A New Strategy for Tumor Antigen Discovery Based on in Vitro Priming of Naïve T Cells with Dendritic Cells

Henry Kao, Andrew A. Amoscato, Pawel Ciborowski, Olivera J. Finn

Research output: Contribution to journalArticle

Abstract

We describe a method for discovery of new tumor antigens that uses dendritic cells (DCs) as antigen-presenting cells to prime autologous naive CD4+ and CD8+ T cells from healthy donors against tumor proteins and peptides. For the identification of HLA class I-restricted tumor antigens, peptides were extracted from tumor HLA class I molecules, fractionated by reverse phase-high performance liquid chromatography, and loaded onto in vitro-generated DCs to prime naïve CD8+ T cells. Our results show that we were able to prime naïve CD8+ T cells in vitro to several peptide fractions and generate specificity for the tumor. Electrospray ionization mass spectrometry was used to confirm that these fractions contained peptides derived from MHC class I molecules, and the primed CD8+ T cells were used to further analyze the immunostimulatory peptide fractions. For the identification of HLA class II-restricted tumor antigens, we fractionated tumor protein extracts using reverse phase-high performance liquid chromatography and loaded individual fractions onto DCs to prime naive CD4+ T cells. Our results show that we were also able to prime naïve CD4+ T cells to several protein fractions and generate specificity for the tumor. These results illustrate the potential of this method to identify new immunostimulatory MHC class I- and class II-restricted tumor antigens.

Original languageEnglish (US)
JournalClinical Cancer Research
Volume7
Issue number3
StatePublished - Jan 1 2001

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Neoplasm Antigens
Dendritic Cells
T-Lymphocytes
Peptides
Neoplasms
High Pressure Liquid Chromatography
Proteins
Electrospray Ionization Mass Spectrometry
Antigen-Presenting Cells
In Vitro Techniques

ASJC Scopus subject areas

  • Oncology
  • Cancer Research

Cite this

A New Strategy for Tumor Antigen Discovery Based on in Vitro Priming of Naïve T Cells with Dendritic Cells. / Kao, Henry; Amoscato, Andrew A.; Ciborowski, Pawel; Finn, Olivera J.

In: Clinical Cancer Research, Vol. 7, No. 3, 01.01.2001.

Research output: Contribution to journalArticle

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