Journal: bioRxiv

Article Title: Strain-Specific Human Natural Killer Cell Recognition of Influenza A Virus

doi: 10.1101/148528

Figure Lengend Snippet: (A) Cytokine concentrations assessed by Luminex®, values displayed represent the mean fluorescence intensity in pH1N1-infected/H3N2 infected conditions (MOI=3). Cytokines elevated by 2.5-fold over the level in mock-infected monocytes are plotted. (B) Impact of cytokine receptor blocking on the NK cell IFN-γ response evaluated by pre-incubating NK cells for 1 hr with blocking antibodies specific to IFNAR2, IL-12R, IL-15R, IL-18R, and IFNGR1 followed by co-culture with infected monocytes. 24 HPI, intracellular cytokine staining was used to assess NK cell IFN-γ + frequency compared to treatment with an isotype control antibody (H3N2: n =2-6, pH1N1: n =9). (C) NK cells were incubated for 1 hr with antibodies specific to IFNAR2 and CD226 or CD54 followed by co-culture with autologous infected monocytes. At 24 HPI, intracellular cytokine staining was used to assess IFN-γ production compared to treatment with an isotype control antibody ( n =6). * P < 0.05, ** P < 0.005, Wilcoxon signed-rank test. (D) Model of strain-specific NK cell recognition of influenza A infection. pH1N1-infection of monocytes virus does not downregulate CD54 and CD112 to the same extent as H3N2-infected monocytes. CD54 expression is retained on Flu-NP + cells while CD112 expression is preferentially retained on exposed, uninfected monocytes. pH1N1 infection of monocytes elicits enhanced IFN-α secretion and blockade of IFNAR2 dampens NK cell anti-pH1N1 IFN-γ production.

Article Snippet: IL-12R β 2 Ab (R&D Systems AF1959-SP) was used at a final concentration of 83.33 μg/mL.

Techniques: Luminex, Fluorescence, Infection, Blocking Assay, Co-Culture Assay, Staining, Control, Incubation, Virus, Expressing