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A Step-By-Step Method to Detect Neutralizing Antibodies Against AAV using a Colorimetric Cell-Based Assay
Published on: December 7, 2021
Measuring immune responses to recombinant AAV gene transfer
Ashley T Martino1, Roland W Herzog, Ignacio Anegon
1Department of Pharmaceutical Sciences, St. John's University, Queens, NY, USA.
Methods in Molecular Biology (Clifton, N.J.)
|October 29, 2011
Summary
Immune responses to adeno-associated virus (AAV) vectors hinder gene therapy. This study details assays like ELISPOT and ELISA to detect anti-capsid immunity, crucial for repeated AAV administration and therapeutic success.
Area of Science:
- Gene therapy
- Immunology
- Virology
Background:
- Adeno-associated virus (AAV)-based gene transfer faces challenges from adaptive immune responses against the vector or transgene.
- Immune responses, including neutralizing antibodies against the AAV capsid and cytotoxic T lymphocytes, impede clinical translation and repeated vector administration.
- Preexisting humoral and cellular immunity in humans, from natural infections, can further complicate gene therapy efficacy.
Purpose of the Study:
- To address the critical need for reliable assays to detect and quantify anti-AAV immune responses.
- To provide detailed methodologies for established immunological assays applicable to AAV gene therapy research.
- To enable monitoring of immune responses against both AAV vectors and therapeutic transgene products.
Main Methods:
- Detailed explanation and provision of ELISPOT assays for cellular immunity detection.
- Description of immunocapture (ELISA) assays for antibody quantification.
- Explanation of neutralization assays to assess functional immune responses against AAV vectors.
Main Results:
- Established methodologies for ELISPOT, ELISA, and neutralization assays are presented.
- These assays are demonstrated to be adaptable for monitoring immune responses in both humans and experimental animals.
- The techniques can quantify immune responses against AAV capsids and therapeutic transgene products.
Conclusions:
- Accurate detection and measurement of anti-AAV immune responses are essential for successful gene therapy.
- The provided assay methodologies offer robust tools for preclinical and clinical monitoring.
- These assays facilitate the advancement of AAV-based gene therapies by managing immune-related obstacles.
