ŧŐţ ǀź ƭƹŻƹżĭř ƽŚƷ Żř ƢŤƄƯ ƩƺƬſ ŚƷ ƽ ƾƴưƿř ŲſŚě ƽźǀĭ ŢƸū

ŧŐţ ǀź ƭƹŻƹżĭř ƽŚƷ Żř ƢŤƄƯ ƩƺƬſ ŚƷ ƽ ƾƴưƿř ŲſŚě ƽźǀĭ ŢƸū

T
*
91/1/25 :
90/11/18 :
:
Multiple Sclerosis
.
.
.
TH2
TH17,TH1
TH17 TH1
:
C57BL/6
.
( µg)
(
)
×
.
(MTT)
.
(IFN- ,IL-17 IL-10 TGF- 1)
.
(3-(4, 5-dimethylthiazolyl-2)-2, 5-diphenyltetrazolium bromide
:
IFN- ,IL-17
(p<0.001)
T
TGF- 1 ,IL-10 (p<0.001)
.
:
.
:
TH17 TH1
:
:
Email: [email protected]
I
.
.( )
(Galectins HLA-G TGF- PD-L1)
(IL-10,TGF- )
(
)
-
...
(
PBS
T75
(
)
cell/cm
. (
–Gibco) DMEM-LG
.
(
2
×
FBS
(PGE2 ,IDO,NO)
rpm)
(
)
– Gibco)
CO2
.( - )
.
.
Confluency>70%
(
EDTA
.
/
-Sigma-Aldrich)
T25
.( )
.( - )
MSC
.
PAS flow cytometer , Partec GmbH, )
.
Germany)
.( )
Sca-1-FITC CD73-PE CD29-FITC
(eBioscience)
)
(eBioscience) CD45-FITC
( )
( )
.( )
(
.
BSA
FBS
DMEM
-
Confluency
( )
in vivo
TCD4
Thery
.(% )
g
. (
)
)
g
)
TCD4
(TH17,TH1
g
.
PBS
.
g
PBS
.( )
-
Femur Tibia
( )
2
Phosphate buffered saline
Fetal bovine serum
4
Differential centrifugation
5
Bovine serum albumin
3
1
Exosome
Zappia
C57Bl/6
.
Thery
:
Inhibition
ratio
of
cell
proliferation:
–
(Lymphocyte+MOG+MSC or Exosome)
.
[OD520
OD520(Lymphocute+MOG)]/
OD520(Lymphocute+MOG) - OD520(MSC or Exosome+MOG)] x100% [
( )
– Invitrogen)
/
.
(
-Merck)
mM
.
(
PBS
C57BL/6
g
.
.
.
( )
CD81-
CD9-FITC
(
Pluchino
.
.
MOG35-55 (AnaSpec(
(
) (eBioscience) FITC
µg
– Sigma-Aldrich)
µl
.
ng
(
µl
– Sigma-Aldrich )
.
.
.
(
(Philips BioTwin, CM100, The Netherlands)
.
(
/
.
80Kv
(3-(4, 5-dimethylthiazolyl-2)-2, 5-diphenyltetrazolium bromide) MTT
Gao
.
(A)
.
.
( )
x
) FBS
(B)
%
RPMI
µg
(
.
l
(
µg)
(
( µg/ml)
.
.
(
µg)
( ×
)
)
µl CO2
MTT
MOG35-55
0
C
(
)
.
µl
.
(
(
(
(
(
(0
(
(
(
.
– Sigma-Aldrich)
Shaker
nm
...
General Linear Model
SPSS
(Repeated Measure)
.
P 0.05
.
.
MOG35-55
-
TGF- 1 ,IFN- IL-17 IL-10
(eBioscience)
.(
(% ) Sca-1 (%
)
.
) CD29(%
)
(
) CD73
(
) CD45 (%
(one way
)
.(
.
MINITAB
ANOVA)
)
:( )
.
:( )
C57BL/6
.
CD45
.
CD73 Sca-1 CD29
.
-
.
) CD9 ,CD81
.(
.(
)
)
(
:( )
.
:( )
.
CD9,CD81
.
-
.p 0.001))
.(p 0.05)
MTT
.
...
Exosome))
:( )
MSC))
(Exosome MSC
)
.
(p 0.01)
.(p 0.05)
.
.
.
(Exosome)
:( )
(MSC)
.
(Control)
.(p 0.05)
.( )
TH1
IL-17
TH17
IFNTH17 TH1
.
.( )
TGF-
.
.
IL-10
(
) IFN- (
MS
:( )
) IL-17
.(Exosome)
(MSC)
EAE
in vitro
(
) IFN-
(
) IL-17
MOG35-55
.
TH1,TH17
.
.
.(P 0.01)
(
) TGF- 1 (
:( )
) IL-10
.(Exosome)
(MSC)
TGF-
EAE
in vitro
(
) TGF- 1 (
.
1,IL-10
.
. (p 0.05)
.(p 0.001) (
) IL-10
MOG35-55
)
IL-10 ,TGF- 1
IL-17
(p 0.001)
IFN-
...
T reg
. (
)
TH17 TH1
.(
IL-10
IFN-
.(
)
)
IL-17
.(p 0.05)
TGF- 1
.
.(
) (p 0.05)
Fas- PD-L2 PD-L1 TGFL
.( )
/
.( - )
) IFN-
(HLA-G TGF- Galectins PD-L1)
.
(TH17
) IL-17 (TH1
IL-10 TGF- ,
T
reg
TH1
.
.
T
/
T
.
.
/
.( )
.( )
Ratajczak
In vivo
.
mRNA
.( )
T
B
.( )
B
.( - )
/
.(
.
)
.
.
.
T
MHC II
.(
)
.
.
"
"
in vivo
.
.
(
)
.
.
in vivo
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