TY - JOUR
T1 - Pitting of malaria parasites in microfluidic devices mimicking spleen interendothelial slits
AU - Elizalde-Torrent, Aleix
AU - Trejo-Soto, Claudia
AU - Méndez-Mora, Lourdes
AU - Nicolau, Marc
AU - Ezama, Oihane
AU - Gualdrón-López, Melisa
AU - Fernández-Becerra, Carmen
AU - Alarcón, Tomás
AU - Hernández-Machado, Aurora
AU - del Portillo, Hernando A.
N1 - Funding Information:
We thank Lorena Martin-Jaular, Isabel Crespo and Marco Fernandez for helpful discussions on flow cytometry. A.E.-T. acknowledges support from the Ministry of Education, Culture and Sport under the program Formación de Profesorado Universitario (AP2012-2145). C. T. S acknowledges support from MINECO (Spain) under project FIS2016 78883 C2 1 P. L.M.-M. acknowledges support from AGAUR (Generalitatde Catalunya) under program Doctorat Industrial (2018 DI 068). MG-L is a postdoctoral fellow supported by ISGlobal (Spain). T.A. acknowledges support from the Ministry of Economy & Competitivity (MINECO) for funding awarded to the Barcelona Graduate School of Mathematics under the “Maria de Maeztu” programme, grant number MDM-2014-0445. T.A. has been partially funded by the CERCA Programme of the Generalitat de Catalunya. T.A. gratefully acknowledges the Spanish MINECO for funding under grants MTM2015-71509-C2-1-R A.H.-M. acknowledges support from Ministerio de Ciencia e Innovación (Spain) under project PID2019-106063GB-100 and AGAUR (Gener-alitat de Catalunya) under project 2017 SGR-1061.I. Work in the laboratory of Carmen Fernández-Becerra and Hernando A del Portillo is funded by the Ministerio Español de Economía y Competitividad (SAF2016 80655-R), by the Ministerio de Ciencia e Innovación (PID2019-111795RB-I00) and “la Caixa” Foundation (LCF/PR/ HR21/52410021). ISGlobal and IGTP are members of the CERCA Programme, Generalitat de Catalunya. CFB and HdP also acknowledge support from the Spanish Ministry of Science and Innovation through the Centro de Excelencia Severo Ochoa 2019-2023” Program (CEX2018-000806-S), and support from the Generalitat de Catalunya through the CERCA Program. This research is part of the ISGlobal’s Program on the Molecular Mechanisms of Malaria which is partially supported by the Fundación Ramón Areces.
Publisher Copyright:
© 2021, The Author(s).
PY - 2021/12
Y1 - 2021/12
N2 - The spleen is a hematopoietic organ that participates in cellular and humoral immunity. It also serves as a quality control mechanism for removing senescent and/or poorly deformable red blood cells (RBCs) from circulation. Pitting is a specialized process by which the spleen extracts particles, including malaria parasites, from within circulating RBCs during their passage through the interendothelial slits (IES) in the splenic cords. To study this physiological function in vitro, we have developed two microfluidic devices modeling the IES, according to the hypothesis that at a certain range of mechanical stress on the RBC, regulated through both slit size and blood flow, would force it undergo the pitting process without affecting the cell integrity. To prove its functionality in replicating pitting of malaria parasites, we have performed a characterization of P. falciparum-infected RBCs (P.f.-RBCs) after their passage through the devices, determining hemolysis and the proportion of once-infected RBCs (O-iRBCs), defined by the presence of a parasite antigen and absence of DAPI staining of parasite DNA using a flow cytometry-based approach. The passage of P.f.-RBCs through the devices at the physiological flow rate did not affect cell integrity and resulted in an increase of the frequency of O-iRBCs. Both microfluidic device models were capable to replicate the pitting of P.f.-RBCs ex vivo by means of mechanical constraints without cellular involvement, shedding new insights on the role of the spleen in the pathophysiology of malaria.
AB - The spleen is a hematopoietic organ that participates in cellular and humoral immunity. It also serves as a quality control mechanism for removing senescent and/or poorly deformable red blood cells (RBCs) from circulation. Pitting is a specialized process by which the spleen extracts particles, including malaria parasites, from within circulating RBCs during their passage through the interendothelial slits (IES) in the splenic cords. To study this physiological function in vitro, we have developed two microfluidic devices modeling the IES, according to the hypothesis that at a certain range of mechanical stress on the RBC, regulated through both slit size and blood flow, would force it undergo the pitting process without affecting the cell integrity. To prove its functionality in replicating pitting of malaria parasites, we have performed a characterization of P. falciparum-infected RBCs (P.f.-RBCs) after their passage through the devices, determining hemolysis and the proportion of once-infected RBCs (O-iRBCs), defined by the presence of a parasite antigen and absence of DAPI staining of parasite DNA using a flow cytometry-based approach. The passage of P.f.-RBCs through the devices at the physiological flow rate did not affect cell integrity and resulted in an increase of the frequency of O-iRBCs. Both microfluidic device models were capable to replicate the pitting of P.f.-RBCs ex vivo by means of mechanical constraints without cellular involvement, shedding new insights on the role of the spleen in the pathophysiology of malaria.
UR - http://www.scopus.com/inward/record.url?scp=85118932475&partnerID=8YFLogxK
U2 - 10.1038/s41598-021-01568-w
DO - 10.1038/s41598-021-01568-w
M3 - Article
AN - SCOPUS:85118932475
VL - 11
JO - Scientific Reports
JF - Scientific Reports
SN - 2045-2322
IS - 1
M1 - 22099
ER -