Fig. 1 | Signal Transduction and Targeted Therapy

Fig. 1

From: Intranasal adenovirus-vectored Omicron vaccine induced nasal immunoglobulin A has superior neutralizing potency than serum antibodies

Fig. 1The alternative text for this image may have been generated using AI.

Analysis of immunoglobulins in nasal mucosal lining fluids (NMLFs). a Neutralizing activities of NMLFs against VSV pseudoviruses bearing spikes of Wildtype (WT), Delta, Omicron BA.1, and BA.5. NMLFs were collected from eight donors before and 3–6 weeks after two intranasal booster doses. Neutralization assay was performed using 100 µl samples containing an equal amount of total IgA (3 μg) and presented as the percentage of inhibition. Data are shown as % inhibition. Individual data are presented (n = 8). b Detection of IgA that binds to spikes of WT, Delta, BA.2, and BA.5 in NMLFs. NMLFs from eight donors were collected before and 3–6 weeks after two intranasal doses. Spike-specific IgA was detected using an electrochemiluminescence method (Meso Scale Discovery). Individual data were normalized to equal amounts of total protein. Data are shown as the geometric mean fold increase (GMFI) with a 95% confidence interval (CI). Individual data are presented (n = 8). c Neutralizing activities of serum samples against VSV pseudoviruses bearing spikes of WT, Delta, BA.1, and BA.5. Serum samples were collected from eight donors before and 3–6 weeks after intranasal vaccination. A neutralization assay was performed using serial dilutions of the samples, and the results are presented as neutralizing titers (NT50). The cutoff value was set at 1:30. Data are shown as the geometric mean titer (GMT) with 95% CI. Individual data are presented (n = 8). d Detection of IgA that binds to the spikes of WT, Delta, BA.2, and BA.5 in serum samples. Serum samples from eight donors were collected before and 3–6 weeks after two intranasal booster doses. Spike-specific IgA was detected using an electrochemiluminescence method (Meso Scale Discovery). Data are shown as GMFI with 95% CI. Individual data are presented (n = 8). e Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS‒PAGE) and western blot analysis of NMLFs. 8 μg total protein in NMLFs of two representative donors were loaded onto an SDS-PAGE gel and stained with Coomassie blue. Purified human serum IgG and IgA were used as references. Western blot analysis was performed with 200 ng total protein in NMLFs, using an anti-human IgA heavy chain (HC) antibody and an anti-human IgG H + L antibody to detect IgA and IgG, respectively. f Nasal sIgA samples were purified and pooled from NMLFs of six donors, then subjected to gel-filtration chromatography (GFC) separation. Each collected fraction was subjected to western blot analysis using an anti-human IgA heavy chain (HC) antibody. g Molecular size distribution of nasal sIgA. Nasal sIgA were purified and pooled from NMLFs of six donors, then subjected to sedimentation velocity-analytical ultracentrifugation (SV-AUC). The data were analyzed with SEDFIT software to obtain sedimentation coefficient distribution C (S)

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