Journal: bioRxiv

Article Title: Targeting the Annexin A1-FPR2/ALX pathway for host-directed therapy in dengue disease

doi: 10.1101/2021.11.02.466887

Figure Lengend Snippet: (A) Experimental design. 8-week-old A129 mice were mock-infected or inoculated with 1×10 3 PFU of DENV-2 by the intravenous (i.v.) route. From day 2, mice, were treated or not twice a day with 150μg of Ac 2-26 by the intraperitoneal (i.p.) route. (B) Mice were culled in the indicated time points after infection, and plasma was collected for AnxA1 quantification by ELISA (n=5). (C) Bodyweight loss was assessed in the indicated time points and expressed as a percentage of initial body weight. Mock (open white circles), Ac 2-26 (open red circles), and DENV-2-infected mice treated with vehicle (black closed circles) or Ac 2-26 (red closed circles); n=4-8. Five days after infection, animals were culled, and blood and tissue collected for the following analysis: (D) platelet counts, shown as the number of platelets × 10 3 /μL of blood (n=5-14); (E) haematocrit levels, shown as % volume occupied by red blood cells (n=8); concentrations of (F) CCL5 and (G) IL-6 in plasma and spleen of mock and DENV-infected mice, treated or not with Ac 2-26 . Concentrations were assessed by ELISA and are shown as pg/mL of plasma or as pg/100mg of the spleen (CCL2, n=4-7; IL-6, n=4-13). (H , I) Liver of control and DENV-2-infected mice, treated or not with the AnxA1 peptide, were collected, formalin-fixed, and processed into paraffin sections. (H) Histopathological scores and (I) representative images of liver sections stained with haematoxylin and eosin. Scale Bar, 100 μm. (J) Plasma ALT activity represented as units/L (H-J, n=4-8). (K) Viral loads recovered from plasma, spleen, and liver of infected mice treated or not with Ac 2-26 , examined by plaque assay in Vero cells. Results are shown as the log of PFU/mL of plasma or as the log of PFU/mg of spleen and liver (n=8). All results are expressed as mean (horizontal bars) ± SD. In B, differences over time and between treatments were compared by one-way ANOVA followed by Tukey’s multiple comparisons test: ****p<0.0001 versus mock-infected animals or comparing the different groups, as indicated in the graph. In C-J, data were analysed by one-way ANOVA followed by Tukey’s multiple comparisons test: *p<0.05, **p<0.01, ***p<0.001 and ****p<0.0001 versus mock-infected group; # p<0.05, ## p<0.01, ### p<0.001 and #### p<0.0001 versus mock-infected group treated with Ac 2-26 . In K, statistical analyses were performed by unpaired Student’s t-tests for each organ. – Source data 1: Raw data for .

Article Snippet: The alanine aminotransferase (ALT) activity was measured in individual serum samples from A129 mice, using a commercially available colourimetric kit (Bioclin, Quibasa, Belo Horizonte, Brazil).

Techniques: Infection, Clinical Proteomics, Enzyme-linked Immunosorbent Assay, Control, Staining, Activity Assay, Plaque Assay

Journal: bioRxiv

Article Title: Targeting the Annexin A1-FPR2/ALX pathway for host-directed therapy in dengue disease

doi: 10.1101/2021.11.02.466887

Figure Lengend Snippet: 8-week-old A129 mice were mock-infected or inoculated with 4 × 10 4 PFU of DENV-1, 1 × 10 3 PFU of DENV-3 or 1 × 10 4 PFU of DENV-4 by the intravenous route (n=5). From day 2, mice were treated with vehicle (black closed circles) or 150μg of Ac 2-26, i.p. twice a day (open red circles). Five days after infection, animals were culled, and blood and tissue were collected for the following analysis: (A) Platelet counts, shown as the number of platelets × 10 3 /μL of blood. (B) Haematocrit levels, shown as % volume occupied by red blood cells. (C) plasma ALT activity represented as units/L. Concentrations of L-6 in (D) plasma and (E) spleen of mock- and DENV-infected mice, treated or not with Ac 2-26 , assessed by ELISA. Concentrations are shown as pg/mL of plasma or as pg/100mg of the spleen. Viral loads recovered from (F) plasma and (G) spleen of mice infected with the three serotypes of DENV and treated or not with Ac 2-26 , evaluated by plaque assay in Vero cells. Results are shown as the log of PFU/mL of plasma or as the log of PFU/mg of spleen and liver. In A-E, data were analysed by two-way ANOVA followed by Dunnett’s (*p<0.05, **p<0.01, ***p<0.001 and ****p<0.0001 versus mock-infected group) or Šídák’s (statistical differences between infected mice treated with vehicle or Ac 2-26 , as indicated in the graphs) multiple comparison test. In F-G, statistical analysis was performed by two-way ANOVA followed by Šídák’s multiple comparison test, and differences between animals treated with vehicle or Ac 2-26 are indicated in the graphs. Horizontal bars represent mean values. – Source data 1: Raw data for .

Article Snippet: The alanine aminotransferase (ALT) activity was measured in individual serum samples from A129 mice, using a commercially available colourimetric kit (Bioclin, Quibasa, Belo Horizonte, Brazil).

Techniques: Infection, Clinical Proteomics, Activity Assay, Enzyme-linked Immunosorbent Assay, Plaque Assay, Comparison