The regulatory outcome of cross-talk between regulatory B cells … · 2019. 2. 26. · Plasma Cell...
Transcript of The regulatory outcome of cross-talk between regulatory B cells … · 2019. 2. 26. · Plasma Cell...
Claudia Mauri
University College London
United Kingdom
The regulatory outcome of cross-talk between regulatory B cells and invariant
natural killer T cells
PlasmaCell
B cell
Pathogenicantib
odi es
“Normal”antib
odi es
PlasmaCell
B cell
T cell
MHCIIpeptide
iNKTcell
CD1diTCR lipid
B cellB cell
iNKTcell
T cell
TCR
Antigen presentation Antibody production
B cells play a pleiotropic role in the immune system
B cell subsets
Bregs
immunosuppression
Francis Lund et al. Nature Immunol 2000
Defense against parasites
Contribute to allergies and asthma
Defense against intracellular pathogens
Contribution in the pathogenesis of autoimmune diseases
Naïve B cells
B effector 1
IFNg Th1
B effector 2
IL-4
Th2
Naïve B cells
B cells can suppress inflammation
Mice lacking IL-10 producing B cells develop an exacerbated arthritis
IL-10-/- B cell mice: lower Treg numbers but higher Th17 and Th1 cells than WT mice
A
100 101 102 103 104100
101
102
103
104
100 101 102 103 104100
101
102
103
104
CD
4
FoxP3
WT B cells IL-10-/- B cells 8.75% 3.94%
Fo
xP
3
WT B cells
100 101 102 103 104100
101
102
103
104
100 101 102 103 104100
101
102
103
104
0.42% 0.16%
CD4
CD4+CD25-
IL-10-/- B cells
Synovia draining LN
Janus face at the Vatican (Rome) Habemus papam!
CD19
CD24hi
CD21hi
CD23hi
AA4+/int
CD1dhi
IgMhi
CD21hi
CD5
CD1dhi
CD24hi
IgMhi
Yan
ab
a K
. 20
08
(B1
0) Ev
ans
J. 2
007
T2-M
ZP
Markers for Bregs
immature memory
mature
;
Sims, G. P. et al. Blood; 2005
Anolik J. et al. Arthritis and Rheum.2007 Carsetti R. Immunol. Rev. 2004 Blair P. et al Immunity. 2010
B cell subsets in periphereal blood mononuclear cells
CHO-CD154
Blair et al. Immunity 2010
Memory
mature
Immature
IL-10 producing B cells are enriched within the CD24hiCD38hi gate
Blair et al Immunity 2010;32(1):129-40. 2010
Flores-Borja et al. Science Trans Med Feb 20;5(173):2013
CD19+CD24hiCD38hiB cells from healthy individuals suppress T helper cell differentiation
T cells are converted into FOXP3+T regs
FoxP
3
Bregs induce regulatory T cells
Flores Borja et al. (Science Trans Med. 2013)
CD4+ +
-
+ +
-
+
++
cp
m (
10
3)
CD25+
CD127-
0
25
50
75
100
125 *
Healthy
8.78.4
100
101
102
103
104
1.4 11.4H
4
no B cells
CD4+ + anti-CD3
CD4+ CD24intCD38int CD24hiCD38hi
4
6
8
10
0
2
CD24int
CD38int
CD24hi
CD38hi
CD
4+F
oxP
3+ (
%)
**
-
+
- +
- -
CD4
Treg
Beff
Th17
Th1Breg
Treg
Breg
Autoantibodies IL-17, IFNg, TNFa
BeffTh17
Th1Breg
Treg
Healthy Autoimmunity
Systemic lupus erythematosus (SLE)
• Systemic autoimmune disease affects approximately
0.1% of overall population
• Extremely diverse with manifestations in major organ
systems
• Treatments range from steroids to cell-targeted
treatments, such as Rituximab, a chimeric monoclonal
antibody targeting and depleting CD20 expressing B cells
HEALTHY
SLE
Blair et al Immunity 2010;32(1):129-40.
CD19+CD24hiCD38hi B cells from SLE patients fail to
suppress IFNg+ /TNFa+CD4+ T cell differentiation
• Immature B cells isolated from SLE are not
numerically different from healthy individuals
• Effector T cells from SLE are not refractory to
suppression
• Intrinsic defect in Bregs in SLE patients
IL-10 production in SLE patients is reduced on immature B cells, but not on other B cell subsets
Paul Blair et al Immunity 2010
B cell IL-10 production
Uns
timulat
ed
anti CD3
0
1
2
3
4
5Healthy controls
SLEIL
-10
%
healthy
SLE
Paul Blair Madhvi Menon
%im
mat
ure
B c
ell
IL-1
0+
CHO-CD40L medium
CD1d expression in SLE patients is reduced on immature B cells, but not on other B cell subsets
Bosma et al Immunity 2012
CD1d
isotype
mature
memory
immature
but not on monocytes
B cells Monocytes
Healthy
SLE
Healthy
SLE
CD1d
% o
f M
ax
Increased CD1d internalisation-rate leads to the reduction of CD1d expression on B cells from SLE
patients
He
alt
hy
S
LE
0 min 5 min
CD1d DAPI
CD1d DAPI
CD1d DAPI
CD1d DAPI
CD1d-lipid antigen presentation to iNKT cell
iNKT cells are important for B cell antibody production and memory
What is the impact of the reduction of CD1d expression in SLE B cells in iNKT cell differentiation and function
iNKT cells are reduced in number in SLE patients
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iNK
T c
ells
(x10
6/L
ite
r)
0
1
2
3
4
51015
Hea
lthy
SLE
CD1d
isotype
mature
memory
immature
Healthy, and not SLE, iNKT cell proliferate in response to a-Gal-Cer stimulation
Bosma et al Immunity 2012
Day0 Day 7 Day0 Day 7
HEALTHY SLE
IL-2
(ng
/ml)
IFN
-g (
ng/m
l)
TN
F-a
(p
g/m
l)
IL-1
0 (
pg
/ml)
0
2
4
6
0
2
4
6
* **
*
0
50
100
150
0
10
20
30
40
50
H SLE H SLE H SLE
IFN-g (ng/ml)0 5 10 15
TNF-a (pg/ml)0 10 20 30 40
***
Healthy PBMC-B cells + Allogeneic healthy B cells
SLE PBMC-B cells + Healthy B cells
Healthy PBMC-B cells + SLE B cells
SLE PBMC-B cells + Allogeneic SLE B cells
Healthy PBMC-B cells + Allogeneic healthy B cells
SLE PBMC-B cells + Healthy B cells
Healthy PBMC-B cells + SLE B cells
SLE PBMC-B cells + Allogeneic SLE B cells
C
Healthy
PBMC -B cells
SLE
PBMC -B cells
Healthy PBMC -B cells
+ SLE B cells
SLE PBMC-B cells
+ Healthy B cells
1 2 3
Regulatory B cells induce higher iNKT cell proliferation than mature B cells
Breg iNKT
cell +
aGal-Cer+IL-2
Beff iNKT
cell +
Bosma et al Immunity 2012
Depletion of immature B cells from PBMCs
impairs iNKT cell expansion
Whole
PBMCs
iNKT cell
expansion (day 7)
a-GalCer
+ IL-2
B cell depleting (rituximab) treated patient groups
RBRnr
RBRr
BCD = B cell depleted; RBRnr = rituximab, B cell repopulated, non-responding (Global BILAG Score after repopulation);
RBRr =rituximab, B cell repopulated, responding (No change in Global BILAG Score after repopulation)
iNKT cell numbers remain low during B cell depletion but is restored upon return of B cells in responding patients
BC
DR
r B
CD
Rn
r
0 min 5 min
CD1d DAPI
CD1d DAPI
CD1d DAPI
CD1d DAPI
CD1d recycling is normalizes in repopulated B cells patients responding to B cell depletion therapy
BCD = B cell depleted BCDRnr = B cell depleted repopulated, non-responding BCDRr = B cell depleted repopulated,responding
RBRr
0.001 0.01 0.1 10.1
1
10
100
r = 0.3770
p = 0.0017
% C
D1d
hi
imm
atu
re B
cells 0
6
BILAG
% iNKT cells
Restoration of CD1d expression on immature B cells correlate with higher number of iNKT cells and
better clinical response to rituximab
Conclusions from studies in SLE patients
• Immature B cells sustain the in vitro expansion and
activation of iNKT cells in healthy individuals via CD1d
• SLE patients present a decreased expression of CD1d hi on
immature B cells
• SLE iNKT cells are numerically and functionally impaired
• iNKT cell, B cell, CD1d defect in SLE patients are
exclusively reversed in patients responding to rituximab
treatment
• Positive correlation between iNKT cell and immature B cell
and amelioration of disease.
UCL Paul Blair Fabian Flores-Borja Anneleen Bosma Azza Abdel Gadir Elizabeth Rosser Madhvi Menon Kiran Nistala Christoper Piper David Isenberg Elizabeth Jury
Washington University David Rawlings University of Oxford Vincenzo Cerundolo INSERM Agnes Lehuen