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 7

ILY specifically ablates reactive astrocytes.

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ILY specifically ablates reactive astrocytes. (A) Brain injury model. Le...
(A) Brain injury model. Left panel indicates the sites in the brain injury model with the local injection of ILY (10 ng/g) or PBS. Left panel adapted with permission from Elsevier Academic Press (32). The costaining of hCD59 (red) and GFAP (green) within the sections of the brain lesion sites is indicated by the white dashed line. The lower pictures are the magnifications of the white box area in the upper image. (B) Spinal cord injury model. The left panel indicates the transected area in the spinal cord at T7 with the local injection of ILY. The area in gray indicates the injury location. Parasagittal sections of the spinal cord around the lesion from a representative mouse in each group (right color images) indicate a high density of GFAP+-reactive astrocytes and scar tissue at/around the lesion site. The images on the right side indicate reactive astrocytes at a higher magnification. Representative images from 5 different mice are shown. Scale bars: 100 μm.