Production of glycosylphosphatidylinositol-anchored proteins for vaccines and directed binding of immunoliposomes to specific cell types
Abstract Background: Liposomes are highly useful carriers for delivering drugs or antigens. The association of glycosylphosphatidylinositol (GPI)-anchored proteins to liposomes potentially enhances the immunogenic effect of vaccine antigens by increasing their surface concentration. Furthermore, the...
Published in: | Journal of Venomous Animals and Toxins including Tropical Diseases |
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ftdoajarticles:oai:doaj.org/article:8cc7df5e18554ca694313ca9b4c16326 2023-05-15T15:16:07+02:00 Production of glycosylphosphatidylinositol-anchored proteins for vaccines and directed binding of immunoliposomes to specific cell types Wesley L. Fotoran Nicole Kleiber Thomas Müntefering Eva Liebau Gerhard Wunderlich 2020-08-01T00:00:00Z https://doi.org/10.1590/1678-9199-jvatitd-2020-0032 https://doaj.org/article/8cc7df5e18554ca694313ca9b4c16326 EN eng SciELO http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1678-91992020000100323&tlng=en http://www.scielo.br/pdf/jvatitd/v26/1678-9199-jvatitd-26-e20200032.pdf https://doaj.org/toc/1678-9199 1678-9199 doi:10.1590/1678-9199-jvatitd-2020-0032 https://doaj.org/article/8cc7df5e18554ca694313ca9b4c16326 Journal of Venomous Animals and Toxins including Tropical Diseases, Vol 26 (2020) Plasmodium Liposome Immune targeting Vaccine Arctic medicine. Tropical medicine RC955-962 Toxicology. Poisons RA1190-1270 Zoology QL1-991 article 2020 ftdoajarticles https://doi.org/10.1590/1678-9199-jvatitd-2020-0032 2022-12-31T08:02:11Z Abstract Background: Liposomes are highly useful carriers for delivering drugs or antigens. The association of glycosylphosphatidylinositol (GPI)-anchored proteins to liposomes potentially enhances the immunogenic effect of vaccine antigens by increasing their surface concentration. Furthermore, the introduction of a universal immunoglobulin-binding domain can make liposomes targetable to virtually any desired receptor for which antibodies exist. Methods: We developed a system for the production of recombinant proteins with GPI anchors and histidine tags and Strep-tags for simplified purification from cells. This system was applied to i) the green fluorescent protein (GFP) as a reporter, ii) the promising Plasmodium falciparum vaccine antigen PfRH5 and iii) a doubled immunoglobulin Fc-binding domain termed ZZ from protein A of Staphylococcus aureus. As the GPI-attachment domain, the C-terminus of murine CD14 was used. After the recovery of these three recombinant proteins from Chinese hamster ovary (CHO) cells and association with liposomes, their vaccine potential and ability to target the CD4 receptor on lymphocytes in ex vivo conditions were tested. Results: Upon immunization in mice, the PfRH5-GPI-loaded liposomes generated antibody titers of 103 to 104, and showed a 45% inhibitory effect on in vitro growth at an IgG concentration of 600 µg/mL in P. falciparum cultures. Using GPI-anchored ZZ to couple anti-CD4 antibodies to liposomes, we created immunoliposomes with a binding efficiency of 75% to CD4+ cells in splenocytes and minimal off-target binding. Conclusions: Proteins are very effectively associated with liposomes via a GPI-anchor to form proteoliposome particles and these are useful for a variety of applications including vaccines and antibody-mediated targeting of liposomes. Importantly, the CHO-cell and GPI-tagged produced PfRH5 elicited invasion-blocking antibodies qualitatively comparable to other approaches. Article in Journal/Newspaper Arctic Directory of Open Access Journals: DOAJ Articles Arctic Journal of Venomous Animals and Toxins including Tropical Diseases 26 |
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Open Polar |
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Directory of Open Access Journals: DOAJ Articles |
op_collection_id |
ftdoajarticles |
language |
English |
topic |
Plasmodium Liposome Immune targeting Vaccine Arctic medicine. Tropical medicine RC955-962 Toxicology. Poisons RA1190-1270 Zoology QL1-991 |
spellingShingle |
Plasmodium Liposome Immune targeting Vaccine Arctic medicine. Tropical medicine RC955-962 Toxicology. Poisons RA1190-1270 Zoology QL1-991 Wesley L. Fotoran Nicole Kleiber Thomas Müntefering Eva Liebau Gerhard Wunderlich Production of glycosylphosphatidylinositol-anchored proteins for vaccines and directed binding of immunoliposomes to specific cell types |
topic_facet |
Plasmodium Liposome Immune targeting Vaccine Arctic medicine. Tropical medicine RC955-962 Toxicology. Poisons RA1190-1270 Zoology QL1-991 |
description |
Abstract Background: Liposomes are highly useful carriers for delivering drugs or antigens. The association of glycosylphosphatidylinositol (GPI)-anchored proteins to liposomes potentially enhances the immunogenic effect of vaccine antigens by increasing their surface concentration. Furthermore, the introduction of a universal immunoglobulin-binding domain can make liposomes targetable to virtually any desired receptor for which antibodies exist. Methods: We developed a system for the production of recombinant proteins with GPI anchors and histidine tags and Strep-tags for simplified purification from cells. This system was applied to i) the green fluorescent protein (GFP) as a reporter, ii) the promising Plasmodium falciparum vaccine antigen PfRH5 and iii) a doubled immunoglobulin Fc-binding domain termed ZZ from protein A of Staphylococcus aureus. As the GPI-attachment domain, the C-terminus of murine CD14 was used. After the recovery of these three recombinant proteins from Chinese hamster ovary (CHO) cells and association with liposomes, their vaccine potential and ability to target the CD4 receptor on lymphocytes in ex vivo conditions were tested. Results: Upon immunization in mice, the PfRH5-GPI-loaded liposomes generated antibody titers of 103 to 104, and showed a 45% inhibitory effect on in vitro growth at an IgG concentration of 600 µg/mL in P. falciparum cultures. Using GPI-anchored ZZ to couple anti-CD4 antibodies to liposomes, we created immunoliposomes with a binding efficiency of 75% to CD4+ cells in splenocytes and minimal off-target binding. Conclusions: Proteins are very effectively associated with liposomes via a GPI-anchor to form proteoliposome particles and these are useful for a variety of applications including vaccines and antibody-mediated targeting of liposomes. Importantly, the CHO-cell and GPI-tagged produced PfRH5 elicited invasion-blocking antibodies qualitatively comparable to other approaches. |
format |
Article in Journal/Newspaper |
author |
Wesley L. Fotoran Nicole Kleiber Thomas Müntefering Eva Liebau Gerhard Wunderlich |
author_facet |
Wesley L. Fotoran Nicole Kleiber Thomas Müntefering Eva Liebau Gerhard Wunderlich |
author_sort |
Wesley L. Fotoran |
title |
Production of glycosylphosphatidylinositol-anchored proteins for vaccines and directed binding of immunoliposomes to specific cell types |
title_short |
Production of glycosylphosphatidylinositol-anchored proteins for vaccines and directed binding of immunoliposomes to specific cell types |
title_full |
Production of glycosylphosphatidylinositol-anchored proteins for vaccines and directed binding of immunoliposomes to specific cell types |
title_fullStr |
Production of glycosylphosphatidylinositol-anchored proteins for vaccines and directed binding of immunoliposomes to specific cell types |
title_full_unstemmed |
Production of glycosylphosphatidylinositol-anchored proteins for vaccines and directed binding of immunoliposomes to specific cell types |
title_sort |
production of glycosylphosphatidylinositol-anchored proteins for vaccines and directed binding of immunoliposomes to specific cell types |
publisher |
SciELO |
publishDate |
2020 |
url |
https://doi.org/10.1590/1678-9199-jvatitd-2020-0032 https://doaj.org/article/8cc7df5e18554ca694313ca9b4c16326 |
geographic |
Arctic |
geographic_facet |
Arctic |
genre |
Arctic |
genre_facet |
Arctic |
op_source |
Journal of Venomous Animals and Toxins including Tropical Diseases, Vol 26 (2020) |
op_relation |
http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1678-91992020000100323&tlng=en http://www.scielo.br/pdf/jvatitd/v26/1678-9199-jvatitd-26-e20200032.pdf https://doaj.org/toc/1678-9199 1678-9199 doi:10.1590/1678-9199-jvatitd-2020-0032 https://doaj.org/article/8cc7df5e18554ca694313ca9b4c16326 |
op_doi |
https://doi.org/10.1590/1678-9199-jvatitd-2020-0032 |
container_title |
Journal of Venomous Animals and Toxins including Tropical Diseases |
container_volume |
26 |
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1766346424253939712 |