infection.

Ibarra-Meneses AV, Mondal D, Alvar J, Moreno J, Carrillo E. Cytokines and chemokines measured in dried SLA-stimulated whole blood spots for asymptomatic Leishmania infantum and Leishmania donovani infection. Sci Rep. 2017 Dec 8;7(1):17266.

PUBMED DOI

Cellular Markers of Active Disease and Cure in Different Forms of Leishmania infantum-Induced Disease.

Botana L, Matía B, San Martin JV, Romero-Maté A, Castro A, Molina L, Fernandez L, Ibarra-Meneses A, Aguado M, Sánchez C, Horrillo L, Chicharro C, Nieto J, Ortega S, Ruiz-Giardin JM, Carrillo E, Moreno J. Cellular Markers of Active Disease and Cure in Different Forms of Leishmania infantum-Induced Disease. Front Cell Infect Microbiol. 2018 Nov 13;8:381.

PUBMED DOI

Carroll MW et al. Temporal and spatial analysis of the 2014-2015 Ebola virus outbreak in West Africa. Nature.

Carroll MW et al. Temporal and spatial analysis of the 2014-2015 Ebola virus outbreak in West Africa. Nature. 2015 Aug 6;524(7563):97-101. doi: 10.1038/nature14594. Epub 2015 Jun 17. PMID: 26083749. ​

Fernandez-Garcia MD, Meertens L, Chazal M, Hafirassou ML, Dejarnac O, Zamborlini A, Despres P, Sauvonnet N, Arenzana-Seisdedos F, Jouvenet N, Amara A. Vaccine and Wild-Type Strains of Yellow Fever Virus Engage Distinct Entry Mechanisms and Differentially Stimulate Antiviral Immune Responses.

Fernandez-Garcia MD, Meertens L, Chazal M, Hafirassou ML, Dejarnac O, Zamborlini A, Despres P, Sauvonnet N, Arenzana-Seisdedos F, Jouvenet N, Amara A. Vaccine and Wild-Type Strains of Yellow Fever Virus Engage Distinct Entry Mechanisms and Differentially Stimulate Antiviral Immune Responses. mBio. 2016 Feb 9;7(1):e01956-15. doi: 10.1128/mBio.01956-15. PMID: 26861019; PMCID:PMC4752603.

Identification and whole-genome characterization of a recombinant Enterovirus B69 isolated from a patient with Acute Flaccid Paralysis in Niger, 2015

Fernandez-Garcia MD, Majumdar M, Kebe O, Ndiaye K, Martin J. Identification and whole-genome characterization of a recombinant Enterovirus B69 isolated from a patient with Acute Flaccid Paralysis in Niger, 2015. Sci Rep. 2018 Feb 1;8(1):2181. doi: 10.1038/s41598-018-20346-9. PMID: 29391547; PMCID: PMC5795009.

Majumdar M, Sharif S, Klapsa D, Wilton T, Alam MM, Fernandez-Garcia MD, Rehman L, Mujtaba G, McAllister G, Harvala H, Templeton K, Mee ET, Asghar H, Ndiaye K, Minor PD, Martin J. Environmental Surveillance Reveals Complex Enterovirus Circulation Patterns in Human Populations. Open Forum Infect Dis. 2018

Majumdar M, Sharif S, Klapsa D, Wilton T, Alam MM, Fernandez-Garcia MD, Rehman L, Mujtaba G, McAllister G, Harvala H, Templeton K, Mee ET, Asghar H, Ndiaye K, Minor PD, Martin J. Environmental Surveillance Reveals Complex Enterovirus Circulation Patterns in Human Populations. Open Forum Infect Dis. 2018 Oct 1;5(10):ofy250. doi: 10.1093/ofid/ofy250. PMID: 30377626; PMCID: PMC6201154.

Fernandez-Garcia MD, Majumdar M, Kebe O, Fall AD, Kone M, Kande M, Dabo M, Sylla MS, Sompare D, Howard W, Faye O, Martin J, Ndiaye K. Emergence of Vaccine-Derived Polioviruses during Ebola Virus Disease Outbreak, Guinea, 2014-2015.

Fernandez-Garcia MD, Majumdar M, Kebe O, Fall AD, Kone M, Kande M, Dabo M, Sylla MS, Sompare D, Howard W, Faye O, Martin J, Ndiaye K. Emergence of Vaccine-Derived Polioviruses during Ebola Virus Disease Outbreak, Guinea, 2014-2015. Emerg Infect Dis. 2018 Jan;24(1):65-74. doi: 10.3201/eid2401.171174. PMID:29260690; PMCID: PMC5749474.

Tarragó, D.; Mateos, M.-L.; Avellón, A.; Pérez-Vázquez, M.-D.; Tenorio, A.2004

Tarragó, D.; Mateos, M.-L.; Avellón, A.; Pérez-Vázquez, M.-D.; Tenorio, A.2004. Quantitation of Cytomegalovirus DNA in Cerebrospinal Fluid and Serum Specimens from AIDS Patients Using a Novel Highly Sensitive Nested Competitive PCR and the Cobas Amplicor CMV Monitor™ Journal of Medical Virology. 72-2, pp.249-256. ISSN 01466615. 10. Tarragó, D.; Quereda, C.; Tenorio, A.2003. Different cytomegalovirus glycoprotein B genotype distribution in serum and cerebrospinal fluid specimens determined by a novel multiplex nested PCR Journal of Clinical Microbiology. 41-7, pp.2872-2877. ISSN 00951137.

Fernandez-Garcia, Maria Dolores (AC); Kebe, Ousmane; Fall, Aichatou D.; Dia, Hamet; Diop, Ousmane M.; Delpeyroux, Francis; Ndiaye, Kader. 2016.

Fernandez-Garcia, Maria Dolores (AC); Kebe, Ousmane; Fall, Aichatou D.; Dia, Hamet; Diop, Ousmane M.; Delpeyroux, Francis; Ndiaye, Kader. (1/ 7). 2016. Enterovirus A71 Genogroups C and E in Children with Acute Flaccid Paralysis, West Africa EMERGING INFECTIOUS DISEASES. 22-4, pp.753-755. ISSN 1080-6040.

Molecular epidemiology of coxsackievirus B3 infection in Spain, 2004-2015.

K Calderón, M Díaz-de Cerio, C Muñoz-Almagro, N Rabella, I Martínez-Rienda, A Moreno-Docón, G Trallero, M Cabrerizo*. Molecular epidemiology of coxsackievirus B3 infection in Spain, 2004-2015. Arch Virol 161: 1365-1370 (2016).

PUBMED DOI

Development and Evaluation of a Serological Assay for the Diagnosis of Tuberculosis in Alpacas and Llamas.

Development and Evaluation of a Serological Assay for the Diagnosis of Tuberculosis in Alpacas and Llamas. Infantes-Lorenzo, Jose A.; Whitehead, Claire E.; Moreno, Inmaculada; et ál..FRONTIERS IN VETERINARY SCIENCE Volumen: 5 Número de artículo: 189 Fecha de publicación: AUG 13 2018

PUBMED DOI

Influence of the Microenvironment in the Transcriptome of Leishmania infantum Promastigotes: Sand Fly versus Culture

Influence of the Microenvironment in the Transcriptome of Leishmania infantum Promastigotes: Sand Fly versus Culture. Alcolea, Pedro J.; Alonso, Ana; Dominguez, Mercedes; et ál..PLOS NEGLECTED TROPICAL DISEASES Volumen: 10 Número: 5 Número de artículo: e0004693 Fecha de publicación: MAY 2016

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Additional Information

La inducción de la tolerancia al aloinjerto sigue siendo una meta por alcanzar en el trasplante de órganos. La mayoría de las estrategias terapéuticas se centran en la inhibición del sistema inmunológico adaptativo, pero datos recientes demuestran que el reconocimiento alogénico de las células mieloides inicia el rechazo al trasplante. Terapias dirigidas hacia las células mieloides “in vivo” representan un objetivo potencial para inducir tolerancia inmunológica, pero permanece inexplorado clínicamente.Nuestro laboratorio utiliza una nanoinmunoterapia revolucionaria de nanopartículas de lipoproteínas de alta densidad (HDL) cargadas con rapamicina (mTORi-HDL) que previenen las modificaciones epigenéticas asociadas con la inmunidad entrenada, un estado funcional de los macrófagos recientemente descubierto. Usando un modelo experimental de trasplante en ratón, nuestros resultados demuestran que la administración de esta inmunoterapia con mTORi-HDL previene la respuesta inmunológica y promueve la tolerancia al órgano trasplantado.Nuestro laboratorio muestra un enfoque de investigación multidisciplinar articulado en tres objetivos diferentes para evaluar la relevancia clínica y los efectos terapéuticos de la inmunoterapia como preparación para un ensayo clínico en trasplante de órganos. Los objetivos generales estarán orientados a confirmar la identificación de la inmunidad entrenada como biomarcador y valor analítico para predecir el riesgo de rechazo en pacientes trasplantados bajo tres condiciones: periodos prolongadas de reperfusión isquémica (IRI) (objetivo 1), alosensibilización (objetivo 2) e infección (objetivo 3).

Induction of allograft tolerance remains a goal to be achieved in organ transplantation. Most therapeutic strategies focus on inhibition of the adaptive immune system, but recent data demonstrate that allogeneic recognition of myeloid cells initiates transplant rejection. Therapies targeting myeloid cells “in vivo” represent a potential target to induce immunological tolerance, but remain clinically unexplored. 

Our laboratory uses a revolutionary nanoimmunotherapy of high-density lipoprotein (HDL) nanoparticles loaded with rapamycin (mTORi-HDL) that prevents epigenetic modifications associated with trained immunity, a recently discovered functional state of macrophages. Using an experimental mouse transplant model, our results demonstrate that the administration of this immunotherapy with mTORi-HDL prevents the immune response and promotes tolerance to the transplanted organ. 

Our laboratory shows a multidisciplinary research approach articulated in three different objectives to evaluate the clinical relevance and therapeutic effects of immunotherapy in preparation for a clinical trial in organ transplantation. The general objectives will be aimed at confirming the identification of trained immunity as a biomarker and analytical value to predict the risk of rejection in transplant patients under three conditions: prolonged periods of ischemic reperfusion (IRI) (objective 1), allosensitization (objective 2) and infection (objective 3).

Induction of allograft tolerance remains a goal to be achieved in organ transplantation. Most therapeutic strategies focus on inhibition of the adaptive immune system, but recent data demonstrate that allogeneic recognition of myeloid cells initiates transplant rejection. Therapies targeting myeloid cells “in vivo” represent a potential target to induce immunological tolerance, but remain clinically unexplored. 

Our laboratory uses a revolutionary nanoimmunotherapy of high-density lipoprotein (HDL) nanoparticles loaded with rapamycin (mTORi-HDL) that prevents epigenetic modifications associated with trained immunity, a recently discovered functional state of macrophages. Using an experimental mouse transplant model, our results demonstrate that the administration of this immunotherapy with mTORi-HDL prevents the immune response and promotes tolerance to the transplanted organ. 

Our laboratory shows a multidisciplinary research approach articulated in three different objectives to evaluate the clinical relevance and therapeutic effects of immunotherapy in preparation for a clinical trial in organ transplantation. The general objectives will be aimed at confirming the identification of trained immunity as a biomarker and analytical value to predict the risk of rejection in transplant patients under three conditions: prolonged periods of ischemic reperfusion (IRI) (objective 1), allosensitization (objective 2) and infection (objective 3).

Content with Investigacion Virología Molecular .