Monitoring of CD4+ and CD8+ T-cell responses after dendritic cell-based immunotherapy using CFSE dye dilution analysis.
CFSE dye dilution analysis and [3H] thymidine incorporation were used side by side to assess proliferative responses of peripheral blood mononuclear cells (PBMCs) after vaccination of renal cell carcinoma patients (n=6) with antigen-loaded dendritic cells. Immune responses against the control antigen keyhole limpet hemocyanin (KLH) were induced in all patients. While [3H] thymidine incorporation revealed a 4 to 977-fold increase in KLH-induced proliferation (mean: 209-fold), CFSE-labeling experiments demonstrated that the KLH-responsive population of postvaccination PBMCs represented 7-53% (mean: 23%). Combining CFSE-labeling with T-cell subset analysis confirmed the presence of CD4+ KLH-reactive T cells but also revealed a substantial population of CD8+ KLH-reactive T cells in one patient as well as minor populations of CD8+ KLH-reactive T cells in three other patients. Our data indicate that CFSE dye dilution analysis is a valuable tool for immune monitoring after dendritic cell vaccination.
Putz
Thomas
T
Department of Urology, Medical University of Innsbruck, Anichstrasse, Innsbruck, Austria.
Ramoner
Reinhold
R
Gander
Hubert
H
Rahm
Andrea
A
Bartsch
Georg
G
Höltl
Lorenz
L
Thurnher
Martin
M
D015496:CD4-Positive T-Lymphocytes / Q000276:immunology*; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology*; D002292:Carcinoma, Renal Cell / Q000628:therapy; D049109:Cell Proliferation; D004396:Coloring Agents; D016207:Cytokines / Q000032:analysis; D003713:Dendritic Cells / Q000276:immunology / Q000637:transplantation*; D005434:Flow Cytometry / Q000379:methods; D006433:Hemocyanins / Q000276:immunology; D006801:Humans; D016219:Immunotherapy, Adoptive / Q000379:methods*; D007963:Leukocytes, Mononuclear / Q000166:cytology; D008722:Methods
D004396:Coloring Agents; D016207:Cytokines; D006433:Hemocyanins; C032808:keyhole-limpet hemocyanin