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. 2019 Oct 9;8:e49163. doi: 10.7554/eLife.49163

Figure 2. Liver Kupffer cells clear CHIKV from the circulation.

(A) WT C57BL/6 mice that underwent a sham or splenectomy surgery were inoculated i.v. with CHIKV and viral genomes in the inoculum and serum at 45 min post-inoculation were quantified by RT-qPCR. Mean ± SD. N = 6, two experiments. Mann-Whitney test; p>0.05. (B) Splenectomized WT C57BL/6 mice were treated i.v. with PLL or CLL. At 42 hr post-treatment, mice were inoculated i.v. with CHIKV and viral genomes in the inoculum (input) and serum at 45 min post-inoculation were quantified by RT-qPCR. Mean ± SD. N = 6, two experiments. Mann-Whitney test; **p<0.01. (C) WT C57BL/6 mice were treated and inoculated as in (B), and viral genomes present at 45 min post-inoculation in the serum or indicated tissues were quantified by RT-qPCR. Mean ± SD. N = 9, two experiments. Mann-Whitney test or Two-tailed t-test; ***p<0.001, ****p<0.0001. (D and E) WT C57BL/6 mice were treated as in (B) and inoculated with CHIKV at 42 hr post-treatment. Livers were collected at 45 min post-inoculation, and RNA Scope in situ hybridization (D) or IHC (E) were performed to visualize viral RNA localization or F4/80+ macrophages, respectively. Brown staining is indicative of viral RNA (D) or F4/80+ macrophages (E). N = 6–7, two experiments. Figure 2—figure supplement 1 shows representative images of CHIKV RNA Scope in situ hybridization and F4/80 IHC for all biological replicates, and Figure 2—figure supplement 2 shows flow cytometry analysis of liver macrophages and dendritic cells (DCs) in PLL- and CLL-treated mice.

Figure 2—source data 1. Raw data for Figure 2A-C.
elife-49163-fig2-data1.xlsx (30.9KB, xlsx)
DOI: 10.7554/eLife.49163.009

Figure 2.

Figure 2—figure supplement 1. CHIKV RNA and F4/80+ cells are detectable in the livers of PLL- but not CLL-treated mice.

Figure 2—figure supplement 1.

CHIKV-specific RNA scope in situ hybridization (A) and F4/80 antigen-specific immunohistochemistry (B) of liver sections from each mouse in the experiment shown in Figure 1D and E to visualize CHIKV RNA and F4/80+ cells (brown staining). N = 6–7, two experiments.
Figure 2—figure supplement 2. Clodronate liposome treatment depletes liver macrophages, but has minimal impact on DCs.

Figure 2—figure supplement 2.

Livers were collected from splenectomized WT C57BL/6 mice in Figure 2B at 45 min post-inoculation and macrophage and DC populations were evaluated by flow cytometry. The representative flow plots (A) and percentage of cells (B) are shown. Mean ± SD. N = 3, one experiment (representative of two additional experiments in WT C57BL/6 mice that had not undergone surgery). Two-tailed, unpaired t-test. **p<0.01.