Tumor-associated antigen-specific T cells with nivolumab are safe and persist in vivo in relapsed/refractory Hodgkin lymphoma

Abstract

Hodgkin lymphoma (HL) Reed Sternberg cells express tumor-associated antigens (TAA) that are potential targets for cellular therapies. We recently demonstrated that TAA-specific T cells (TAA-Ts) targeting WT1, PRAME, and Survivin were safe and associated with prolonged time to progression in solid tumors. Hence, we evaluated whether TAA-Ts when given alone or with nivolumab were safe and could elicit antitumor effects in vivo in patients with relapsed/refractory (r/r) HL. Ten patients were infused with TAA-Ts (8 autologous and 2 allogeneic) for active HL (n = 8) or as adjuvant therapy after hematopoietic stem cell transplant (n = 2). Six patients received nivolumab priming before TAA-Ts and continued until disease progression or unacceptable toxicity. All 10 products recognized 1 or more TAAs and were polyfunctional. Patients were monitored for safety for 6 weeks after the TAA-Ts and for response until disease progression. The infusions were safe with no clear dose-limiting toxicities. Patients receiving TAA-Ts as adjuvant therapy remain in continued remission at 3+ years. Of the 8 patients with active disease, 1 patient had a complete response and 7 had stable disease at 3 months, 3 of whom remain with stable disease at 1 year. Antigen spreading and long-term persistence of TAA-Ts in vivo were observed in responding patients. Nivolumab priming impacted TAA-T recognition and persistence. In conclusion, treatment of patients with r/r HL with TAA-Ts alone or in combination with nivolumab was safe and produced promising results. This trial was registered at www.clinicaltrials.gov as #NCT022039303 and #NCT03843294.

Trial registration: ClinicalTrials.gov NCT03843294 NCT22039303.

© 2022 by The American Society of Hematology. Licensed under Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International (CC BY-NC-ND 4.0), permitting only noncommercial, nonderivative use with attribution. All other rights reserved.

Figures

Graphical abstract

Figure 1.

Phenotype of TAA-T products. (A) Fold expansion (n = 12). (B) Immunophenotype of the TAA-T products (n = 10). (C) Polyclonality of TAA-T as assessed by TCR-Vβ deep sequencing (n = 6). (D) Lack of cytotoxicity of TAA-T (effectors) against non–antigen-pulsed PHA blasts (targets) at an various effector to target ratios (n = 10). (E) Exhaustion markers on TAA-T products (n = 6). PHA, phytohaemagglutinin

Figure 2.

Functional characterization of TAA-T products. (A) Tumor antigen specificity as measured by IFNγ ELISPOT assay of the 12 infused products after overnight restimulation with overlapping 15mer pepmixes of actin (irrelevant control antigen), WT1, PRAME, and Survivin. (B) Polyfunctionality as assessed by the release of IFNγ and TNFα by CD3+, CD4+, and CD8+ TAA-Ts in response to irrelevant antigen. Actin and TAA (PRAME) shown in a representative dot plot of TAA-T product and summarized for 6 TAA-T products (C). * and ** denote P < .01 for difference between actin and PRAME.

Figure 3.

Clinical outcomes. Swimmers plot showing the outcome of patients after receiving TAA-Ts with or without nivolumab. (A) Patients in remission at the time of TAA-T infusion. (B) Patients with measurable disease at the time of TAA-T infusion. The timing of nivolumab is indicated by the blue bar in relation to the TAA-T infusion, which is indicated by an asterisk.

Figure 4.

Recognition of tumor specific antigens over time. Recovery of antigen-specific T cells and antigen spreading as detected by IFNγ ELISPOT assay in patients receiving TAA-T alone in responders (A-B) and nonresponders (C-D). For each figure, reactivity to the targeted antigens WT1, PRAME, and Survivin is shown in panel i and nontargeted antigens MAGE family, SSX2, and SOX2 is in panel ii.

Figure 5.

Recognition of tumor specific antigens over time. Recovery of antigen-specific T cells and antigen spreading as detected by IFNγ ELISPOT assay in patients receiving TAA-T and nivolumab in responders (A-D) and nonresponders (E-F). For each figure, reactivity to the targeted antigens WT1, PRAME, and Survivin is shown in panel i and nontargeted antigens MAGE family, SSX2, and SOX2 in panel ii.

Figure 6.

Impact of nivolumab on persistence of functional TAA-T cells. (A) Comparison of T-cell reactivity to the targeted antigens as detected by IFNγ ELISPOT assay after nivolumab priming and before TAA-T infusion and at 3 months between responding (R) and nonresponding patients (NR). (B) Persistence of polyfunctional TAA-T cells over time in responders. (C) Persistence of polyfunctional TAA-T cells over time in nonresponders. (D) Representative plot of patient demonstrating recovery of in vivo polyfunctional CD4+ and CD8+ TAA-T cells secreting both IFNγ and TNFα was detected after brief ex vivo expansion after restimulation with antigens at several follow-up time points.

Figure 7.

Persistence of new TAA-T clones expanded in the product that were not present at baseline. (A) Using TCRVβ deep sequencing, new clones detected in the TAA-T product were identified and tracked in PB over time. (B) Longitudinal tracking of new clones expanded in product over time for patients receiving TAA-T alone. Multi-institutional Prospective Research of Expanded Multi-antigen Specifically Oriented Lymphocytes for the Treatment of VEry High Risk Hematopoietic Malignancies (RESOLVE) and TAA-T with nivolumab [TAA-T with nivolumab on Phase I Study Utilizing Tumor Associated Antigen Specific T cells (TAA-T) with PD1 Inhibitor Nivolumab for Relapsed/Refractory Lymphoma (SUSTAIN) trial]. (C) Tracking of clones enriched in TAA-T product and not present at baseline in patient 6.