studi di immunogenicità in modelli murini laura pisapia giovanna del pozzo napoli, 27 giugno 2013
TRANSCRIPT
Studi di immunogenicità in modelli murini
Laura Pisapia
Giovanna Del Pozzo
Napoli, 27 Giugno 2013
Corso C. et al. EBP1 and DRBP76/NF90 binding proteins are included in the Major Histocompatibility Complex Class II RNA operon. 2011 N.A.R Sep 1;39(16):7263-75.
Pisapia L. et al. Co-regulated expression of alpha and beta mRNAs encoding HLA-DR surface heterodimers is mediated by the MHCII RNA operon. 2013 N.A.R. Apr1;41(6):3772-86
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MHC class II RNA Operon
EBP1
Identified as ErbB3 binding protein, is
the human homologue of mouse cell-cycle regulated
protein p38-2G4
In carcinoma cell lines p48 isoform functions as tumor
suppressor
In PC12 and glioblastoma p48 works as oncogene, while p42 isoform is a tumor suppressor
Translational control:• in the cytoplasm is associated to
80S ribosomes and polysomes• inhibits translation of AR mRNA
Transcriptional repressor:Inhibits transcription of E2F regulated promoters
by recruiting HDAC2/Rb/Sin3A complex
RNA binding protein:• influences bcl2 stability,
promotes decay of androgen receptor mRNA,• binds UTRs of MHCII
p48 and p42 isoforms
• p48 isoform binds 5’ and 3’UTR of MHCII mRNAs
• The over expression of p48 isoform determines an up- regulation of MHC Class II in different cancer cell lines
Ctrl EBP1 p48 EBP1 p42
0
4
8
12
16M14
HLA
-DR
MFI fo
ld v
ari
ati
on
Ctrl EBP1 p48 EBP1 p42
0
4
8
12
16U87
HLA
-DR
MFI fo
ld v
ari
ati
on
Ctrl EBP1 p48 EP1 p42
0
1
2
3
MCF7
HLA
-DR
MFI fo
ld v
ari
ati
on
Ctrl EBP1 p48 EP1 p42
0
1
2
3
B16 OVAH
LA-D
R M
FI f
old
va
ria
tio
n
Over-expression of p42 and p48 EBP1 influences MHCII amount
OT-II MOUSEOT-I MOUSE
Spleen Spleen
T CD4+ magnetic selectionT CD8+ magnetic selection
B16-OVAa and CD8+ co-culture
B16-OVA and CD4+ co-culture
Melanoma B16-OVACD8+
T cell
CD4+ T cell
CD8 OVA257–264
(SIINFEKL) peptide
CD4 OVA323–339
(ISQAVHAAHAEINEAGR) peptide
CD8+ CFSE labelling
CD4+ CFSE labelling
Ctrl EBP1 p48 EBP1 p42
0
0.5
1
T CD8+ proliferation assay
CFS
E M
FI
fold
vari
ati
on
Ctrl EBP1 p48 EBP1 p42
0
0.5
1
T CD4+ proliferation assay
CFS
E M
FI
fold
vari
ati
on
T cells activation and proliferation are
demonstrated by the decrease of CFSE labelling
CFSE