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. 2019 Mar 4;216(3):527–538. doi: 10.1084/jem.20181765

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© 2019 Rowe et al.

This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms/). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 4.0 International license, as described at https://creativecommons.org/licenses/by-nc-sa/4.0/).

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Figure 4. — Ccl5 regulates lineage of MLL-AF9 leukemia in vivo. (A) Neonatal leukemia cells were transduced with a lentivirus bearing a full-length Ccl5 cDNA and engrafted into sublethally irradiated neonatal recipients. When recipients became moribund, mice were euthanized, and survival data are presented. Results are combined from two independent transplantation experiments. n = 13 mice each in control and Ccl5 group. z = 2.75, P = 0.0006 by log-rank test. (B and C) Neonatal leukemia cells expressing either ectopic Ccl5 or transduced with empty vector were engrafted into neonatal recipients, and the percentage of lineage⁻ (Gr-1⁻/Mac-1⁻) B220⁺ cells in the resultant leukemia was quantified. Results are aggregated over two independently transplanted cohorts analyzed by Student’s t test (mean ± SEM is presented; n = 7 control and 3 Ccl5 mice obtained for analysis; *, P = 0.14). Numbers on plots indicate the percentage of cells in each gate. (D) Representative histology including H&E staining (left) immunohistochemistry for CD11b and B220 (arrows indicate B220⁺ foci, shown at higher magnification in inset; scale bars, 100 µm (10 µm in the inset). (E) Serially transplanted neonatal mixed-lineage leukemia cells (either tertiary or secondary transplant in two independent transplant experiments) were engrafted into 6–8-wk-old Ccl5 KO mice or age-matched, congenic recipients. Representative flow cytometry profile of the input leukemia is shown, as well as flow cytometry profiles of the resultant leukemia in the indicated recipients. Numbers on plots indicate the percentage of cells in each gate. (F) Quantification of the percentage of B220⁺ leukemia cells in the indicated recipients. Results are compiled from two independent transplantation experiments with different leukemias used for engraftment in each experiment (analysis by Student’s t test; results are presented as mean ± SEM; n = 6 mice obtained for analysis for both the wild-type and Ccl5 KO conditions; *, P = 0.03).