studi di immunogenicità in modelli murini laura pisapia giovanna del pozzo
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Studi di immunogenicità in modelli murini Laura Pisapia Giovanna Del Pozzo. Napoli, 27 Giugno 2013. - PowerPoint PPT PresentationTRANSCRIPT
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 p420
4
8
12
16 M14
HLA
-DR
MFI
fold
var
iatio
n
Ctrl EBP1 p48 EBP1 p420
4
8
12
16U87
HLA
-DR
MFI
fold
var
iatio
n
Ctrl EBP1 p48 EP1 p420
1
2
3MCF7
HLA
-DR
MFI
fold
var
iatio
n
Ctrl EBP1 p48 EP1 p420
1
2
3
B16 OVAHL
A-DR
MFI
fold
var
iatio
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 p420
0.5
1
T CD8+ proliferation assay
CFSE
MFI
fold
var
iati
on
Ctrl EBP1 p48 EBP1 p420
0.5
1
T CD4+ proliferation assay
CFSE
MFI
fold
var
iati
on
T cells activation and proliferation are
demonstrated by the decrease of CFSE labelling
CFSE