Therapy with autologous T cells that have been gene-engineered to express chimeric antigen receptors (CAR) or T cell receptors (TCR) provides a feasible and broadly applicable treatment for cancer patients. In a clinical study in advanced renal cell carcinoma (RCC) patients with CAR T cells specific for carbonic anhydrase IX (CAIX), we observed toxicities that (most likely) indicated in vivo function of CAR T cells as well as low T cell persistence and clinical response rates. The latter observations were confirmed by later clinical trials in other solid tumor types and other gene-modified T cells. To improve the efficacy of T cell therapy, we have redefined in vitro conditions to generate T cells with young phenotype, a key correlate with clinical outcome. For their impact on gene-modified T cell phenotype and function, we have tested various anti-CD3/CD28 mAb-based T cell activation and expansion conditions as well as several cytokines prior to and/or after gene transfer using two different receptors: CAIX CAR and MAGE-C2(ALK)/HLA-A2 TCR. In a total set of 16 healthy donors, we observed that T cell activation with soluble anti-CD3/CD28 mAbs in the presence of both IL15 and IL21 prior to TCR gene transfer resulted in enhanced proportions of gene-modified T cells with a preferred in vitro phenotype and better function. T cells generated according to these processing methods demonstrated enhanced binding of pMHC, and an enhanced proportion of CD8+, CD27+, CD62L+, CD45RA+T cells. These new conditions will be translated into a GMP protocol in preparation of a clinical adoptive therapy trial to treat patients with MAGE-C2-positive tumors.
Get full access to this article
View all access options for this article.
References
1.
BondanzaA., ValtolinaV., MagnaniZ., et al. (2006). Suicide gene therapy of graft-versus-host disease induced by central memory human T lymphocytes. Blood, 107, 1828–1836.
2.
BrentjensR.J., DavilaM.L., RiviereI., et al. (2013). CD19-targeted T cells rapidly induce molecular remissions in adults with chemotherapy-refractory acute lymphoblastic leukemia. Sci. Transl. Med., 5, 177ra38.
3.
BrentjensR.J., RiviereI., ParkJ.H., et al. (2011). Safety and persistence of adoptively transferred autologous CD19-targeted T cells in patients with relapsed or chemotherapy refractory B-cell leukemias. Blood, 118, 4817–4828.
4.
ButlerM.O., FriedlanderP., MilsteinM.I., et al. (2011). Establishment of antitumor memory in humans using in vitro-educated CD8+ T cells. Sci. Transl. Med., 3, 80ra34.
5.
CieriN., CamisaB., CocchiarellaF., et al. (2013). IL-7 and IL-15 instruct the generation of human memory stem T cells from naive precursors. Blood, 121, 573–584.
6.
DavilaM.L., RiviereI., WangX., et al. (2014). Efficacy and toxicity management of 19-28z CAR T cell therapy in B cell acute lymphoblastic leukemia. Sci. Transl. Med., 6, 224ra25.
7.
DuarteR.F., ChenF.E., LowdellM.W., et al. (2002). Functional impairment of human T-lymphocytes following PHA-induced expansion and retroviral transduction: implications for gene therapy. Gene Ther., 9, 1359–1368.
8.
GattinoniL., and RestifoN.P. (2013). Moving T memory stem cells to the clinic. Blood, 121, 567–568.
9.
GattinoniL., KlebanoffC.A., PalmerD.C., et al. (2005). Acquisition of full effector function in vitro paradoxically impairs the in vivo antitumor efficacy of adoptively transferred CD8+ T cells. J. Clin. Invest., 115, 1616–1626.
10.
GattinoniL., PowellD.J.Jr., RosenbergS.A., and RestifoN.P. (2006). Adoptive immunotherapy for cancer: building on success. Nat. Rev. Immunol., 6, 383–393.
11.
GattinoniL., ZhongX.S., PalmerD.C., et al. (2009). Wnt signaling arrests effector T cell differentiation and generates CD8+ memory stem cells. Nat. Med., 15, 808–813.
12.
GattinoniL., LugliE., JiY., et al. (2011). A human memory T cell subset with stem cell-like properties. Nat. Med., 17, 1290–1297.
13.
GilhamD.E., DebetsR., PuleM., et al. (2012). CAR-T cells and solid tumors: tuning T cells to challenge an inveterate foe. Trends Mol. Med., 18, 377–384.
14.
GruppS.A., KalosM., BarrettD., et al. (2013). Chimeric antigen receptor-modified T cells for acute lymphoid leukemia. N. Engl. J. Med., 368, 1509–1518.
15.
HinrichsC.S., SpolskiR., PaulosC.M., et al. (2008). IL-2 and IL-21 confer opposing differentiation programs to CD8+ T cells for adoptive immunotherapy. Blood, 111, 5326–5333.
16.
HinrichsC.S., BormanZ.A., CassardL., et al. (2009). Adoptively transferred effector cells derived from naive rather than central memory CD8+ T cells mediate superior antitumor immunity. Proc. Natl. Acad. Sci. USA, 106, 17469–17474.
17.
HinrichsC.S., BormanZ.A., GattinoniL., et al. (2011). Human effector CD8+ T cells derived from naive rather than memory subsets possess superior traits for adoptive immunotherapy. Blood, 117, 808–814.
18.
HollymanD., StefanskiJ., PrzybylowskiM., et al. (2009). Manufacturing validation of biologically functional T cells targeted to CD19 antigen for autologous adoptive cell therapy. J. Immunother., 32, 169–180.
19.
HuangJ., KhongH.T., DudleyM.E., et al. (2005). Survival, persistence, and progressive differentiation of adoptively transferred tumor-reactive T cells associated with tumor regression. J. Immunother., 28, 258–267.
20.
HuarteE., FisherJ., TurkM.J., et al. (2009). Ex vivo expansion of tumor specific lymphocytes with IL-15 and IL-21 for adoptive immunotherapy in melanoma. Cancer Lett., 285, 80–88.
21.
JohnsonL.A., MorganR.A., DudleyM.E., 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.
22.
KalosM., LevineB.L., PorterD.L., et al. (2011). T cells with chimeric antigen receptors have potent antitumor effects and can establish memory in patients with advanced Leukemia. Sci Transl Med, 3, 95ra73.
23.
KanekoS., MastaglioS., BondanzaA., et al. (2009). IL-7 and IL-15 allow the generation of suicide gene-modified alloreactive self-renewing central memory human T lymphocytes. Blood, 113, 1006–1015.
24.
KershawM.H., WestwoodJ.A., ParkerL.L., et al. (2006). A phase I study on adoptive immunotherapy using gene-modified T cells for ovarian cancer. Clin. Cancer Res., 12, 6106–6115.
25.
KlebanoffC.A., FinkelsteinS.E., SurmanD.R., et al. (2004). IL-15 enhances the in vivo antitumor activity of tumor-reactive CD8+ T cells. Proc. Natl. Acad. Sci. USA, 101, 1969–1974.
26.
KlebanoffC.A., GattinoniL., Torabi-PariziP., et al. (2005). Central memory self/tumor-reactive CD8+ T cells confer superior antitumor immunity compared with effector memory T cells. Proc. Natl. Acad. Sci. USA, 102, 9571–9576.
27.
KochenderferJ.N., DudleyM.E., FeldmanS.A., et al. (2012). B-cell depletion and remissions of malignancy along with cytokine-associated toxicity in a clinical trial of anti-CD19 chimeric-antigen-receptor-transduced T cells. Blood, 119, 2709–2720.
28.
KunertA., StraetemansT., GoversC., et al. (2013). TCR-engineered T cells meet new challenges to treat solid tumors: choice of antigen, T cell fitness, and sensitization of tumor milieu. Front Immunol, 4, 363.
29.
LamersC.H., Van De GriendR.J., BraakmanE., et al. (1992). Optimization of culture conditions for activation and large-scale expansion of human T lymphocytes for bispecific antibody-directed cellular immunotherapy. Int. J. Cancer, 51, 973–979.
30.
LamersC.H., WillemsenR.A., LuiderB.A., et al. (2002). Protocol for gene transduction and expansion of human T lymphocytes for clinical immunogene therapy of cancer. Cancer Gene Ther., 9, 613–623.
31.
LamersC., Van ElzakkerP., LangeveldS., et al. (2006a). Process validation and clinical evaluation of a protocol to generate gene-modified T lymphocytes for imunogene therapy for metastatic renal cell carcinoma: GMP-controlled transduction and expansion of patient's T lymphocytes using a carboxy anhydrase IX-specific scFv transgene. Cytotherapy, 8, 542–553.
32.
LamersC.H., SleijferS., VultoA.G., et al. (2006b). Treatment of metastatic renal cell carcinoma with autologous T-lymphocytes genetically retargeted against carbonic anhydrase IX: first clinical experience. J. Clin. Oncol., 24, e20–e22.
33.
LamersC.H., WillemsenR.A., Van ElzakkerP., et al. (2006c). Phoenix-ampho outperforms PG13 as retroviral packaging cells to transduce human T cells with tumor-specific receptors: implications for clinical immunogene therapy of cancer. Cancer Gene Ther., 13, 503–509.
34.
LamersC.H., WillemsenR., Van ElzakkerP., et al. (2011). Immune responses to transgene and retroviral vector in patients treated with ex vivo-engineered T cells. Blood, 117, 72–82.
35.
LamersC.H., SleijferS., Van SteenbergenS., et al. (2013a). Treatment of Metastatic Renal Cell Carcinoma With CAIX CAR-engineered T cells: Clinical Evaluation and Management of On-target Toxicity. Mol. Ther., 21, 904–912.
36.
LamersC.H., Van ElzakkerP., Van SteenbergenS.C., et al. (2013b). Long-term stability of T-cell activation and transduction components critical to the processing of clinical batches of gene-engineered T cells. Cytotherapy, 15, 620–626.
37.
LeeD.W., KochenderferJ.N., Stetler-StevensonM., et al. (2014). T cells expressing CD19 chimeric antigen receptors for acute lymphoblastic leukaemia in children and young adults: a phase 1 dose-escalation trial. Lancet. Early Online Publication. DOI: 10.1016/S0140-6736(14)61403-3
38.
LiuS., RileyJ., RosenbergS., and ParkhurstM. (2006). Comparison of common gamma-chain cytokines, interleukin-2, interleukin-7, and interleukin-15 for the in vitro generation of human tumor-reactive T lymphocytes for adoptive cell transfer therapy. J. Immunother., 29, 284–293.
39.
LurquinC., LetheB., De PlaenE., et al. (2005). Contrasting frequencies of antitumor and anti-vaccine T cells in metastases of a melanoma patient vaccinated with a MAGE tumor antigen. J. Exp. Med., 201, 249–257.
40.
MarkleyJ.C., and SadelainM. (2010). IL-7 and IL-21 are superior to IL-2 and IL-15 in promoting human T cell-mediated rejection of systemic lymphoma in immunodeficient mice. Blood, 115, 3508–3519.
41.
MaudeS.L., FreyN., ShawP.A., et al. (2014). Chimeric antigen receptor T cells for sustained remissions in leukemia. N. Engl. J. Med., 371, 1507–1517.
42.
MorganR.A., DudleyM.E., WunderlichJ.R., et al. (2006). Cancer regression in patients after transfer of genetically engineered lymphocytes. Science, 314, 126–129.
43.
PouwN., Treffers-WesterlakenE., KraanJ., et al. (2010a). Combination of IL-21 and IL-15 enhances tumour-specific cytotoxicity and cytokine production of TCR-transduced primary T cells. Cancer Immunol. Immunother., 59, 921–931.
44.
PouwN., Treffers-WesterlakenE., MondinoA., et al. (2010b). TCR gene-engineered T cell: Limited T cell activation and combined use of IL-15 and IL-21 ensure minimal differentiation and maximal antigen-specificity. Mol. Immunol., 47, 1411–1420.
45.
RobbinsP.F., DudleyM.E., WunderlichJ., 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.
46.
RobbinsP.F., MorganR.A., FeldmanS.A., et al. (2011). Tumor regression in patients with metastatic synovial cell sarcoma and melanoma using genetically engineered lymphocytes reactive with NY-ESO-1. J. Clin. Oncol., 29, 917–924.
47.
RosenbergS.A., YangJ.C., SherryR.M., et al. (2011). Durable complete responses in heavily pretreated patients with metastatic melanoma using T cell transfer immunotherapy. Clin. Cancer Res., 17, 4550–4557.
48.
SchneidersF.L., De BruinR.C., Van Den EertweghA.J., et al. (2012). Circulating invariant natural killer T-cell numbers predict outcome in head and neck squamous cell carcinoma: updated analysis with 10-year follow-up. J. Clin. Oncol., 30, 567–570.
49.
StraetemansT., Van BrakelM., Van SteenbergenS., et al. (2012). TCR gene transfer: MAGE-C2/HLA-A2 and MAGE-A3/HLA-DP4 epitopes as melanoma-specific immune targets. Clin. Dev. Immunol., 2012, 586314.
50.
TillB.G., JensenM.C., WangJ., et al. (2008). Adoptive immunotherapy for indolent non-Hodgkin lymphoma and mantle cell lymphoma using genetically modified autologous CD20-specific T cells. Blood, 112, 2261–2271.
51.
UemuraH., OkajimaE., DebruyneF.M.J., and OosterwijkE. (1994). Internal image antiidiotype antibodies related to renal-cell carcinoma-associated antigen G250. Int. J. Cancer, 56, 609–614.
52.
WangX., BergerC., WongC.W., et al. (2011). Engraftment of human central memory-derived effector CD8+ T cells in immunodeficient mice. Blood, 117, 1888–1898.
53.
YangS., JiY., GattinoniL., et al. (2013). Modulating the differentiation status of ex vivo-cultured anti-tumor T cells using cytokine cocktails. Cancer Immunol. Immunother., 62, 727–736.
Supplementary Material
Please find the following supplemental material available below.
For Open Access articles published under a Creative Commons License, all supplemental material carries the same license as the article it is associated with.
For non-Open Access articles published, all supplemental material carries a non-exclusive license, and permission requests for re-use of supplemental material or any part of supplemental material shall be sent directly to the copyright owner as specified in the copyright notice associated with the article.
