The Plasmodium vivax and Extracellular Vesicles Research Group is co-led by two principal investigators: Hernando A. del Portillo, ICREA Research Professor, and Carmen Fernandez-Becerra, associated research professor at Barcelona Institute for Global Health (ISGlobal). They joined IGTP in November 2015 through a collaborative agreement between ISGlobal and IGTP.

This research group is recognised as a Consolidated Research Group by the AGAUR (Agència de Gestió i Ajuts Universitaris i de Recerca) (2021 SGR 01554).

The group brings their expertise in malaria and exosome research, extracellular vesicles (EVs) of endocytic origin, with a very strong motivation to produce translational impact. The group uses a series of molecular and cellular biology technologies combined with immune-epidemiological studies in endemic regions of vivax malaria (i) to study the biology of the parasite, (ii) to study the role of circulating EVs form natural infections in host-parasite intercellular communication, to unveil mechanistic insights into cryptic erythrocytic infections in the spleen and bone marrow, and (iii) to discover new biomarkers of parasitic infections associated with circulating EVs. In addition, the group is pioneering the use of reticulocyte-derived exosomes as a new vaccine and platform against malaria. To this end, Dr. del Portillo has co-founded INNOVEX THERAPEUTICS SL, the first Spanish spin-off devoted to the use of exosomes as new therapeutic agents and diagnostic tools. It is important to note that the spin-off is presently located at IGTP.

Additionally, the group has implemented small-scale soluble protein production in the cell-free wheat germ system for multiplex immunological assay, in vivo imaging of malaria parasites, CRISPR-Cas9 technology for functional assays with malaria parasites, humanized mouse models and organs-on-a-chip technology to advance on their studies of cryptic asymptomatic infections and development of a culture system for P. vivax. These research areas and technologies are therefore amenable for collaborations with groups working on extracellular vesicles, vaccines and hematological disorders at IGTP.

The research of the group is fully compatible with the possibility of creating a future horizontal Institutional project to develop vaccines against HIV, TB and malaria, together known as the "big three" and responsible for a staggering 5.6 million deaths annually.

Keywords: Plasmodium vivax, exosomes, extracellular vesicles, malaria, vaccines, hematological disorders.

Plasmodium vivax and Exosome Research (PVREX)

Group leader

  • Hernando A. Del Portillo, PhD

    Hernando A. Del Portillo, PhD

    Hernando del Portillo studied molecular aspects of parasites at University of Georgia, USA, where he received his PhD in 1985, followed by two WHO postdoctoral trainings at New York University Medical Centre (USA) and Institut Pasteur (France) where he specialised in molecular biology of malaria parasites. Since then, he has pursued molecular studies in malaria to unveil the mechanisms of virulence and to develop rational control strategies. He consolidated his independent scientific career as an assistant/associate professor at the Department of Parasitology, University of Sao Paulo, Brazil and as a visiting professor at the Centre for Molecular Biology (ZMBH), Heidelberg, Germany. He joined ISGlobal as an ICREA research professor in 2007 and co-ascribed to the Germans Trias i Pujol Research Institute (IGTP) in 2015. For six years until 2022, he was the president of the Grupo Español de Investigación e Innovación en Vesículas Extracelulares (GEIVEX) and the Coordinator of the Spanish Network of Excellency on Extracellular Vesicles Research. Currently, he is the co-founder of Innovex Therapeutics SL, the first Spanish company using Extracellular Vesicles as vaccines against infectious diseases.

    Contact: hernandoa.delportillo(ELIMINAR)@isglobal.org
    ORCID: 0000-0002-5278-3452
    ICREA webpage: https://www.icrea.cat/Web/ScientificStaff/Hernando-A-del-Portillo-Obando-438
    ISGlobal webpage: https://www.isglobal.org/en/our-team/-/profiles/11800

  • Carmen Fernández Becerra, PhD

    Carmen Fernández Becerra, PhD

    Dr Carmen Fernández-Becerra earned her PhD in Biological Sciences from the University of Granada in 1996. She pursued her postdoctoral research in Brazil (University of Sao Paulo) and Germany (Zentrum für Molekulare Biologie), focusing on biological aspects of Plasmodium vivax and Toxoplasma gondii.

    Currently, she is an associate research professor at Barcelona Institute for Global Health (ISGlobal), which she joined in 2007, and has also been affiliated with Germans Trias i Pujol Research Institute (IGTP) since 2015. Her research group studies the molecular basis of malaria pathology caused by P. vivax, contributing to significant discoveries such as the identification of the first multigene variant subtelomeric family involved in pathology and the physiological role of extracellular vesicles (EVs) produced by P. vivax-infected reticulocytes. In recent years, her research has expanded into EVs as a potential tool for identifying new biomarkers of parasitic infections, such as Chagas disease. Her research has received funding from various national and international agencies, resulting in numerous high-impact scientific publications.

    Contact: carmen.fernandez(ELIMINAR)@isglobal.org
    ORCID: 0000-0001-5154-0013
    ISGlobal webpage: https://www.isglobal.org/en/our-team/-/profiles/2100

Research lines

Exosome-mediated mechanisms of non-hypnozoite cryptic infections in P. vivax malaria

Strong support of non-hypnozoite cryptic infections, mainly in the spleen and bone marrow, has been published. Noticeably, extracellular vesicles in the size range of exosomes obtained from human patients facilitated binding to the human spleen. The group thus hypothesizes that EVs, specifically reticulocyte-derived exosomes, from P. vivax infections mediate bone marrow defects on erythropoiesis and spleen extramedullary erythropoiesis facilitating cryptic infections likely responsible for asymptomatic transmission. As this species cannot be continuously culture in vitro, proving this hypothesis represents a major technical challenge. Two different approaches implementing new human science technological frontiers are being pursued, organs-on-a-chip and humanized mouse models of these human organs, thus facilitating to stringently test the role of exosomes as intercellular communicators in a space and velocity that will facilitate their uptake by cells in these hemopoietic tissues. 

Reticulocyte-derived exosomes (Rex) vaccines against P. vivax

T-cell responses were believed not to play a major role in natural immunity against blood stages of malaria as P. falciparum, the most virulent human malaria parasite, predominantly invades mature red blood cells which lack antigen presenting machinery. However, the group's recent study demonstrated that reticulocyte exosomes contain HLA Class I antigens actively up taken by dendritic cells and that circulating extracellular vesicles directly obtained from patients contained novel parasite antigens 4. These results support further studies of reticulocyte exosomes from vivax infections to discover new antigens for vaccination and of human reticulocyte exosomes as a potential vaccine delivery platform for eliciting cytotoxic T-cell responses against vivax malaria.

Hypnozoite biomarker discovery

The team is using human liver-chimeric FRG-KO-(huHep) mice treated with schizonticidal drugs to determine the proteome composition of plasma-derived exosomes from animals exclusively infected with P. vivax hypnozoites. Moreover, their previous proteome analysis showed the presence of unique human proteins that are presently being used to highly enriched exclusively for exosomes coming from human hepatocytes; thus, increasing the signal for biomarker discovery. They will produce 3-5 unique parasite proteins and monoclonal antibodies against them to facilitate the construction of microfluidic devices with the ultimate goal of developing POC diagnostic device easily deployable in the field to contribute to the elimination of vivax malaria.

Functional analysis of subtelomeric variant genes

As infected reticulocytes with mature developmental stages of P. vivax are detected in the peripheral circulation, it has been amply accepted that there is no cytoadherence, hence no sequestration in this species. Against this dogma, this research group identified the largest subtelomeric multigene family of human malaria parasites, the vir multigene family with expressed VIR proteins, likely involved in vivax pathology through cytoadherence of (pRetics). Proof-of-principle of vivax pRetics cytoadherence partly mediated by VIR proteins was later demonstrated in vitro using endothelial, placental and brain cells, and more recently the group demonstrated that a P. falciparum transgenic line expressing a VIR protein was able to cytoadhere to human spleen fibroblasts. Its researchers are performing functional binding assay of variant surface proteins expressed in the human bone marrow and spleen during natural infections using CRISPR/cas-9 technologies to produce new transgenic lines of P. falciparum.

Extracellular vesicles (EVs) as potential new biomarkers in parasitic infections

The fact that EVs have been found in most biological fluids makes them an important target for the identification of new biomarkers in parasitic infections. In Chagas disease, the group is exploring EVs for identifying novel biomarkers specifically in the context of therapeutic response and disease progression during the chronic phase of the disease. In the field of Leishmaniasis, PvREX has recently started a collaborative project, planned to characterize EVs recovered from plasma in different clinical groups of human leishmaniasis (symptomatic, asymptomatic, treated). In parallel, the group also intends to evaluate the potential of extracellular vesicles as biomarkers for canine leishmaniasis. More recently, it has started a new collaborative project to identifying EV-Biomarkers in the context of severe P. falciparum malaria in children.

Active projects

Plasmodium vivax and Extracellular Vesicles Research Group (PVREX) recognised as a Consolidated Research Group (GRC)

PI: Carmen Fernández Becerra, Hernando A. Del Portillo
Funding agency: Agència de Gestió d'Ajuts Universitaris i de Recerca (AGAUR)
Agency code: 2021 SGR 01554 G65341695
Start date: 2023
End date: 2025

Novel organ-on-a-chip technology to study extracellular vesicles-mediated cryptic infections in Plasmodium vivax malaria

PI: Hernando A. Del Portillo
Funding agency: Fundación "laCaixa"
Agency code: LCF/PR/HR21/52410021
Start date: 2021
End date: 2024

CIBER en Enfermedades Infecciosas (CIBERINFEC)

PI: Joaquim Gascon
Funding agency: Instituto de Salud Carlos III (ISCIII)
Agency code: INFECG21/2022
Start date: 2021
End date: 2024

Uso de Vesículas extracelulares (EVs) como herramienta para la identificación de nuevos biomarcadores en la enfermedad de Chagas

PI: Carmen Fernández Becerra, Joaquim Gascon
Funding agency: Mundo Sano
Date: 2027

Extracellular Vesicles as Intercellular Communicators and Biomarkers of Cryptic Erythrocytic Infections in Plasmodium vivax malaria (VivaxEVTalk)

PIs: Carmen Fernández Becerra, Hernando A. Del Portillo
Funding agency: MICINN
Agency code: PID2022-142908OB-I00
Start date: 2023
End date: 2026

Past projects

Plasmodium vivax: a cryptic parasite challenging malaria elimination

PI: Carmen Fernández Becerra, Hernando A. Del Portillo
Funding agency: Ministerio de Ciencia e Innovación Español (MICINN)
Agency code: PID2019-111795RB-I00
Start date: 2020
End date: 2023

Extracellular vesicles as novel prognostic biomarkers for Chagas disease, an emerging chronic infection in Catalonia

PI: Carmen Fernández Becerra
Funding agency: La Marató de TV3
Agency code: 566/U/2018
Start date: 2019
End date: 2023

Scientific publications

Highlighted 2023 publications

Fernandez-Becerra C, Xander P, Alfandari D, Dong G, Aparici-Herraiz I, Rosenhek-Goldian I, Shokouhy M, Gualdron-Lopez M, Lozano N, Cortes-Serra N, Karam PA, Meneghetti P, Madeira RP, Porat Z, Soares RP, Costa AO, Rafati S, da Silva AC, Santarém N, Fernandez-Prada C, Ramirez MI, Bernal D, Marcilla A, Pereira-Chioccola VL, Ronalte Alves L, Del Portillo H, Regev-Rudzki N, Correia de Almeida I, Schenkman S, Olivier N, Torrecilhas AC. Guidelines for the Purification and Characterization of Extracellular Vesicles of Parasites. Journal of Extracellular Biology. 2023 Oct 19;2:e117. DOI: 10.1002/jex2.117

Esteves S, Lima C, Costa I, Osório H, Fernandez-Becerra C, Santarém N, Cordeiro-da-Silva A. Characterization and Proteomic Analysis of Plasma EVs Recovered from Healthy and Diseased Dogs with Canine Leishmaniosis. Int J Mol Sci. 2023 Mar 13;24(6):5490. DOI: 10.3390/ijms24065490.


Additional information

Particle Metrix (ZetaView®) equipment for studies on Extracellular Vesicles (EVs)

Thanks to the support of ISGlobal and IGTP, the Plasmodium vivax and Exosomes Research Group (PvREX), led by Carmen Fernández Becerra and Hernando A. del Portillo, has recently acquired a new Particle Metrix (ZetaView®) equipment for their studies on Extracellular Vesicles (EVs).

"Nanoparticle Tracking Analysis (NTA)" is a technique used to characterise nanoparticles (EVs, liposomes, VLPs) in suspension ranging between 10 and 2000 nm. This equipment, compared to more conventional ones, allows real-time visualisation of particles in suspension, thus calculating their size, dispersion, and fluorescence simultaneously. One of the new improvements the ZetaView® offers is the addition of the Zeta Potential, which enables the characterisation of the surface charge of nanoparticles.

The PvREX group, located at Germans Trias i Pujol Research Institute (IGTP) in Badalona, offers the use of this equipment as a laboratory-based service to all groups working on EVs and nanoparticles. This service will have a specialised technician responsible for performing the requested services.

Through the website ZetaView-ISGlobal, all relevant service requests can be made and all available prices can be consulted. For more information, do not hesitate to contact PvREX.

Doctoral theses

Title: Evaluation of Extracellular Vesicles Potential as Biomarkers for Canine Leishmaniasis
Author: Sofia Sampaio Esteves
Supervisors: Carmen Fernández Becerra (co-supervisor)
University: Universidade de Porto
Date of defence: 19/04/2023

Title: Extracellular vesicles as tool for the identification of new biomarkers in Chagas disease
Author: Núria Cortés Serra
Supervisors: Carmen Fernández Becerra
University: Universitat de Barcelona
Date of defence: 24/11/2022

Title: Extracellular vesicles as intercellular communicators in cryptic erythrocytic infections in Plasmodium vivax malaria
Author: Iris Aparici Herraiz
Supervisors: Hernando A. del Portillo, Carmen Fernández Becerra 
University: Universitat de Barcelona
Date of defence: 15/09/2022


- Investigación

Científicos muestran el papel clave del bazo y las vesículas extracelulares en las infecciones crípticas en la malaria

Un reciente estudio liderado por Carmen Fernández-Becerra y Hernando A del Portillo, destacados investigadores de ISGlobal y del IGTP, ha aportado nuevas perspectivas sobre el papel del bazo en la malaria, específicamente en las infecciones causadas por Plasmodium vivax.

- Investigación

Plasmodium vivax, un parásito de la malaria resistente a la eliminación

¿Por qué resulta tan difícil eliminar el parásito de la malaria Plasmodium vivax, el "olvidado"? El grupo de imnvestigación de IsGlobal e IGTP PvREX presenta datos científicos recientes.

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