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Published on: December 7, 2021
Analyzing cellular immunity to AAV in a canine model using ELISPOT assay.
Zejing Wang1, Rainer Storb, Stephen J Tapscott
1Program in Transplantation Biology, Division of Clinical Research, Fred Hutchinson Cancer Research Center, Seattle, WA, USA. zwang@fhcrc.org
Methods in Molecular Biology (Clifton, N.J.)
|September 30, 2011
Summary
Researchers developed a canine-specific ELISPOT assay to detect immune responses to adeno-associated viral (AAV) vectors. This method identifies T-cell epitopes in dogs receiving AAV gene therapy, aiding future treatments.
Area of Science:
- Biotechnology
- Immunology
- Gene Therapy
Background:
- Adeno-associated viral (AAV) vectors are promising for gene replacement therapy.
- Host immune responses to AAV vectors can hinder in vivo gene delivery.
- Cellular immunity against AAV capsid proteins is observed in animal models and human trials.
Purpose of the Study:
- To develop and detail a canine-specific ELISPOT assay for detecting T-cell immunity to AAV vectors.
- To identify T-cell epitopes in dogs treated with AAV via intramuscular injection.
Main Methods:
- Development of a canine-specific ELISPOT assay.
- Utilized a panel of overlapping peptides spanning the AAV VP1 capsid protein sequence.
- Assayed peripheral blood T-cell responses post-AAV intramuscular injection.
Main Results:
- The assay successfully detected specific T-cell responses in dogs treated with AAV.
- Identified T-cell epitopes within the AAV capsid protein.
Conclusions:
- The developed ELISPOT assay is a high-throughput method for detecting canine T-cell immunity to AAV.
- This assay facilitates epitope identification without requiring large cell numbers or MHC-matched cell lines.
- The method supports the advancement of AAV gene therapy by monitoring immune responses.
