Journal: Vaccines
Article Title: Mucosal Delivery of Recombinant SARS-CoV-2 Spike Receptor-Binding Domain Antigen Containing Immune-Stimulating Peptides Induces Protective Immune Responses Against Viral Infection in huACE2 Mice
doi: 10.3390/vaccines14050421
Figure Lengend Snippet: Interaction between Co4B and Siglec F. ( A ) Similarity between the Co4B and VAP-1 amino acid sequences. ( B ) EGFP and EGFP-Co4B were incubated on plates coated with recombinant Siglec F, and binding was detected using ELISA. ( C ) Binding of recombinant Siglec F to immobilized EGFP–Co4B was quantified by ELISA. ( D ) Binding of Siglec F to immobilized Co4B peptides was assessed by ELISA using serial dilutions of Siglec F. ( E ) NALTs were collected and sectioned after administration of EGFP-Co4B. Colocalization of EGFP-Co4B and Siglec F on M cells (identified using NKM 16-2-4, red) in NALT was assessed based on fluorescence signal overlap. Scale bars represent 50 μm. ( F ) The expression of Siglec F (purple) on M cells (green) in a Peyer’s patch (PP) was visualized by confocal laser-scanning microscopy, demonstrating colocalization of EGFP-Co4B with Siglec F on M cells. Scale bar represents 10 μm. Data represent the mean ± SDs of at least two independent experiments. Statistical significance in ( B , C ) was determined using an unpaired t -test, and in ( D ) using two-way ANOVA (* p < 0.05, *** p < 0.001, **** p < 0.0001).
Article Snippet: To investigate interactions between Co4B and Siglec F, recombinant EGFP-Co4B, EGFP alone, and Co4B peptide alone (Mimotopes, Mulgrave, VIC, Australia) were individually coated onto ELISA plates.
Techniques: Incubation, Recombinant, Binding Assay, Enzyme-linked Immunosorbent Assay, Fluorescence, Expressing, Confocal Laser Scanning Microscopy