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GUIDELINES FOR THE PURIFICATION AND CHARACTERIZATION OF EXTRACELLULAR VESICLES OF PARASITES
extracellular vesicles
helminth
host-parasite interaction
infection
protocols
protozoan parasites
Author
Fernandez-Becerra, Carmen
Xander, Patrícia
Alfandari, Daniel
Dong, George
Aparici-Herraiz, Iris
Rosenhek-Goldian, Irit
Shokouhy, Mehrdad
Gualdron-Lopez, Melisa
Lozano, Nicholy
Cortes-Serra, Nuria
Abou Karam, Paula
Meneghetti, Paula
Madeira, Rafael Pedro
Porat, Ziv
Soares, Rodrigo Pedro Pinto
Costa, Adriana Oliveira
Rafati, Sima
ilva, Anabela-Cordeiro da S
Santarém, Nuno
Fernandez-Prada, Christopher
Ramirez, Marcel I.
Bernal, Dolores
Marcilla, Antonio
Pereira-Chioccola, Vera Lucia
Alves, Lysangela Ronalte
Portillo, Hernando Del
Regev-Rudzki, Neta
Almeida, Igor Correia de
Schenkman, Sergio
Olivier, Martin
Torrecilhas, Ana Claudia
Xander, Patrícia
Alfandari, Daniel
Dong, George
Aparici-Herraiz, Iris
Rosenhek-Goldian, Irit
Shokouhy, Mehrdad
Gualdron-Lopez, Melisa
Lozano, Nicholy
Cortes-Serra, Nuria
Abou Karam, Paula
Meneghetti, Paula
Madeira, Rafael Pedro
Porat, Ziv
Soares, Rodrigo Pedro Pinto
Costa, Adriana Oliveira
Rafati, Sima
ilva, Anabela-Cordeiro da S
Santarém, Nuno
Fernandez-Prada, Christopher
Ramirez, Marcel I.
Bernal, Dolores
Marcilla, Antonio
Pereira-Chioccola, Vera Lucia
Alves, Lysangela Ronalte
Portillo, Hernando Del
Regev-Rudzki, Neta
Almeida, Igor Correia de
Schenkman, Sergio
Olivier, Martin
Torrecilhas, Ana Claudia
Affilliation
ISGlobal, Barcelona Institute for Global Health. Hospital Clínic-Universitat de Barcelona. Barcelona, Spain/IGTP Institut d’Investigació Germans Trias i Pujol. Badalona. Spain/CIBERINFEC, ISCIII-CIBER de Enfermedades Infecciosas. Instituto de Salud Carlos III. Madrid, Spain
Universidade Federal de São Paulo. Instituto de Ciências Ambientais, Químicas e Farmacêuticas. Departamento de Ciências Farmacêuticas. Laboratório de Imunologia Celular e Bioquímica de Fungos e Protozoários. São Paulo, SP, Brazil
Department of Biomolecular Sciences. Weizmann Institute of Science. Rehovot, Israel
The Research Institute of the McGill University Health Centre. McGill University. Montréal, Québec, Canada
ISGlobal, Barcelona Institute for Global Health. Hospital Clínic-Universitat de Barcelona. Barcelona, Spain
ISGlobal, Barcelona Institute for Global Health. Hospital Clínic-Universitat de Barcelona. Barcelona, Spain
Department of Immunotherapy and Leishmania Vaccine Research. Pasteur Institute of Iran. Tehran, Iran
ISGlobal, Barcelona Institute for Global Health. Hospital Clínic-Universitat de Barcelona. Barcelona, Spain
Universidade Federal de São Paulo. Instituto de Ciências Ambientais, Químicas e Farmacêuticas. Departamento de Ciências Farmacêuticas. Laboratório de Imunologia Celular e Bioquímica de Fungos e Protozoários. São Paulo, SP, Brazil
ISGlobal, Barcelona Institute for Global Health. Hospital Clínic-Universitat de Barcelona. Barcelona, Spain
Department of Biomolecular Sciences. Weizmann Institute of Science. Rehovot, Israel
Universidade Federal de São Paulo. Instituto de Ciências Ambientais, Químicas e Farmacêuticas. Departamento de Ciências Farmacêuticas. Laboratório de Imunologia Celular e Bioquímica de Fungos e Protozoários. São Paulo, SP, Brazil
Universidade Federal de São Paulo. Instituto de Ciências Ambientais, Químicas e Farmacêuticas. Departamento de Ciências Farmacêuticas. Laboratório de Imunologia Celular e Bioquímica de Fungos e Protozoários. São Paulo, SP, Brazil
Flow Cytometry Unit. Life Sciences Core Facilities. WIS. Rehovot, Israel
Fundação Oswaldo Cruz. Instituto René Rachou. Belo Horizonte, MG, Brazil
Universidade Federal de Minas Gerais. Faculdade de Farmácia. Departamento de Análises Clínicas e Toxicológicas. Belo Horizonte, MG, Brasil
Department of Immunotherapy and Leishmania Vaccine Research. Pasteur Institute of Iran. Tehran, Iran
Host-Parasite Interactions Group. Institute of Research and Innovation in Health, University of Porto. Porto, Portugal/ Department of Biological Sciences. Faculty of Pharmacy. University of Porto. Porto, Portugal
Host-Parasite Interactions Group. Institute of Research and Innovation in Health, University of Porto. Porto, Portugal/ Department of Biological Sciences. Faculty of Pharmacy. University of Porto. Porto, Portugal
Faculty of Veterinary Medicine. Université de Montréal. Québec, Canada
Fundação Oswaldo Cruz. Instituto Carlos Chagas. Curitiba, PR, Brasil Laboratório de Biologia Molecular e Sistemática de Tripanossomatideos. Extracellular Vesicles and Host-Parasite Interactions Research Group.
Departament de Bioquímica i Biologia Molecular. Facultat de Ciències Biològiques. Universitat de València. Burjassot, Valencia, Spain
Àrea de Parasitologia. Departament de Farmàcia i Tecnologia Farmacèutica i Parasitologia. Universitat de València. Burjassot, Valencia, Spain
Instituto Adolfo Lutz. Centro de Parasitologia e Micologia Laboratório de Biologia Molecular de Parasitas e Fungos. São Paulo, SP, Brasil
Fundação Oswaldo Cruz. Instituto Carlos Chagas. Laboratório de Regulação da Expressão Gênica. Curitiba, PR, Brazil/Center in Infectious Diseases. Division of Infectious Disease and Immunity CHU de Quebec Research Center. Department of Microbiology, Infectious Disease and Immunology. Faculty of Medicine. University Laval. Quebec City, Quebec, Canada
ISGlobal, Barcelona Institute for Global Health. Hospital Clínic-Universitat de Barcelona. Barcelona, Spain/ IGTP Institut d’Investigació Germans Trias i Pujol. Badalona. Spain/ICREA Institució Catalana de Recerca i Estudis Avanc ̧ats. Barcelona, Spain
Department of Biomolecular Sciences. Weizmann Institute of Science. Rehovot, Israel
Department of Biological Sciences. Border Biomedical Research Center. The University of Texas at El Paso. El Paso, TX, USA
Universidade Federal de São Paulo. Departamento de Microbiologia, Imunologia e Parasitologia. São Paulo, SP, Brazil
The Research Institute of the McGill University Health Centre. McGill University. Montréal, Québec, Canada
Universidade Federal de São Paulo. Instituto de Ciências Ambientais, Químicas e Farmacêuticas. Departamento de Ciências Farmacêuticas. Laboratório de Imunologia Celular e Bioquímica de Fungos e Protozoários. São Paulo, SP, Brazil
Universidade Federal de São Paulo. Instituto de Ciências Ambientais, Químicas e Farmacêuticas. Departamento de Ciências Farmacêuticas. Laboratório de Imunologia Celular e Bioquímica de Fungos e Protozoários. São Paulo, SP, Brazil
Department of Biomolecular Sciences. Weizmann Institute of Science. Rehovot, Israel
The Research Institute of the McGill University Health Centre. McGill University. Montréal, Québec, Canada
ISGlobal, Barcelona Institute for Global Health. Hospital Clínic-Universitat de Barcelona. Barcelona, Spain
ISGlobal, Barcelona Institute for Global Health. Hospital Clínic-Universitat de Barcelona. Barcelona, Spain
Department of Immunotherapy and Leishmania Vaccine Research. Pasteur Institute of Iran. Tehran, Iran
ISGlobal, Barcelona Institute for Global Health. Hospital Clínic-Universitat de Barcelona. Barcelona, Spain
Universidade Federal de São Paulo. Instituto de Ciências Ambientais, Químicas e Farmacêuticas. Departamento de Ciências Farmacêuticas. Laboratório de Imunologia Celular e Bioquímica de Fungos e Protozoários. São Paulo, SP, Brazil
ISGlobal, Barcelona Institute for Global Health. Hospital Clínic-Universitat de Barcelona. Barcelona, Spain
Department of Biomolecular Sciences. Weizmann Institute of Science. Rehovot, Israel
Universidade Federal de São Paulo. Instituto de Ciências Ambientais, Químicas e Farmacêuticas. Departamento de Ciências Farmacêuticas. Laboratório de Imunologia Celular e Bioquímica de Fungos e Protozoários. São Paulo, SP, Brazil
Universidade Federal de São Paulo. Instituto de Ciências Ambientais, Químicas e Farmacêuticas. Departamento de Ciências Farmacêuticas. Laboratório de Imunologia Celular e Bioquímica de Fungos e Protozoários. São Paulo, SP, Brazil
Flow Cytometry Unit. Life Sciences Core Facilities. WIS. Rehovot, Israel
Fundação Oswaldo Cruz. Instituto René Rachou. Belo Horizonte, MG, Brazil
Universidade Federal de Minas Gerais. Faculdade de Farmácia. Departamento de Análises Clínicas e Toxicológicas. Belo Horizonte, MG, Brasil
Department of Immunotherapy and Leishmania Vaccine Research. Pasteur Institute of Iran. Tehran, Iran
Host-Parasite Interactions Group. Institute of Research and Innovation in Health, University of Porto. Porto, Portugal/ Department of Biological Sciences. Faculty of Pharmacy. University of Porto. Porto, Portugal
Host-Parasite Interactions Group. Institute of Research and Innovation in Health, University of Porto. Porto, Portugal/ Department of Biological Sciences. Faculty of Pharmacy. University of Porto. Porto, Portugal
Faculty of Veterinary Medicine. Université de Montréal. Québec, Canada
Fundação Oswaldo Cruz. Instituto Carlos Chagas. Curitiba, PR, Brasil Laboratório de Biologia Molecular e Sistemática de Tripanossomatideos. Extracellular Vesicles and Host-Parasite Interactions Research Group.
Departament de Bioquímica i Biologia Molecular. Facultat de Ciències Biològiques. Universitat de València. Burjassot, Valencia, Spain
Àrea de Parasitologia. Departament de Farmàcia i Tecnologia Farmacèutica i Parasitologia. Universitat de València. Burjassot, Valencia, Spain
Instituto Adolfo Lutz. Centro de Parasitologia e Micologia Laboratório de Biologia Molecular de Parasitas e Fungos. São Paulo, SP, Brasil
Fundação Oswaldo Cruz. Instituto Carlos Chagas. Laboratório de Regulação da Expressão Gênica. Curitiba, PR, Brazil/Center in Infectious Diseases. Division of Infectious Disease and Immunity CHU de Quebec Research Center. Department of Microbiology, Infectious Disease and Immunology. Faculty of Medicine. University Laval. Quebec City, Quebec, Canada
ISGlobal, Barcelona Institute for Global Health. Hospital Clínic-Universitat de Barcelona. Barcelona, Spain/ IGTP Institut d’Investigació Germans Trias i Pujol. Badalona. Spain/ICREA Institució Catalana de Recerca i Estudis Avanc ̧ats. Barcelona, Spain
Department of Biomolecular Sciences. Weizmann Institute of Science. Rehovot, Israel
Department of Biological Sciences. Border Biomedical Research Center. The University of Texas at El Paso. El Paso, TX, USA
Universidade Federal de São Paulo. Departamento de Microbiologia, Imunologia e Parasitologia. São Paulo, SP, Brazil
The Research Institute of the McGill University Health Centre. McGill University. Montréal, Québec, Canada
Universidade Federal de São Paulo. Instituto de Ciências Ambientais, Químicas e Farmacêuticas. Departamento de Ciências Farmacêuticas. Laboratório de Imunologia Celular e Bioquímica de Fungos e Protozoários. São Paulo, SP, Brazil
Abstract
Parasites are responsible for the most neglected tropical diseases, affecting over a billion people worldwide (WHO, 2015) and accounting for billions of cases a year and responsible for several millions of deaths. Research on extracellular vesicles (EVs) has increased in recent years and demonstrated that EVs shed by pathogenic parasites interact with host cells playing an important role in the parasite's survival, such as facilitation of infection, immunomodulation, parasite adaptation to the host environment and the transfer of drug resistance factors. Thus, EVs released by parasites mediate parasite-parasite and parasite-host intercellular communication. In addition, they are being explored as biomarkers of asymptomatic infections and disease prognosis after drug treatment. However, most current protocols used for the isolation, size determination, quantification and characterization of molecular cargo of EVs lack greater rigor, standardization, and adequate quality controls to certify the enrichment or purity of the ensuing bioproducts. We are now initiating major guidelines based on the evolution of collective knowledge in recent years. The main points covered in this position paper are methods for the isolation and molecular characterization of EVs obtained from parasite-infected cell cultures, experimental animals, and patients. The guideline also includes a discussion of suggested protocols and functional assays in host cells
Keywords
EVs methodologyextracellular vesicles
helminth
host-parasite interaction
infection
protocols
protozoan parasites
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