Expression of CD103 identifies human regulatory T cell subsets
Zoulfia Allakhverdi1, David Fitzpatrick2, Marika Sarfati 1 and Guy Delespesse1
1CHUM 2Amgen
Research Center, Notre-Dame Hospital, Montreal, Quebec, Canada Inc, Seattle, WA
Introduction
Naturally occurring CD4+CD25+ Treg have an essential role in physiological as well as pathological conditions including autoimmunity, pregnancy, allergy, infections, transplantation and cancer.
Studying these cells in the context of ongoing disease has been problematic, largely because Treg and activated CD4+ T cells express the same surface molecules, such as CD25, OX40, GITR, CTLA-4. The transcriptional repressor FoxP3 is a selective marker for the identification of mouse and human CD4+ nTreg. However, FoxP3 is an intracellular but not a surface marker.
CD103 is a specific marker of mouse CD4+ nTreg subsets, allowing their classification into 3 groups defined by the co-expression of CD25, i.e. CD103-CD25+, CD103 +CD25+ and CD103+CD25-.
Lehmann J et al. 2002. Proc Natl Acad Sci USA 99(20): 13031.
These 3 subsets express similar amounts of FoxP3 mRNA and display the same suppressive activity in vivo and in vitro.
Is CD103 a marker of human CD4+ Treg subsets?
I. Expression of CD103 and CD25 on freshly isolated CD4+ T cells
Cord Blood
0.13 0.01
Tonsils
2.96 0.32
3.7
9.8
Adult Blood
0.56 0.30
60
FoxP3 mRNA relative expression
CD103
9.3
50 40 30 20 10 0
R2
R3
R4
R5
CD25
Lymph Node
0.08 0.03
2.1
CD25
T - +T + +T - +T 5 5 D2 - D25 - CD2 + C C 03 103 103 1 CD CD CD
CD103
+ 5 25 25 D2 - D + D 3 C 03- C 103 C 10 D1 CD CD C
10 9 8 7 6 5 4 3 2 1 0
3H,
cpm x1000
-
II. In vitro induction of CD103+CD25+ cells in response to allogeneic antigen
iDC
6.1
14.5
CD103+ cells (%)
20 16 12 8 4 0
% of CD103+ cells
mDC
iDC mDC 20 16 12 8 4 Day 0 Day 3 Day 7 Day 14
**
CD103
CD25
iDC mDC
30 25 20 15 10 5 0
2500
cpm x 1000
IL-2, pg/ml
2000 1500 1000 500
3H,
+ ) ) 3 03 (1:1 (1:4 10 - + T 1 CD 103 CD + T + + T + 3 3 CD 10 D10 CD C
T
0
+ T 03 1:1) 1:4) 03 - +T 1 D1 T ( T ( CD 103 C+ + + + 3 3 CD 10 D10 CD C
III. Induction of CD103+ suppressor cells upon CD3/B7.1 stimulation in the presence of TGF-β
cpm x 1000
37 25 20 15 10 5 0
FoxP3 mRNA relative expression
CD103
CD25
) ) T :1 :4 - + (1 (1 3 T T 10 + + + + 3 3 CD 10 10 CD CD
T
7 6 5 4 3 2 1 0
3H,
4 CD C D4
+ +C
3 10 D C
+ +C
3 10 D
-
D4
IL-2
8000 1600 1200 800 400 0 4000 2000 6000
IFN-γ
5000 4000 3000 2000 1000
IL-10
T 1) 4) 1: (1: - + ( T T 103 + + + + CD 03 103 1 CD CD T
T 1:1) 1:4) T - + ( ( 3 T T 10 + + + + 3 3 CD 10 D10 CD C
0
4) T :1) 1: - + T 1 ( ( 3 T +T 10 + + + 3 3 CD 10 D10 CD C
0
Conclusion I:
Co-expression of CD25 and CD103 identifies 3 subsets of human Treg (i.e. CD25+CD103-, CD25-CD103+ and CD25+CD103+)
CD103+CD4+ T cells are suppressor cells regardless of CD25 expression
IV. Differential induction of CD103 on neonatal and adult CD4+ T cells
Cord Blood
iDC
5.1 1.9 20
Adult Blood
**
Cord Blood Adult Blood
140 120 100 80 60 40 4 0 20 0
CD103+ cells (MFI)
CD103+ cells (%)
16 12 8
mDC
15.2 CD103 3.3
CD25
IV. Endogenous TGF-β and IL12 regulate the induction of CD103
mDC
18.5
mDC + rhLAP
3.8
mDC + ctr Ab mDC + anti IL-12
12.5 29
mDC + ctr Ab mDC + anti IL-4
14 14.1
mDC + ctr Ab mDC + anti IL-10
15 14.7
CD103
V. Mechanisms of differential expression of CD103 on neonatal and adult CD4+ T cells Is it due to IL-12?
Cord Blood
15.9
Adult Blood
mDC + ctr Ab 2.9
mDC + anti-IL-12 36.5 CD103 6.9
CD25
VI. Mechanisms of differential expression of CD103 on neonatal and adult CD4+ T cells Is it due to TGF-β?
Cord Blood
mDC 16.5 2.4
Adult Blood
mDC + TGF-β 36.7 CD103 15.2
CD25
VII. Mechanisms of differential expression of CD103 on neonatal and adult CD4+ T cells
Cord Blood
CD3/B7.1 + TGF-β
35.6
Adult Blood
16.5
CD103
CD25
VIII. Differential induction of CD103 on neonatal and adult CD4+ T cells Neonatal and adult CD103+ T cells display similar suppressive activities on a per cell basis
Cord Blood 45 40 Adult Blood
cpm x 1000
3H,
35 30 25 20 15 10 5 0
T
1) 4) 1: (1: - + ( 03 + + T + T + D1 03 3 C 1 0 D D1 C C
T
1) 4) 1: (1: - + ( 03 + + T + + T 1 CD 103 103 CD CD T
IX. Differential induction of CD103 on neonatal and adult CD4+ T cells
Increased numbers of CD103 suppressor cells in mixed lymphocyte cultures of neonatal T cells result in enhanced suppression compared with adults
Cord Blood Adult Blood
45 40
*** ** ** ** ***
***
cpm x 1000
3H,
35 30 25 20 15 10 5 0
1ry culture
2ry culture
2ry culture CD103-
Conclusion II
The induction of CD103 suppressor lymphocytes in response to allogeneic cells is greater in neonatal than in adult blood This differential CD103 expression results from intrinsic T cell factors and not from different production of endogenous cytokines On a per cell basis neonatal and adult CD103+ T cells display similar suppressive activity The induction of greater numbers of CD103+ Treg cells by alloantigen stimulated cord blood may be related to its successful clinical utilization as an alternative of adult bone marrow transplantation