We are analyzing https://link.springer.com/article/10.1007/s00262-013-1481-z.

Title:
CD8+ T-cell priming and boosting: more antigen-presenting DC, or more antigen per DC? | Cancer Immunology, Immunotherapy
Description:
RNA transfection is a standard method to load dendritic cells (DC) with antigen for therapeutic cancer vaccination. While electroporation yields high transfection efficiency and satisfying expression levels, lipofection results in only few cells expressing high amounts of antigen. We compared antigen loading of human monocyte-derived DC by MelanA RNA electroporation and lipofection. No differences in phenotype or migrational capacity were detected, but lipofected DC induced stronger cytokine secretion by antigen-specific T cells and were superior in priming and boosting of MelanA-specific CD8+ T cells. Interestingly, T cells stimulated with the differently transfected DC did not differ in their functional avidity. To determine whether the amount of antigen per cell is indeed responsible for the superiority of the lipofected DC, we increased the amount of MelanA RNA fivefold and mixed those DC with mock-electroporated ones to mimic the antigen distribution of lipofected cells. This significantly improved the stimulatory capacity, indicating that indeed the amount of antigen per cell seems to be the responsible feature for the observed superiority of lipofected DCs. These data suggest that a few DC that express high amounts of antigen are more immunogenic than many DC expressing lower amounts, although this needs to be tested in a two-armed immunogenicity trial.
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Keywords {🔍}

cells, article, pubmed, google, scholar, dendritic, cas, antigen, van, rna, cell, electroporation, gene, immunol, cancer, human, mrna, schaft, schuler, delivery, ther, immunotherapy, transfection, lipofection, access, vaccines, privacy, cookies, content, monocytederived, transfected, blood, dorrie, kampgen, tumor, bonehill, thielemans, immunother, cytotoxic, lymphocytes, gilboa, data, publish, research, search, wellner, wohn, high, loading, lipofected,

Topics {✒️}

friedrich-alexander-university erlangen-nuremberg specific tumor-reactive ctl dendritic cell-based vaccines high-affinity tumor-reactive cytotoxic month download article/chapter dfg-german research foundation monocyte-derived dendritic cells wild-type viral mrna dc-based cancer vaccines human monocyte-derived dc generate cytolytic tumor-specific tumor-antigen-encoding mrna gerold schuler & jan dörrie effective clinical-scale production cd8+ t-cell priming high-level antigen expression mrna-based gene transfer dendritic cell vaccines antigen-presenting dc high-efficiency gene transfer cell therapy article cancer immunology tumor-derived rna dendritic cells transfected human dendritic cells original article published full article pdf privacy choices/manage cookies melana-specific cd8+ high transfection efficiency monocyte elutriation directly cell responses load dendritic cells mature dendritic cells dendritic cells leads target dendritic cells dendritic cells nucleofected dendritic cells pulsed engineering dendritic cells class ii molecules express high amounts melana rna fivefold van der bp human hematopoietic cells sustained antigen presentation tumor antigen loading differently transfected dc autologous melanoma model van bockstaele dr melana rna electroporation

Questions {❓}

CD8+ T-cell priming and boosting: more antigen-presenting DC, or more antigen per DC?
CD8+ T-cell priming and boosting: more antigen-presenting DC, or more antigen per DC?
Lesterhuis WJ, Aarntzen EH, de Vries IJ, Schuurhuis DH, Figdor CG, Adema GJ, Punt CJ (2008) Dendritic cell vaccines in melanoma: from promise to proof?

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         description: RNA transfection is a standard method to load dendritic cells (DC) with antigen for therapeutic cancer vaccination. While electroporation yields high transfection efficiency and satisfying expression levels, lipofection results in only few cells expressing high amounts of antigen. We compared antigen loading of human monocyte-derived DC by MelanA RNA electroporation and lipofection. No differences in phenotype or migrational capacity were detected, but lipofected DC induced stronger cytokine secretion by antigen-specific T cells and were superior in priming and boosting of MelanA-specific CD8+ T cells. Interestingly, T cells stimulated with the differently transfected DC did not differ in their functional avidity. To determine whether the amount of antigen per cell is indeed responsible for the superiority of the lipofected DC, we increased the amount of MelanA RNA fivefold and mixed those DC with mock-electroporated ones to mimic the antigen distribution of lipofected cells. This significantly improved the stimulatory capacity, indicating that indeed the amount of antigen per cell seems to be the responsible feature for the observed superiority of lipofected DCs. These data suggest that a few DC that express high amounts of antigen are more immunogenic than many DC expressing lower amounts, although this needs to be tested in a two-armed immunogenicity trial.
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