scholarly journals 101 Expansion of tumor-infiltrating lymphocytes (TIL) using static bag for the clinical manufacturing rapid expansion protocol (REP) process

2021 ◽  
Vol 9 (Suppl 3) ◽  
pp. A110-A110
Author(s):  
Kenneth Onimus ◽  
Adrian Wells ◽  
Nermin Gerges ◽  
Courtney Herman ◽  
Shwetha Lakshmipathi ◽  
...  
Keyword(s):  
Cell Therapy ◽  
Clin Oncol ◽  
Tumor Infiltrating Lymphocytes ◽  
Full Scale ◽  
Rapid Expansion ◽  
Effector Memory ◽  
Small Scale ◽  
Autologous Tumor ◽  
List Type ◽  
Infiltrating Lymphocytes

BackgroundLifileucel (LN-144) and LN-145, adoptive cell therapies using autologous tumor-infiltrating lymphocytes (TIL), have demonstrated encouraging efficacy with acceptable safety in a variety of tumor types.1–4 The lifileucel Gen 2 clinical manufacturing process uses gas-permeable rapid expansion (G-Rex®, Wilson Wolf, Saint Paul, MN) bioreactors for TIL expansion.5 Static gas-permeable cell culture bags (EXP-Pak™, Charter Medical, Winston-Salem, NC) are alternate bioreactors that have been used for clinical manufacturing of T cells.6 In this study, TIL product characteristics were compared after expansion at small- and full-scale using EXP-Pak bags and G-Rex bioreactors.MethodsCryopreserved pre-REP TIL were cultured with OKT-3 and irradiated peripheral blood mononuclear cells in either small-scale (G-Rex 5M flasks or EXP-50 bags) or full-scale (G-Rex 500MCS or EXP-5L bags) conditions. The same culture media formulation was used throughout the process. Final harvested TIL products were characterized for the following quality attributes: total viable cells (TVC), purity (% viability), identity (% CD45+CD3+), and activity (IFN-gamma release). Additional characterization was performed to determine the TIL differentiation, central and effector memory subsets, activation, exhaustion status, and impurities using multi-color flow cytometry. The T-cell receptor (TCR) repertoires of the final products were assessed for unique CDR3 (uCDR3) counts and shared clones using RNA-seq.ResultsThe median (range) TVC from 6 small-scale runs of G-Rex condition was 90.8×109 (26.9×109–98.6×109) cells, and the corresponding TVC of EXP-Pak bag condition was 119×109 (63.2×109–141×109) cells. Full-scale runs (n=3) yielded similar TVC for G-Rex and EXP-Pak bag conditions. Both conditions had comparable purity, identity, and activity (table 1). 91–99% of TCR Vbeta clones of EXP-Pak bag TIL were present in G-Rex (table 2). Cell proliferation, cell cycle, and mitochondrial function of TIL generated from EXP-Pak bags and G-Rex flask methodologies will be presented.ConclusionsThe final TIL product generated in the EXP-Pak bag condition did not differ from cells produced in the G-Rex flask functionally or phenotypically and demonstrated similar growth profiles. These data support further evaluation of EXP-Pak bags for potential use in clinical and potential commercial TIL cell therapy manufacturing applications.AcknowledgementsThis study and analysis were funded by Iovance Biotherapeutics, Inc. (San Carlos, CA, USA). Editorial support was provided by Amanda Kelly (Iovance).ReferencesSarnaik AA, et al. J Clin Oncol. 2021; doi: 10.1200/JCO.21.00612.Thomas SS, et al. J Clin Oncol. 2021;39 (suppl; abstract 9537).Jazaeri A, et al. J Clin Oncol. 2019;37 (suppl; abstract 2538).Jimeno A, et al. J Immunother Cancer. 2020;8 (suppl; abstract A378).Wardell S, et al. J Immunother Cancer. 2019;7 (suppl 1; abstract P226).BioProcess International. 2014 [white paper]. https://bioprocessintl.com/sponsored-content/efficacy-utility-new-exp-pak-closed-system-disposable-cell-expansion-bags-designed-cell-therapy-applications/Accessed July 23, 2021.Abstract 101 Table 1Summary of final TIL product attributesAbstract 101 Table 2Common uCDR3 clones and% overlap of TCR clones

The impact of checkpoint blockade prior to adoptive cell therapy using tumor-infiltrating lymphocytes for metastatic melanoma: An update from MD Anderson Cancer Center.

2017 ◽  
Vol 35 (7_suppl) ◽  
pp. 138-138 ◽  
Author(s):  
Marie-Andree Forget ◽  
Cara L. Haymaker ◽  
Kenneth R. Hess ◽  
Jason Roszik ◽  
Scott Eric Woodman ◽  
...  
Keyword(s):  
Cell Therapy ◽  
Metastatic Melanoma ◽  
Adoptive Cell Therapy ◽  
Tumor Infiltrating Lymphocytes ◽  
Checkpoint Blockade ◽  
High Dose ◽  
Autologous Tumor ◽  
Initial Report ◽  
Infiltrating Lymphocytes ◽  
The Impact

138 Background: Adoptive cell therapy (ACT) of autologous tumor-infiltrating lymphocytes (TIL) can produce long lasting treatment responses in patients (pts) with metastatic melanoma. In our initial report of 31 treated pts, we demonstrated overall response rate (ORR) of 48%. Due to the evolution of treatment options for metastatic melanoma with heavy reliance on immunomodulatory therapies, we sought to re-evaluate responses in the era of checkpoint blockade. Methods: Pts receive treatment consisting of 7 days of lymphodepleting chemotherapy consisting of cyclophosphamide and fludarabine. High dose interleukin-2 (720,000 IU/kg IV q 8 hrs up to 15 doses) was infused after TIL infusion. Responses were assessed by imaging per irRC. Results: A total of 74 pts have been treated in our initial TIL study with an updated ORR assessment of 43%. Stratification of pts according to their checkpoint blockade immune-modulator therapy prior to TIL ACT, demonstrated that prior Ipilimumab (Ipi) decreased ORR to 33% compared to 51% in checkpoint naïve pts and decreased overall survival (OS) post-TIL therapy (median 7.7 vs 24.6 months respectively). There were too few pts to assess the impact of anti-PD1 as a single agent. A multiparametric analysis revealed that LDH levels at the time of therapy mainly influences OS and ORR to TIL but still could not invalidate the impact of Ipi prior to TIL ACT. The durability of the response was also found to be different between the 2 groups (30% for Ipi prior and 50% No Ipi prior). This new reality also impacted previously reported parameters that correlated with ORR to TIL ACT such as the expression of B-and-T lymphocyte attenuator (BTLA) on CD8+ TIL. Interestingly, assessment of mutation load (ML) of the tumors prior to TIL ACT showed that the check point naïve group display a high ML ( > 100) within their tumor whereas some patients who had Ipi prior to TIL have a low ML ( < 100). Conclusions: Our preliminary analysis shows that pre-treatment with Ipi decreases ORR to TIL ACT. Understanding how prior ipi modifies TIL, the tumor and microenvironment will help to define the full extent of the impact and how to best treat Ipi refractory pts with TIL. Clinical trial information: NCT00338377.

scholarly journals Characterization of Ex Vivo Expanded Tumor Infiltrating Lymphocytes from Patients with Malignant Melanoma for Clinical Application

2011 ◽  
Vol 2011 ◽  
pp. 1-6 ◽  
Author(s):  
Niels Junker ◽  
Per thor Straten ◽  
Mads Hald Andersen ◽  
Inge Marie Svane
Keyword(s):  
Malignant Melanoma ◽  
Specific Activity ◽  
Ex Vivo ◽  
Tumor Infiltrating Lymphocytes ◽  
Effector Memory ◽  
Autologous Tumor ◽  
Advanced Malignant Melanoma ◽  
Clinical Responses ◽  
Infiltrating Lymphocytes

Clinical trials of adoptive transfer of autologous tumor infiltrating lymphocytes (TILs) to patients with advanced malignant melanoma have shown remarkable results with objective clinical responses in 50% of the treated patients. In order to initiate a clinical trial in melanoma, we have established a method for expanding TILs to clinical relevant quantities in two steps with in 8 weeks. Further characterization of expanded TILs revealed an oligoclonal composition of T-cells with an effector memory like phenotype. When autologous tumor was available, TILs showed specific activity in all patients tested. TIL cultures contained specificity towards tumor cells as well as peptides derived from tumor-associated antigens (TAAs) during expansion procedures.

scholarly journals Randomized Selection Design Trial Evaluating CD8+-Enriched Versus Unselected Tumor-Infiltrating Lymphocytes for Adoptive Cell Therapy for Patients With Melanoma

2013 ◽  
Vol 31 (17) ◽  
pp. 2152-2159 ◽  
Author(s):  
Mark E. Dudley ◽  
Colin A. Gross ◽  
Robert P.T. Somerville ◽  
Young Hong ◽  
Nicholas P. Schaub ◽  
...  
Keyword(s):  
Cell Therapy ◽  
Adoptive Cell Therapy ◽  
Interleukin 2 ◽  
Evaluation Criteria ◽  
Tumor Infiltrating Lymphocytes ◽  
Peripheral Blood Lymphocytes ◽  
Response To Treatment ◽  
High Dose ◽  
Autologous Tumor ◽  
Infiltrating Lymphocytes

Purpose Adoptive cell therapy (ACT) with autologous tumor-infiltrating lymphocytes (TILs) and high-dose interleukin-2 (IL-2) administered to lymphodepleted patients with melanoma can cause durable tumor regressions. The optimal TIL product for ACT is unknown. Patients and Methods Patients with metastatic melanoma were prospectively assigned to receive unselected young TILs versus CD8+-enriched TILs. All patients received lymphodepleting chemotherapy and high-dose IL-2 therapy and were assessed for response, toxicity, survival, and immunologic end points. Results Thirty-four patients received unselected young TILs with a median of 8.0% CD4+ lymphocytes, and 35 patients received CD8+-enriched TILs with a median of 0.3% CD4+ lymphocytes. One month after TIL infusion, patients who received CD8+-enriched TILs had significantly fewer CD4+ peripheral blood lymphocytes (P = .01). Twelve patients responded to therapy with unselected young TILs (according to Response Evaluation Criteria in Solid Tumors [RECIST]), and seven patients responded to CD8+-enriched TILs (35% v 20%; not significant). Retrospective studies showed a significant association between response to treatment and interferon gamma secretion by the infused TILs in response to autologous tumor (P = .04), and in the subgroup of patients who received TILs from subcutaneous tumors, eight of 15 patients receiving unselected young TILs responded but none of eight patients receiving CD8+-enriched TILs responded. Conclusion A randomized selection design trial was feasible for improving individualized TIL therapy. Since the evidence indicates that CD8+-enriched TILs are not more potent therapeutically and they are more laborious to prepare, future studies should focus on unselected young TILs.

scholarly journals Adoptive Cell Therapy with Tumor-Infiltrating Lymphocytes in Advanced Melanoma Patients

2018 ◽  
Vol 2018 ◽  
pp. 1-10 ◽  
Author(s):  
Mélanie Saint-Jean ◽  
Anne-Chantal Knol ◽  
Christelle Volteau ◽  
Gaëlle Quéreux ◽  
Lucie Peuvrel ◽  
...  
Keyword(s):  
Cell Therapy ◽  
Adoptive Cell Therapy ◽  
Interleukin 2 ◽  
Tumor Infiltrating Lymphocytes ◽  
Advanced Melanoma ◽  
Autologous Tumor ◽  
Unresectable Stage ◽  
Melanoma Patients ◽  
Infiltrating Lymphocytes

Immunotherapy for melanoma includes adoptive cell therapy with autologous tumor-infiltrating lymphocytes (TILs). This monocenter retrospective study was undertaken to evaluate the efficacy and safety of this treatment of patients with advanced melanoma. All advanced melanoma patients treated with TILs using the same TIL expansion methodology and same treatment interleukin-2 (IL-2) regimen between 2009 and 2012 were included. After sterile intralesional excision of a cutaneous or subcutaneous metastasis, TILs were produced according to a previously described method and then infused into the patient who also received a complementary subcutaneous IL-2 regimen. Nine women and 1 man were treated for unresectable stage IIIC (n=4) or IV (n=6) melanoma. All but 1 patient with unresectable stage III melanoma (1st line) had received at least 2 previous treatments, including anti-CTLA-4 antibody for 4. The number of TILs infused ranged from 0.23 × 109 to 22.9 × 109. Regarding safety, no serious adverse effect was reported. Therapeutic responses included a complete remission, a partial remission, 2 stabilizations, and 6 progressions. Among these 4 patients with clinical benefit, 1 is still alive with 9 years of follow-up and 1 died from another cause after 8 years of follow-up. Notably, patients treated with high percentages of CD4 + CD25 + CD127lowFoxp3+ T cells among their TILs had significantly shorter OS. The therapeutic effect of combining TILs with new immunotherapies needs further investigation.

Adoptive cell therapy for melanoma patients using tumor-infiltrating lymphocytes, lymphodepleting chemotherapy, and low-dose interleukin-2.

2012 ◽  
Vol 30 (15_suppl) ◽  
pp. 2574-2574
Author(s):  
Eva Ellebaek ◽  
Trine Zeeberg Iversen ◽  
Niels Junker ◽  
Marco Donia ◽  
Lotte Engell-Noerregaard ◽  
...  
Keyword(s):  
Cell Therapy ◽  
Low Dose ◽  
Adoptive Cell Therapy ◽  
Pilot Trial ◽  
Interleukin 2 ◽  
Tumor Infiltrating Lymphocytes ◽  
Complete Response ◽  
High Dose ◽  
Autologous Tumor ◽  
Infiltrating Lymphocytes

2574 Background: Adoptive T cell therapy is based on isolation of tumor infiltrating lymphocytes (TIL) from autologous tumor, in vitro activation and expansion, and the reinfusion of these cells into a lymphodepleted patient. Together with high-dose interleukin (IL)-2 this treatment has in a few other countries successively been given to patients with advanced malignant melanoma and an impressive 50% response rate has been observed. Here we report the experience from a Danish Translational Research Center using low-dose subcutaneous IL-2 injections. Methods: A pilot trial including patients with metastatic melanoma, PS ≤1, age <70, measurable and progressive disease, and no CNS involvement. Six patients were treated with lymphodepleting chemotherapy, TIL infusion, and 14 days of low-dose IL-2, 2 MIU/day. Results: Low-dose IL-2 considerably decreased the toxicity of the treatment with no grade 3-4 events related to this drug. Two of the 6 treated patients have an ongoing complete response (30+ and 7+ months), 2 patients had stable disease (4.5 and 5 months) and 2 patients progressed shortly after treatment. Five patients went through surgery but were not treated because of rapid disease progression (2), development of brain metastases (2) or the inability to grow cells (1). Tumor-reactivity of the infused cells and peripheral monocytes before and after therapy were analyzed. High tumor responses in the infusion products were correlated to clinical response and also an induction of tumor reactive T cells were seen in the peripheral blood for 1 patient in complete response. Conclusions: Complete and durable responses are induced after treatment with adoptive cell transfer in combination with low-dose IL-2 questioning the need for high and toxic doses of IL-2.

Sign in / Sign up

Export Citation Format

Share Document