Optimization and validation of a robust human T-cell culture method for monitoring phenotypic and polyfunctional antigen-specific CD4 and CD8 T-cell responses.
OBJECTIVE
Monitoring cellular immune responses is one prerequisite for the rational development of cancer vaccines.


METHODS
We describe an extensive effort to optimize and validate quantitatively an in vitro T-cell culture method by determining the phenotype and function of both CD4(+) and CD8(+) T cells, including measurement of the phenotype markers CCR7, CD45RA, CD28 and CD27 and the functional markers interferon (IFN)-gamma, interleukin (IL)-2, macrophage inflammatory protein (MIP)-1beta, tumor necrosis factor (TNF)-alpha and CD107a.


RESULTS
Autologous peripheral blood mononuclear cells (PBMC) were potent stimulators that expanded antigen (Ag)-specific CD8(+) T cells during short-term culture with the addition of IL-2 and IL-15 cytokines. Polyfunctional Ag-specific CD4(+) and CD8(+) T cells were detectable using this method.


CONCLUSIONS
Our culture system represents a robust human T-cell culture protocol that permits phenotypic, quantitative and qualitative evaluation of vaccine-induced CD4(+) and CD8(+) T-cell responses.
Lin
Yun
Y
Ludwig Center for Cancer Immunotherapy, Immunology Program, Sloan-Kettering Institute, Newy York, New York 10021, USA.
Gallardo
Humilidad F
HF
Ku
Geoffrey Y
GY
Li
Hao
H
Manukian
Gregor
G
Rasalan
Teresa S
TS
Xu
Yinyan
Y
Terzulli
Stephanie L
SL
Old
Lloyd J
LJ
Allison
James P
JP
Houghton
Alan N
AN
Wolchok
Jedd D
JD
Yuan
Jianda
J
D000941:Antigens / Q000276:immunology; D015703:Antigens, CD / Q000378:metabolism; D000943:Antigens, Differentiation / Q000378:metabolism; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology / Q000378:metabolism* / Q000473:pathology; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology / Q000378:metabolism* / Q000473:pathology; D019496:Cancer Vaccines; D018929:Cell Culture Techniques; D002478:Cells, Cultured; D016207:Cytokines / Q000378:metabolism; D005240:Feasibility Studies; D006801:Humans; D009369:Neoplasms / Q000276:immunology* / Q000628:therapy
D000941:Antigens; D015703:Antigens, CD; D000943:Antigens, Differentiation; D019496:Cancer Vaccines; D016207:Cytokines