Cre-inducible human CD59 mediates rapid cell ablation after intermedilysin administration
Dechun Feng ... Xuebin Qin, Bin Gao
Dechun Feng ... Xuebin Qin, Bin Gao
Published June 1, 2016
Citation Information: J Clin Invest. 2016;126(6):2321-2333. doi:10.1172/JCI84921.
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Categories: Technical Advance Immunology

Cre-inducible human CD59 mediates rapid cell ablation after intermedilysin administration

Abstract

Cell ablation is a powerful tool for studying cell lineage and/or function; however, current cell-ablation models have limitations. Intermedilysin (ILY), a cytolytic pore-forming toxin that is secreted by Streptococcus intermedius, lyses human cells exclusively by binding to the human complement regulator CD59 (hCD59), but does not react with CD59 from nonprimates. Here, we took advantage of this feature of ILY and developed a model of conditional and targeted cell ablation by generating floxed STOP-CD59 knockin mice (ihCD59), in which expression of human CD59 only occurs after Cre-mediated recombination. The administration of ILY to ihCD59+ mice crossed with various Cre-driver lines resulted in the rapid and specific ablation of immune, epithelial, or neural cells without off-target effects. ILY had a large pharmacological window, which allowed us to perform dose-dependent studies. Finally, the ILY/ihCD59-mediated cell-ablation method was tested in several disease models to study immune cell functionalities, hepatocyte and/or biliary epithelial damage and regeneration, and neural cell damage. Together, the results of this study demonstrate the utility of the ihCD59 mouse model for studying the effects of cell ablation in specific organ systems in a variety of developmental and disease states.

Authors

Dechun Feng, Shen Dai, Fengming Liu, Yosuke Ohtake, Zhou Zhou, Hua Wang, Yonggang Zhang, Alison Kearns, Xiao Peng, Faliang Zhu, Umar Hayat, Man Li, Yong He, Mingjiang Xu, Chunling Zhao, Min Cheng, Lining Zhang, Hong Wang, Xiaofeng Yang, Cynthia Ju, Elizabeth C. Bryda, Jennifer Gordon, Kamel Khalili, Wenhui Hu, Shuxin Li, Xuebin Qin, Bin Gao

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Generation of ihCD59 knockin mice. (A) Map of the pBT378-CAG-LSL-hCD59 v...

Figure 1

Generation of ihCD59 knockin mice.

(A) Map of the pBT378-CAG-LSL-hCD59 vector for pronuclear injection. (B) General strategies for the generation of ihCD59 mice. The STOP cassette, which prohibits transgene expression, is removed by crossing the inducible transgenic strain to a cell-specific Cre-expressing mouse strain. The consequent expression of the transgene renders the respective tissues sensitive to rapid cell lysis induced by the injection of ILY. (C and D) Representative FACS analyses of hCD59 expression on T cells in Lck-Cre+ihCD59+ mice (C) and on monocytes/neutrophils in Lysm-Cre+ihCD59+ mice (D). (E and F) Splenocytes that were isolated from Lck-Cre+ihCD59+ mice were incubated with ILY in vitro for 10 minutes. FACS analyses were performed. E shows 29% live hCD59+ spleen cells and 0.057% live hCD59+ spleen cells before and after ILY incubation, respectively. F shows live/dead T and B cells. In C and D, the representative graphs from 5 mice are shown. In E and F, the representative graphs from 4 mice are shown.