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Measuring Influenza Neutralizing Antibody Responses to A(H3N2) Viruses in Human Sera by Microneutralization Assays Using MDCK-SIAT1 Cells
Published on: November 22, 2017
Fluorometric high-throughput assay for measuring chlamydial neutralizing antibody
Timothy Southern1, Leah Bess, Jillian Harmon
1Laboratory of Intracellular Parasites, Rocky Mountain Laboratories, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Hamilton, Montana, USA.
Clinical and Vaccine Immunology : CVI
|September 28, 2012
Summary
A new high-throughput assay accurately measures Chlamydia trachomatis-neutralizing antibodies in human sera. This tool aids in developing a broadly protective vaccine against this common bacterial sexually transmitted infection and cause of blindness.
Area of Science:
- Immunology
- Microbiology
- Vaccinology
Background:
- Chlamydia trachomatis is a major cause of preventable blindness (trachoma) and bacterial sexually transmitted infections globally.
- Current treatments involve antibiotics, but a broadly protective vaccine is needed to control C. trachomatis infections.
- Identifying chlamydial antigens that induce pan-neutralizing antibodies is crucial for vaccine development.
Purpose of the Study:
- To describe an objective, high-throughput in vitro assay for measuring C. trachomatis-neutralizing antibodies.
- To enable the screening of human sera for neutralization activity to identify key vaccine antigens.
- To support vaccine development and epidemiological studies of C. trachomatis infection.
Main Methods:
- Developed a 96-well microtiter format assay for antibody-mediated neutralization of C. trachomatis infection.
- Quantified neutralization using immunostaining of fixed cells followed by automated fluorometric analysis.
- Validated the assay by comparing fluorometric results to manual inclusion counts and assessing serovar-specific neutralization.
Main Results:
- Automated fluorometric analysis demonstrated a direct correlation with labor-intensive manual inclusion counts.
- The assay successfully identified C. trachomatis serovar- and serocomplex-specific neutralization.
- The assay provides an objective, high-throughput method for analyzing serum neutralization.
Conclusions:
- The developed fluorometric assay is a reliable and efficient tool for quantifying C. trachomatis-neutralizing antibodies.
- This assay facilitates the identification of antigens that elicit broadly neutralizing antibodies for vaccine development.
- The assay is suitable for epidemiological studies, clinical vaccine trials, and preclinical research on Chlamydia infections.
