TY - JOUR
T1 - Co-Stimulation through 4-1BB/CD137 Improves the Expansion and Function of CD8+ Melanoma Tumor-Infiltrating Lymphocytes for Adoptive T-Cell Therapy
AU - Chacon, Jessica Ann
AU - Wu, Richard C.
AU - Sukhumalchandra, Pariya
AU - Molldrem, Jeffrey J.
AU - Sarnaik, Amod
AU - Pilon-Thomas, Shari
AU - Weber, Jeffrey
AU - Hwu, Patrick
AU - Radvanyi, Laszlo
N1 - Funding Information:
We are grateful to Bristol Myers Squibb (Drs. Marie Jure-Kunkel and Stacey Goldberg) for their generous gift of the agonistic anti-4-1BB antibody (BMS-663513). We also thank Dr. Hanspeter Pircher from the University of Freiburg (Freiburg, Germany) for providing the anti-KLRG-1 antibody. Jessica Ann Chacon gratefully acknowledges support from the UT Health Innovation for Cancer Prevention Research Pre-Doctoral Fellowship, The University of Texas School of Public Health-Cancer Prevention and Research Institute of Texas grant #RP101503. We are also thankful to the TIL Lab (Chris Toth, Renjith Ramachandran, Seth Wardell, Audrey Gonzalez, Orenthial Fulbright, and Rahmatu Mansaray) at M.D. Anderson Cancer Center for the processing of melanoma tumors and initial culturing of TIL for our experiments. We thank Drs. Willem Overwijk, Chantale Bernatchez, and Cara Haymaker for critical reading of the manuscript and their comments.
PY - 2013/4/1
Y1 - 2013/4/1
N2 - Adoptive T-cell therapy (ACT) using tumor-infiltrating lymphocytes (TIL) can induce tumor regression in up to 50% or more of patients with unresectable metastatic melanoma. However, current methods to expand melanoma TIL, especially the "rapid expansion protocol" (REP) were not designed to enhance the generation of optimal effector-memory CD8+ T cells for infusion. One approach to this problem is to manipulate specific co-stimulatory signaling pathways to enhance CD8+ effector-memory T-cell expansion. In this study, we determined the effects of activating the TNF-R family member 4-1BB/CD137, specifically induced in activated CD8+ T cells, on the yield, phenotype, and functional activity of expanded CD8+ T cells during the REP. We found that CD8+ TIL up-regulate 4-1BB expression early during the REP after initial TCR stimulation, but neither the PBMC feeder cells in the REP or the activated TIL expressed 4-1BB ligand. However, addition of an exogenous agonistic anti-4-1BB IgG4 (BMS 663513) to the REP significantly enhanced the frequency and total yield of CD8+ T cells as well as their maintenance of CD28 and increased their anti-tumor CTL activity. Gene expression analysis found an increase in bcl-2 and survivin expression induced by 4-1BB that was associated with an enhanced survival capability of CD8+ post-REP TIL when re-cultured in the absence or presence of cytokines. Our findings suggest that adding an agonistic anti-4-1BB antibody during the time of TIL REP initiation produces a CD8+ T cell population capable of improved effector function and survival. This may greatly improve TIL persistence and anti-tumor activity in vivo after adoptive transfer into patients.
AB - Adoptive T-cell therapy (ACT) using tumor-infiltrating lymphocytes (TIL) can induce tumor regression in up to 50% or more of patients with unresectable metastatic melanoma. However, current methods to expand melanoma TIL, especially the "rapid expansion protocol" (REP) were not designed to enhance the generation of optimal effector-memory CD8+ T cells for infusion. One approach to this problem is to manipulate specific co-stimulatory signaling pathways to enhance CD8+ effector-memory T-cell expansion. In this study, we determined the effects of activating the TNF-R family member 4-1BB/CD137, specifically induced in activated CD8+ T cells, on the yield, phenotype, and functional activity of expanded CD8+ T cells during the REP. We found that CD8+ TIL up-regulate 4-1BB expression early during the REP after initial TCR stimulation, but neither the PBMC feeder cells in the REP or the activated TIL expressed 4-1BB ligand. However, addition of an exogenous agonistic anti-4-1BB IgG4 (BMS 663513) to the REP significantly enhanced the frequency and total yield of CD8+ T cells as well as their maintenance of CD28 and increased their anti-tumor CTL activity. Gene expression analysis found an increase in bcl-2 and survivin expression induced by 4-1BB that was associated with an enhanced survival capability of CD8+ post-REP TIL when re-cultured in the absence or presence of cytokines. Our findings suggest that adding an agonistic anti-4-1BB antibody during the time of TIL REP initiation produces a CD8+ T cell population capable of improved effector function and survival. This may greatly improve TIL persistence and anti-tumor activity in vivo after adoptive transfer into patients.
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U2 - 10.1371/journal.pone.0060031
DO - 10.1371/journal.pone.0060031
M3 - Article
C2 - 23560068
AN - SCOPUS:84875663346
SN - 1932-6203
VL - 8
JO - PloS one
JF - PloS one
IS - 4
M1 - e60031
ER -