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185 Ex vivo efficacy of adoptive T cell therapy using autologous CD8+ T cells isolated by PD-1 positivity from peripheral blood mononuclear cells in solid tumors
  1. Sungkyu Lee1 and
  2. Yong Wha Moon2
  1. 1Seoul National University, Seoul, Korea, Republic of
  2. 2CHA University, Seongnam, Korea, Republic of


Background Tumor-infiltrating lymphocytes have been shown to display an antitumor activity in solid tumors including melanoma. Previously the proof of concept was developed that CD8+ T cells isolated by programmed cell death-1 (PD-1) receptor positivity from peripheral blood mononuclear cells (PBMCs), recognized tumor-specific antigens and neoantigens, and thus were reactive to tumors. We evaluated ex vivo efficacy of autologous, tumor-reactive CD8+ T cells isolated by PD-1 positivity from PBMCs of breast cancer and leiomyosarcoma patients.

Methods Fresh tumor tissues were cultured in RPMI supplemented with 20% FBS. PD-1 positive CD8+ T cells were isolated from PBMCs of the patients and expanded with Dynabeads human T-Activator CD3/CD28 in RPMI supplemented with 10% FBS and IL-2. Antitumor activity of PD-1-positive CD8+ T cells was tested by performing cytotoxic T lymphocyte (CTL) assay with the cultured autologous tumor cell targets. Specifically, the CTL assay was carried out by culturing tumor cells in multi-well plates overnight, and then adding the PD-1+CD8+ T cells to the plates. After 2 days, tumor cells were harvested from the wells and stained for CD45, CD8 and Live/Dead (ThermoFisher Scientific) to determine the killing activity of PD-1+CD8+ T cells. The results were compared to ones obtained with PD-1-CD8+ T cells.

Results We have successfully established ex vivo models for adoptive cell therapy using autologous PD-1+CD8+ T cells from three breast cancer and one leiomyosarcoma patients. Two breast cancer patients were hormone-receptor-positive and one was triple-negative. Stages were IA (n=1) or IIB (n=2). One leiomyosarcoma patient was in stage IV. All tumor and peripheral blood samples were obtained at chemotherapy-naïve status. CTL assays demonstrated that PD-1+CD8+ T cells efficiently killed target cells compared to PD-1-CD8+ T cells in all the breast cancer and leiomyosarcoma models. More and detailed results will be presented and discussed.

Conclusions Autologous PD-1+CD8+ T cells isolated from PBMCs of a few cancer patients displayed efficient antitumor activity in our established ex vivo models for adoptive cell therapy. Our results warrant further clinical development for adoptive T cell therapy in various types of cancers.

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