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Published on: November 7, 2014
Inhibition of p38 MAPK activity in B-NHL Raji cells by treatment with engineered CD20-specific T cells
1Laboratory of Internal Medicine, The First Affiliated Hospital of Wenzhou Medical College, Wenzhou, Zhejiang 325000, P.R. China.
Abstract:
Adoptive immunotherapy with T cells expressing CD20-specific chimeric T-cell receptors is a promising approach to lymphoma therapy. However, modification of the cellular signaling pathways in target tumor cells by treatment with engineered CD20-specific T cells has yet to be fully elucidated. In this study, the non-Hodgkin's lymphoma Raji cell line was co-cultured with T cells that were genetically modified with anti-CD20scFvFc/CD28/CD3ζ or anti-CD20scFvFc gene. The cytolytic activity of engineered CD20-specific T cells and IL-10 secretion was quantitated by Cytotoxicity and ELISA assays, respectively. The engineered CD20-specific T cells and Raji cells were sorted using flow cytomety for the Western blot analysis. Treatment of Raji cells with T cells genetically modified with anti-CD20scFvFc/CD28/CD3ζ chimera (compared to anti-CD20scFvFc) yielded a higher cytotoxicity against Raji cells in vitro. Additionally, we found that engineered CD20-specific T cells caused a decrease in IL-10 secretion and inhibition of phosphor-STAT3 and Bcl-2 expression in Raji cells, possibly through the down-regulation of p38 MAPK and NF-κB activity. These results indicate that the treatment of Raji cells with engineered CD20-specific T cells inhibited the cellular p38 MAPK signaling pathways, which enhanced its antitumor activities against CD20-positive tumor cells.
Insights
Engineered T cells targeting CD20 enhanced lymphoma treatment by inhibiting key signaling pathways. This adoptive immunotherapy approach shows promise for treating CD20-positive tumors.
Area of Science:
- Immunology
- Oncology
- Cell Biology
Background:
- Adoptive immunotherapy using engineered T cells is a promising lymphoma treatment.
- The precise cellular mechanisms by which CD20-specific T cells impact tumor signaling require further investigation.
Purpose of the Study:
- To investigate the effects of CD20-specific chimeric antigen receptor T cells on cellular signaling pathways in non-Hodgkin's lymphoma cells.
- To evaluate the antitumor activity of engineered T cells against CD20-positive lymphoma cells.
Main Methods:
- Co-culture of Raji lymphoma cells with T cells genetically modified with anti-CD20scFvFc/CD28/CD3ζ or anti-CD20scFvFc.
- Quantification of T cell cytolytic activity and IL-10 secretion via Cytotoxicity and ELISA assays.
- Western blot analysis to assess signaling pathway modulation (p38 MAPK, NF-κB, STAT3, Bcl-2).
Main Results:
- T cells expressing anti-CD20scFvFc/CD28/CD3ζ demonstrated superior in vitro cytotoxicity against Raji cells compared to anti-CD20scFvFc.
- Engineered T cells reduced IL-10 secretion and inhibited phosphor-STAT3 and Bcl-2 expression in Raji cells.
- These effects were associated with the down-regulation of p38 MAPK and NF-κB signaling pathways.
Conclusions:
- Engineered CD20-specific T cells effectively inhibit p38 MAPK signaling in lymphoma cells.
- This inhibition enhances the antitumor efficacy of adoptive immunotherapy against CD20-positive tumors.
