A new immunotherapy strategy targeted CD30 in peripheral T-cell lymphomas: CAR-modified T-cell therapy based on CD30 mAb

Yang Wu,Rong Ma,Shu-Chen Dong,Wei-Yan Tang,Jian-Zhong Wu,Dan Chen,Ji-Feng Feng,Jian-Qiu Wu,Ya Lu

doi:10.1038/s41417-021-00295-8

Abstract

Chimeric antigen receptor T-cell immunotherapy (CAR-T) has shown remarkable efficacy in treating tumors of lymphopoietic origin. Herein, we demonstrate an effective CAR-T cell treatment for recurrent and malignant CD30-positive peripheral T-cell lymphomas (PTCL) has been demonstrated. The extracellular fragment gene sequences of CD30 were obtained from tumor tissues of PTCL patients and cloned into a plasmid vector to express the CD30 antigen. The CD30 targeting single-chain antibody fragment (scFv) was obtained from CD30-positive monoclonal hybridoma cells, which were obtained from CD30 antigen immunized mice. After a second-generation of CAR lentiviral construction, CD30 CAR T cells were produced and used to determine the cytotoxicity of this construct toward Karpas 299 cells. The results of CD30 CAR T-mediated cell lysis show that 9C11-2 CAR T cells could significantly promote the lysis of CD30-positive Karpas 299 cells in both LDH and real-time cell electronic sensing (RTCA) assays. In vivo data show that 9C11-2 CAR T cells effectively suppress the tumor growth in a Karpas 299 cell xenograft NCG mouse model. The CD30 CAR T cells exhibited an efficient cytotoxic effect after being co-cultured with the target cells and they also exhibited a significant tumor-inhibiting ability after being intravenously injected into PTCL xenograft tumors; these observations suggest that the new CD30 CAR-T cell may be a promising therapeutic candidate for cancer therapy.

Highlights

These authors contributed : Yang Wu, Dan Chen
The use of mature CAR-T cell immunotherapy outlined in this study undoubtedly provides a new CAR-T cell therapy based on anti-CD 30 mAb for the treatment of refractory or recurrent peripheral T-cell lymphomas (PTCL)
The expression of the CD30 antigen in PTCL tumor tissue was shown by IHC, and the results revealed that CD30positive PTCL tissue had much more brown or light brown staining (Fig. 1A, right) than CD30-negative tissue (Fig. 1A, left)

Summary

1234567890();,: 1234567890();,: Introduction

The discovery of new and effective treatment strategies for PTCL is of the utmost importance for clinicians and patients of this form of NHL. CAR-T cells targeting CD19 or CD20 have achieved remarkable results in the treatment of acute hepatitis B leukemia, chronic lymphocytic leukemia, and NHL [16,17,18,19,20], indicating that CAR-T has attractive clinical application prospects for lymphoma treatment. The use of mature CAR-T cell immunotherapy outlined in this study undoubtedly provides a new CAR-T cell therapy based on anti-CD 30 mAb for the treatment of refractory or recurrent PTCL. Recombinant CD30 proteins were expressed by transforming the recombinant plasmid pET28a-CD30 into E. coli DH5α cells incubated in LB medium with kanamycin (100 μg/mL, Sigma, USA). The purified CD30 protein was verified using Western blot analysis and mass spectrum identification

Material and method

Results

Discussion

Conclusion

Compliance with ethical standards