Immunofluorescence study of cytoskeleton in endothelial cells induced with malaria sera
<p><strong>Background:</strong> Endothelial cells (ECs) play a major role in malaria pathogenesis, as a point of direct contact of parasitized red blood cells to the blood vessel wall. The study of cytoskeleton structures of ECs, whose main functions are to maintain shape...
| Main Authors: | , , , , , , , , , |
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| Format: | Journal article |
| Language: | English |
| Published: |
BioMed Central
2024
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| _version_ | 1797113207733616640 |
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| author | Wongsawat, M Glaharn, S Srisook, C Dechkhajorn, W Chaisri, U Punsawad, C Techarang, T Chotivanich, K Krudsood, S Viriyavejakul, P |
| author_facet | Wongsawat, M Glaharn, S Srisook, C Dechkhajorn, W Chaisri, U Punsawad, C Techarang, T Chotivanich, K Krudsood, S Viriyavejakul, P |
| author_sort | Wongsawat, M |
| collection | OXFORD |
| description | <p><strong>Background:</strong> Endothelial cells (ECs) play a major role in malaria pathogenesis, as a point of direct contact of parasitized red blood cells to the blood vessel wall. The study of cytoskeleton structures of ECs, whose main functions are to maintain shape and provide strength to the EC membrane is important in determining the severe sequelae of <em>Plasmodium falciparum</em> malaria. The work investigated the cytoskeletal changes (microfilaments-actin, microtubules-tubulin and intermediate filaments-vimentin) in ECs induced by malaria sera (<em>Plasmodium vivax</em>, uncomplicated <em>P. falciparum</em> and complicated <em>P. falciparum</em>), in relation to the levels of pro-inflammatory cytokines.</p>
<p><strong>Methods:</strong> Morphology and fluorescence intensity of EC cytoskeleton stimulated with malaria sera were evaluated using immunofluorescence technique. Levels of tumour necrosis factor (TNF) and interferon (IFN)-gamma (γ) were determined using enzyme-linked immunosorbent assay (ELISA). Control experimental groups included ECs incubated with media alone and non-malaria patient sera. Experimental groups consisted of ECs incubated with malaria sera from <em>P. vivax</em>, uncomplicated <em>P. falciparum</em> and complicated <em>P. falciparum</em>. Morphological scores of cytoskeletal alterations and fluorescence intensity were compared across each experiment group, and correlated with TNF and IFN-γ.</p>
<p><strong>Results:</strong> The four morphological changes of cytoskeleton included (1) shrinkage of cytoskeleton and ECs with cortical condensation, (2) appearance of eccentric nuclei, (3) presence of “spiking pattern” of cytoskeleton and EC membrane, and (4) fragmentation and discontinuity of cytoskeleton and ECs. Significant damages were noted in actin filaments compared to tubulin and vimentin filaments in ECs stimulated with sera from complicated <em>P. falciparum</em> malaria. Morphological damages to cytoskeleton was positively correlated with fluorescence intensity and the levels of TNF and IFN-γ.</p>
<p><strong>Conclusions:</strong> ECs stimulated with sera from complicated <em>P. falciparum</em> malaria showed cytoskeletal alterations and increased in fluorescence intensity, which was associated with high levels of TNF and IFN-γ. Cytoskeletal changes of ECs incubated with complicated <em>P. falciparum</em> malaria sera can lead to EC junctional alteration and permeability changes, which is mediated through apoptotic pathway. The findings can serve as a basis to explore measures to strengthen EC cytoskeleton and alleviate severe malaria complications such as pulmonary oedema and cerebral malaria. In addition, immunofluorescence intensity of cytoskeleton could be investigated as potential prognostic indicator for malaria severity.</p> |
| first_indexed | 2024-04-23T08:25:09Z |
| format | Journal article |
| id | oxford-uuid:25393ba1-c8f5-4a0d-8adb-bdf2cbd3236a |
| institution | University of Oxford |
| language | English |
| last_indexed | 2024-04-23T08:25:09Z |
| publishDate | 2024 |
| publisher | BioMed Central |
| record_format | dspace |
| spelling | oxford-uuid:25393ba1-c8f5-4a0d-8adb-bdf2cbd3236a2024-04-16T06:56:34ZImmunofluorescence study of cytoskeleton in endothelial cells induced with malaria seraJournal articlehttp://purl.org/coar/resource_type/c_dcae04bcuuid:25393ba1-c8f5-4a0d-8adb-bdf2cbd3236aEnglishSymplectic ElementsBioMed Central2024Wongsawat, MGlaharn, SSrisook, CDechkhajorn, WChaisri, UPunsawad, CTecharang, TChotivanich, KKrudsood, SViriyavejakul, P<p><strong>Background:</strong> Endothelial cells (ECs) play a major role in malaria pathogenesis, as a point of direct contact of parasitized red blood cells to the blood vessel wall. The study of cytoskeleton structures of ECs, whose main functions are to maintain shape and provide strength to the EC membrane is important in determining the severe sequelae of <em>Plasmodium falciparum</em> malaria. The work investigated the cytoskeletal changes (microfilaments-actin, microtubules-tubulin and intermediate filaments-vimentin) in ECs induced by malaria sera (<em>Plasmodium vivax</em>, uncomplicated <em>P. falciparum</em> and complicated <em>P. falciparum</em>), in relation to the levels of pro-inflammatory cytokines.</p> <p><strong>Methods:</strong> Morphology and fluorescence intensity of EC cytoskeleton stimulated with malaria sera were evaluated using immunofluorescence technique. Levels of tumour necrosis factor (TNF) and interferon (IFN)-gamma (γ) were determined using enzyme-linked immunosorbent assay (ELISA). Control experimental groups included ECs incubated with media alone and non-malaria patient sera. Experimental groups consisted of ECs incubated with malaria sera from <em>P. vivax</em>, uncomplicated <em>P. falciparum</em> and complicated <em>P. falciparum</em>. Morphological scores of cytoskeletal alterations and fluorescence intensity were compared across each experiment group, and correlated with TNF and IFN-γ.</p> <p><strong>Results:</strong> The four morphological changes of cytoskeleton included (1) shrinkage of cytoskeleton and ECs with cortical condensation, (2) appearance of eccentric nuclei, (3) presence of “spiking pattern” of cytoskeleton and EC membrane, and (4) fragmentation and discontinuity of cytoskeleton and ECs. Significant damages were noted in actin filaments compared to tubulin and vimentin filaments in ECs stimulated with sera from complicated <em>P. falciparum</em> malaria. Morphological damages to cytoskeleton was positively correlated with fluorescence intensity and the levels of TNF and IFN-γ.</p> <p><strong>Conclusions:</strong> ECs stimulated with sera from complicated <em>P. falciparum</em> malaria showed cytoskeletal alterations and increased in fluorescence intensity, which was associated with high levels of TNF and IFN-γ. Cytoskeletal changes of ECs incubated with complicated <em>P. falciparum</em> malaria sera can lead to EC junctional alteration and permeability changes, which is mediated through apoptotic pathway. The findings can serve as a basis to explore measures to strengthen EC cytoskeleton and alleviate severe malaria complications such as pulmonary oedema and cerebral malaria. In addition, immunofluorescence intensity of cytoskeleton could be investigated as potential prognostic indicator for malaria severity.</p> |
| spellingShingle | Wongsawat, M Glaharn, S Srisook, C Dechkhajorn, W Chaisri, U Punsawad, C Techarang, T Chotivanich, K Krudsood, S Viriyavejakul, P Immunofluorescence study of cytoskeleton in endothelial cells induced with malaria sera |
| title | Immunofluorescence study of cytoskeleton in endothelial cells induced with malaria sera |
| title_full | Immunofluorescence study of cytoskeleton in endothelial cells induced with malaria sera |
| title_fullStr | Immunofluorescence study of cytoskeleton in endothelial cells induced with malaria sera |
| title_full_unstemmed | Immunofluorescence study of cytoskeleton in endothelial cells induced with malaria sera |
| title_short | Immunofluorescence study of cytoskeleton in endothelial cells induced with malaria sera |
| title_sort | immunofluorescence study of cytoskeleton in endothelial cells induced with malaria sera |
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