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

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

Research output: Contribution to journalArticlepeer-review

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; CD62Lhi/ CD44low) and central memory (TCM; CD62Lhi/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.

Original languageEnglish (US)
Pages (from-to)343-352
Number of pages10
JournalJournal of Immunotherapy
Volume34
Issue number4
DOIs
StatePublished - May 2011

Keywords

  • HIV
  • adoptive transfer
  • cancer
  • gene therapy
  • immunotherapy
  • lentivirus
  • melanoma
  • retrovirus

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology
  • Pharmacology
  • Cancer Research

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