Sensitive detection of human papillomavirus type 16 E7-specific T cells by ELISPOT after multiple in vitro stimulations of CD8+ T cells with peptide-pulsed autologous dendritic cells

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• 作者：Nathalie Cools (1)
  Peter Ponsaerts (1)
  Marc Lenjou (1)
  Griet Nijs (1)
  Dirk R Van Bockstaele (1)
  Viggo FI Van Tendeloo (1)
  Zwi N Berneman (1)
  
• 刊名：Molecular Cancer
• 出版年：2006
• 出版时间：December 2006
• 年：2006
• 卷：5
• 期：1
• 全文大小：1723KB
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• 作者单位：Nathalie Cools (1)
  Peter Ponsaerts (1)
  Marc Lenjou (1)
  Griet Nijs (1)
  Dirk R Van Bockstaele (1)
  Viggo FI Van Tendeloo (1)
  Zwi N Berneman (1)
  
  1. Laboratory of Experimental Hematology, Antwerp University, Faculty of Medicine, Antwerp University Hospital, Belgium
  
• ISSN：1476-4598

文摘

Background Cervical cancer is the second most common gynecological cancer amongst women world-wide. Despite optimized protocols, standard treatments still face several disadvantages. Therefore, research aims at the development of immune-based strategies using tumor antigen-loaded dendritic cells for the induction of cellular anti-tumor immunity. Results In this study, we used dendritic cells loaded with the HLA-A2-restricted HPV type 16 E711-0 peptide in order to induce an in vitro CD8+ T cell response. For this purpose, peptide-pulsed dendritic cells were co-cultured with autologous CD8+ T cells. After 5 weekly stimulations with peptide-pulsed mature dendritic cells, cultured T cells were analyzed for antigen specificity by an IFN-γ ELISPOT assay. Using this ELISPOT assay, we were able to detect E7-specific IFN-γ-secreting CD8+ T cells in 5/5 healthy donors. Conclusion We show that peptide-pulsed mature dendritic cells are able to stimulate a HPV type 16 E7 peptide-specific immune response in vitro. These experiments describe an efficient culture protocol for antigen-specific T cells for use in pre-clinical vaccination research and confirm the need for sensitive T cell assays for detection of tumor-specific immune responses in vitro.