Type I interferon underlies severe disease associated with Junín virus infection in mice
- Department of Animal, Dairy and Veterinary Sciences, Utah State University, United States
- Utah Veterinary Diagnostic Laboratory, Utah State University, United States
- Division of Surgical Oncology, Department of Surgery, David Geffen School of Medicine at University of California, Los Angeles (UCLA), United States
- Department of Microbiology, Immunology and Molecular Genetics, David Geffen School of Medicine at UCLA, United States
- UCLA Molecular Biology Institute, United States
- UCLA Jonsson Comprehensive Cancer Center, United States
- UCLA AIDS Institute, United States
Figures
Figure 1
Expression of hTfR1 results in severe disease following JUNV infection.
Three-week-old WT, hTfR1 HET or hTfR1 HOM mice (n = 8–9/group) were infected i.p. with 105 CCID50 of JUNV and monitored daily. (A) Survival and (B) weight change of animals relative to the day of virus challenge (group mean and standard error of the mean; SEM) are shown. Sham-infected controls (n = 6) consisted of a mix of WT, hTfR1 HET and hTfR1 HOM hTfR1 mice. Aggregate data from two independent experiments. ***p=0.0003 and ****p<0.0001 compared to hTfR1 HOM mice.
Figure 2
Age-dependent susceptibility of hTfR1 HOM mice to lethal JUNV disease.
Shown are (A) survival (n = 6/group for the 3- and 4-week-old mice, n = 3/group for the 5- and 6-week-old mice), (B) weight change in 3 to 4 week-old mice, (C) weight change in 5 to 6 week-old mice and D) clinical disease scores for each group infected i.p. with 105 CCID50 of JUNV. Open symbols in the weight change graphs indicate age-matched sham-infected controls (n = 3). The weight data are represented as the group mean and SEM of the percent change in weight of animals relative to their starting weights on the day of virus challenge. Clinical scoring is expressed as group mean and SEM. *p=0.0448 compared to age-matched sham-infected controls.
Figure 3
JUNV lethal dose determination in 3-week-old hTfR1 HOM mice.
Shown are (A) survival (n = 7/JUNV challenge group, n = 3 sham-infected controls), (B) weight change relative to the day of virus challenge (group mean and SEM) and (C) clinical disease scores (group mean and SEM) for groups of mice infected i.p. with 105, 104 or 103 CCID50 of JUNV or sham-infected. LD90 = 3793; LD50 <1000 CCID50.
Figure 4
Temporal analysis of weight change, clinical disease signs, IFN-α and viral burden during the course of JUNV infection in 3-week-old hTfR1 HOM mice.
Animals (n = 28) were infected i.p. with 104 CCID50 of JUNV and subsets of 4 mice were designated for euthanasia on days 2, 4, 6, 8, 10, 12 and 14 p.i. for blood and tissue collection and analysis. Due to death prior to sample collection on days 12 and 14, only 2 mice were available on day 12. Four sham-infected animals were included as controls and 3 (one per day) were euthanized on days 2, 6 and 10 for sample collection. (A) Weight change of animals relative to the day of virus challenge, (B) clinical disease scores (mean and SEM), (C) serum IFN-α concentrations and (D) tissue and serum viral titers (the x-axis represents the limit of detection) are shown. **p=0.0060, ****p<0.0001 compared to sham-infected normal controls. SI, sham-infected.
Figure 5
Histopathology in 3-week-old hTfR1 HOM mice infected with JUNV.
Representative sections of brain (cerebral cortex) from (A and C) a sham-infected mouse and (B and D) a JUNV-infected mouse at day 12 p.i. (B) Neutrophilic encephalitis characterized by a perivascular cuff of inflammatory cells (arrow) and neutrophilic infiltration in the neuropil (arrowhead). (D) Higher magnification image showing necrotic cells (arrows) and neutrophils (arrowhead) within the neuropil. (E and G) Spleen (white pulp) from a sham-infected control mouse. (F and H) Spleen (white pulp) of a JUNV-infected mouse at day 12 p.i. (F) Tingible body macrophages (arrow) and individual lymphocyte cell death (arrowhead) are scattered within a periarteriolar lymphoid sheath. (H) Higher magnification image showing tingible body macrophages with cytoplasmic engulfed apoptotic debris (arrow) and individual lymphocyte cell death (arrowhead). Hematoxylin and eosin stain. A, B, E and F: 400 × magnification, bar = 50 μm. C, D, G and H: 1000 × magnification, bar = 20 μm.
Figure 6
Immunohistochemistry for JUNV antigen in 3-week-old hTfR1 HOM mice.
Representative sections of brain (midbrain) from (A and C) a sham-infected mouse and (B and D) a JUNV-infected mouse at day 12 p.i. Note the presence of the virus in neurons (brown staining). (E and G) Spleen (white pulp) from a sham-infected control mouse. (F and H) Virus antigen in mononuclear cells in the spleen (white pulp) of a JUNV-infected mouse at day 12 p.i. Hematoxylin counterstain. A, B, E and F: 400 × magnification, bar = 50 μm. C, D, G and H: 1000 × magnification, bar = 20 μm.
Figure 7
Effect of type I and II IFN response on JUNV infection and disease outcome in 3-week-old hTfR1 HOM mice.
Mice (n = 3/group) of different phenotypic backgrounds were challenged with 105 CCID50 of JUNV and monitored daily for (A) survival, (B) weight change relative to the day of virus challenge (group mean and SEM), (C) clinical disease (group mean and SEM) and (D) tissue viral loads present on day 28 p.i. in surviving animals (virus was not detected in serum or intestine or in any WT mice). The x-axis represents the virus titer assay limit of detection. *p=0.0246 compared to WT mice or all other phenotypes.
Figure 8
Effect of anti-IFN-α/βR mAb treatment on JUNV pathogenicity in 3-week-old hTfR1 HOM mice.
Animals (n = 3–4/group) were treated with a single 500 µg dose of IFN-α/βR-blocking mAb (with or without additional 250 µg maintenance doses every other day for one or two weeks) and infected i.p. the following day with 105 CCID50 of JUNV. The mice were monitored daily for (A) survival, (B) weight change relative to the day of virus challenge (mean and SEM), (C) clinical disease (mean and SEM) and (D) tissue viral loads in surviving animals on day 28 p.i. (virus was undetectable in serum, liver, lung, heart and intestine). The x-axis represents the virus titer assay limit of detection. *p=0.0114 comparing the two-week placebo treatment to the single and one-week mAb treatments; *p=0.0238 comparing the two-week placebo treatment to the two-week mAb treatment. Tx, treatment.
Tables
Key resources table
| Reagent type (species) or resource | Designation | Source or reference | Identifiers | Additional information |
|---|---|---|---|---|
| Genetic reagent (Mus musculus) | C57BL/6 hTfR1 knock-in mice (human TFRC replacing the mouse Tfrc) | Genentech | ||
| Genetic reagent (Mus musculus) | AG129 mice deficient in IFN-α/β receptor (R) and IFN-γR (Ifnar-/-; Ifngr-/-) | Washington University Medical School | ||
| Genetic reagent (Mus musculus) | Wild-type (WT) mice (hybrid C57BL/6 × 129 background) | This paper | See Materials and methods | |
| Genetic reagent (Mus musculus) | hTfR1 heterozygous (HET) mice (hybrid C57BL/6 × 129 background) | This paper | See Materials and methods | |
| Genetic reagent (Mus musculus) | hTfR1 homozygous (HOM) mice (hybrid C57BL/6 × 129 background) | This paper | See Materials and methods | |
| Genetic reagent (Mus musculus) | IFN-α/βR-deficient mice (hybrid C57BL/6 × 129 background) | This paper | See Materials and methods | |
| Genetic reagent (Mus musculus) | IFN-α/β and -γR-deficient mice (hybrid C57BL/6 × 129 background) | This paper | See Materials and methods | |
| Genetic reagent (Mus musculus) | hTfR1 HOM–IFN-α/βR-deficient mice (hybrid C57BL/6 × 129 background) | This paper | See Materials and methods | |
| Genetic reagent (Mus musculus) | hTfR1 HOM–IFN-α/β and -γR-deficient mice (hybrid C57BL/6 × 129 background) | This paper | See Materials and methods | |
| Strain, strain background (Junín virus) | Recombinant JUNV Romero strain | University of Texas Medical Branch | ||
| Cell line (Cercopithecus aethiops) | Vero | ATCC | Cat# CCL-81 RRID:CVCL_0059 | |
| Commercial assay or kit | VeriKine Mouse Interferon Alpha ELISA Kit | PBL Assay Science | Cat# 42115–1 | |
| Antibody | Goat anti-mouse IgG1 Fab (polyclonal) | Jackson ImmunoResearch Laboratories | Cat# 115-007-185 RRID:AB_2632498 | (1:10) |
| Antibody | Anti-JUNV nucleoprotein antibody (QC03-BF11) | BEI Resources | Cat# NR-43775 | (1:100) |
| Antibody | Goat anti-mouse IgG (H+L) - HRP secondary antibody (polyclonal) | Thermo Fisher Scientific | Cat# G-21040 RRID:AB_2536527 | (1:100) |
| Antibody | Anti-mouse IFNAR-1 (anti-IFN-α/βR) monoclonal antibody (MAR1-5A3) | Bio X Cell | Cat# BE0241 RRID:AB_2687723 | (500 µg primary dose; 250 µg maintenance dose) |
| Software | GraphPad Prism software | GraphPad Prism (https://www.graphpad.com) | RRID_SCR_002798 | Version 8.4.1 |
Additional files
Download links
A two-part list of links to download the article, or parts of the article, in various formats.
Downloads (link to download the article as PDF)
Open citations (links to open the citations from this article in various online reference manager services)
Cite this article (links to download the citations from this article in formats compatible with various reference manager tools)
Type I interferon underlies severe disease associated with Junín virus infection in mice
eLife 9:e55352.
https://doi.org/10.7554/eLife.55352
