A Method to Assess Fc-mediated Effector Functions Induced by Influenza Hemagglutinin Specific Antibodies

Mark J. Bailey; Felix Broecker; Paul E. Leon; Gene S. Tan

doi:10.3791/56256

A Method to Assess Fc-mediated Effector Functions Induced by Influenza Hemagglutinin Specific Ant...

JoVE Journal
Immunology and Infection

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JoVE Journal Immunology and Infection

A Method to Assess Fc-mediated Effector Functions Induced by Influenza Hemagglutinin Specific Antibodies

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04:47 min

February 23, 2018

DOI: 10.3791/56256-v

Mark J. Bailey^1,2, Felix Broecker¹, Paul E. Leon^1,2, Gene S. Tan³

¹Department of Microbiology,Icahn School of Medicine at Mount Sinai, ²Graduate School of Biomedical Sciences,Icahn School of Medicine at Mount Sinai, ³Department of Infectious Disease,J. Craig Venter Institute

Overview

This article describes a method for measuring the activation of Fc-mediated effector functions by antibodies targeting the influenza virus hemagglutinin. The assay can be adapted for monoclonal antibodies or polyclonal sera against other viral glycoproteins.

Key Study Components

Area of Science

Immunology
Vaccinology

Background

Fc-mediated immunity plays a crucial role in the immune response.
Understanding antibody interactions with viral proteins is essential for vaccine development.
This method allows for rapid assessment of antibody efficacy.

Purpose of Study

To evaluate the ability of antibodies to activate Fc-mediated effector functions.
To provide a protocol that can be completed within 24 hours.
To facilitate research in immunology and vaccine efficacy.

Methods Used

Transfection of human embryonic kidney 293 cells with DNA coding for viral hemagglutinin.
Measurement of antibody-dependent cellular cytotoxicity.
Use of a 96-well plate format for high-throughput analysis.
Data analysis conducted within 24 hours post-experiment.

Main Results

The method successfully measures Fc-mediated effector functions.
Results can be obtained quickly, enhancing research efficiency.
Adaptable for various antibodies targeting different viral glycoproteins.

Conclusions

This assay provides a valuable tool for assessing antibody efficacy.
It contributes to the understanding of Fc-mediated immunity.
The rapid turnaround time supports timely research advancements.

Frequently Asked Questions

What is Fc-mediated immunity?

Fc-mediated immunity refers to the immune responses triggered by the Fc region of antibodies, which can activate various effector functions.

How long does the assay take?

The entire assay can be completed within 24 hours, from material preparation to data analysis.

Can this method be used for other viruses?

Yes, the assay can be adapted to assess antibodies targeting other viral surface glycoproteins.

What type of cells are used in this protocol?

Human embryonic kidney 293 cells are used for transfection in this protocol.

Who demonstrated this procedure?

Mark Bailey, a graduate student from Peter Palese's laboratory, demonstrated this procedure.

What is the main advantage of this technique?

The main advantage is the rapid assessment of antibody efficacy, allowing for timely research outcomes.

We describe a method to measure the activation of Fc-mediated effector functions by antibodies that target the influenza virus hemagglutinin. This assay can also be adapted to assess the ability of monoclonal antibodies or polyclonal sera targeting other viral surface glycoproteins to induce Fc-mediated immunity.

The overall goal of this protocol is to assess the ability of an antibody or a serum sample to activate Fc-mediated effector function. This method can help answer key questions in the immunology and vaccinology fields by measuring the activation of antibody-dependent Fc-mediated immunity. The main advantage of this technique is that the experiment can be done within 24 hours from the material preparation to the data analysis.

Demonstrating this procedure will be Mark Bailey, graduate student from the laboratory of Peter Palese. To induce influenza virus hemagglutinin expression via transfection, first plate human embryonic kidney 293 cells at a two times 10 to the fourth cells per well concentration in a white tissue culture treated 96-well plate. After four hours at 37 degrees Celsius and 5%CO2, transfect the cells with 100 nanograms of DNA coding for viral hemagglutinin and 0.2 microliters of transfection reagent per well and a total volume of 50 microliters of 1X optimum-reduced serum medium.

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