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 6

Highly selective ablation of hepatocytes and/or BECs in Cre+ihCD59+ mice after ILY injection.

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Highly selective ablation of hepatocytes and/or BECs in Cre+ihCD59+ mice...
(A–C) Three lines of Cre+ihCD59+ mice were generated. Representative immunostaining images (from 9 mice in each group) on the left show hCD59 protein expression in both hepatocytes and BECs in Alb-Cre+ihCD59+ mice (brown staining in A), hCD59 expression in hepatocytes only in Ad-Alb-Cre+ihCD59+ mice (red staining in B), and hCD59 protein expression in BECs in Sox9-CreERT+ihCD59+ mice (C, CK19 and CD59 costaining) after tamoxifen treatment. These mice were given a single ILY injection (150 ng/g, i.v.), followed by the measurement of serum ALT and bilirubin. BD, bile ducts; PV, portal vein. (D–F) Liver regeneration after ILY-mediated ablation of hepatocytes and/or BECs. Cre+ihCD59+ mice (n = 7 in each group) received 150 ng/g ILY (i.v.) and were euthanized 40 or 48 hours after ILY injection. BrdU (50 mg/kg) was given 2 hours before sacrifice. Representative immunohistochemical staining of BrdU (D, n = 9) or immunofluorescence staining of BrdU and CK19 (E, n = 9, arrows indicate double-positive cells) in the livers of ILY-treated mice (40 hours after ILY injection). The percentages of BrdU+ hepatocytes and BECs from 40 and 48 hours after ILY treatment were determined (F). Scale bars: 100 μm. Values represent mean ± SD (n = 9). **P < 0.01; ***P < 0.001. Comparisons between groups as indicated in the figure were determined by 1-way ANOVA followed by Bonferroni’s post-hoc test.