ABSTRACT
Purpose
To develop a liposome formulation incorporating antigen-presenting cells (APCs) membrane microdomains with enriched epitope/MHC complexes to evaluate the activities of these liposomes (RAFTsomes) to activate T cells and prime immune responses.
Methods
We isolated membrane microdomain structures that contained the epitope/MHC complexes from ovalbumin (OVA) primed dendritic cells (DCs), and reconstituted them on liposomes surface by detergent dialysis. The resulted RAFTsomes were purified by density gradient centrifugation. Their T cell activation functions were evaluated by IL-2 secreting and proliferation assays in vitro. In vivo immune responses and the protective effect against OVA expressing EG.7 tumor challenge were also examined.
Results
Membrane microdomains containing enriched epitope/MHC complexes can be reconstituted into liposomes with defined size and composition. The integrity and activities of these complexes after reconstitution were confirmed by in vitro T cell assays. OVA epitope loaded RAFTsomes injected in vivo resulted in high anti-OVA IgG production (predominantly IgG1). The immunized mice were protected from EG.7 tumor cell inoculation challenge.
Conclusions
Based on these findings, we propose that RAFTsomes can be prepared with unique properties that may be used as an antigen delivery system for immunotherapeutic applications.
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Abbreviations
- APCs:
-
antigen-presenting cells
- DCs:
-
dendritic cells
- DOPC:
-
1,2-dioleoyl-sn-glycero-3-phosphocholine
- MHC:
-
major histocompatibility complex
- OVA:
-
ovalbumin
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ACKNOWLEDGMENTS AND DISCLOSURES
We like to thank Prof. Xuetao Cao for the generous gift of MF2.2D9 and EG.7 cell lines and Prof. Tao Xi for assisting with the figures preparation. The TEM and LC-MS/MS analysis were performed at the Instrumental Analysis Center of Shanghai Jiao Tong University. This study was supported by grants from the Natural Science Foundation of China No. 30571721 and 30825045.
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A table (Table SI) of proteomics study with lipid rafts extracted from mDCs.
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Ding, Q., Chen, J., Wei, X. et al. RAFTsomes Containing Epitope-MHC-II Complexes Mediated CD4+ T Cell Activation and Antigen-Specific Immune Responses. Pharm Res 30, 60–69 (2013). https://doi.org/10.1007/s11095-012-0849-7
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DOI: https://doi.org/10.1007/s11095-012-0849-7