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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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 2

LV gene–modified HSCs functionally engraft and repopulate transplant recipients over the long term.

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LV gene–modified HSCs functionally engraft and repopulate transplant rec...
(A) Overview of transplant procedure. Lin– cells were isolated and transduced with LV-TCR. Competitive reconstitution was performed using transduced Lin– cells (CD45.1, 5 × 104) combined with congenic BM cells (CD45.2, 2.5 × 105) from DR4 Tg mice, then transplanted into irradiated DR4 Tg recipients. A parallel arm involving Lin– cells (5 × 104) from TCR Tg mice combined with unfractionated BM (2.5 × 105) from DR4 transgenic mice was transplanted into similarly irradiated hosts. (B) Flow cytometry analysis of PB at 6 and 12 months demonstrated high-level TCR gene expression: CD45+CD3+CD4+Tyrp1 Ultimer+ staining of LV-TCR and TCR Tg transplant groups. (C) Flow cytometry analysis of PB at 12 months demonstrated low-level TCR gene expression within CD8 compartment: CD45+CD3+CD8+Tyrp1 Ultimer+ staining of LV-TCR and TCR Tg transplant groups. (D) Flow cytometry analysis of PB at 12 months demonstrated high-level co-expression of the murine TCRβ chain in both CD4+Ultimer+ and CD4+Ultimer– subpopulations. Data are representative of flow cytometry studies done on 4 separate transplant experiments.