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Review
. 2025 Jan 19;26(2):816.
doi: 10.3390/ijms26020816.

Challenges in Humoral Immune Response to Adeno-Associated Viruses Determination

Daria A Naumova  1   2 , Tatyana Krokunova  1   2 , Denis Maksimov  1   2 , Olga N Mityaeva  1   2 , Ekaterina A Astakhova  1   2 , Pavel Yu Volchkov  1   2
Affiliations
Review

Challenges in Humoral Immune Response to Adeno-Associated Viruses Determination

Daria A Naumova et al. Int J Mol Sci. .

Abstract

Adeno-associated viruses (AAVs) are non-pathogenic, replication-deficient viruses that have gained widespread attention for their application as gene therapy vectors. While these vectors offer high transduction efficiency and long-term gene expression, the host immune response poses a significant challenge to their clinical success. This review focuses on the obstacles to evaluating the humoral response to AAVs. We discuss the problems with the validation of in vitro tests and the possible approaches to overcome them. Using published data on neutralizing titers of AAV serotypes, we built the first antigenic maps of AAVs in order to visualize the antigenic relationships between varying serotypes.

Keywords: AAV; NHP models; anti-AAV antibodies; antigenic cartography; humanized models; mice models; neutralizing antibodies.

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Conflict of interest statement

The authors declare no conflicts of interest.

Figures

Figure 1
Figure 1
The present test systems for detecting anti-adeno-associated virus (AAV) neutralizing antibodies (NAbs).
Figure 2
Figure 2
A figure illustrating the importance of considering the number of full and empty capsids in neutralization tests. Serotype A (subfigure (A)) and serotype B (subfigure (B)) take in equal numbers of viral particles, as measured by viral genomes. However, the serotype B sample contains a lower percentage of full capsids than serotype A. Adeno-associated virus (AAV) neutralizing antibodies (NAbs) bind not only full capsids but also empty capsids, so the number of cells transduced by serotype B is higher.
Figure 3
Figure 3
A figure illustrating the importance of the variance in the multiplicity of infections (MOIs) of adeno-associated virus (AAV) serotypes. Chosen doses of serotype A (subfigure (A)) and serotype B (subfigure (B)) transduce an equal number of target cells in the absence of serum antibodies. However, in the presence of an equal volume/number of specific AAV antibodies from serum, the levels of transduced target cells are different, which reflects the lower obtained viral dosage of serotype A, rather than the difference in antigenic characteristics between the two serotypes.
Figure 4
Figure 4
Antigenic map of adeno-associated viruses (AAVs) constructed on published titer neutralization data. AAV serotypes are represented by areas, and sera by circles. Areas considered experimental and the statistical uncertainty of their location. One grid line corresponds to one antigenic unit [59,96,97,98].
See this image and copyright information in PMC

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