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

SP Kerkar, L Sanchez-Perez, S Yang… - Journal of …, 2011 - journals.lww.com
SP Kerkar, L Sanchez-Perez, S Yang, ZA Borman, P Muranski, Y Ji, D Chinnasamy…
Journal of immunotherapy, 2011journals.lww.com
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 …
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.
Lippincott Williams & Wilkins