title: Transgenic overexpression of toxin-related ecto-ADP-ribosyltransferase ART2.2 sensitizes T cells but not B cells to NAD-induced cell death.
abstract: T cells constitutively express low amounts of a toxin-related ADP-ribosylating ecto-enzyme, ART2.2. In inflammatory settings, cells release NAD, the substrate for ART2.2. The ART2.2 catalyzed ADP-ribosylation of cell surface proteins induces cell death. However, the low expression levels of ART2.2 have hampered analysis of ART2.2 in physiological settings. Here we report the generation of transgenic mice over-expressing ART2.2 under the control of the H2K promoter and Igμ enhancer. ART2.2 transgenic mice were healthy and fertile and exhibited normal development of the major lymphocyte subsets. Most T cells and a small subpopulation of B cells from transgenic mice showed more than 10-fold higher levels of ART2.2 expression than their wild-type counterparts. Exposure of ART2.2-transgenic T cells to low, submicromolar concentrations of NAD caused cell membrane alterations including uptake of propidium iodide, externalization of phosphatidylserine, and shedding of CD62L, while ART2.2-transgenic B cells were resistant to NAD. The ART2.2-overexpressing animals described here confirm that ART2.2 is an essential component for the regulation of T-cell functions by extracellular NAD and provide a useful tool to further elucidate the function of ART2.2 in vivo.
authors.affiliation: Institute of Immunology, University Medical Center, Martinistrasse 52, 20246 Hamburg, Germany.
authors.forename: Peter
authors.initials: P
authors.lastname: Bannas
authors.forename: Felix
authors.initials: F
authors.lastname: Scheuplein
authors.forename: Lennart
authors.initials: L
authors.lastname: Well
authors.forename: Irm
authors.initials: I
authors.lastname: Hermans-Borgmeyer
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 / Q000276:immunology / Q000378:metabolism*; D000818:Animals; D001402:B-Lymphocytes / Q000276:immunology / Q000378:metabolism*; D015139:Blotting, Southern; D016923:Cell Death / Q000276:immunology; D002469:Cell Separation; D005434:Flow Cytometry; D015870:Gene Expression; D005786:Gene Expression Regulation / Q000276:immunology*; D006183:H-2 Antigens / Q000235:genetics; D016130:Immunophenotyping; D016131:Lymphocyte Subsets / Q000276:immunology* / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D008822:Mice, Transgenic / Q000276:immunology* / Q000378:metabolism; D009243:NAD / Q000378:metabolism; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic; D013601:T-Lymphocytes / Q000276:immunology / Q000378:metabolism*
chemical_list: D006183:H-2 Antigens; C024755:H-2K(K) antigen; D009243:NAD; D036002:ADP Ribose Transferases; C449339:Art2b protein, mouse