eoe sub articles

EoE Sub Articles

All procedures involving human participants have been performed in compliance with the institutional, national, and international guidelines for human welfare and have been reviewed by the local institutional review board.
1. Serum Collection
<ol>
	<li>Collect serum samples from humans at time points of interest. For this study, we collected sera at days 0 (time of influenza vaccination), +7, +30, +60, and +180 after vaccination.</li>
	<li>To obtain the serum, centrifuge the...

the hemagglutination inhibition assay to detect serum antibodies against a target antigen

Source: Kaufmann, L. et al., <a href="https://app.jove.com/v/55833">An Optimized Hemagglutination Inhibition (HI) Assay to Quantify Influenza-specific Antibody Titers.</a> J. Vis. Exp. (2017)
The video demonstrates a hemagglutination inhibition assay for detecting serum antibodies against specific antigens. Confirmation of hemagglutination inhibition in the test well and hemagglutination in the control well is achieved through distinct visual patterns. This assay plays a crucial role in assessing immune response, vaccine effectiveness, and studying viral infections in virology and immunology.

<img alt="Figure 1" src="/files/ftp_upload/21787/21787_Figure1.jpg" /> 
Figure 1: Principle of hemagglutination and hemagglutination inhibition.&#160;No hemagglutination occurs in a negative control situation without viruses and antibodies (left column), and erythrocytes hemagglutinate only in the presence of influenza virus (middle column). However, when the hemagglutinin of the influenza virus is blocked by virus-specific antibodies then no hemagglutination can occ...

The Hemagglutination Inhibition Assay to Detect Serum Antibodies Against a Target Antigen

All procedures involving human participants have been performed in compliance with the institutional, national, and international guidelines for human ...

Begin with a V-shaped bottom multi-well plate and add diluted serum from an influenza-vaccinated human containing influenza-specific antibodies, to the test well.
Introduce viral hemagglutinin &#8212; an influenza-specific antigen in the test and control wells.
During incubation, in the test well, influenza-specific antibodies interact with antigens, forming complexes. While in the control well, antigens remain unbound.
Incubate the wells with avian red blood cells or RBCs.
In the control well, free antigens interact with RBC&#39;s surface receptors specific to viral hemagglutinin, causing RBCs to clump, leading to hemagglutination, and forming a compact pattern at the well&#39;s bottom.
However, in the test well, the presence of antigen-antibody complexes and the absence of free antigens inhibit hemagglutination, settling RBCs at the bottom.
Post-assay, tilt the plate. The control well exhibits a small, compact circular pattern with a hazy appearance &#8212; indicative of hemagglutination.
In contrast, the test well shows a tear-like diffused pattern, suggesting hemagglutination inhibition.