Fig 1.
Loss of TIM-1 reduces mortality following EBOV GP/rVSV and EBOV GP ΔO/rVSV infection, but not G/rVSV.
A and B. Female BALB/c Ifnar-/- (control) and BALB/c Ifnar -/-/TIM-1-/- (TIM-1-/-) mice infected with 105 iu EBOV GP/rVSV (A; n = 5 mice per group) or EBOV GP ΔO/rVSV (B; n = 13–17 mice per group) by intravenous (i.v.) injection. C. Female BALB/c Ifnar -/- (control) and BALB/c Ifnar -/-/TIM-1-/- (TIM-1-/-) mice infected with 105 iu G/rVSV (n = 10 mice per group) by i.v. infection. D. Similar G/rVSV challenge studies as shown in panel C, but mice were challenged with 101 iu (n = 5 mice per group) of G/rVSV. Survival was assessed following infection for all mouse studies. Significance for survival curve was determined by Log Rank (Mantel-Cox) test, * p< 0.05, **p < 0.01. LT50 = median lethal time until death; NC, noncalculable; ns, not significant.
Fig 2.
Reduced viremia and virus titers in a variety of organs of TIM-1-/- mice at late time points following i.v. EBOV GP ΔO/rVSV infection.
Serum and organs were harvested from BALB/c Ifnar -/- (Control) and BALB/c Ifnar -/-/TIM-1-/- (TIM-1-/-) mice at days 1, 3 and 5 following infection with 105 iu of EBOV GP ΔO /rVSV by i.v. injection. Titers were determined by endpoint dilution of serum or homogenized organ samples on Vero cells. Solid lines indicate geometric mean for each data set. Dotted line indicates the level of detection. Adrenal gland titers are displayed as per gland homogenized in 1 ml of PBS. Significance was calculated by Mann-Whitney test to compare control to TIM-1-/- mice at each time point; *p < 0.05; **p < 0.01; ns, not significant.
Fig 3.
Cytokine expression in liver, spleen and kidney of EBOV GPΔO/rVSV-infected Ifnar-/- and Ifnar -/-/TIM-1-/- mice.
Tissues were harvested from uninfected and infected BALB/c Ifnar -/- (control) and BALB/c Ifnar -/-/TIM-1-/- (TIM-1-/-) mice. In infected mice, tissues were harvested at 3 or 5 days following infection with 105 iu of EBOV GPΔO/rVSV by i.v. injection. RNA was isolated from the organs and expression of mouse TNF, IL-6, IL-10 and IL-12, was quantified by qRT-PCR. Results represent cytokines expression relative to murine HPRT for at least three independent livers, spleens and kidneys. Data points represent values for individual mice. Solid lines indicate the mean for each data set. Statistical significance was determined by Student’s t-test compared the control mice for each time point and is only shown for those comparisons observed to differ. *p<0.05.
Fig 4.
Chemokine CXCL10 and CCL2 expression in the liver, spleen and kidney of EBOV GPΔO/rVSV-infected control and TIM-1-/- mice.
Tissues were harvested from uninfected and infected BALB/c Ifnar-/- (control) and BALB/c Ifnar -/-/TIM-1-/- (TIM-1-/-) mice. In infected mice, tissues were harvested at 3 or 5 days following infection with 105 iu of EBOV GPΔO/rVSV by i.v. injection. RNA was isolated from the organs and expression of proinflammatory chemokines, mouse CXCL10 and CCL2, was quantified by qRT-PCR. Results represent chemokine expression relative to murine HPRT for at least three independent livers, spleens and kidneys. Data points represent values for individual mice. Solid lines indicate the mean for each data set. Statistical significance was determined by Student’s t-test compared the control mice for each time point and is only shown for those comparisons observed to differ. *p<0.05.
Fig 5.
T cell depletion does not alter the survival protection conferred by the loss of TIM-1 expression.
A. Intraperitoneal injection of α-CD8 mAb, clone 2.43, and α-CD4 mAb, clone GK1.5, treatment at days -1 and 2 systemically depleted T cell populations in female BALB/c Ifnar-/- (control) and BALB/c Ifnar-/- /TIM-1-/- (TIM-1-/-) mice as determined by α-CD90 mAb staining of peripheral blood mononuclear cells at day 5 following EBOV GP/rVSV infection. CD90.2 overlay depicts the subset of cells gated in the panel on the left. B. Survival was assessed following infection with 7x102 iu of EBOV GP/rVSV administered by intravenous infection (n = 10 mice per group) and two treatments of α-CD8 mAb and α-CD4 mAb at Day -1 and 2 from infection. Significance for survival curve was determined by Log Rank (Mantel-Cox) test. LT50 = median lethal time until death; ***p < 0.001.