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My next paper

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on 6 June 2016

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Transcript of My next paper

Primary lymphoid organs
Tertiary Lymphoid Structures
My next paper !?!
Lupus nephritis
SLE
The Pathogenesis of Lupus Nephritis
Deposition of autoantibodies
Tertiary lymphoid Structure
Secondary lymphoid organs
Lymphatic system
Aim
LTi
LTo
Which cells play LTo cells role in TLS development?
... "Drawing on studies of
lymph nodes
, Peyer’s patches and nasal-associated lymphoid tissue, an integrative model of lymphoid-tissue development, involving adhesion molecules, cytokines and chemokines, which emphasizes the role of interactions between
CD3–CD4+CD45+ ‘inducer’ cells
and
VCAM1+ICAM1+ stromal ‘organizer’ cells
is presented." ...
... ?
... "
Lymphoid tissue–inducer cells
accumulate in the developing lymph node, forming small clusters with
resident stromal organizer cells
, to initiate a cascade of intracellular and intercellular events that lead to the maturation of the primordial lymph node." ...
Immunomodulator
Immunostimulatory
Design of Experiments
Mesenchymal Stem Cells stimulation
Stimulating MSCs with different
proinflammatory
cytokines at different concentrations for
18h
IL-1B TNF-a IFN-a IFN-y IL-10 IL-6



CXCL12
CXCL13
CCR5
CCR7
RANKL
TNF-a
IL-1B
VCAM1
ICAM1
CCL19
CCL21
PDPN
LTBr
LTB
IL-4
IL-7
Genes involved in TLS development
T cells
&
MSCs
co-culture
co-culture stimulated MSCs with
T cells from healthy doner

1:1 & 1:100 ratio
PBMCs
&
MSCs
co-culture
Optimize Starvation
3% FBS

18h
before and
0.1% FBS
at time of stimulation

0% FBS

4h
before and
normal FBS
at time of stimulation

Normal

FBS
(7.3%)
all the
time
Time course
MSCs Stimulated with
IL-1B & TNF-a 8 ng/ml

1h 3h 6h 12h 24h 48h 72h
Cytokines selection
MSCs Stimulated with diffrent cytokines combinations
2ng 4ng 8ng 16ng 20ng 25ng 35ng
IL-1B TNF-a IFN-a IFN-y 8ng/ml

IL-1B TNF-a 8ng/ml
18h
IL-1B TNF-a IFN-a IFN-y 8ng/ml
18h
Next steps
co-culture stimulated MSCs with
PBMCs from healthy doner

1:1 & 1:100 ratio
co-culture MSCs & T cells in the transwell system

T cells differentiation after co-culturing

Genes of interest expression in MSCs & T cells after co-culturing

Perform all experiments with immune cells from SLE patients (ten patients)

Locating MSCs in TLS in Lupu-prone mice kidney
KF
SED
PK
VCAM1
ICAM1
CXCL12
CXCL13
Evaluated Genes
CCL19
CCL21
IL-4
IL-7
IL-1B
TNF-a
TLS group
More group members are loading
Tanks !

CXCL12
CXCL13
CCR5
CCR7
RANKL
TNF-a
IL-1B
VCAM1
ICAM1
CCL19
CCL21
PDPN
LTBr
LTB
IL-4
IL-7
Thought line
MSCs contribute to
lymphoid organs development

MSCs are
immunomodulator

MSCs can be
immunostimulator
in specific condition


Low population ratio
compares to immune cells


Optical Density
Optical Density
Optical Density
We defined:

Optimal pro-inflammatory cytokines combination
Optimal starvation setup
Optimal cytokines concentration
Optimal time point

We showed:

PBMCs & T-cells proliferation increase within co-culture with stimulated MSCs at high cell ratio.
Concluding Remarks
(MSCs:PBMCs/T-cells 1:100)
Full transcript