Single domain antibodies from llama effectively and specifically block T cell ecto-ADP-ribosyltransferase ART2.2 in vivo

Friedrich Koch-Nolte, Jan Reyelt, Britta Schößow, Nicole Schwarz, Felix Scheuplein, Stefan Rothenburg, Friedrich Haag, Vanina Alzogaray, Ana Cauerhff, Fernando A. Goldbaum

Research output: Contribution to journalArticle

69 Scopus citations

Abstract

The purpose of our study was to develop a tool for blocking the function of a specific leukocyte ecto-enzyme in vivo. ART2.2 is a toxin-related ectoenzyme that transfers the ADP-ribose moiety from NAD onto other cell surface proteins. ART2.2 induces T cell death by activating the cytolytic P2 X 7 purinoceptor via ADP-ribosylation. Here, we report the generation of ART2.2-blocking single domain antibodies from an immunized llama. The variable domain of heavy-chain antibodies (VHH domain) represents the smallest known antigen-binding unit generated by adaptive immune responses. Their long CDR3 endows VHH domains with the extraordinary capacity to extend into and block molecular clefts. Following intravenous injection, the ART2.2-specific VHH domains effectively shut off the enzymatic and cytotoxic activities of ART2.2 in lymphatic organs. This blockade was highly specific (blocking ART2.2 but not the related enzymes ART1 or ART2.1), rapid (within 15 min after injection), and reversible (24 h after injection). Our findings constitute a proof of principle that opens up a new avenue for targeting leukocyte ecto-enzymes in vivo and that can serve as a model also for developing new antidotes against ADP-ribosylating toxins.

Original languageEnglish (US)
Pages (from-to)3490-3498
Number of pages9
JournalFASEB Journal
Volume21
Issue number13
DOIs
StatePublished - Nov 2007
Externally publishedYes

Keywords

  • ADP-ribosylation
  • Enzyme inhibitors
  • Leukocyte ecto-enzymes
  • Recombinant antibodies

ASJC Scopus subject areas

  • Medicine(all)

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    Koch-Nolte, F., Reyelt, J., Schößow, B., Schwarz, N., Scheuplein, F., Rothenburg, S., Haag, F., Alzogaray, V., Cauerhff, A., & Goldbaum, F. A. (2007). Single domain antibodies from llama effectively and specifically block T cell ecto-ADP-ribosyltransferase ART2.2 in vivo. FASEB Journal, 21(13), 3490-3498. https://doi.org/10.1096/fj.07-8661com