title: 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.
authors.affiliation: Surgery Branch, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892, USA.
authors.forename: Sid P
authors.initials: SP
authors.lastname: Kerkar
authors.forename: Luis
authors.initials: L
authors.lastname: Sanchez-Perez
authors.forename: Shicheng
authors.initials: S
authors.lastname: Yang
authors.forename: Zachary A
authors.initials: ZA
authors.lastname: Borman
authors.forename: Pawel
authors.initials: P
authors.lastname: Muranski
authors.forename: Yun
authors.initials: Y
authors.lastname: Ji
authors.forename: Dhanalakshmi
authors.initials: D
authors.lastname: Chinnasamy
authors.forename: Andrew D M
authors.initials: AD
authors.lastname: Kaiser
authors.forename: Christian S
authors.initials: CS
authors.lastname: Hinrichs
authors.forename: Christopher A
authors.initials: CA
authors.lastname: Klebanoff
authors.forename: Christopher D
authors.initials: CD
authors.lastname: Scott
authors.forename: Luca
authors.initials: L
authors.lastname: Gattinoni
authors.forename: Richard A
authors.initials: RA
authors.lastname: Morgan
authors.forename: Steven A
authors.initials: SA
authors.lastname: Rosenberg
authors.forename: Nicholas P
authors.initials: NP
authors.lastname: Restifo
mesh_terms: D000818:Animals; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology*; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology*; D018014:Gene Transfer Techniques; D005818:Genetic Engineering; D005822:Genetic Vectors / Q000235:genetics; D057809:HEK293 Cells; D006801:Humans; D016219:Immunotherapy, Adoptive; D019169:Jurkat Cells; D008546:Melanoma, Experimental / Q000276:immunology / Q000628:therapy; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D041681:NIH 3T3 Cells; D011948:Receptors, Antigen, T-Cell / Q000235:genetics / Q000276:immunology; D012190:Retroviridae / Q000235:genetics; D014161:Transduction, Genetic; D058951:gp100 Melanoma Antigen / Q000235:genetics / Q000276:immunology
chemical_list: D011948:Receptors, Antigen, T-Cell; D058951:gp100 Melanoma Antigen