4-1BB-mediated expansion affords superior detection of in vivo primed effector memory CD8+ T cells from melanoma sentinel lymph nodes

B J R Sluijter; M F C M van den Hout; A G M Stam; S M Lougheed; M M Suhoski; A J M van den Eertwegh; M P van den Tol; P A M van Leeuwen; S Meijer; R J Scheper; C H June; T D de Gruijl; S J A M Santegoets

doi:10.1016/j.clim.2010.07.009

4-1BB-mediated expansion affords superior detection of in vivo primed effector memory CD8+ T cells from melanoma sentinel lymph nodes

Clin Immunol. 2010 Nov;137(2):221-33. doi: 10.1016/j.clim.2010.07.009. Epub 2010 Aug 13.

Authors

B J R Sluijter¹, M F C M van den Hout, A G M Stam, S M Lougheed, M M Suhoski, A J M van den Eertwegh, M P van den Tol, P A M van Leeuwen, S Meijer, R J Scheper, C H June, T D de Gruijl, S J A M Santegoets

Affiliation

¹ Department of Surgical Oncology, VU University Medical Center, Cancer Center Amsterdam, Amsterdam, The Netherlands.

PMID: 20708974
DOI: 10.1016/j.clim.2010.07.009

Abstract

We have been studying the re-activation of tumor-associated antigen (TAA)-specific CD8(+) T cells in sentinel lymph nodes (SLN) of melanoma patients upon intradermal administration of the CpG-B oligodeoxynucleotide PF-3512676. To facilitate functional testing of T cells from small SLN samples, high-efficiency polyclonal T cell expansion is required. In this study, SLN cells were expanded via classic methodologies with plate- or bead-bound anti-CD3/CD28 antibodies and with the K562/CD32/4-1BBL artificial APC system (K32/4-1BBL aAPC) and analyzed for responsiveness to common recall or TAA-derived peptides. K32/4-1BBL-expanded T cell populations contained significantly more effector/memory CD8(+) T cells. Moreover, recall and melanoma antigen-specific CD8(+) T cells were more frequently detected in K32/4-1BBL-expanded samples as compared with anti-CD3/CD28-expanded samples. We conclude that K32/4-1BBL aAPC are superior to anti-CD3/CD28 antibodies for the expansion of in vivo-primed specific CD8(+) T cells and that their use facilitates the sensitive monitoring of functional anti-tumor T cell immunity in SLN.

MeSH terms

4-1BB Ligand / genetics
Antibodies, Monoclonal / immunology
Antigen-Presenting Cells / immunology
Antigens, CD / metabolism
CD8-Positive T-Lymphocytes / cytology
CD8-Positive T-Lymphocytes / drug effects
CD8-Positive T-Lymphocytes / immunology*
CD8-Positive T-Lymphocytes / metabolism
Cell Count
Cell Proliferation
Epitopes, T-Lymphocyte / immunology
Humans
Immunologic Memory / immunology*
Interferon-gamma / metabolism
Interleukins / metabolism
K562 Cells
Lymph Nodes / drug effects
Lymph Nodes / immunology*
Lymph Nodes / surgery
Lymphocyte Activation / drug effects
Lymphocyte Activation / immunology*
Lysosomal-Associated Membrane Protein 1 / metabolism
Melanoma / immunology*
Melanoma-Specific Antigens / immunology
Oligodeoxyribonucleotides / administration & dosage
Oligodeoxyribonucleotides / pharmacology
Receptors, IgG / genetics
Sentinel Lymph Node Biopsy*
T-Lymphocyte Subsets / cytology
T-Lymphocyte Subsets / drug effects
T-Lymphocyte Subsets / immunology
T-Lymphocyte Subsets / metabolism
Transfection
Tumor Necrosis Factor Receptor Superfamily, Member 9 / metabolism*
Tumor Necrosis Factor-alpha / metabolism

Substances

4-1BB Ligand
Antibodies, Monoclonal
Antigens, CD
Epitopes, T-Lymphocyte
Interleukins
Lysosomal-Associated Membrane Protein 1
Melanoma-Specific Antigens
Oligodeoxyribonucleotides
ProMune
Receptors, IgG
TNFRSF9 protein, human
Tumor Necrosis Factor Receptor Superfamily, Member 9
Tumor Necrosis Factor-alpha
Interferon-gamma