Microencapsulation of inorganic nanocrystals into PLGA microsphere vaccines enables their intracellular localization in dendritic cells by electron and fluorescence microscopy
Biodegradable poly-(D,L-lactide-co-glycolide) microspheres (PLGA-MS) are approved as a drug delivery system in humans and represent a promising antigen delivery device for immunotherapy against cancer. Immune responses following PLGA-MS vaccination require cross-presentation of encapsulated antigen...
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Published in | Journal of controlled release Vol. 151; no. 3; pp. 278 - 285 |
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Main Authors | , , , , , , |
Format | Journal Article |
Language | English |
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10.05.2011
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Abstract | Biodegradable poly-(D,L-lactide-co-glycolide) microspheres (PLGA-MS) are approved as a drug delivery system in humans and represent a promising antigen delivery device for immunotherapy against cancer. Immune responses following PLGA-MS vaccination require cross-presentation of encapsulated antigen by professional antigen presenting cells (APCs). While the potential of PLGA-MS as vaccine formulations is well established, the intracellular pathway of cross-presentation following phagocytosis of PLGA-MS is still under debate. A part of the controversy stems from the difficulty in unambiguously identifying PLGA-MS within cells. Here we show a novel strategy for the efficient encapsulation of inorganic nanocrystals (NCs) into PLGA-MS as a tool to study their intracellular localization. We microencapsulated NCs as an electron dense marker to study the intracellular localization of PLGA-MS by transmission electron microscopy (TEM) and as fluorescent labels for confocal laser scanning microscopy. Using this method, we found PLGA-MS to be rapidly taken up by dendritic cells and macrophages. Co-localization with the lysosomal marker LAMP1 showed a lysosomal storage of PLGA-MS for over two days after uptake, long after the initiation of cross-presentation had occurred. Our data argue against an escape of PLGA-MS from the endosome as has previously been suggested as a mechanism to facilitate cross-presentation of PLGA-MS encapsulated antigen.
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AbstractList | Biodegradable poly-(D,L-lactide-co-glycolide) microspheres (PLGA-MS) are approved as a drug delivery system in humans and represent a promising antigen delivery device for immunotherapy against cancer. Immune responses following PLGA-MS vaccination require cross-presentation of encapsulated antigen by professional antigen presenting cells (APCs). While the potential of PLGA-MS as vaccine formulations is well established, the intracellular pathway of cross-presentation following phagocytosis of PLGA-MS is still under debate. A part of the controversy stems from the difficulty in unambiguously identifying PLGA-MS within cells. Here we show a novel strategy for the efficient encapsulation of inorganic nanocrystals (NCs) into PLGA-MS as a tool to study their intracellular localization. We microencapsulated NCs as an electron dense marker to study the intracellular localization of PLGA-MS by transmission electron microscopy (TEM) and as fluorescent labels for confocal laser scanning microscopy. Using this method, we found PLGA-MS to be rapidly taken up by dendritic cells and macrophages. Co-localization with the lysosomal marker LAMP1 showed a lysosomal storage of PLGA-MS for over two days after uptake, long after the initiation of cross-presentation had occurred. Our data argue against an escape of PLGA-MS from the endosome as has previously been suggested as a mechanism to facilitate cross-presentation of PLGA-MS encapsulated antigen.
[Display omitted] Biodegradable poly-(D,L-lactide-co-glycolide) microspheres (PLGA-MS) are approved as a drug delivery system in humans and represent a promising antigen delivery device for immunotherapy against cancer. Immune responses following PLGA-MS vaccination require cross-presentation of encapsulated antigen by professional antigen presenting cells (APCs). While the potential of PLGA-MS as vaccine formulations is well established, the intracellular pathway of cross-presentation following phagocytosis of PLGA-MS is still under debate. A part of the controversy stems from the difficulty in unambiguously identifying PLGA-MS within cells. Here we show a novel strategy for the efficient encapsulation of inorganic nanocrystals (NCs) into PLGA-MS as a tool to study their intracellular localization. We microencapsulated NCs as an electron dense marker to study the intracellular localization of PLGA-MS by transmission electron microscopy (TEM) and as fluorescent labels for confocal laser scanning microscopy. Using this method, we found PLGA-MS to be rapidly taken up by dendritic cells and macrophages. Co-localization with the lysosomal marker LAMP1 showed a lysosomal storage of PLGA-MS for over two days after uptake, long after the initiation of cross-presentation had occurred. Our data argue against an escape of PLGA-MS from the endosome as has previously been suggested as a mechanism to facilitate cross-presentation of PLGA-MS encapsulated antigen. Biodegradable poly-(D,L-lactide-co-glycolide) microspheres (PLGA-MS) are approved as a drug delivery system in humans and represent a promising antigen delivery device for immunotherapy against cancer. Immune responses following PLGA-MS vaccination require cross-presentation of encapsulated antigen by professional antigen presenting cells (APCs). While the potential of PLGA-MS as vaccine formulations is well established, the intracellular pathway of cross-presentation following phagocytosis of PLGA-MS is still under debate. A part of the controversy stems from the difficulty in unambiguously identifying PLGA-MS within cells. Here we show a novel strategy for the efficient encapsulation of inorganic nanocrystals (NCs) into PLGA-MS as a tool to study their intracellular localization. We microencapsulated NCs as an electron dense marker to study the intracellular localization of PLGA-MS by transmission electron microscopy (TEM) and as fluorescent labels for confocal laser scanning microscopy. Using this method, we found PLGA-MS to be rapidly taken up by dendritic cells and macrophages. Co-localization with the lysosomal marker LAMP1 showed a lysosomal storage of PLGA-MS for over two days after uptake, long after the initiation of cross-presentation had occurred. Our data argue against an escape of PLGA-MS from the endosome as has previously been suggested as a mechanism to facilitate cross-presentation of PLGA-MS encapsulated antigen. AB: |
Author | Thiry, Marc Tromsdorf, Ulrich I. Groettrup, Marcus Hentschel, Joachim Schliehe, Christopher Schliehe, Constanze Weller, Horst |
Author_xml | – sequence: 1 givenname: Christopher surname: Schliehe fullname: Schliehe, Christopher organization: Division of Immunology, Department of Biology, Constance University, 78457 Konstanz, Germany – sequence: 2 givenname: Constanze surname: Schliehe fullname: Schliehe, Constanze organization: Institute of Physical Chemistry, Hamburg University, 20146 Hamburg, Germany – sequence: 3 givenname: Marc surname: Thiry fullname: Thiry, Marc organization: Institute of Physical Chemistry, Hamburg University, 20146 Hamburg, Germany – sequence: 4 givenname: Ulrich I. surname: Tromsdorf fullname: Tromsdorf, Ulrich I. organization: Institute of Physical Chemistry, Hamburg University, 20146 Hamburg, Germany – sequence: 5 givenname: Joachim surname: Hentschel fullname: Hentschel, Joachim organization: Electron Microscopy Facility, Department of Biology, Constance University, 78457 Konstanz, Germany – sequence: 6 givenname: Horst surname: Weller fullname: Weller, Horst organization: Institute of Physical Chemistry, Hamburg University, 20146 Hamburg, Germany – sequence: 7 givenname: Marcus surname: Groettrup fullname: Groettrup, Marcus email: Marcus.Groettrup@uni-konstanz.de organization: Division of Immunology, Department of Biology, Constance University, 78457 Konstanz, Germany |
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Keywords | Biodegradation Microencapsulation Immunostimulation Microsphere Antigenicity Polylactic acid Dendritic cell Pharmaceutical technology Biodegradability Vaccine Glycolic acid polymer Electron microscopy Lactone polymer Lactic acid polymer Aliphatic polymer Microparticle Copolymer Fluorescence microscopy Nanocrystal |
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Snippet | Biodegradable poly-(D,L-lactide-co-glycolide) microspheres (PLGA-MS) are approved as a drug delivery system in humans and represent a promising antigen... |
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SubjectTerms | Animals Antigenicity Biodegradation Biological and medical sciences Biological Transport Cells, Cultured Cross-Priming - immunology Dendritic Cells - metabolism Dendritic Cells - ultrastructure Drug Carriers - chemistry Drug Compounding Endosomes - metabolism Endosomes - ultrastructure General pharmacology Immunostimulation Lactic Acid - chemistry Lead - chemistry Medical sciences Mice Mice, Inbred C57BL Microencapsulation Microscopy, Electron, Transmission Microscopy, Fluorescence Microsphere Microspheres Nanoparticles - chemistry Pharmaceutical technology. Pharmaceutical industry Pharmacology. Drug treatments Polyglycolic Acid - chemistry Polylactic acid Quantum Dots Sulfides - chemistry Vaccines - administration & dosage Vaccines - pharmacokinetics |
Title | Microencapsulation of inorganic nanocrystals into PLGA microsphere vaccines enables their intracellular localization in dendritic cells by electron and fluorescence microscopy |
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