Journal: bioRxiv

Article Title: Poor sleep impairs immune responses and influenza vaccine protection

doi: 10.1101/2025.05.07.652783

Figure Lengend Snippet: a) Serological titers post-vaccination. Serological titers in the mice blood at 35 days post-vaccination (dpv) but before challenge, were determined by hemagglutination inhibition (HAI) and microneutralization assays. b) Avidities of vaccine-specific antibodies. The avidity index was calculated based on the area under the entire antibody titration curve using GraphPad Prism (version 10.1.2). The antibody index equals to an AUC ratio of treated and untreated reactions. All samples were assessed in duplicate, and mean titers were calculated . c) quantities of total IgG and subclass IgGs. d ) Aera under curves (AUC) for total IgG and subclass IgGs. e) ELISpot analysis of HA- and vaccine-specific antibody-secreting B-cells and virus-specific IFN-γ–producing T-cells. Splenocytes were collected from CSF-treated (gSF) and normal-sleep control (gSC) mice one week prior to viral challenge. The frequency of antigenic-specific IgG-secreting B-cells was assessed by ELISpot following in vitro stimulation with recombinant HA protein of CA/07 and the purified vaccine virus. The frequency of virus-specific IFN-γ–producing T-cells was measured by ELISpot after stimulation with whole CA/07 virus. Data are shown as the number of spot-forming cells per million splenocytes.

Article Snippet: Goat anti-Mouse IgM Secondary Antibody (Cat. 31440, Invitrogen, Waltham, MA, USA), Goat anti-Mouse IgG Fc Secondary Antibody (Cat. 31437, Invitrogen, Waltham, MA, USA), Goat Anti-Mouse IgG3 (Cat. 75952S, Cell Signaling Technology, Danvers, MA, USA), Goat Anti-Mouse IgG2b (Cat. 43593S, Cell Signaling Technology, Danvers, MA, USA), Goat Anti-Mouse IgG1 (Cat. 96714S, Cell Signaling Technology, Danvers, MA, USA), Goat Anti-Mouse IgG2c (Cat. 56970S, Cell Signaling Technology, Danvers, MA, USA).

Techniques: HI Assay, Titration, Enzyme-linked Immunospot, Virus, Control, In Vitro, Recombinant, Purification