Monitoring CD4+ T cell responses against viral and tumor antigens using T cells as novel target APC.
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.
Atanackovic
Djordje
D
Ludwig Institute for Cancer Research, Branch at Memorial Sloan-Kettering Cancer Center, 1275 York Avenue, Box 32, New York, NY 10021, USA.
Matsuo
Mitsutoshi
M
Ritter
Erika
E
Mazzara
Gail
G
Ritter
Gerd
G
Jäger
Elke
E
Knuth
Alexander
A
Old
Lloyd J
LJ
Gnjatic
Sacha
S
D000818:Animals; D017951:Antigen Presentation / Q000276:immunology; D000938:Antigen-Presenting Cells / Q000276:immunology*; D000951:Antigens, Neoplasm / Q000276:immunology; D000956:Antigens, Viral / Q000276:immunology; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology*; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology; D002289:Carcinoma, Non-Small-Cell Lung / Q000276:immunology; D006801:Humans; D007118:Immunoassay / Q000379:methods*; D018189:Immunomagnetic Separation; D008175:Lung Neoplasms; D008213:Lymphocyte Activation / Q000276:immunology*; D008545:Melanoma / Q000276:immunology; D016176:T-Lymphocyte Subsets / Q000276:immunology*
D000951:Antigens, Neoplasm; D000956:Antigens, Viral