FRET microscopy autologous tumor lysate processing in mature dendritic cell vaccine therapy
<p>Abstract</p> <p>Background</p> <p>Antigen processing by dendritic cells (DC) exposed to specific stimuli has been well characterized in biological studies. Nonetheless, the question of whether autologous whole tumor lysates (as used in clinical trials) are similarly...
| Main Authors: | , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
BMC
2010-06-01
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| Series: | Journal of Translational Medicine |
| Online Access: | http://www.translational-medicine.com/content/8/1/52 |
| _version_ | 1811284415121195008 |
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| author | Ridolfi Ruggero Ridolfi Laura Pancisi Elena Granato Anna Petrini Massimiliano Knutson Jay R Rosales Tilman Ancarani Valentina Fiammenghi Laura Riccobon Angela Neyroz Paolo |
| author_facet | Ridolfi Ruggero Ridolfi Laura Pancisi Elena Granato Anna Petrini Massimiliano Knutson Jay R Rosales Tilman Ancarani Valentina Fiammenghi Laura Riccobon Angela Neyroz Paolo |
| author_sort | Ridolfi Ruggero |
| collection | DOAJ |
| description | <p>Abstract</p> <p>Background</p> <p>Antigen processing by dendritic cells (DC) exposed to specific stimuli has been well characterized in biological studies. Nonetheless, the question of whether autologous whole tumor lysates (as used in clinical trials) are similarly processed by these cells has not yet been resolved.</p> <p>Methods</p> <p>In this study, we examined the transfer of peptides from whole tumor lysates to major histocompatibility complex class II molecules (MHC II) in mature dendritic cells (mDC) from a patient with advanced melanoma. Tumor antigenic peptides-MHC II proximity was revealed by Förster Resonance Energy Transfer (FRET) measurements, which effectively extends the application of fluorescence microscopy to the molecular level (<100Å). Tumor lysates were labelled with Alexa-488, as the donor, and mDC MHC II HLA-DR molecules were labelled with Alexa-546-conjugated IgG, as the acceptor.</p> <p>Results</p> <p>We detected significant energy transfer between donor and acceptor-labelled antibodies against HLA-DR at the membrane surface of mDC. FRET data indicated that fluorescent peptide-loaded MHC II molecules start to accumulate on mDC membranes at 16 hr from the maturation stimulus, steeply increasing at 22 hr with sustained higher FRET detected up to 46 hr.</p> <p>Conclusions</p> <p>The results obtained imply that the patient mDC correctly processed the tumor specific antigens and their display on the mDC surface may be effective for several days. These observations support the rationale for immunogenic efficacy of autologous tumor lysates.</p> |
| first_indexed | 2024-04-13T02:28:17Z |
| format | Article |
| id | doaj.art-3f92897e0d8149b6bc782b49c5a9a52a |
| institution | Directory Open Access Journal |
| issn | 1479-5876 |
| language | English |
| last_indexed | 2024-04-13T02:28:17Z |
| publishDate | 2010-06-01 |
| publisher | BMC |
| record_format | Article |
| series | Journal of Translational Medicine |
| spelling | doaj.art-3f92897e0d8149b6bc782b49c5a9a52a2022-12-22T03:06:41ZengBMCJournal of Translational Medicine1479-58762010-06-01815210.1186/1479-5876-8-52FRET microscopy autologous tumor lysate processing in mature dendritic cell vaccine therapyRidolfi RuggeroRidolfi LauraPancisi ElenaGranato AnnaPetrini MassimilianoKnutson Jay RRosales TilmanAncarani ValentinaFiammenghi LauraRiccobon AngelaNeyroz Paolo<p>Abstract</p> <p>Background</p> <p>Antigen processing by dendritic cells (DC) exposed to specific stimuli has been well characterized in biological studies. Nonetheless, the question of whether autologous whole tumor lysates (as used in clinical trials) are similarly processed by these cells has not yet been resolved.</p> <p>Methods</p> <p>In this study, we examined the transfer of peptides from whole tumor lysates to major histocompatibility complex class II molecules (MHC II) in mature dendritic cells (mDC) from a patient with advanced melanoma. Tumor antigenic peptides-MHC II proximity was revealed by Förster Resonance Energy Transfer (FRET) measurements, which effectively extends the application of fluorescence microscopy to the molecular level (<100Å). Tumor lysates were labelled with Alexa-488, as the donor, and mDC MHC II HLA-DR molecules were labelled with Alexa-546-conjugated IgG, as the acceptor.</p> <p>Results</p> <p>We detected significant energy transfer between donor and acceptor-labelled antibodies against HLA-DR at the membrane surface of mDC. FRET data indicated that fluorescent peptide-loaded MHC II molecules start to accumulate on mDC membranes at 16 hr from the maturation stimulus, steeply increasing at 22 hr with sustained higher FRET detected up to 46 hr.</p> <p>Conclusions</p> <p>The results obtained imply that the patient mDC correctly processed the tumor specific antigens and their display on the mDC surface may be effective for several days. These observations support the rationale for immunogenic efficacy of autologous tumor lysates.</p>http://www.translational-medicine.com/content/8/1/52 |
| spellingShingle | Ridolfi Ruggero Ridolfi Laura Pancisi Elena Granato Anna Petrini Massimiliano Knutson Jay R Rosales Tilman Ancarani Valentina Fiammenghi Laura Riccobon Angela Neyroz Paolo FRET microscopy autologous tumor lysate processing in mature dendritic cell vaccine therapy Journal of Translational Medicine |
| title | FRET microscopy autologous tumor lysate processing in mature dendritic cell vaccine therapy |
| title_full | FRET microscopy autologous tumor lysate processing in mature dendritic cell vaccine therapy |
| title_fullStr | FRET microscopy autologous tumor lysate processing in mature dendritic cell vaccine therapy |
| title_full_unstemmed | FRET microscopy autologous tumor lysate processing in mature dendritic cell vaccine therapy |
| title_short | FRET microscopy autologous tumor lysate processing in mature dendritic cell vaccine therapy |
| title_sort | fret microscopy autologous tumor lysate processing in mature dendritic cell vaccine therapy |
| url | http://www.translational-medicine.com/content/8/1/52 |
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