Abstract |
T-cell receptor (TCR) gene therapy enables for the rapid creation of antigen-specific T cells from mice of any strain and represents a valuable tool for preclinical immunotherapy studies. Here, we describe the superiority of γ-retroviral vectors compared with lentiviral vectors for transduction of murine T cells and surprisingly illustrate robust gene-transfer into phenotypically naive/memory-stem cell like (TN/TSCM; CD62L(hi)/CD44(low)) and central memory (TCM; CD62L(hi)/CD44(hi)) CD8+ T cells using murine stem cell-based γ-retroviral vectors (MSGV1). We created MSGV1 vectors for a major histocompatibility complex-class I-restricted TCR specific for the melanocyte- differentiation antigen, glycoprotein 100 (MSGV1-pmel-1), and a major histocompatibility complex-class II-restricted TCR specific for tyrosinase-related protein-1 (MSGV1-TRP-1), and found that robust gene expression required codon optimization of TCR sequences for the pmel-1 TCR. To test for functionality, we adoptively transferred TCR-engineered T cells into mice bearing B16 melanomas and observed delayed growth of established tumors with pmel-1 TCR engineered CD8+ T cells and significant tumor regression with TRP-1 TCR transduced CD4 T cells. We simultaneously created lentiviral vectors encoding the pmel-1 TCR, but found that these vectors mediated low TCR expression in murine T cells, but robust gene expression in other murine and human cell lines. These results indicate that preclinical murine models of adoptive immunotherapies are more practical using γ-retroviral rather than lentiviral vectors.
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Authors | Sid P Kerkar, Luis Sanchez-Perez, Shicheng Yang, Zachary A Borman, Pawel Muranski, Yun Ji, Dhanalakshmi Chinnasamy, Andrew D M Kaiser, Christian S Hinrichs, Christopher A Klebanoff, Christopher D Scott, Luca Gattinoni, Richard A Morgan, Steven A Rosenberg, Nicholas P Restifo |
Journal | Journal of immunotherapy (Hagerstown, Md. : 1997)
(J Immunother)
Vol. 34
Issue 4
Pg. 343-52
(May 2011)
ISSN: 1537-4513 [Electronic] United States |
PMID | 21499127
(Publication Type: Journal Article, Research Support, N.I.H., Intramural)
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Chemical References |
- Receptors, Antigen, T-Cell
- gp100 Melanoma Antigen
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Topics |
- Animals
- CD4-Positive T-Lymphocytes
(immunology)
- CD8-Positive T-Lymphocytes
(immunology)
- Gene Transfer Techniques
- Genetic Engineering
- Genetic Vectors
(genetics)
- HEK293 Cells
- Humans
- Immunotherapy, Adoptive
- Jurkat Cells
- Melanoma, Experimental
(immunology, therapy)
- Mice
- Mice, Inbred C57BL
- Mice, Knockout
- NIH 3T3 Cells
- Receptors, Antigen, T-Cell
(genetics, immunology)
- Retroviridae
(genetics)
- Transduction, Genetic
- gp100 Melanoma Antigen
(genetics, immunology)
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