Measuring melanoma-specific cytotoxic T lymphocytes elicited by dendritic cell vaccines with a tumor inhibition assay in vitro.
Improving cancer vaccines depends on assays measuring elicited tumor-specific T-cell immunity. Cytotoxic effector cells are essential for tumor clearance and are commonly evaluated using 51Cr release from labeled target cells after a short (4 hours) incubation with T cells. The authors used a tumor inhibition assay (TIA) that assesses the capacity of cytotoxic T lymphocytes (CTLs) to control the survival/growth of EGFP-labeled tumor cell lines. TIA was validated using CD8+ T cells primed in vitro against melanoma and breast cancer cells. TIA was then used to assess the CTL function of cultured CD8+ T cells isolated from patients with metastatic melanoma who underwent vaccination with peptide-pulsed CD34+ HPCs-derived DCs. After the DC vaccination, T cells from six of eight patients yielded CTLs that could inhibit the survival/growth of melanoma cells. The results of TIA correlated with killing of tumor cells in a standard 4-hour 51Cr release assay, yet TIA allowed detection of CTL activities that appeared marginal in the 51Cr release assay. Thus, TIA might prove valuable for measuring spontaneous and induced antigen-specific cytotoxic T cells.
Paczesny
Sophie
S
Baylor Institute for Immunology Research, Baylor University Medical Center, Dallas, Texas 75204, USA.
Shi
Honhgzhen
H
Saito
Hiroaki
H
Mannoni
Patrice
P
Fay
Joseph
J
Banchereau
Jacques
J
Palucka
A Karolina
AK
D018952:Antigens, CD34 / Q000096:biosynthesis; D018414:CD8-Positive T-Lymphocytes / Q000166:cytology / Q000276:immunology; D019496:Cancer Vaccines; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D002857:Chromium / Q000378:metabolism; D002860:Chromium Radioisotopes; D018920:Coculture Techniques; D003470:Culture Media / Q000494:pharmacology; D003713:Dendritic Cells / Q000166:cytology*; D005434:Flow Cytometry; D049452:Green Fluorescent Proteins / Q000378:metabolism; D015234:HLA-A Antigens / Q000737:chemistry; D015789:HLA-A2 Antigen; D006801:Humans; D020014:K562 Cells; D008545:Melanoma / Q000097:blood / Q000276:immunology* / Q000628:therapy*; D018613:Microscopy, Confocal; D008856:Microscopy, Fluorescence; D009000:Monocytes / Q000166:cytology; D010455:Peptides / Q000737:chemistry; D013601:T-Lymphocytes / Q000276:immunology; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology*; D013997:Time Factors
D018952:Antigens, CD34; D019496:Cancer Vaccines; D002860:Chromium Radioisotopes; D003470:Culture Media; D015234:HLA-A Antigens; C435939:HLA-A*02:01 antigen; D015789:HLA-A2 Antigen; D010455:Peptides; D002857:Chromium; D049452:Green Fluorescent Proteins