anti il 22 pe  (R&D Systems)

Journal: bioRxiv

Article Title: Mitochondrial cyclophilin D promotes disease tolerance by licensing NK cell development and IL-22 production against influenza virus

doi: 10.1101/2021.05.28.445832

Figure Lengend Snippet: (A) Levels of IFN-γ in the BAL of infected WT and CypD -/- mice as determined by ELISA. (B) Intracellular cytokine staining of IFN-γ in NK cells. FACS plots are representative of the quantification on the right and are gated against the FMO. (C) Levels of IL-22 in the BAL of infected WT and CypD -/- mice as determined by ELISA. (D) Intracellular cytokine staining for IL-22 by NK cells in the BAL. FACS plots on the left are representative of the quantification on the right. An IL-22-deficient mouse was used as a staining control for specificity. (E) Model of recombinant IL-22 experiments as used in F-I. (F) Mice were infected with LD 50 90 PFU, administered recombinant IL-22 or PBS on day 5 p.i. and survival monitored. (G-I) a sublethal dosage of 50 PFU was used and mice were administered IL-22 or PBS as before. At day 7, viral loads were quantified (G), pulmonary inflammation and fibrosis were assessed by hematoxylin and eosin or Masson’s Trichrome staining, respectively (H; scale bar = 30µM) or erythrocytes in the BAL were enumerated (I). (J) Schematic of NK cell transfer experiments performed in K-N. On day 7 post-infection, day 2 post-transfer, protein (L), erythrocyte (M) and leukocyte levels in the BAL or lung viral loads (N) were assessed. In panels A, C, F, I, K, L and N data are combinations of two or three independent experiments. In B, D, G and M data are from one experiment that is representative of two or three independent experiments. Micrographs in H are representative of 3 or 4 individual mice. In A-D, G, I and K-N, each symbol represents data from one individual mouse, while in H total n values are displayed in the panel legend. Statistical analyses were performed as follows: in A-D Two-way ANOVA followed by Sidak’s multiple comparisons test; F Log Rank Test; G, I Two-way ANOVA followed by Tukey’s multiple comparisons test; K-N One-way ANOVA followed by Dunnett’s multiple comparisons test, Kruskal-Wallis test. See also Figure S5 .

Article Snippet: Cells were then stained extracellularly, fixed and permeabilized using BD Cytofix/Cytoperm (BD Biosciences), before being stained intracellularly for PE-conjugated anti-IL-22 and APC-conjugated anti-IFN-γ (BD Biosciences).

Techniques: Infection, Enzyme-linked Immunosorbent Assay, Staining, Recombinant

Journal: bioRxiv

Article Title: Mitochondrial cyclophilin D promotes disease tolerance by licensing NK cell development and IL-22 production against influenza virus

doi: 10.1101/2021.05.28.445832

Figure Lengend Snippet: (A-K) Mice were infected with 50 PFU of IAV. (A) The frequency of IFN-γ-producing splenic NK cells at day 5 post-infection. (B-D) WT and Ifngr -/- mice were infected and the levels of protein (B), erythrocytes (C) and total cells (D) in the BAL at 7 days post-infection were determined. Following administration of recombinant IFN-γ, levels of protein (E) and erythrocytes (F) were assessed in the BAL. (G-I) WT and IL-22-deficient mice were infected and amount of protein (G) and number of cells (H) were enumerated, as well as pulmonary inflammation by hematoxylin and eosin staining (I; scale bar = 30µM). (J) Frequency of IL-22-producing NK cells in the spleens of WT and CypD-deficient mice at 5 and 7 days post-IAV infection. In all panels except I and K, symbols represent an individual mouse. In I, micrographs are a representative image taken from one of four mice. In K quantification is done from 10 random micrographs and each symbol represents one micrograph. In A and J, data are taken from one experiment that is representative of three. In B-H, panels are a compilation of two individual experiments. Differences were determined as follows: in A-D, G-H Two-tailed Student’s T-test; in E, F, K Two-way ANOVA followed by Tukey’s multiple comparisons test or Sidak’s (J). Refers to .

Article Snippet: Cells were then stained extracellularly, fixed and permeabilized using BD Cytofix/Cytoperm (BD Biosciences), before being stained intracellularly for PE-conjugated anti-IL-22 and APC-conjugated anti-IFN-γ (BD Biosciences).

Techniques: Infection, Recombinant, Staining, Two Tailed Test