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Genetic engineering of murine CD8+ and CD4+ T cells for preclinical adoptive immunotherapy studies.

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.
AuthorsSid 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
JournalJournal of immunotherapy (Hagerstown, Md. : 1997) (J Immunother) Vol. 34 Issue 4 Pg. 343-52 (May 2011) ISSN: 1537-4513 [Electronic] United States
PMID21499127 (Publication Type: Journal Article, Research Support, N.I.H., Intramural)
Chemical References
  • Receptors, Antigen, T-Cell
  • gp100 Melanoma Antigen
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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