HAL Id: inserm-00375651
https://www.hal.inserm.fr/inserm-00375651
Submitted on 5 May 2009HAL is a multi-disciplinary open access archive for the deposit and dissemination of sci-entific research documents, whether they are pub-lished or not. The documents may come from teaching and research institutions in France or abroad, or from public or private research centers.
L’archive ouverte pluridisciplinaire HAL, est destinée au dépôt et à la diffusion de documents scientifiques de niveau recherche, publiés ou non, émanant des établissements d’enseignement et de recherche français ou étrangers, des laboratoires publics ou privés.
activation-induced cell death.
Lior Mayo, Jasmine Jacob-Hirsch, Ninette Amariglio, Gideon Rechavi,
Marie-Jo Moutin, Frances Lund, Reuven Stein
To cite this version:
Lior Mayo, Jasmine Jacob-Hirsch, Ninette Amariglio, Gideon Rechavi, Marie-Jo Moutin, et al.. Dual role of CD38 in microglial activation and activation-induced cell death.. Journal of Immunology, Publisher : Baltimore : Williams & Wilkins, c1950-. Latest Publisher : Bethesda, MD : American Association of Immunologists, 2008, 181 (1), pp.92-103. �inserm-00375651�
Fig 1.
A
0 5 10 15 20 25 0 30 60 90 120*
*
*
*
*
AMD (ng/ml) % SurvivalB
0 30 60 90 120 0 50 100 CHX (ng/ml)*
*
% SurvivalA
B
0.2 0.4 0 0.6 0.81 1.2 Control LPS IFN g LPS/IFN g (1) (0.4) (0) (0.4) Cas-11 A.U. 0.2 0.4 0 0.6 0.81 1.2 Control LPS IFN g LPS/IFN g (1) (0) (0) (0.1) Nos2 A.U. CD38 Control LPS IFN g LPS/IFN g 1 2 3 4 0 (1) (1.3) (4.3) (1.8) A.U. IFNg LPS/IFNg Control LPS PE-CD38 CountsFig 3
A
PE-CD38 Counts WT Cd38-/-B
LPS/IFNg Control CD38 expression (A.U.) 0 1 2*
0 10 20 30 100 101 102 103 104 0 10 20 30 100 101 102 103 104Time (h) 40 20 0 60 Con 24 48 72
*
*
Cd38-/-WT% Cell death (LDH release)
50 30 10 0 48 24 40 20 60 72 Time (h) Cd38-/-WT
*
*
% Survival (MTT assay)A
B
80 100 120 ConFig. 5
Ba/F3 CD38neg Ba/F3 CD38exp
B
10 30 40 50 0 20% Cell death (LDH release)
IgG aCD38 (90) aCD38 (NIM-R5)
-
+ + + + + + + + + +
Secodary Ab F(ab)2 LPS/IFNg Control 0 0 1 5 25 1 5 25 Primary Ab (mg/ml) 1 5 25A
% Survival (MTT assay)* *
Isotype control (IgG)
aCD38 (90) aCD38 (NIM-R5)
80 60 40 20 0 100 Mock
Ba/F3 CD38neg Ba/F3 CD38exp
PE-CD38 Counts
D
C
Fluorescence (420nM) LPS/IFNg Control*
0 250 500 750 1000 Cd38-/-WT LPS/IFNg Control*
*
0 2 4 6 cADPR (pmol/mg)LPS/IFNg 8-Br-cADPR N(8-Br-A)D+
% Cell death (LDH release)
-+
+
+
--
+
*
+
- Cd38-/-WT 50 40 30 20 0* * *
Ryanodine-
-
-+
-+
-10Fig 7
0 1 2 3 SNAP (mM) 0 20 40 60E
Cd38-/-WT% Cell death (LDH release)
A
iNOS Caspase-11*
0 0.4 0.8 1.2 A.U. Cd38-/-WT iNOS Caspase-11 b-tubulin Cd38-/-WT LPS/IFNg - + - + Time (h) 80 Nitrite (mM / MTT) 60 40 20 0 60 40 20 80*
*
*
WT control Cd38-/- control WT LPS/IFNg Cd38-/- LPS/IFNgB
D
40 80 100 120 0 60 0 0 1 5 25 1 5 25 Primary Ab (mg/ml) 1 5 25-
+ + + + + + + + + +
Secodary Ab F(ab)2 LPS/IFNg Control 20 % NitriteIgG aCD38 (90) aCD38 (NIM-R5) LPS/IFNg 8-Br-cADPR N(8-Br-A)D+ + -+ -+ + -+ -+ 8 0 2 4 6
Nitrite production (fold increase)
Cd38-/-WT
* * * *
+ -Ryanodine - - - +-C
50 100 200 [Ca 2+ ]i (nM) LPS/IFNg 8-Br-cADPR - + -- + -+ + - Cd38-/-WT 0 150
*
#*
#Fig 9
C
0 25 50 75 100 % IL-12 secretion*
Cd38-/-WT*
* *
N(8-Br-A)D+ LPS/IFNg 8-Br-cADPR-
-+
+
+
--
+
+
-Ryanodine-
-
-+
-+
-N(8-Br-A)D+ LPS/IFNg 8-Br-cADPR-
-Ryanodine -0 25 50 75 100 %TNF a secretion+
-+
+
-+
-+
-+
-+
-*
Cd38-/-WT*
*
*
40 80 120 0 0 0 1 5 25 1 5 25 Primary Ab (mg/ml) 1 5 25 - + + + + + + + + + + Secodary Ab F(ab)2 IgG aCD38(clone 90) (NIM-R5)aCD38
% IL-12 secretion LPS/IFNg Control
D
40 80 120 0 0 0 1 5 25 1 5 25 Primary Ab (mg/ml) 1 5 25 - + + + + + + + + + + Secodary Ab F(ab)2 IgG aCD38(clone 90) (NIM-R5)aCD38
% TNF a secretion LPS/IFNg Control LPS/IFNg 8-Br-cADPR + -- + -+ + - - Cd38-/-WT
