Abstract

BackgroundAnti-CD20 monoclonal antibody treatment has not only increased survival and cure rates in many non-Hodgkin lymphomas, but also has prompted an explosion in the development of novel antibodies and biologically active substances with specific cellular targets in the field of malignancies treatment. Since the robust immune responses are elicited by the gene-modified T cells, gene based T cell therapy may also provide a powerful tool for cancer immunotherapy.MethodsIn this study, we developed a vector construction encoding a chimeric T cell receptor that recognizes the CD20 antigen and delivers co-stimulatory signals to achieve T cell activation. One non-Hodgkin lymphoma cell line Raji cells co-cultured with peripheral blood-derived T cells were stably transfected with anti-CD20scFvFc/CD28/CD3zeta gene or anti-CD20scFvFc gene. T cells expressing anti-CD20scFvFc/CD28/CD3zeta or anti-CD20scFvFc gene co-cultured with CD20 positive Raji cells for different times. Cell lysis assay was carried by [3H]TdR release assay. The expressions of Fas, Bcl-2 and Caspase-3 of Raji cells were detected by flow cytometric. The secretion of IFN-gamma and IL-2 in co-culture medium was tested by ELISA assay. Activity of AP-1 was analyzed by EMSA.ResultsFollowing efficient transduction of peripheral blood-derived T cells with anti-CD20scFvFc/CD28/CD3zeta gene, an obvious cell lysis of Raji cells was observed in co-culture. T cells transduced anti-CD20scFvFc/CD28/CD3zeta gene had superior secretion of IFN-gamma and IL-2 compared to T cells transduced anti-CD20scFvFc gene. Also it led to a much stronger Fas-induced apoptosis signaling transduction in target cancer cells.ConclusionSo adoptively T cells transduced anti-CD20scFvFc/CD28/CD3zeta gene mediates enhanced anti-tumor activities against CD20 positive tumor cells, suggesting a potential of gene-based immunotherapy for non-Hodgkin lymphoma.

Highlights

  • Anti-CD20 monoclonal antibody treatment has increased survival and cure rates in many non-Hodgkin lymphomas, and has prompted an explosion in the development of novel antibodies and biologically active substances with specific cellular targets in the field of malignancies treatment

  • Experimental protocol The subjects were assigned into three groups: blank group, control group (T cells transduced with anti-CD20scFvFc receptor co-cultured with Raji cells), and experimental group (T cells transduced with anti-CD20scFvFc/CD28/ CD3ζ receptor co-cultured with Raji cells)

  • Expression of anti-CD20scFvFc/CD28/CD3ζ in Peripheral Blood Mononuclear Cell (PBMC) T Lymphocyte Subsets of PBMCs was analyzed by flow cytometry

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Summary

Introduction

Anti-CD20 monoclonal antibody treatment has increased survival and cure rates in many non-Hodgkin lymphomas, and has prompted an explosion in the development of novel antibodies and biologically active substances with specific cellular targets in the field of malignancies treatment. Since the robust immune responses are elicited by the gene-modified T cells, gene based T cell therapy may provide a powerful tool for cancer immunotherapy. Non-Hodgkin’s lymphoma, known as one of hematologic malignancies, is aggressive tumor with a poor prognosis. The clinical outcome of the patients has improved dramatically with combination chemotherapy (CHOP and other standard protocols) and anti-CD20 monoclonal antibody therapy, non-Hodgkin’s lymphoma has been proved to be refractory or relapse, and is failure to standard treatments [1]. As a means of immune escape, tumors do not express or down-regulate co-stimulatory ligands [4]. It allowed T cells to become activated, escape pro-apoptotic conditions, and preferentially expand in culture compared to unmodified cells [5]

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