Date of Award

2017

Degree Type

Thesis

Degree Name

Master of Science (MS)

Department

Microbiology and Immunology

Abstract

Lymphangioleiomyomatosis (LAM) is a low-grade neoplastic disease affecting primarily women. It is characterized by cystic lung disease as well as renal and retroperitoneal tumors called angiomyolipomas and lymphangiomas. Tumor cells have smooth muscle features as well as neuroendocrine cell surface markers, and the disease can be diagnosed by HMB45 staining of tumor cells. We questioned whether expression of melanocytic antigens, specifically gp100, creates an opportunity to treat LAM by adoptive T cell transfer. LAM lung lesions demonstrate poor immune surveillance, therefore adoptive T cell transfer could offer benefits in this disease.

A link was made between melanoma and depigmenting vitiligo lesions as a source of gp100-reactive T cells. These T cells were first isolated from a vitiligo patient and the resulting gp100-reactive T cell receptor-bearing T cells were measured for reactivity towards relevant targets. The vitiligo lesion-derived T cell receptor, SILv44, was cloned into a lentiviral vector and introduced into magnetically sorted primary human CD8+ T cells. Transgenic T cells were combined 5:1 with melanoma cells, and LAM10224, a patient-derived LAM cell line, and responses were measured by ELISA detecting IFN-γ, after 48 hrs. Cytotoxicity was measured by CD107a expression. Transgenic T cells conferred specific reactivity to HLA-A2+, gp100+ cells including LAM10224 HLA-A2 transfected cells in vitro. The SILv44 T cell receptor demonstrated specific IFN-γ release in response to gp100, HLA-A2+ LAM10224 and melanoma cells, as well as comparable CD107a expression to that of PMA-ionomycin stimulated cells with minimal off target reactivity. Because LAM10224 has been shown to form tumors in immunodeficient mice, we challenged groups of 2 SCID mice with LAM10224 cells transfected to express HLA-A2. One group was treated with SILv44 transgenic T cells, the remaining two groups were treated with SILv44 transgenic T cells in combination with anti-PD1 antibody and untransduced T cells respectively. Both SILv44 alone and in combination with antiPD1 effectively reduced tumor burden in mice when compared to control. These data provide pre-clinical evidence for the efficacy of adoptive T cell as a treatment, combined with checkpoint inhibition, to curtail the burden of LAM disease in human patients.

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Creative Commons License
This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 3.0 License.

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