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

Published

Journal Article

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.

Full Text

Duke Authors

Cited Authors

  • Kerkar, SP; Sanchez-Perez, L; Yang, S; Borman, ZA; Muranski, P; Ji, Y; Chinnasamy, D; Kaiser, ADM; Hinrichs, CS; Klebanoff, CA; Scott, CD; Gattinoni, L; Morgan, RA; Rosenberg, SA; Restifo, NP

Published Date

  • May 2011

Published In

Volume / Issue

  • 34 / 4

Start / End Page

  • 343 - 352

PubMed ID

  • 21499127

Pubmed Central ID

  • 21499127

Electronic International Standard Serial Number (EISSN)

  • 1537-4513

Digital Object Identifier (DOI)

  • 10.1097/CJI.0b013e3182187600

Language

  • eng

Conference Location

  • United States