A method for visualizing surface-exposed and internal PfEMP1 adhesion antigens in <it>Plasmodium falciparum </it>infected erythrocytes
<p>Abstract</p> <p>Background</p> <p>The insertion of parasite antigens into the host erythrocyte membrane and the structure and distribution of <it>Plasmodium falciparum </it>adhesion receptors on that membrane are poorly understood. Laser scanning confocal...
| Main Authors: | , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
BMC
2008-06-01
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| Series: | Malaria Journal |
| Online Access: | http://www.malariajournal.com/content/7/1/101 |
| _version_ | 1818565822418780160 |
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| author | Arnot David E Theander Thor G Turner Louise Joergensen Louise Jensen Anja TR Salanti Ali Sowa Kordai M Bengtsson Dominique |
| author_facet | Arnot David E Theander Thor G Turner Louise Joergensen Louise Jensen Anja TR Salanti Ali Sowa Kordai M Bengtsson Dominique |
| author_sort | Arnot David E |
| collection | DOAJ |
| description | <p>Abstract</p> <p>Background</p> <p>The insertion of parasite antigens into the host erythrocyte membrane and the structure and distribution of <it>Plasmodium falciparum </it>adhesion receptors on that membrane are poorly understood. Laser scanning confocal microscopy (LSCM) and a novel labelling and fixation method have been used to obtain high resolution immuno-fluorescent images of erythrocyte surface PfEMP1 and internal antigens which allow analysis of the accumulation of PfEMP1 on the erythrocyte membrane during asexual development.</p> <p>Methods</p> <p>A novel staining technique has been developed which permits distinction between erythrocyte surface PfEMP1 and intracellular PfEMP1, in parasites whose nuclear material is exceptionally well resolved. Primary antibody detection by fluorescence is carried out on the live parasitized erythrocyte. The surface labelled cells are then fixed using paraformaldehyde and permeabilized with a non-ionic detergent to permit access of antibodies to internal parasite antigens. Differentiation between surface and internal antigens is achieved using antibodies labelled with different fluorochromes and confocal microscopy</p> <p>Results</p> <p>Surface exposed PfEMP1 is first detectable by antibodies at the trophozoite stage of intracellular parasite development although the improved detection method indicates that there are differences between different laboratory isolates in the kinetics of accumulation of surface-exposed PfEMP1.</p> <p>Conclusion</p> <p>A sensitive method for labelling surface and internal PfEMP1 with up to three different fluorochromes has been developed for laser scanning confocal optical microscopy and the analysis of the developmental expression of malaria adhesion antigens.</p> |
| first_indexed | 2024-12-14T01:45:52Z |
| format | Article |
| id | doaj.art-bc82b131763442dfa562ad17192101a5 |
| institution | Directory Open Access Journal |
| issn | 1475-2875 |
| language | English |
| last_indexed | 2024-12-14T01:45:52Z |
| publishDate | 2008-06-01 |
| publisher | BMC |
| record_format | Article |
| series | Malaria Journal |
| spelling | doaj.art-bc82b131763442dfa562ad17192101a52022-12-21T23:21:34ZengBMCMalaria Journal1475-28752008-06-017110110.1186/1475-2875-7-101A method for visualizing surface-exposed and internal PfEMP1 adhesion antigens in <it>Plasmodium falciparum </it>infected erythrocytesArnot David ETheander Thor GTurner LouiseJoergensen LouiseJensen Anja TRSalanti AliSowa Kordai MBengtsson Dominique<p>Abstract</p> <p>Background</p> <p>The insertion of parasite antigens into the host erythrocyte membrane and the structure and distribution of <it>Plasmodium falciparum </it>adhesion receptors on that membrane are poorly understood. Laser scanning confocal microscopy (LSCM) and a novel labelling and fixation method have been used to obtain high resolution immuno-fluorescent images of erythrocyte surface PfEMP1 and internal antigens which allow analysis of the accumulation of PfEMP1 on the erythrocyte membrane during asexual development.</p> <p>Methods</p> <p>A novel staining technique has been developed which permits distinction between erythrocyte surface PfEMP1 and intracellular PfEMP1, in parasites whose nuclear material is exceptionally well resolved. Primary antibody detection by fluorescence is carried out on the live parasitized erythrocyte. The surface labelled cells are then fixed using paraformaldehyde and permeabilized with a non-ionic detergent to permit access of antibodies to internal parasite antigens. Differentiation between surface and internal antigens is achieved using antibodies labelled with different fluorochromes and confocal microscopy</p> <p>Results</p> <p>Surface exposed PfEMP1 is first detectable by antibodies at the trophozoite stage of intracellular parasite development although the improved detection method indicates that there are differences between different laboratory isolates in the kinetics of accumulation of surface-exposed PfEMP1.</p> <p>Conclusion</p> <p>A sensitive method for labelling surface and internal PfEMP1 with up to three different fluorochromes has been developed for laser scanning confocal optical microscopy and the analysis of the developmental expression of malaria adhesion antigens.</p>http://www.malariajournal.com/content/7/1/101 |
| spellingShingle | Arnot David E Theander Thor G Turner Louise Joergensen Louise Jensen Anja TR Salanti Ali Sowa Kordai M Bengtsson Dominique A method for visualizing surface-exposed and internal PfEMP1 adhesion antigens in <it>Plasmodium falciparum </it>infected erythrocytes Malaria Journal |
| title | A method for visualizing surface-exposed and internal PfEMP1 adhesion antigens in <it>Plasmodium falciparum </it>infected erythrocytes |
| title_full | A method for visualizing surface-exposed and internal PfEMP1 adhesion antigens in <it>Plasmodium falciparum </it>infected erythrocytes |
| title_fullStr | A method for visualizing surface-exposed and internal PfEMP1 adhesion antigens in <it>Plasmodium falciparum </it>infected erythrocytes |
| title_full_unstemmed | A method for visualizing surface-exposed and internal PfEMP1 adhesion antigens in <it>Plasmodium falciparum </it>infected erythrocytes |
| title_short | A method for visualizing surface-exposed and internal PfEMP1 adhesion antigens in <it>Plasmodium falciparum </it>infected erythrocytes |
| title_sort | method for visualizing surface exposed and internal pfemp1 adhesion antigens in it plasmodium falciparum it infected erythrocytes |
| url | http://www.malariajournal.com/content/7/1/101 |
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