Enhanced receptor expression and in vitro effector function of a murine-human hybrid MART-1-reactive T cell receptor following a rapid expansion |
| |
Authors: | Stephanie L Goff Laura A Johnson Mary A Black Hui Xu Zhili Zheng Cyrille J Cohen Richard A Morgan Steven A Rosenberg Steven A Feldman |
| |
Institution: | (1) Present address: Department of Surgery, College of Physicians and Surgeons, Columbia University, New York, NY, USA;(2) Present address: Division of Neurosurgery, Department of Surgery, Duke University Medical Center, Durham, NC, USA;(3) Surgery Branch, National Cancer Institute, National Institutes of Health, 10 Center Drive, Bethesda, MD 20892-1201, USA;(4) Present address: Laboratory of Tumor Immunology and Immunotherapy, The Mina and Everard Goodman Faculty of Life Sciences, Bar-llan University, Ramat Gan, Israel;(5) Surgery Branch, National Cancer Institute, National Institutes of Health, ACRF, 1B37A, 10 Center Drive, Bethesda, MD 20892-1201, USA; |
| |
Abstract: | Peripheral blood lymphocytes (PBL) genetically modified to express T cell receptors (TCR) specific to known melanoma antigens,
such as melanoma antigen recognized by T cells-1 (MART-1), and gp100 can elicit objective tumor regression when administered
to patients with metastatic melanoma. It has also been demonstrated that modifications within the constant regions of a fully
human TCR can enhance surface expression and stability without altering antigen specificity. In this study, we evaluated the
substitution of murine constant regions for their human counterpart within the DMF5 MART-1-specific TCR. Unlike previous studies,
all modified TCRs were inserted into retroviral vectors and analyzed for expression and function following a clinical transduction
protocol. PBL were transduced with retroviral supernatant generated from stable packaging lines encoding melanoma-specific
TCRs. This protocol resulted in high levels of antigen-specific T cells without the need for additional peptide stimulation
and selection. Both the human and murinized TCR efficiently transduced PBL; however, the murinized TCR exhibited significantly
higher tetramer binding, mean fluorescence intensity, as well as, increased in vitro effector function following our clinical
transduction and expansion protocol. Additional TCR modifications including insertion of a second disulfide bond or the linker
modifications evaluated herein did not significantly enhance TCR expression or subsequent in vitro effector function. We conclude
that the substitution of a human constant region with a murine constant region was sufficient to increase receptor expression
and tetramer binding as well as antitumor activity of the DMF5 TCR and could be a tool to augment other antigen-specific TCR. |
| |
Keywords: | |
本文献已被 SpringerLink 等数据库收录! |
|