Treatment with inhaled aerosolised ethanol reduces viral load and potentiates macrophage responses in an established influenza mouse model

Figures & data

Table 1. Baseline exposure measurements.

	Saline Control	40% Ethanol	60% Ethanol	80% Ethanol
Pre-Exposure Weight (g)	18.5 ± 0.9	18.6 ± 1.0	18.1 ± 1.3	18.3 ± 0.9
Post-Exposure Weight (g)	18.8 ± 1.2	18.8 ± 1.0	18.4 ± 1.3	18.0 ± 0.9
Total Cells (x10^4 cells/mL)	7.0(1.9-15.3)	6.1(3.4-11.3)	3.8(1.2-5.5)	3.9(2.8-7.1)
Macrophages (x10^4 cells/mL)	5.5(1.9-10.3)	5.6(3.4-11.3)	3.8(1.2-5.5)	3.9(2.8-7.1)
Neutrophils (x10^3 cells/mL)	1.1(0-55.1)	0.0(0.0-5.0)	0.0(0.0-0.4)	0.0(0.0-0.1)*
Lymphocytes (cells/mL)	0.0 (0.0-0.0)	0.0 (0.0-313.3)	0.0 (0.0-0.0)	0.0(0.0-0.0)
BAL cell viability (%)	91.8 ± 3.9	92.5 ± 4.5	93.0 ± 7.2	92.9 ± 8.6
Epithelial thickness (√area/Pbm)	0.035 ± 0.006	–	–	0.031 ± 0.005
Airway smooth muscle mass (√area/Pbm)	0.200 ± 0.036	–	–	0.184 ± 0.024
Lumen (√area/Pbm)	0.042 ± 0.005	–	–	0.039 ± 0.005
IL-6	2.7(2.7-4.1)	2.7(2.7-4.3)	2.7(2.7-2.7)	2.7(2.7-9.8)
TNFα	5.0(5.0-34.6)	5.0(5.0-5.0)	5.0(5.0-5.0)	5.0(5.0-5.0)

Baseline assessments from naïve BALB/c mice (n = 10/group) exposed to 3x30minute exposures with saline (control) or three different concentrations of ethanol (40%,60%,80%). data are presented as mean ± standard deviation or median (range). *statistically significant difference (p < 0.05) relative to control. IL-6 = interleukin-6, TNFα = tumour necrosis factor alpha, p_bm = perimeter of the basement membrane.

Figure 1. Efficacy of inhaled ethanol in treating established influenza infection. Female BALB/c mice received 10^4.5 plaque-forming units of immediate virulence “Mem71” influenza. 48-h after infection, mice were exposed to 3 days of 3x30minute exposures to saline (control) or 40%, 60%, or 80% ethanol and analyzed 24-h after the last exposure. (A) Viral load in whole lung tissue was measured by quantitative polymerase chain reaction (n = 10/group). (B) Mice were weighed daily, with weights normalized relative to starting weight (100%). bronchoalveolar lavage samples were collected by washing with chilled saline and then centrifuged to separate the cell pellet and supernatant. (C) Differential counts in the cell pellet were obtained using light microscopy after Rapid-Stain. (D) Interleukin-6 (IL-6) and tumour necrosis factor alpha (TNFα) levels in the supernatant were measured by ELISA. * = holm adjusted p < 0.05 (calculated with t-test in a and Mann-Whitney U-tests in C,D with the control group as reference).

Data availability statement

The authors confirm that the data supporting the findings of this study are available within the article.