Immune-mediated specific depletion of intestinal stem cells

Abstract

Functional studies of specific stem cell populations often require depletion of tissue-specific stem cells in an in vivo model to allow for the interrogation of their contribution to the maintenance and/or regeneration of their home tissue. Depletion methods need an exquisite specificity to uniquely eliminate the target cell type. To achieve such specificity, a commonly used approach has been murine models with expression of the Diphtheria Toxin Receptor (DTR) in the cell of interest. The major caveat of using these DTR-expressing transgenic mice is the need to generate new DTR models for every new cell population of interest. While DTR-expressing models are limited, the number of available GFP-expressing mice is large. To take advantage of this plethora of cell type-specific GFP-reporter mice, we sought to exploit the body’s own killer cells as a depletion tool. Thus, we generated a mouse model whose cytotoxic T cells recognize and kill GFP-expressing cells, called the Jedi (Agudo et al., Nat Biotechnol 33:1287–1292, 2015). Jedi T cells now enable the depletion of virtually almost any cell type by using a suitable GFP-expressing transgenic mouse (Agudo et al., Nat Biotechnol 33:1287–1292, 2015; Chen et al., J Clin Invest 128(8):3413–3424, 2018). Here, we explain in detail how to achieve depletion of Lgr5+ stem cells in the intestine with a single injection of Jedi T cells (Agudo et al., Immunity 48:271–285.e5, 2018) with a methodology that can be extrapolated to any other GFP-expressing cell.