Abstract
Adoptive cell transfer therapy with reactive T cells is one of the most promising immunotherapeutic modalities for metastatic melanoma patients. Homing of the transferred T cells to all tumor sites in sufficient numbers is of great importance. Here, we seek to exploit endogenous chemotactic signals in order to manipulate and enhance the directional trafficking of transferred T cells toward melanoma. Chemokine profiling of 15 melanoma cultures shows that CXCL1 and CXCL8 are abundantly expressed and secreted from melanoma cultures. However, the complimentary analysis on 40 melanoma patient-derived tumor-infiltrating lymphocytes (TIL) proves that the corresponding chemokine receptors are either not expressed (CXCR2) or expressed at low levels (CXCR1). Using the in vitro transwell system, we demonstrate that TIL cells preferentially migrate toward melanoma and that endogenously expressing CXCR1 TIL cells are significantly enriched among the migrating lymphocytes. The role of the chemokines CXCL1 and CXCL8 is demonstrated by partial abrogation of this enrichment with anti-CXCL1 and anti-CXCL8 neutralizing antibodies. The role of the chemokine receptor CXCR1 is validated by the enhanced migration of CXCR1-engineered TIL cells toward melanoma or recombinant CXCL8. Cytotoxicity and IFNγ secretion activity are unaltered by CXCR1 expression profile. Taken together, these results mark CXCR1 as a candidate for genetic manipulations to enhance trafficking of adoptively transferred T cells. This approach is complimentary and potentially synergistic with other genetic strategies designed to enhance anti-tumor potency.
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References
Vicari AP, Caux C (2002) Chemokines in cancer. Cytokine Growth Factor Rev 13:143–154
Harlin H, Meng Y, Peterson AC, Zha Y, Tretiakova M et al (2009) Chemokine expression in melanoma metastases associated with CD8+ T-cell recruitment. Cancer Res 69:3077–3085
Viola A, Molon B, Contento RL (2008) Chemokines: coded messages for T-cell missions. Front Biosci 13:6341–6353
Balkwill F (2004) Cancer and the chemokine network. Nat Rev Cancer 4:540–550
Jin T, Xu X, Hereld D (2008) Chemotaxis, chemokine receptors and human disease. Cytokine 44:1
Zhang T, Somasundaram R, Berencsi K, Caputo L, Gimotty P et al (2006) Migration of cytotoxic T lymphocytes toward melanoma cells in three-dimensional organotypic culture is dependent on CCL2 and CCR4. Eur J Immunol 36:457–467
Kunz M, Toksoy A, Goebeler M, Engelhardt E, Brocker E et al (1999) Strong expression of the lymphoattractant C-X-C chemokine Mig is associated with heavy infiltration of T cells in human malignant melanoma. J Pathol 189:552–558
Zhang T, Somasundaram R, Berencsi K, Caputo L, Rani P et al (2005) CXC Chemokine Ligand 12 (Stromal Cell-Derived Factor 1-±) and CXCR4-Dependent Migration of CTLs toward Melanoma Cells in Organotypic Culture. J Immunol 174:5856
Vianello F, Papeta N, Chen T, Kraft P, White N et al (2006) Murine B16 Melanomas Expressing High Levels of the Chemokine Stromal-Derived Factor-1/CXCL12 Induce Tumor-Specific T Cell Chemorepulsion and Escape from Immune Control. J Immunol 176:2902
Dudley ME, Yang JC, Sherry R, Hughes MS, Royal R et al (2008) Adoptive Cell Therapy for Patients With Metastatic Melanoma: Evaluation of Intensive Myeloablative Chemoradiation Preparative Regimens. J Clin Oncol 26:5233
Dudley ME, Wunderlich JR, Yang JC, Sherry RM, Topalian SL et al (2005) Adoptive cell transfer therapy following non-myeloablative but lymphodepleting chemotherapy for the treatment of patients with refractory metastatic melanoma. J Clin Oncol 23:2346–2357
Rosenberg SA, Restifo NP, Yang JC, Morgan RA, Dudley ME (2008) Adoptive cell transfer: a clinical path to effective cancer immunotherapy. Nat Rev Cancer 8:299–308
Mackensen A, Meidenbauer N, Vogl S, Laumer M, Berger J et al (2006) Phase I study of adoptive T-cell therapy using antigen-specific CD8+ T cells for the treatment of patients with metastatic melanoma. J Clin Oncol 24:5060–5069
Khammari A, Labarriere N, Vignard V, Nguyen JM, Pandolfino MC et al (2009) Treatment of metastatic melanoma with autologous Melan-A/MART-1-specific cytotoxic T lymphocyte clones. J Invest Dermatol 129:2835–2842
Brimnes MK, Gang AO, Donia M, Thor Straten P, Svane IM, et al. (2012) Generation of autologous tumor-specific T cells for adoptive transfer based on vaccination, in vitro restimulation and CD3/CD28 dynabead-induced T cell expansion. Cancer Immunol Immunother
Robbins PF, Dudley ME, Wunderlich J, El-Gamil M, Li YF et al (2004) Cutting edge: persistence of transferred lymphocyte clonotypes correlates with cancer regression in patients receiving cell transfer therapy. J Immunol 173:7125–7130
Zhou J, Dudley ME, Rosenberg SA, Robbins PF (2005) Persistence of multiple tumor-specific T-cell clones is associated with complete tumor regression in a melanoma patient receiving adoptive cell transfer therapy. J Immunother 28:53–62
Peterson AC, Harlin H, Gajewski TF (2003) Immunization with Melan-A peptide-pulsed peripheral blood mononuclear cells plus recombinant human interleukin-12 induces clinical activity and T-cell responses in advanced melanoma. J Clin Oncol 21:2342–2348
Cormier JN, Salgaller ML, Prevette T, Barracchini KC, Rivoltini L et al (1997) Enhancement of cellular immunity in melanoma patients immunized with a peptide from MART-1/Melan A. Cancer J Sci Am 3:37–44
Schadendorf D, Ugurel S, Schuler-Thurner B, Nestle FO, Enk A et al (2006) Dacarbazine (DTIC) versus vaccination with autologous peptide-pulsed dendritic cells (DC) in first-line treatment of patients with metastatic melanoma: a randomized phase III trial of the DC study group of the DeCOG. Ann Oncol 17:563–570
Rosenberg SA, Sherry RM, Morton KE, Scharfman WJ, Yang JC et al (2005) Tumor progression can occur despite the induction of very high levels of self/tumor antigen-specific CD8 + T cells in patients with melanoma. J Immunol 175:6169–6176
Morgan RA, Dudley ME, Wunderlich JR, Hughes MS, Yang JC et al (2006) Cancer regression in patients after transfer of genetically engineered lymphocytes. Science 314:126–129
Johnson LA, Morgan RA, Dudley ME, Cassard L, Yang JC et al (2009) Gene therapy with human and mouse T-cell receptors mediates cancer regression and targets normal tissues expressing cognate antigen. Blood 114:535–546
Rosenberg SA, Dudley ME (2009) Adoptive cell therapy for the treatment of patients with metastatic melanoma. Curr Opin Immunol 21:233–240
Peng W, Ye Y, Rabinovich BA, Liu C, Lou Y et al (2010) Transduction of tumor-specific T cells with CXCR2 chemokine receptor improves migration to tumor and antitumor immune responses. Clin Cancer Res 16:5458–5468
Fisher B, Packard BS, Read EJ, Carrasquillo JA, Carter CS et al (1989) Tumor localization of adoptively transferred indium-111 labeled tumor infiltrating lymphocytes in patients with metastatic melanoma. J Clin Oncol 7:250–261
Gajewski TF (2007) Failure at the effector phase: immune barriers at the level of the melanoma tumor microenvironment. Clin Cancer Res 13:5256–5261
Gajewski TF, Meng Y, Blank C, Brown I, Kacha A et al (2006) Immune resistance orchestrated by the tumor microenvironment. Immunol Rev 213:131
Markel G, Seidman R, Cohen Y, Besser MJ, Sinai TC et al (2009) Dynamic expression of protective CEACAM1 on melanoma cells during specific immune attack. Immunology 126:186–200
Besser MJ, Shapira-Frommer R, Treves AJ, Zippel D, Itzhaki O et al (2010) Clinical responses in a phase II study using adoptive transfer of short-term cultured tumor infiltration lymphocytes in metastatic melanoma patients. Clin Cancer Res 16:2646–2655
Markel G, Seidman R, Stern N, Cohen-Sinai T, Izhaki O et al (2006) Inhibition of human tumor-infiltrating lymphocyte effector functions by the homophilic carcinoembryonic cell adhesion molecule 1 interactions. J Immunol 177:6062–6071
Markel G, Ortenberg R, Seidman R, Sapoznik S, Koren-Morag N, et al. (2010) Systemic dysregulation of CEACAM1 in melanoma patients. Cancer Immunol Immunother 59(2):215–230
Svec J, Ergang P, Mandys V, Kment M, Pacha J (2010) Expression profiles of proliferative and antiapoptotic genes in sporadic and colitis-related mouse colon cancer models. Int J Exp Pathol 91:44–53
Orchard PJ, Blazar BR, Burger S, Levine B, Basso L et al (2002) Clinical-scale selection of anti-CD3/CD28-activated T cells after transduction with a retroviral vector expressing herpes simplex virus thymidine kinase and truncated nerve growth factor receptor. Hum Gene Ther 13:979–988
Bialer G, Horovitz-Fried M, Ya’acobi S, Morgan RA, Cohen CJ (2010) Selected murine residues endow human TCR with enhanced tumor recognition. J Immunol 184:6232–6241
Zhao Y, Zheng Z, Cohen CJ, Gattinoni L, Palmer DC et al (2006) High-efficiency transfection of primary human and mouse T lymphocytes using RNA electroporation. Mol Ther 13:151–159
Besser MJ, Shapira-Frommer R, Treves AJ, Zippel D, Itzhaki O et al (2009) Minimally cultured or selected autologous tumor-infiltrating lymphocytes after a lympho-depleting chemotherapy regimen in metastatic melanoma patients. J Immunother 32:415–423
Eikawa S, Ohue Y, Kitaoka K, Aji T, Uenaka A et al (2010) Enrichment of Foxp3+ CD4 regulatory T cells in migrated T cells to IL-6- and IL-8-expressing tumors through predominant induction of CXCR1 by IL-6. J Immunol 185:6734–6740
Kershaw MH, Wang G, Westwood JA, Pachynski RK, Tiffany HL et al (2002) Redirecting migration of T cells to chemokine secreted from tumors by genetic modification with CXCR2. Hum Gene Ther 13:1971–1980
Hess C, Means TK, Autissier P, Woodberry T, Altfeld M et al (2004) IL-8 responsiveness defines a subset of CD8 T cells poised to kill. Blood 104:3463–3471
Yang EV, Kim SJ, Donovan EL, Chen M, Gross AC et al (2009) Norepinephrine upregulates VEGF, IL-8, and IL-6 expression in human melanoma tumor cell lines: implications for stress-related enhancement of tumor progression. Brain Behav Immun 23:267–275
Ludwig A, Petersen F, Zahn S, Gotze O, Schroder JM et al (1997) The CXC-chemokine neutrophil-activating peptide-2 induces two distinct optima of neutrophil chemotaxis by differential interaction with interleukin-8 receptors CXCR-1 and CXCR-2. Blood 90:4588–4597
Rajagopalan L, Rajarathnam K (2004) Ligand selectivity and affinity of chemokine receptor CXCR1. Role of N-terminal domain. J Biol Chem 279:30000–30008
Gasser O, Missiou A, Eken C, Hess C (2005) Human CD8+ T cells store CXCR1 in a distinct intracellular compartment and up-regulate it rapidly to the cell surface upon activation. Blood 106:3718–3724
Gasser O, Schmid TA, Zenhaeusern G, Hess C (2006) Cyclooxygenase regulates cell surface expression of CXCR3/1-storing granules in human CD4+ T cells. J Immunol 177:8806–8812
Eikawa S, Ohue Y, Kitaoka K, Aji T, Uenaka A et al (2010) Enrichment of Foxp3+ CD4 regulatory T cells in migrated T cells to IL-6- and IL-8-expressing tumors through predominant induction of CXCR1 by IL-6. J Immunol 185:6734–6740
Shioda T, Nakayama EE, Tanaka Y, Xin X, Liu H et al (2001) Naturally occurring deletional mutation in the C-terminal cytoplasmic tail of CCR5 affects surface trafficking of CCR5. J Virol 75:3462–3468
Quereux G, Pandolfino MC, Knol AC, Khammari A, Volteau C et al (2007) Tissue prognostic markers for adoptive immunotherapy in melanoma. Eur J Dermatol 17:295–301
Meidenbauer N, Marienhagen J, Laumer M, Vogl S, Heymann J et al (2003) Survival and tumor localization of adoptively transferred Melan-A-specific T cells in melanoma patients. J Immunol 170:2161–2169
Richmond A, Thomas HG (1988) Melanoma growth stimulatory activity: isolation from human melanoma tumors and characterization of tissue distribution. J Cell Biochem 36:185–198
Rofstad EK, Halsor EF (2000) Vascular endothelial growth factor, interleukin 8, platelet-derived endothelial cell growth factor, and basic fibroblast growth factor promote angiogenesis and metastasis in human melanoma xenografts. Cancer Res 60:4932–4938
Cheong HS, Shin HD, Lee SO, Park BL, Choi YH et al (2006) Polymorphisms in interleukin 8 and its receptors (IL8, IL8RA and IL8RB) and association of common IL8 receptor variants with peripheral blood eosinophil counts. J Hum Genet 51:781
Dumitrascu D (1996) Mast cells as potent inflammatory cells. Rom J Intern Med 34:159–172
Chuntharapai A, Lee J, Hebert CA, Kim KJ (1994) Monoclonal antibodies detect different distribution patterns of IL-8 receptor A and IL-8 receptor B on human peripheral blood leukocytes. J Immunol 153:5682–5688
Inngjerdingen M, Damaj B, Maghazachi AA (2001) Expression and regulation of chemokine receptors in human natural killer cells. Blood 97:367–375
Smith ML, Olson TS, Ley K (2004) CXCR2- and E-selectin-induced neutrophil arrest during inflammation in vivo. J Exp Med 200:935–939
Peng W, Ye Y, Rabinovich BA, Liu C, Lou Y et al (2010) Transduction of tumor-specific T Cells with CXCR2 chemokine receptor improves migration to tumor and antitumor immune responses. Clin Cancer Res 16:5458–5468
Pule MA, Savoldo B, Myers GD, Rossig C, Russell HV et al (2008) Virus-specific T cells engineered to coexpress tumor-specific receptors: persistence and antitumor activity in individuals with neuroblastoma. Nat Med 14:1264–1270
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Many special thanks to Haya and Nehemia Lemelbaum for the enormous support that enabled the authors to conduct these studies.
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Sapoznik, S., Ortenberg, R., Galore-Haskel, G. et al. CXCR1 as a novel target for directing reactive T cells toward melanoma: implications for adoptive cell transfer immunotherapy. Cancer Immunol Immunother 61, 1833–1847 (2012). https://doi.org/10.1007/s00262-012-1245-1
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DOI: https://doi.org/10.1007/s00262-012-1245-1