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Construction of an anti-programmed death-ligand 1 chimeric antigen receptor and determination of its antitumor

Jiasen Xie1,2,3, Zishan Zhou2,3, Shunchang Jiao2

  • 1Department of Biochemistry, College of Basic Medical Sciences, Jilin University, Changchun, Jilin 130021, P.R. China.

Oncology Letters
|June 22, 2018
PubMed

Insights

Researchers designed a programmed death-ligand 1 (PD-L1)-specific chimeric antigen receptor (CAR) for T cells. This novel CAR demonstrated mild cytotoxicity against PD-L1-positive cancer cells, indicating potential for solid tumor treatment.

Area of Science:

  • Immunology
  • Oncology
  • Biotechnology

Background:

  • Chimeric antigen receptors (CARs) are engineered proteins for targeted cancer immunotherapy.
  • Programmed death-ligand 1 (PD-L1) is a key immune checkpoint protein often overexpressed in solid tumors.
  • Developing CARs targeting PD-L1 offers a potential strategy to overcome tumor-induced immunosuppression.

Purpose of the Study:

  • To design and construct a novel PD-L1-specific CAR.
  • To evaluate the transduction efficiency and expression of the PD-L1 CAR on T cells.
  • To assess the anti-tumor efficacy and cytokine production of PD-L1 CAR-transduced T cells against solid tumor cells.

Main Methods:

  • A PD-L1-specific CAR was engineered using a single-chain variable fragment (scFv), CD28, 4-1BB, and CD3ζ signaling domains.
  • The CAR construct was cloned into a lentiviral vector (pLVX) for T cell transduction.
  • Flow cytometry was used to confirm CAR expression on transduced peripheral blood mononuclear cells (PBMCs).
  • Cytotoxicity assays were performed using PD-L1-positive NCI-H358 and PD-L1-low A549 cancer cell lines.
  • Enzyme-linked immunosorbent assay (ELISA) was employed to measure cytokine release (IFN-γ, IL-2, TNF-α).

Main Results:

  • Efficient transduction and successful expression of the PD-L1 CAR on T cells were confirmed by flow cytometry.
  • Mild cytotoxicity of CAR-transduced T cells was observed against PD-L1-positive NCI-H358 cells at an effector-to-target ratio of 10:1.
  • A significant increase in interferon-γ (IFN-γ) release was detected upon co-culture with PD-L1-positive tumor cells.
  • Interleukin-2 (IL-2) and tumor necrosis factor-α (TNF-α) levels remained unchanged.

Conclusions:

  • The engineered PD-L1-specific CAR (scFv-28Bz) can be successfully expressed on T cells.
  • The CAR-transduced T cells exhibit modest cytotoxic activity and enhanced IFN-γ production against PD-L1-expressing cancer cells.
  • This study presents a potential CAR-based immunotherapy approach for treating solid tumors, particularly those expressing PD-L1.
Keywords:
A549NCI-H358chimeric antigen receptorprogrammed death-ligand 1

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