title: Technical Advance: a new cell preparation strategy that greatly improves the yield of vital and functional Tregs and NKT cells.
abstract: Release of NAD(+) during preparation of murine lymphocytes causes enzymatic ADP-ribosylation of cell-surface proteins on T cells, catalyzed by toxin-related ecto-ADP-ribosyltransferase, ARTC2. ADP-riboslyation activates the cytolytic P2X7 ion channel and affects, in particular, the vitality and function of Tregs and NKT cells. Here, we describe a simple method-injection of an ARTC2-blocking nanobody-to greatly improve Treg and NKT cell vitality and to preserve their function during in vitro assays and in adoptive-transfer experiments. Moreover, we present a method for the sorting of functional, primary NKT cells, based on coexpression of ARTC2 and NK1.1. Our results pave the way for the efficient ex vivo proliferation of Tregs and NKT cells and for new experimental and therapeutic uses of these important regulatory cells.
authors.affiliation: 1.Institute of Immunology, Martinistrasse 52, 20246 Hamburg, Germany. nolte@uke.de.
authors.forename: Björn
authors.initials: B
authors.lastname: Rissiek
authors.forename: Welbeck
authors.initials: W
authors.lastname: Danquah
authors.forename: Friedrich
authors.initials: F
authors.lastname: Haag
authors.forename: Friedrich
authors.initials: F
authors.lastname: Koch-Nolte
mesh_terms: D036002:ADP Ribose Transferases / Q000037:antagonists & inhibitors; D019264:Adoptive Transfer / Q000379:methods; D000818:Animals; D019138:Antibodies, Blocking; D002469:Cell Separation / Q000379:methods*; D002470:Cell Survival; D005434:Flow Cytometry / Q000379:methods*; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D055611:Natural Killer T-Cells / Q000737:chemistry*; D061905:Single-Domain Antibodies; D050378:T-Lymphocytes, Regulatory / Q000166:cytology*
chemical_list: D019138:Antibodies, Blocking; D061905:Single-Domain Antibodies; D036002:ADP Ribose Transferases
keywords: ARTC2; NAD+; cell preparation; natural killer T cells; regulatory T cells