Journal article Open Access

Tetramer-Based Enrichment of Preexisting Anti-AAV8 CD8+ T Cells in Human Donors Allows the Detection of a TEMRA Subpopulation

Vandamme Céline; Xicluna Rebecca; Hesnard Leslie; Devaux Marie; Jaulin Nicolas; Guilbaud Mickaël; Le Duff Johanne; Couzinié Célia; Moullier Philippe; Saulquin Xavier; Adjali Oumeya


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        <foaf:name>Adjali Oumeya</foaf:name>
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    <dct:title>Tetramer-Based Enrichment of Preexisting Anti-AAV8 CD8+ T Cells in Human Donors Allows the Detection of a TEMRA Subpopulation</dct:title>
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    <dct:description>&lt;p&gt;&lt;strong&gt;Abstract&lt;/strong&gt;&lt;/p&gt; &lt;p&gt;Pre-existing immunity to AAV capsid may compromise the safety and efficiency of rAAV-mediated gene transfer in patients. Anti-capsid cytotoxic immune responses have proven to be a challenge to characterize because of the scarcity of circulating AAV-specific CD8+&amp;nbsp;T lymphocytes which can seldom be detected with conventional flow cytometry or ELISpot assays. Here, we used fluorescent MHC class I tetramers combined with magnetic enrichment to detect and phenotype AAV8-specific CD8+&amp;nbsp;T cells in human PBMCs without prior amplification. We showed that all healthy individuals tested carried a pool of AAV8-specific CD8+&amp;nbsp;T cells with a CD45RA+CCR7&amp;minus;&amp;nbsp;terminally-differentiated effector memory cell (TEMRA) fraction.&amp;nbsp;&lt;em&gt;Ex vivo&lt;/em&gt;&amp;nbsp;frequencies of total AAV-specific CD8+&amp;nbsp;T cells were not predictive of IFN&amp;gamma; ELISpot responses but interestingly we evidenced a correlation between the proportion of TEMRA&amp;nbsp;cells and IFN&amp;gamma; ELISpot positive responses. TEMRA&amp;nbsp;cells may then play a role in recombinant AAV-mediated cytotoxicity in patients with preexisting immunity. Overall, our results encourage the development of new methods combining increased detection sensitivity of AAV-specific T cells and their poly-functional assessment to better characterize and monitor AAV capsid-specific cellular immune responses in the perspective of rAAV-mediated clinical trials.&lt;/p&gt;</dct:description>
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