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ORIGINAL RESEARCH article

Front. Immunol.
Sec. T Cell Biology
Volume 15 - 2024 | doi: 10.3389/fimmu.2024.1450524

Polyfunctional T cells and unique cytokine clusters imprint the anti rAAV2/rAAV9 vector immune response

Provisionally accepted
Stephan Jonas Holtkamp Stephan Jonas Holtkamp 1*Florian R. Lagoda Florian R. Lagoda 1Adam Lister Adam Lister 2Pradeep Harish Pradeep Harish 2Ulrike Kleymann Ulrike Kleymann 1Theresa Pesch Theresa Pesch 1Chai Fen Chai Fen 1Munir Pirmohamed Munir Pirmohamed 2,3Dean Naisbitt Dean Naisbitt 2,3Mark Trautwein Mark Trautwein 1
  • 1 Bayer (Germany), Wuppertal, Germany
  • 2 Institute of Integrative Biology, University of Liverpool, Liverpool, North West England, United Kingdom
  • 3 Wolfson Centre for Personalised Medicine, Institute of Translational Medicine, University of Liverpool, Liverpool, North West England, United Kingdom

The final, formatted version of the article will be published soon.

    Polyfunctional T cells programmed to perform activities such as degranulation of lytic enzymes and simultaneous production of multiple cytokines are associated with more effective control of viral infections. Immune responses to recombinant adeno-associated virus (rAAV) vector delivery systems can critically influence therapeutic efficacy and safety of gene therapy. However, knowledge of polyfunctional T cells in anti-AAV immune responses is scarce. To bridge this knowledge gap, we have investigated the polyfunctionality of primary human CD4 T cells from healthy donors after in-vitro exposure to rAAV2 or rAAV9 vectors. By performing proliferation assays of co-cultured T cells and rAAV pulsed monocyte-derived dendritic cells from healthy donors we demonstrate T cell reactivity of 43% and 50% to rAAV2 and rAAV9 vectors, respectively. We validated this frequency in a second screen using another set of healthy donors measuring CD25 and CD71 T cell activation. Single T cell secretome analysis of reactive donors uncovered a Th1 pro-inflammatory, cytolytic and chemoattractive cytokine release profile after stimulation with rAAV2 or rAAV9 vectors. 12.4% and 9.6% of the stimulated T cells displayed a polyfunctional cytokine response, respectively, including elevated polyfunctional inflammatory indices. These responses were characterized by cytokine clusters such as Granzyme B, MIP1-α and TNF-α released in combination by single T cells. Overall, our results provide insights into adaptive immunity with rAAV vector serotypes which will be important in advancing gene therapy safety, vector selection, immunogenicity assessment and better patient selection for AAV gene therapy.

    Keywords: adeno-associated virus, Immunogenicity, CD4 T cell immunity, Gene Therapy, polyfunctional T cells, T cell secretome

    Received: 17 Jun 2024; Accepted: 04 Nov 2024.

    Copyright: © 2024 Holtkamp, Lagoda, Lister, Harish, Kleymann, Pesch, Fen, Pirmohamed, Naisbitt and Trautwein. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

    * Correspondence: Stephan Jonas Holtkamp, Bayer (Germany), Wuppertal, Germany

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