Dye dilution proliferation assay: application of the DDPA to identify tumor-specific T cell precursor frequencies in clinical trials.
A better understanding of immune effector and regulatory pathways has led to innovative, and complex, immunotherapy strategies. CD8(+) cytolytic T lymphocytes (CTL) provide one common pathway of tumor cell destruction. The peripheral blood CTL compartment typically comprises a minority of anti-tumor CD8(+) lymphocytes and the determination of their number during clinical trials is the focus of various laboratory methods. We have monitored tumor specific CD8(+) as well as CD4(+) lymphocyte precursor frequencies in the peripheral blood using a Dye Dilution Proliferation Assay (DDPA). We summarize our experience applying DDPA in a multi-parameter, antigen-specific assay, detailing some of its complexities and advantages. We provide examples of our clinical trial results showing tumor-specific CD8(+) and CD4(+) precursor frequency (PF) data in patients being treated on novel immunotherapy trials.
Schwaab
Thomas
T
Medical Oncology Immunotherapy Program, Dartmouth Medical School, Lebanon, New Hampshire, USA.
Fisher
Jan L
JL
Meehan
Kenneth R
KR
Fadul
Camilo E
CE
Givan
Alice L
AL
Ernstoff
Marc S
MS
D001681:Biological Assay; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology*; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology*; D002986:Clinical Trials as Topic; D004396:Coloring Agents; D006801:Humans; D054504:Precursor Cells, T-Lymphoid / Q000276:immunology / Q000378:metabolism*
D004396:Coloring Agents