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Chimeric antigen receptor T cell targeting EGFRvIII for metastatic lung cancer therapy

  • Zhao Zhang
  • Jun Jiang
  • Xiaodong Wu
  • Mengyao Zhang
  • Dan Luo
  • Renyu Zhang
  • Shiyou LiEmail author
  • Youwen HeEmail author
  • Huijie BianEmail author
  • Zhinan ChenEmail author
Open Access
Research Article
  • 62 Downloads

Abstract

Lung cancer is the most common incident cancer and the leading cause of cancer death. In recent years, the development of tumor immunotherapy especially chimeric antigen receptor T (CAR-T) cell has shown a promising future. Epidermal growth factor receptor variant III (EGFRvIII) is a tumor-specific mutation expressed in various types of tumors and has been detected in non-small cell lung cancer with a mutation rate of 10%. Thus, EGFRvIII is a potential antigen for targeted lung cancer therapy. In this study, CAR vectors were constructed and transfected into virus-packaging cells. Then, activated T cells were infected with retrovirus harvested from stable virus-producing single clone cell lines. CAR expression on the surfaces of the T cells was detected by flow cytometry andWestern blot. The function of CAR-T targeting EGFRvIII was then evaluated. The EGFRvIII-CAR vector was successfully constructed and confirmed by DNA sequencing. A stable virus-producing cell line was produced from a single clone by limited dilution. The culture conditions for the cell line, including cell density, temperature, and culture medium were optimized. After infection with retrovirus, CAR was expressed on more than 90% of the T cells. The proliferation of CAR-T cells were induced by cytokine and specific antigen in vitro. More importantly, EGFRvIII-CART specifically and efficiently recognized and killed A549-EGFRvIII cells with an effector/target ratio of 10:1 by expressing and releasing cytokines, including perforin, granzyme B, IFN-g, and TNF-α. The in vivo study indicated that the metastasis of A549-EGFRvIII cells in mice were inhibited by EGFRvIII-CART cells, and the survival of the mice was significantly prolonged with no serious side effects. EGFRvIII-CART showed significantly efficient antitumor activity against lung cancer cells expressing EGFRvIII in vivo and in vitro. Therefore, CAR-T targeting EGFRvIII is a potential therapeutic strategy in preventing recurrence and metastasis of lung cancer after surgery.

Keywords

chimeric antigen receptor T cells epidermal growth factor receptor lung cancer immunotherapy tumor immunology 

Notes

Acknowledgements

This study was supported by National Key Basic Research and Development Plan (No. 2015CB553701).

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Copyright information

© The Author(s) 2019

Open Access This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http:// creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided the appropriate credit is given to the original author(s) and the source, and a link is provided to the Creative Commons license, indicating if changes were made.

Authors and Affiliations

  1. 1.National Translational Science Center for Molecular MedicineXi’anChina
  2. 2.Fourth Military Medical UniversityXi’anChina
  3. 3.Beijing Institute of GenomicsChinese Academy of ScienceBeijingChina
  4. 4.Beijing Institute of BiotechnologyAcademy of Military Medical SciencesBeijingChina
  5. 5.Department of ImmunologyDuke University Medical CenterDurhamUSA

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