Chimeric antigen receptor containing ICOS signaling domain mediates specific and efficient antitumor effect of T cells against EGFRvIII expressing glioma

Journal of Hematology & Oncology

Shuang-Yin Han¹  Albert J Wong³  Puja Gupta³  Jian Cui³  Li-Ming Zhao¹  Ying-Ying Zhao¹  Yi Wang¹  Yun-Fei Wang¹  Na Cao¹  Ethan Q Han²  Yu-Xiu Yang¹  Chan-Juan Shen¹ 

[1] Translational Research Center, Zhengzhou University People’s Hospital, #7 Weiwu Road, Zhengzhou, Henan 450003, China;Drexel University College of Medicine, Philadelphia, PA 19129, USA;Department of Neurosurgery, Brain Tumor Research Laboratories, Stanford University Medical Center, Stanford, CA 94305, USA

关键词: Glioma;    Chimeric antigen receptor;    Adoptive immunotherapy;   

Others  :  806364
DOI  :  10.1186/1756-8722-6-33

 received in 2013-02-19, accepted in 2013-05-04,  发布年份 2013

【 摘 要 】

Background

Adoptive transfer of chimeric antigen receptor (CAR)-modified T cells appears to be a promising immunotherapeutic strategy. CAR combines the specificity of antibody and cytotoxicity of cytotoxic T lymphocytes, enhancing T cells’ ability to specifically target antigens and to effectively kill cancer cells. Recent efforts have been made to integrate the costimulatory signals in the CAR to improve the antitumor efficacy. Epidermal growth factor receptor variant III (EGFRvIII) is an attractive therapeutic target as it frequently expresses in glioma and many other types of cancers. Our current study aimed to investigate the specific and efficient antitumor effect of T cells modified with CAR containing inducible costimulator (ICOS) signaling domain.

Methods

A second generation of EGFRvIII/CAR was generated and it contained the EGFRvIII single chain variable fragment, ICOS signaling domain and CD3ζ chain. Lentiviral EGFRvIII/CAR was prepared and human CD3⁺ T cells were infected by lentivirus encoding EGFRvIII/CAR. The expression of EGFRvIII/CAR on CD3⁺ T cells was confirmed by flow cytometry and Western blot. The functions of EGFRvIII/CAR⁺ T cells were evaluated using in vitro and in vivo methods including cytotoxicity assay, cytokine release assay and xenograft tumor mouse model.

Results

Chimeric EGFRvIIIscFv-ICOS-CD3ζ (EGFRvIII/CAR) was constructed and lentiviral EGFRvIII/CAR were made to titer of 10⁶ TU/ml. The transduction efficiency of lentiviral EGFRvIII/CAR on T cells reached around 70% and expression of EGFRvIII/CAR protein was verified by immunoblotting as a band of about 57 kDa. Four hour ⁵¹Cr release assays demonstrated specific and efficient cytotoxicity of EGFRvIII/CAR⁺ T cells against EGFRvIII expressing U87 cells. A robust increase in the IFN-γ secretion was detected in the co-culture supernatant of the EGFRvIII/CAR⁺ T cells and the EGFRvIII expressing U87 cells. Intravenous and intratumor injection of EGFRvIII/CAR⁺ T cells inhibited the in vivo growth of the EGFRvIII expressing glioma cells.

Conclusions

Our study demonstrates that the EGFRvIII/CAR-modified T cells can destroy glioma cells efficiently in an EGFRvIII specific manner and release IFN-γ in an antigen dependent manner. The specific recognition and effective killing activity of the EGFRvIII-directed T cells with ICOS signaling domain lays a foundation for us to employ such approach in future cancer treatment.

【 授权许可】

   
2013 Shen et al.; licensee BioMed Central Ltd.

【 图 表 】

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