Cre-inducible human CD59 mediates rapid cell ablation after intermedilysin administration
Dechun Feng, … , Xuebin Qin, Bin Gao
Dechun Feng, … , Xuebin Qin, Bin Gao
Published May 9, 2016
Citation Information: J Clin Invest. 2016;126(6):2321-2333. https://doi.org/10.1172/JCI84921.
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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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Figure 3

ILY treatment depletes liver Kupffer cells in Lysm-Cre+ihCD59+ mice: a model to study Kupffer cell repopulation.

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ILY treatment depletes liver Kupffer cells in Lysm-Cre+ihCD59+ mice: a m...
(A) Representative immunofluorescence staining of hCD59 and F4/80 in livers from 4 different Lysm-Cre+ihCD59+ mice. Arrows indicate the coexpression of F4/80 (green) and hCD59 (red) in the Kupffer cells. (B) Representative F4/80 immunostaining and double immunofluorescence with F4/80 and BrdU in livers from 3 different Lysm-Cre+ihCD59+ mice that were treated with ILY (75 ng/g, i.v.). BrdU (50 mg/kg) was given 2 hours before sacrifice, and mice were euthanized at various time points after ILY injection. Arrows indicate BrdU+F4/80+ Kupffer cells. Scale bars: 100 μm. (C) Percentages of F4/80+ area and F4/80+BrdU+ cells from B were quantified. Values represent mean ± SD (n = 3 for 0 hours and n = 6 for all other time points). (D) A total of 7.5 × 106 GFP+ monocytes from ROSA26-EGFP transgenic mice were adoptively transferred into ILY-treated Lysm-Cre+ihCD59+ mice in which the Kupffer cells were depleted. Three days later, liver macrophages were isolated and subjected to FACS analyses. Kupffer cell–like GFP+CD11blo F4/80hi cells were identified (n = 4).