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![Page 1: Peripheral Control of Alloreactive B cells Anita Chong Section of Transplantation Department of Surgery.](https://reader035.fdocuments.net/reader035/viewer/2022062421/56649cd75503460f9499fbef/html5/thumbnails/1.jpg)
Peripheral Control of Alloreactive B cells
Anita ChongSection of Transplantation
Department of Surgery
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Chong Lab
YiJin Li Dengping Yin
LianLi Ma Jamie Kim
JiKun Shen Anncy Varghese
Jingwen Ding
Tongmin Wang
Audrea Troutman
NextranGerry Byrne
Hui Xu
U ColoradoRoberta Pelanda
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Alloreactive B cells andLong-term Allograft
Survival
• Alloantibodies mediate acute and chronic rejection
• Alloantibody-alloantigen complexes induce maturation of APC
• B cells can activate alloreactive T cells
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Mechanisms of B cell tolerance
Calame, Nat Immunol 2001:1103
AntigenCD40L
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Nature Immunology 2, 1126 - 1132 (2001)
B cells and professional APCs recruit regulatory T cells via CCL4Richard S. Bystry1, Varuna Aluvihare2, Katie A. Welch1, Marinos Kallikourdis1 & Alexander G. Betz1
Using gene expression profiling, we show here that activation of B cells and professional antigen-presenting cells (APCs) induces the expression of common chemokines. Among these, CCL4 was the most potent chemoattractant of a CD4+CD25+ T cell population, which is a characteristic phenotype of regulatory T cells. Depletion of either regulatory T cells or CCL4 resulted in a deregulated humoral response, which culminated in the production of autoantibodies. This suggested that the recruitment of regulatory T cells to B cells and APCs by CCL4 plays a central role in the normal initiation of T cell and humoral responses, and failure to do this leads to autoimmune activation.
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Two-Phase model of
T cell-dependent allo-specific B cell activation
Modified from Baumgarth 2000 Immunol Rev.
SecretedIgM
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Extrinsic control of alloreactive B cells
• Inhibiting the development of effector T helper cells
• Inhibiting effective T-B cell interaction
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Tolerance Model
Day 0C57BL/6
Day 60+ C57BL/6
Anti-CD40L (1 mg MR1/mouse; D-7, 0, 7) + DST (B/6)
C57BL/6 (H-2b) into BALB/c (H-2d) mice
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Anti-CD40L + DSTinduces allograft tolerance
Treatment Graft Survival
None 8 (x7), 9
+ Anti-CD40L > 160 (x11)
+ 2nd heart >60 (x5), >100 (x3)
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•Tolerance can be dominantly transferred
•Tolerant mice reject third-party C3H hearts, exhibit linked tolerance to F1 (B/6 x C3H) hearts.
•A role of CD25+ T cells
•Allograft tolerance is associated with inhibited alloantibody production
Features of Allograft Tolerance
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Peripheral B cell Tolerance
m
Donor-specific (B/6) heart
+ 3.83 BCR-Ki B cells
B/c recipients of B/6 heart
Day 60-80 Day 0: DST + MR1
Naïve
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3.83 BCR-Ki B cells recognize Kb antigens
m
H2-K(b) H2-K(d)
Tetramer
CD19
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m
Peripheral B cell Tolerance: 3.83 Cell Transfer + B/6 heart
Days Post-B Cell Transfer
0
0.25
0.5
0.75
1
1.25O
.D.
0 5 10 15 20
Anti-Id-IgM
Naïve - No TxpTolRej
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Peripheral B cell Tolerance: Lack of T cell help
m
Third-party C3H heart + 3.83 BCR-Ki B cells
Day 60-80 Day 0: DST +
MR1
Naive
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3.83 BCR-Ki B cells have dual
alloreactivity
m
H2-K(b) H2-K(d)H2-K(k)
Tetramer
CD19
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Peripheral B cell Tolerance: Lack of T cell help
m
Third-party C3H heart + 3.83 BCR-Ki B cells
Day 60-80 Day 0: DST +
MR1
Naive
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m
Days Post-B cell Transfer
Peripheral B cell Tolerance: Inhibition of T cell help
-0.25
0
0.25
0.5
0.75
1
1.25O
.D.
0 7 14 21
c) Anti-Id IgM
TolNaive
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Peripheral B cell Tolerance: Inhibition of T cell help
m
B/6 heart + 3.83 BCR-Ki B cells +
Anti-CD40
Day 60-80 Day 0: DST +
MR1
Naive
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m
Control of B cell Responses by limiting CD40-CD40L signals:
0
0.5
1
1.5
O.D
.
0 7 14 21 28
Days Post-B Cell Transfer
Anti-Id IgM
Tol + Anti-CD40
Tol
Rej
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Peripheral B cell Tolerance: Regulation of T
cell help
m
F1 (B/6 x C3H) heart +3.83 BCR-Ki B cells
Day 60-80 Day 0 DST + MR1
Naive
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m
Linked Regulation of T cell Helper Responses
Days Post-B Cell Transfer
-0.25
0
0.25
0.5
0.75
1O
.D.
0 5 10 15 20
Anti-Id IgM
TolNaive
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Cells Maintaining Peripheral B cell Tolerance
m
Spleen Cells from:i. Naïveii. Tolerant (d60-120)iii. CD25-Depl Tolerant
Scid recipients of B/6 heart
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m
Peripheral B cell Tolerance: Role of CD25 Cells
0
50
100
150
All
o-I
gG
0 10 20 30
Tolerant + Htx
Naive + Htx
Naive-No Txp
0
25
50
75
100
All
o I
gG
0 10 20 30
CD25 Depl Tolerant
Tolerant
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Peripheral suppression of alloantibody production:
1.Inhibiting the generation of T cell help
2.Dependent on CD25+ cells
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m
Regulation of T cell help
C3H-TH cell
Recpt. APC
F1 antigensB/6xC3H
3.83 B cell
B/6-TTol/Reg
C3H-TH cell
No T cell Help
T cell Zone
B cell ZoneB/6-TTol/Reg
Phase 1
Phase 2
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Anti-carbohydrate Antibodies in Allograft
transplantation • Naturally occurring and elicited
anti-A or anti-B antibodies
• Donor-specific B-cell tolerance after ABO-incompatible infant heart transplantation (Fan et al. 2004)
• Can peripheral mechanisms control anti-carbohydrate antibody responses?
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Structure of Gal1,3Gal and Blood Group A and B antigens
CH2OH
ROH
O O O
OO
CH2OH CH2OH
NHCOCH3
OH
OH
OH
OH O
H
CH2OH
ROH
O O O
OO
CH2OH CH2OH
NHCOCH3
OH
OH
OH O
H
O
OH
OH
OH
C3H
O
CH2OH
ROH
O O O
OO
CH2OH CH2OH
NHCOCH3
OH
NHCOCH3
OH O
H
O
OH
OH
OH
C3H
O
Gal1,3Gal-ß1,4GlcNAc
Blood Group BGal1,3(Fuc1,2)Gal
Blood Group AGalNAc1,3(Fuc1,2)Gal
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Tolerance Model
Day 0Gal+BALB/c Heart +Intact Active Bone
Day 60Gal+BALB/c Heart
Anti-CD40L (1 mg MR1/mouse; D 0, 7, 14)
C57BL/6-Gal-/- recipients tolerant to BALB/c-Gal+/+ grafts
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IAB and anti-CD40L therapy induces allograft tolerance
Treatment Mean
None 7 d
Anti-CD40L + IAB - 1st BALB/c heart > 130 dAnti-CD40L + IAB - 2nd BALB/c heart > 60 d
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Features of IAB/Anti-CD40L-induced tolerance
•Tolerance can be dominantly transferred
•Tolerant mice reject third-party C3H hearts, exhibit linked tolerance to F1 (B/c x C3H) hearts.
•A role of CD25+ T cells
•Allograft tolerance is associated with suppressed alloantibody production
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Anti-Gal IgM production in IAB/Anti-CD40L-induced tolerant
mice
0
0.25
0.5
0.75
1
1.25
O.D
.
0 30 60 90
Days Post-Transplant
Anti-Gal IgMb
None
MR1 + IAB
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Peripheral B cell Tolerance
m
Donor-specific (B/c) heartOr Third-Party C3H heart
+ Anti-Gal BCR-Tg B cells
B/6 GT-/- recipient of B/c heart
Day 60-80 Day 0: IAB + MR1
Naïve
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IgMa+ CD43+ CD21+
Madcam-1IgMa
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Anti-Gal IgMa production following challenge with BALB/c
hearts
00.5
1
1.52
0 10 20 30
f) Anti-Gal IgMa
Days Post-B cell Transfer
Naive
Tolerant
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Anti-Gal IgMa production following challenge with C3H
hearts
0
0.25
0.5
0.75
1O
.D.
0 7 14
b) Anti-Gal IgMa
Days Post-B cell Transfer
Naive
Tolerant
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Conclusion
•Anti-Gal cell responses are inhibited even in the presence of effector T helper cells
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m
Regulation of B cell responses
C3H-TH cell
Recpt. APC
Balb/c-Galantigens
T cell Help
B/c-TTol/Reg C3H-Gal
antigens
Recpt. APC
Gal-B cell
B/c-TTol/Reg
C3H-TH cell
T cell Zone
B cell Zone
Phase 1
Phase 2
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Extrinsic control of alloreactive B cells
• Inhibiting the generation of effector T helper cells
• Inhibiting the development of effector B cells
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Histology of Spleen
0.0
20.0
40.0
60.0
80.0
Native B/6 C3H F1
2nd Graft
% G
erm
inal
Cen
ter
B/6
C3H
F1 FDC-M1B220
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Peripheral B cell Tolerance:
Effect of TI-B cell stimulation
m
Donor-specific (B/c) heart+ Anti-Gal BCR-Tg B cells
+ CpG
B/6 GT-/- recipient of B/c heart
Day 60-80 Day 0: IAB + MR1
Naïve
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Inhibition of Anti-Gal IgMa production is reversed by CpG
0
0.5
1
1.5
O.D
.
0 7
Days Post-B cell Transfer
Anti-Gal IgMa
Tol + CpG
Tol
Naive-No Txp