Multiparameter Flow Cytometry Analysis of the Human Spleen Applied to Studies of Plasma-Derived EVs From Plasmodium vivax Patients
The spleen is a secondary lymphoid organ with multiple functions including the removal of senescent red blood cells and the coordination of immune responses against blood-borne pathogens, such as malaria parasites. Despite the major role of the spleen, the study of its function in humans is limited...
Main Authors: | , , , , , , , , , |
---|---|
Format: | Article |
Language: | English |
Published: |
Frontiers Media S.A.
2021-03-01
|
Series: | Frontiers in Cellular and Infection Microbiology |
Subjects: | |
Online Access: | https://www.frontiersin.org/articles/10.3389/fcimb.2021.596104/full |
id |
doaj-99c92a404a64443d937957e715457495 |
---|---|
record_format |
Article |
spelling |
doaj-99c92a404a64443d937957e7154574952021-03-01T06:00:41ZengFrontiers Media S.A.Frontiers in Cellular and Infection Microbiology2235-29882021-03-011110.3389/fcimb.2021.596104596104Multiparameter Flow Cytometry Analysis of the Human Spleen Applied to Studies of Plasma-Derived EVs From Plasmodium vivax PatientsMelisa Gualdrón-López0Melisa Gualdrón-López1Míriam Díaz-Varela2Haruka Toda3Iris Aparici-Herraiz4Laura Pedró-Cos5Ricardo Lauzurica6Marcus V. G. Lacerda7Marcus V. G. Lacerda8Marco Antonio Fernández-Sanmartín9Carmen Fernandez-Becerra10Carmen Fernandez-Becerra11Hernando A. del Portillo12Hernando A. del Portillo13Hernando A. del Portillo14ISGlobal, Hospital Clinic - Universitat de Barcelona, Barcelona, SpainIGTP: Germans Trias i Pujol Research Institute, Barcelona, SpainISGlobal, Hospital Clinic - Universitat de Barcelona, Barcelona, SpainISGlobal, Hospital Clinic - Universitat de Barcelona, Barcelona, SpainISGlobal, Hospital Clinic - Universitat de Barcelona, Barcelona, SpainIGTP: Germans Trias i Pujol Research Institute, Barcelona, SpainNephrology Service, Germans Trias i Pujol University Hospital, Badalona, SpainFundaçao de Medicina Tropical Dr. Heitor Vieira Dourado, Manaus, BrazilInstituto Leônidas & Maria Deane (ILMD), Fiocruz, Manaus, BrazilIGTP: Germans Trias i Pujol Research Institute, Barcelona, SpainISGlobal, Hospital Clinic - Universitat de Barcelona, Barcelona, SpainIGTP: Germans Trias i Pujol Research Institute, Barcelona, SpainISGlobal, Hospital Clinic - Universitat de Barcelona, Barcelona, SpainIGTP: Germans Trias i Pujol Research Institute, Barcelona, SpainICREA: Catalan Institution for Research and Advanced Studies, Barcelona, SpainThe spleen is a secondary lymphoid organ with multiple functions including the removal of senescent red blood cells and the coordination of immune responses against blood-borne pathogens, such as malaria parasites. Despite the major role of the spleen, the study of its function in humans is limited by ethical implications to access human tissues. Here, we employed multiparameter flow cytometry combined with cell purification techniques to determine human spleen cell populations from transplantation donors. Spleen immuno-phenotyping showed that CD45+ cells included B (30%), CD4+ T (16%), CD8+ T (10%), NK (6%) and NKT (2%) lymphocytes. Myeloid cells comprised neutrophils (16%), monocytes (2%) and DCs (0.3%). Erythrocytes represented 70%, reticulocytes 0.7% and hematopoietic stem cells 0.02%. Extracellular vesicles (EVs) are membrane-bound nanoparticles involved in intercellular communication and secreted by almost all cell types. EVs play several roles in malaria that range from modulation of immune responses to vascular alterations. To investigate interactions of plasma-derived EVs from Plasmodium vivax infected patients (PvEVs) with human spleen cells, we used size-exclusion chromatography (SEC) to separate EVs from the bulk of soluble plasma proteins and stained isolated EVs with fluorescent lipophilic dyes. The integrated cellular analysis of the human spleen and the methodology employed here allowed in vitro interaction studies of human spleen cells and EVs that showed an increased proportion of T cells (CD4+ 3 fold and CD8+ 4 fold), monocytes (1.51 fold), B cells (2.3 fold) and erythrocytes (3 fold) interacting with PvEVs as compared to plasma-derived EVs from healthy volunteers (hEVs). Future functional studies of these interactions can contribute to unveil pathophysiological processes involving the spleen in vivax malaria.https://www.frontiersin.org/articles/10.3389/fcimb.2021.596104/fullPlasmodium vivaxhuman spleenextracellular vesiclesmultiparameter flow cytometryinteraction |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Melisa Gualdrón-López Melisa Gualdrón-López Míriam Díaz-Varela Haruka Toda Iris Aparici-Herraiz Laura Pedró-Cos Ricardo Lauzurica Marcus V. G. Lacerda Marcus V. G. Lacerda Marco Antonio Fernández-Sanmartín Carmen Fernandez-Becerra Carmen Fernandez-Becerra Hernando A. del Portillo Hernando A. del Portillo Hernando A. del Portillo |
spellingShingle |
Melisa Gualdrón-López Melisa Gualdrón-López Míriam Díaz-Varela Haruka Toda Iris Aparici-Herraiz Laura Pedró-Cos Ricardo Lauzurica Marcus V. G. Lacerda Marcus V. G. Lacerda Marco Antonio Fernández-Sanmartín Carmen Fernandez-Becerra Carmen Fernandez-Becerra Hernando A. del Portillo Hernando A. del Portillo Hernando A. del Portillo Multiparameter Flow Cytometry Analysis of the Human Spleen Applied to Studies of Plasma-Derived EVs From Plasmodium vivax Patients Frontiers in Cellular and Infection Microbiology Plasmodium vivax human spleen extracellular vesicles multiparameter flow cytometry interaction |
author_facet |
Melisa Gualdrón-López Melisa Gualdrón-López Míriam Díaz-Varela Haruka Toda Iris Aparici-Herraiz Laura Pedró-Cos Ricardo Lauzurica Marcus V. G. Lacerda Marcus V. G. Lacerda Marco Antonio Fernández-Sanmartín Carmen Fernandez-Becerra Carmen Fernandez-Becerra Hernando A. del Portillo Hernando A. del Portillo Hernando A. del Portillo |
author_sort |
Melisa Gualdrón-López |
title |
Multiparameter Flow Cytometry Analysis of the Human Spleen Applied to Studies of Plasma-Derived EVs From Plasmodium vivax Patients |
title_short |
Multiparameter Flow Cytometry Analysis of the Human Spleen Applied to Studies of Plasma-Derived EVs From Plasmodium vivax Patients |
title_full |
Multiparameter Flow Cytometry Analysis of the Human Spleen Applied to Studies of Plasma-Derived EVs From Plasmodium vivax Patients |
title_fullStr |
Multiparameter Flow Cytometry Analysis of the Human Spleen Applied to Studies of Plasma-Derived EVs From Plasmodium vivax Patients |
title_full_unstemmed |
Multiparameter Flow Cytometry Analysis of the Human Spleen Applied to Studies of Plasma-Derived EVs From Plasmodium vivax Patients |
title_sort |
multiparameter flow cytometry analysis of the human spleen applied to studies of plasma-derived evs from plasmodium vivax patients |
publisher |
Frontiers Media S.A. |
series |
Frontiers in Cellular and Infection Microbiology |
issn |
2235-2988 |
publishDate |
2021-03-01 |
description |
The spleen is a secondary lymphoid organ with multiple functions including the removal of senescent red blood cells and the coordination of immune responses against blood-borne pathogens, such as malaria parasites. Despite the major role of the spleen, the study of its function in humans is limited by ethical implications to access human tissues. Here, we employed multiparameter flow cytometry combined with cell purification techniques to determine human spleen cell populations from transplantation donors. Spleen immuno-phenotyping showed that CD45+ cells included B (30%), CD4+ T (16%), CD8+ T (10%), NK (6%) and NKT (2%) lymphocytes. Myeloid cells comprised neutrophils (16%), monocytes (2%) and DCs (0.3%). Erythrocytes represented 70%, reticulocytes 0.7% and hematopoietic stem cells 0.02%. Extracellular vesicles (EVs) are membrane-bound nanoparticles involved in intercellular communication and secreted by almost all cell types. EVs play several roles in malaria that range from modulation of immune responses to vascular alterations. To investigate interactions of plasma-derived EVs from Plasmodium vivax infected patients (PvEVs) with human spleen cells, we used size-exclusion chromatography (SEC) to separate EVs from the bulk of soluble plasma proteins and stained isolated EVs with fluorescent lipophilic dyes. The integrated cellular analysis of the human spleen and the methodology employed here allowed in vitro interaction studies of human spleen cells and EVs that showed an increased proportion of T cells (CD4+ 3 fold and CD8+ 4 fold), monocytes (1.51 fold), B cells (2.3 fold) and erythrocytes (3 fold) interacting with PvEVs as compared to plasma-derived EVs from healthy volunteers (hEVs). Future functional studies of these interactions can contribute to unveil pathophysiological processes involving the spleen in vivax malaria. |
topic |
Plasmodium vivax human spleen extracellular vesicles multiparameter flow cytometry interaction |
url |
https://www.frontiersin.org/articles/10.3389/fcimb.2021.596104/full |
work_keys_str_mv |
AT melisagualdronlopez multiparameterflowcytometryanalysisofthehumanspleenappliedtostudiesofplasmaderivedevsfromplasmodiumvivaxpatients AT melisagualdronlopez multiparameterflowcytometryanalysisofthehumanspleenappliedtostudiesofplasmaderivedevsfromplasmodiumvivaxpatients AT miriamdiazvarela multiparameterflowcytometryanalysisofthehumanspleenappliedtostudiesofplasmaderivedevsfromplasmodiumvivaxpatients AT harukatoda multiparameterflowcytometryanalysisofthehumanspleenappliedtostudiesofplasmaderivedevsfromplasmodiumvivaxpatients AT irisapariciherraiz multiparameterflowcytometryanalysisofthehumanspleenappliedtostudiesofplasmaderivedevsfromplasmodiumvivaxpatients AT laurapedrocos multiparameterflowcytometryanalysisofthehumanspleenappliedtostudiesofplasmaderivedevsfromplasmodiumvivaxpatients AT ricardolauzurica multiparameterflowcytometryanalysisofthehumanspleenappliedtostudiesofplasmaderivedevsfromplasmodiumvivaxpatients AT marcusvglacerda multiparameterflowcytometryanalysisofthehumanspleenappliedtostudiesofplasmaderivedevsfromplasmodiumvivaxpatients AT marcusvglacerda multiparameterflowcytometryanalysisofthehumanspleenappliedtostudiesofplasmaderivedevsfromplasmodiumvivaxpatients AT marcoantoniofernandezsanmartin multiparameterflowcytometryanalysisofthehumanspleenappliedtostudiesofplasmaderivedevsfromplasmodiumvivaxpatients AT carmenfernandezbecerra multiparameterflowcytometryanalysisofthehumanspleenappliedtostudiesofplasmaderivedevsfromplasmodiumvivaxpatients AT carmenfernandezbecerra multiparameterflowcytometryanalysisofthehumanspleenappliedtostudiesofplasmaderivedevsfromplasmodiumvivaxpatients AT hernandoadelportillo multiparameterflowcytometryanalysisofthehumanspleenappliedtostudiesofplasmaderivedevsfromplasmodiumvivaxpatients AT hernandoadelportillo multiparameterflowcytometryanalysisofthehumanspleenappliedtostudiesofplasmaderivedevsfromplasmodiumvivaxpatients AT hernandoadelportillo multiparameterflowcytometryanalysisofthehumanspleenappliedtostudiesofplasmaderivedevsfromplasmodiumvivaxpatients |
_version_ |
1724246943018778624 |