Monitoring CD4+ T cell responses against viral and tumor antigens using T cells as novel target APC

J Immunol Methods. 2003 Jul;278(1-2):57-66. doi: 10.1016/s0022-1759(03)00209-6.


CD4+ T cells play an important role in the induction and maintenance of an effective antiviral and antitumor immune response. However, standardized monitoring of antigen-specific CD4+ T cells has not been established at the single-cell level. We now present a sensitive, specific, and simple methodology in which purified memory CD4+ T cells are expanded from PBMC in a single cycle of antigen-driven stimulation and quantitatively assayed by interferon-gamma ELISPOT. Issues of nonspecific background in assays were resolved with the use of innovative target cells, autologous PHA-expanded CD4+ T cells (T-APC). Remarkably, T-APC could not only present peptide epitopes from model antigens NY-ESO-1 and influenza nucleoprotein, but could also process full-length antigen endogenously expressed from recombinant fowlpox vector. This approach makes it possible to monitor CD4+ T cells in large series of patients, regardless of HLA haplotype, against the full peptide repertoire of a given antigen.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antigen Presentation / immunology
  • Antigen-Presenting Cells / immunology*
  • Antigens, Neoplasm / immunology
  • Antigens, Viral / immunology
  • CD4-Positive T-Lymphocytes / immunology*
  • CD8-Positive T-Lymphocytes / immunology
  • Carcinoma, Non-Small-Cell Lung / immunology
  • Humans
  • Immunoassay / methods*
  • Immunomagnetic Separation
  • Lung Neoplasms
  • Lymphocyte Activation / immunology*
  • Melanoma / immunology
  • T-Lymphocyte Subsets / immunology*


  • Antigens, Neoplasm
  • Antigens, Viral