cellbased anticancer vaccines trough glycan engineering T Ferro 15 M Silva 1 Z Silva 15 G Marques 1 H Crespo 1 H Soares 1 S van Vliet 2 A Fernandes ID: 796779
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Slide1
Improved
dendritic
cell-based anti-cancer vaccines trough glycan engineering
T Ferro1,5, M Silva1, Z Silva1,5, G Marques1, H Crespo1, H Soares1, S van Vliet2, A Fernandes3,5, Y van Kooyk2, T Matos4, J A Ferreira6,7, P Videira1,5
Introduction
www.ucibio.pt
Lack of sialic acid induces superior anti-
tumoral immune responses
Deciphering
molecular aspects: the role of MHC-I in antigen presentation
Conclusions
Maturation
S
ignals
Higher
levels
of
sialylation
Immature
DC
Lower
levels
ofsialylation
Mature DC
Dendritic cells (DCs) are the most potent antigen presenting cellsOriginating either from the lymphoid or myeloid lineage, DCs interact and partipate in both innate and adaptive immune responses
Sialic
acid
removal
improves co-stimulatory molecules expression
Increased secretion of cytokines
T cells primed by desialylated DCsshow higher degranulation
Author affiliations: 1. CEDOC, NOVA Medical School / Faculdade de Ciências Médicas, Universidade NOVA de Lisboa, Portugal 2. Department of Molecular Cell Biology and Immunology, VU University Medical Center, Amsterdam, The Netherlands 3. CQE, Centro de Química Estrutural, Instituto Superior Técnico, ULisboa, Portugal 4. StemLab, Cantanhede, Portugal 5. UCIBIO, Departamento Ciências da Vida, Faculdade de Ciências e Tecnologia, Universidade NOVA de Lisboa, Portugal 6. QOPNA, Mass Spectrometry Center, Department of Chemistry, University of Aveiro, Aveiro, Portugal 7. Experimental Pathology and Therapeutics Group, Portuguese Institute of Oncology, Porto, Portugal
DC
maturation
induces loss of sialic acidMaturation can be also achieved by sialic acid shortage
Dendritic cells show increased expression of MHC-I after desialylation
MHC-I
is glicosylated and sialylated: target for sialidase enzymes
Cell
lysates were treated with sialidase and probed for MHC-I by Western Blot.MHC-I shows decreased molecular weight upon desialylation.
Desialylation
improves MHC-I stability on cell surface
CMVpp65
peptide matches HLA-A02*01 pocket.
MHC-I is more stable after sialidase treatment. Extrinsic sialylation by ST6Gal1 counteracts this effect. Control cells in black.
A MHC-I decay assay was performed on T2 cells following sialidase treatment.MHC-I is stable on the membrane for up to 3 hours after sialidase treatment, when Golgi export is blocked by brefeldin A.
Sialic acid as a key modulator of DCs biology
- Immune response is dependent of sialic acid content at cell surface
- Desialylated human DCs show increased hability to activate T cells and promote cytotoxic responses
- MHC-I as a molecular effector of anti-tumoral responses
This work was supported by the Applied Molecular Biosciences Unit - UCIBIO which is financed by national funds from FCT/MEC (UID/Multi/04378/2013) and co-financed by the ERDF under the PT2020 Partnership Agreement (POCI-01-0145-FEDER-007728).
DC
contains
high
levels of the monossacharide sialic acid at the termini of glycans of certain cell surface glycoproteins
Asn
Maturation
T
H
1 phenotype
T cell activationTH1 phenotypeImproved IFN-γsecretion
DCs
Tumor
lysates
DCs
DCs
Sialidase
treated + Tumor lysates
T +
DCs
Tumor
lysates
T
T +
DCs
Sialidase
treated
+ Tumor lysates
T +
DCs
DCs
were
pulsed with MCF-7 lysatesCytotoxic activity was evaluated by tumor cell viability following interaction with primed T cellsDegranulation evaluated by CD107 expression
T +
DCs
Tumor
lysates
T +
DCs
Sialidase treated + Tumor lysates
Glycoprotein
Sialic
acid
N-
acetyl
glucosamine
Galactose
Manose
Fucose
MHC-I
SNA
α
2,6 sialic
acid
binding
lectin
To
understand
the
molecular
mechanisms
behind
this
phenomena
, sialylated
proteins
on
DCs
surface
were
identified
by
a
sialic
acid
binding
lectin (
SNA)
pull
down
,
followed
by
mass
spectrometry
.
The MHC-I was identified and hypothesised as a possible candidate to be modulated by sialic acid content.
In
fact
,
sialidase
treated
human
DCs
show
higher
ability
to
activate
anti-tumor
activity
on
T
cells
and
increased
expression
of
MHC-I.
T2
cell
line
: experimental
model
to
study
MHC-I (HLA-A02*01)
turnover
upon
desialylation
References
: 1.
Oncotarget
. 2016; 7:41053-41066.
doi
:
10.18632/oncotarget.9419
2.
Patent
PCT/IB2016/053901
(
A VIABLE CELL POPULATION, METHOD FOR PRODUCTION AND
USES THEREOF)
- Sialic
acid
may
influence
MHC-I turnover
at
cell
surface