Transplantation of mouse HSCs genetically modified to express a CD4-restricted TCR results in long-term immunity that destroys tumors and initiates spontaneous autoimmunity
Sung P. Ha, … , Hal E. Broxmeyer, Christopher E. Touloukian
Sung P. Ha, … , Hal E. Broxmeyer, Christopher E. Touloukian
Published November 15, 2010
Citation Information: J Clin Invest. 2010;120(12):4273-4288. https://doi.org/10.1172/JCI43274.
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Research Article Genetics

Transplantation of mouse HSCs genetically modified to express a CD4-restricted TCR results in long-term immunity that destroys tumors and initiates spontaneous autoimmunity

Abstract

The development of effective cancer immunotherapies has been consistently hampered by several factors, including an inability to instigate long-term effective functional antitumor immunity. This is particularly true for immunotherapies that focus on the adoptive transfer of activated or genetically modified mature CD8+ T cells. In this study, we sought to alter and enhance long-term host immunity by genetically modifying, then transplanting, mouse HSCs. We first cloned a previously identified tumor-reactive HLA-DR4–restricted CD4+ TCR specific for the melanocyte differentiation antigen tyrosinase-related protein 1 (Tyrp1), then constructed both a high-expression lentivirus vector and a TCR-transgenic mouse expressing the genes encoding this TCR. Using these tools, we demonstrated that both mouse and human HSCs established durable, high-efficiency TCR gene transfer following long-term transplantation into lethally irradiated mice transgenic for HLA-DR4. Recipients of genetically modified mouse HSCs developed spontaneous autoimmune vitiligo that was associated with the presence of a Th1-polarized memory effector CD4+ T cell population that expressed the Tyrp1-specific TCR. Most importantly, large numbers of CD4+ T cells expressing the Tyrp1-specific TCR were detected in secondary HLA-DR4–transgenic transplant recipients, and these mice were able to destroy subcutaneously administered melanoma cells without the aid of vaccination, immune modulation, or cytokine administration. These results demonstrate the creation of what we believe to be a novel translational model of durable lentiviral gene transfer that results in long-term effective immunity.

Authors

Sung P. Ha, Nicholas D. Klemen, Garrett H. Kinnebrew, Andrew G. Brandmaier, Jon Marsh, Giao Hangoc, Douglas C. Palmer, Nicholas P. Restifo, Kenneth Cornetta, Hal E. Broxmeyer, Christopher E. Touloukian

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Figure 10

Six-month secondary TCR transplant recipients reject subcutaneous melanoma.

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Six-month secondary TCR transplant recipients reject subcutaneous melano...
(A) LV-TCR and control transplants (5 mice per group) were subcutaneously injected with 5 × 105 B16-DR4 tumor cells. Three days after injection, tumor size was recorded. Statistical significance between groups was based on tumor size. Data are displayed as mean ± SEM with P value; n = 5 per group. Experiments were performed in a blinded, randomized fashion and executed independently 3 times. (B and C) Tumors from LV-TCR transplants were necrotic. IHC staining of paraffin-embedded tumor (control and LV-TCR) specimens: H&E staining; original magnification, ×400. (D–F) Tumors from LV-TCR transplants were associated with a dense TCR gene–specific infiltration: CD3 and human TCR-β staining; original magnification, ×400. NovaRED substrate was used instead of DAB (previously used on skin) to more effectively differentiate between necrotic tumor/pigment and cellular infiltrate. IHC was performed on 3 independent samples per group with similar results.