Rapid induction of naive T cell apoptosis by ecto-nicotinamide adenine dinucleotide: requirement for mono(ADP-ribosyl)transferase 2 and a downstream effector.
Lymphocytes express a number of NAD-metabolizing ectoenzymes, including mono(ADP-ribosyl)transferases (ART) and ADP ribosylcyclases. These enzymes may regulate lymphocyte functions following the release of NAD in injured or inflammatory tissues We report here that extracellular NAD induces apoptosis in BALB/c splenic T cells with an IC(50) of 3-5 microM. Annexin V staining of cells was observed already 10 min after treatment with NAD in the absence of any additional signal. Removal of GPI-anchored cell surface proteins by phosphatidylinositol-specific phospholipase C treatment rendered cells resistant to NAD-mediated apoptosis. RT-PCR analyses revealed that resting BALB/c T cells expressed the genes for GPI-anchored ART2.1 and ART2.2 but not ART1. ART2-specific antisera blocked radiolabeling of cell surface proteins with both [(32)P]NAD and NAD-mediated apoptosis. Further analyses revealed that natural knockout mice for Art2.a (C57BL/6) or Art2.b (NZW) were resistant to NAD-mediated apoptosis. Labeling with [(32)P]NAD revealed strong cell surface ART activity on T cells of C57BL/6 and little if any activity on cells of NZW mice. T cells of (C57BL/6 x NZW)F(1) animals showed strong cell surface ART activity and were very sensitive to NAD-induced apoptosis. As in BALB/c T cells, ART2-specific antisera blocked cell surface ART activity and apoptosis in (C57BL/6 x NZW)F(1) T cells. The fact that T cells of F(1) animals are sensitive to rapid NAD-induced apoptosis suggests that this effect requires the complementation of (at least) two genetic components. We propose that one of these is cell surface ART2.2 activity (defective in the NZW parent), the other a downstream effector of ADP-ribosylation (defective in the C57BL/6 parent).
Adriouch
S
S
Laboratoire d'Immunodifferenciation, Université Denis-Diderot Paris 7, Paris, France.
Ohlrogge
W
W
Haag
F
F
Koch-Nolte
F
F
Seman
M
M
D036002:ADP Ribose Transferases; D000246:Adenosine Diphosphate Ribose / Q000378:metabolism; D000818:Animals; D000945:Antigens, Differentiation, T-Lymphocyte; D017209:Apoptosis / Q000187:drug effects*; D005110:Extracellular Space / Q000502:physiology; D005260:Female; D017261:Glycosylphosphatidylinositols / Q000378:metabolism / Q000502:physiology; D006649:Histocompatibility Antigens / Q000096:biosynthesis / Q000502:physiology*; D007166:Immunosuppressive Agents / Q000494:pharmacology; D007399:Interphase / Q000276:immunology; D007527:Isoenzymes / Q000096:biosynthesis / Q000502:physiology; D008213:Lymphocyte Activation; D008297:Male; D008562:Membrane Glycoproteins; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D008814:Mice, Inbred NZB; D018345:Mice, Knockout; D009243:NAD / Q000502:physiology*; D011065:Poly(ADP-ribose) Polymerases / Q000096:biosynthesis / Q000502:physiology*; D011817:Rabbits; D051381:Rats; D013601:T-Lymphocytes / Q000166:cytology* / Q000201:enzymology* / Q000276:immunology
D000945:Antigens, Differentiation, T-Lymphocyte; D017261:Glycosylphosphatidylinositols; D006649:Histocompatibility Antigens; D007166:Immunosuppressive Agents; D007527:Isoenzymes; D008562:Membrane Glycoproteins; D009243:NAD; D000246:Adenosine Diphosphate Ribose; D036002:ADP Ribose Transferases; D011065:Poly(ADP-ribose) Polymerases; C449322:Art2a protein, mouse; C449339:Art2b protein, mouse; C090163:Art2b protein, rat