Activation and propagation of tumor-infiltrating lymphocytes on clinical-grade designer artificial antigen-presenting cells for adoptive immunotherapy of melanoma

Marie Andrée Forget, Shruti Malu, Hui Liu, Christopher Toth, Sourindra Maiti, Charuta Kale, Cara Haymaker, Chantale Bernatchez, Helen Huls, Ena Wang, Francesco M. Marincola, Patrick Hwu, Laurence J N Cooper, Laszlo G. Radvanyi

Research output: Contribution to journalArticle

23 Citations (Scopus)

Abstract

Purpose: Adoptive cell therapy with autologous tumor-infiltrating lymphocytes (TIL) is a therapy for metastatic melanoma with response rates of up to 50%. However, the generation of the TIL transfer product is challenging, requiring pooled allogeneic normal donor peripheral blood mononuclear cells (PBMC) used in vitro as "feeders" to support a rapid-expansion protocol. Here, we optimized a platform to propagate TIL to a clinical scale using K56 cells genetically modified to express costimulatory molecules such as CD86, CD137-ligand, and membrane-bound IL-15 to function as artificial antigen-presenting cells (aAPC) as an alternative to using PBMC feeders.

Experimental Design: We used aAPC or γ-irradiated PBMC feeders to propagate TIL and measured rates of expansion. The activation and differentiation state was evaluated by flow cytometry and differential gene expression analyses. Clonal diversity was assessed on the basis of the pattern of T-cell receptor usage. T-cell effector function was measured by evaluation of cytotoxic granule content and killing of target cells.

Results: The aAPC propagated TIL at numbers equivalent to that found with PBMC feeders, whereas increasing the frequency of CD8+ T-cell expansion with a comparable effector-memory phenotype. mRNA profiling revealed an upregulation of genes in the Wnt and stem-cell pathways with the aAPC. The aAPC platform did not skew clonal diversity, and CD8+ T cells showed comparable antitumor function as those expanded with PBMC feeders.

Conclusions: TIL can be rapidly expanded with aAPC to clinical scale generating T cells with similar phenotypic and effector profiles as with PBMC feeders. These data support the clinical application of aAPC to manufacture TIL for the treatment of melanoma.

Original languageEnglish
Pages (from-to)448-460
Number of pages13
JournalJournal of Immunotherapy
Volume37
Issue number9
Publication statusPublished - 10 Dec 2014

Fingerprint

Tumor-Infiltrating Lymphocytes
Adoptive Immunotherapy
Antigen-Presenting Cells
Melanoma
Blood Cells
T-Lymphocytes
4-1BB Ligand
Interleukin-15
Lymphocyte Count
Cell- and Tissue-Based Therapy
T-Cell Antigen Receptor
Flow Cytometry
Research Design
Up-Regulation
Stem Cells
Phenotype
Gene Expression
Messenger RNA
Membranes

Keywords

  • Adoptive T-cell therapy
  • Artificial antigen-presenting cells
  • Costimulation
  • K562
  • Melanoma
  • Tumor-infiltrating lymphocytes

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology
  • Pharmacology
  • Cancer Research

Cite this

Activation and propagation of tumor-infiltrating lymphocytes on clinical-grade designer artificial antigen-presenting cells for adoptive immunotherapy of melanoma. / Forget, Marie Andrée; Malu, Shruti; Liu, Hui; Toth, Christopher; Maiti, Sourindra; Kale, Charuta; Haymaker, Cara; Bernatchez, Chantale; Huls, Helen; Wang, Ena; Marincola, Francesco M.; Hwu, Patrick; Cooper, Laurence J N; Radvanyi, Laszlo G.

In: Journal of Immunotherapy, Vol. 37, No. 9, 10.12.2014, p. 448-460.

Research output: Contribution to journalArticle

Forget, MA, Malu, S, Liu, H, Toth, C, Maiti, S, Kale, C, Haymaker, C, Bernatchez, C, Huls, H, Wang, E, Marincola, FM, Hwu, P, Cooper, LJN & Radvanyi, LG 2014, 'Activation and propagation of tumor-infiltrating lymphocytes on clinical-grade designer artificial antigen-presenting cells for adoptive immunotherapy of melanoma', Journal of Immunotherapy, vol. 37, no. 9, pp. 448-460.
Forget, Marie Andrée ; Malu, Shruti ; Liu, Hui ; Toth, Christopher ; Maiti, Sourindra ; Kale, Charuta ; Haymaker, Cara ; Bernatchez, Chantale ; Huls, Helen ; Wang, Ena ; Marincola, Francesco M. ; Hwu, Patrick ; Cooper, Laurence J N ; Radvanyi, Laszlo G. / Activation and propagation of tumor-infiltrating lymphocytes on clinical-grade designer artificial antigen-presenting cells for adoptive immunotherapy of melanoma. In: Journal of Immunotherapy. 2014 ; Vol. 37, No. 9. pp. 448-460.
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AU - Forget, Marie Andrée

AU - Malu, Shruti

AU - Liu, Hui

AU - Toth, Christopher

AU - Maiti, Sourindra

AU - Kale, Charuta

AU - Haymaker, Cara

AU - Bernatchez, Chantale

AU - Huls, Helen

AU - Wang, Ena

AU - Marincola, Francesco M.

AU - Hwu, Patrick

AU - Cooper, Laurence J N

AU - Radvanyi, Laszlo G.

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N2 - Purpose: Adoptive cell therapy with autologous tumor-infiltrating lymphocytes (TIL) is a therapy for metastatic melanoma with response rates of up to 50%. However, the generation of the TIL transfer product is challenging, requiring pooled allogeneic normal donor peripheral blood mononuclear cells (PBMC) used in vitro as "feeders" to support a rapid-expansion protocol. Here, we optimized a platform to propagate TIL to a clinical scale using K56 cells genetically modified to express costimulatory molecules such as CD86, CD137-ligand, and membrane-bound IL-15 to function as artificial antigen-presenting cells (aAPC) as an alternative to using PBMC feeders.Experimental Design: We used aAPC or γ-irradiated PBMC feeders to propagate TIL and measured rates of expansion. The activation and differentiation state was evaluated by flow cytometry and differential gene expression analyses. Clonal diversity was assessed on the basis of the pattern of T-cell receptor usage. T-cell effector function was measured by evaluation of cytotoxic granule content and killing of target cells.Results: The aAPC propagated TIL at numbers equivalent to that found with PBMC feeders, whereas increasing the frequency of CD8+ T-cell expansion with a comparable effector-memory phenotype. mRNA profiling revealed an upregulation of genes in the Wnt and stem-cell pathways with the aAPC. The aAPC platform did not skew clonal diversity, and CD8+ T cells showed comparable antitumor function as those expanded with PBMC feeders.Conclusions: TIL can be rapidly expanded with aAPC to clinical scale generating T cells with similar phenotypic and effector profiles as with PBMC feeders. These data support the clinical application of aAPC to manufacture TIL for the treatment of melanoma.

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