Different role of Apaf-1 in positive selection, negative selection and death by neglect in foetal thymic organ culture.
Apoptotic protease-activating factor 1 (Apaf-1) is a component of the apoptosome which is required for the activation of procaspase-9. As Apaf-1 knockout (KO) (Apaf-1-/-) mice die before birth, the role of Apaf-1 during thymic selection was investigated using 5 day foetal thymic organ culture (FTOC) of thymi obtained at gestational day 15. There was a lower ratio of CD4 single-positive (SP) to CD8 SP cells and decreased apoptosis of CD4+CD8+ (DP) thymocytes from Apaf-1-/- mice compared with wild-type. To determine if these defects resulted in increased production of neglected thymocytes, the Apaf-1-/- mice were crossed with the T-cell receptor (TCR)-alpha-chain KO mice. There was no difference in thymocyte development in the thymi of TCR-alpha-/-Apaf-1-/- and TCR-alpha-/-Apaf-1+/+ mice 5 days after FTOC. To determine if Apaf-1 is involved in apoptosis during death by negative or positive selection, FTOC of the thymus of Apaf-1-/- Db/HY TCR-alphabeta transgenic (Tg) mice was carried out. There was decreased apoptosis of the HY clonal-specific M33+ thymocytes and an increased percentage of the autoreactive CD8+M33+ thymocytes in male, but not female Apaf-1-/- Db/HY TCR Tg mice. Our data suggest that Apaf-1 is not involved in positive selection or death by neglect, but may have a partial role in negative selection during early thymic T-cell development.
Department of Immulogy and Parasitology, National Defense Medical College, Saitama, Japan.
Y
Y
Matsuki
H G
HG
Zhang
H C
HC
Hsu
P A
PA
Yang
T
T
Zhou
C H
CH
Dodd
F
F
Cecconi
P
P
Gruss
T
T
Tadakuma
J D
JD
Mountz
D000818:Animals; D017209:Apoptosis / Q000276:immunology; D053477:Apoptotic Protease-Activating Factor 1; D015551:Autoimmunity; D005260:Female; D005333:Fetus / Q000166:cytology / Q000276:immunology; D019673:Genes, T-Cell Receptor alpha; D019674:Genes, T-Cell Receptor beta; D008297:Male; D051379:Mice; D018345:Mice, Knockout; D008822:Mice, Transgenic; D009924:Organ Culture Techniques; D011506:Proteins / Q000235:genetics / Q000276:immunology*; D016176:T-Lymphocyte Subsets / Q000166:cytology / Q000276:immunology; D013950:Thymus Gland / Q000166:cytology / Q000196:embryology* / Q000276:immunology
C506593:Apaf1 protein, mouse; D053477:Apoptotic Protease-Activating Factor 1; D011506:Proteins