Authors: | Atanackovic, D.; Matsuo, M.; Ritter, E.; Mazzara, G.; Ritter, G.; Jäger, E.; Knuth, A.; Old, L. J.; Gnjatic, S. |
Article Title: | Monitoring CD4+ T cell responses against viral and tumor antigens using T cells as novel target APC |
Abstract: | 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-γ 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. © 2003 Elsevier B.V. All rights reserved. |
Keywords: | controlled study; human cell; nonhuman; antigen expression; t lymphocyte; cd8-positive t-lymphocytes; animals; melanoma; carcinoma, non-small-cell lung; lung neoplasms; peptide; tumor antigen; antigen presentation; lymphocyte activation; mononuclear cell; antigens, neoplasm; gamma interferon; quantitative analysis; immunomagnetic separation; hla dr antigen; major histocompatibility antigen class 2; cd4-positive t-lymphocytes; epitope; virus antigen; immunoassay; immunostimulation; t-lymphocyte subsets; enzyme linked immunospot assay; memory cell; antigen-presenting cells; virus protein; virus vector; expression vector; antigens, viral; cd4+ t cell; fowlpox virus; nucleoprotein; humans; human; priority journal; article; antigen presentation and processing; immuno-monitoring; viral and tumor antigen; immunological technique |
Journal Title: | Journal of Immunological Methods |
Volume: | 278 |
Issue: | 1-2 |
ISSN: | 0022-1759 |
Publisher: | Elsevier Science, Inc. |
Date Published: | 2003-01-01 |
Start Page: | 57 |
End Page: | 66 |
Language: | English |
DOI: | 10.1016/s0022-1759(03)00209-6 |
PUBMED: | 12957396 |
PROVIDER: | scopus |
DOI/URL: | |
Notes: | Export Date: 12 September 2014 -- Source: Scopus |