免疫学 研究生课程

FL2-H -->
10
FP THY GRAFT
10
FL3-H -->
FL3-H -->
2
10
7.7
10
1
1
1
10
2
10
2.6
84.7
2.3
82.7 8.8
3
10
10
2
10
3
10
4
10
3
10
1
10
2
10
3
10
4
FL2-H -->
FL2-H -->
CD4
Contribution of mouse MHC to negative but not positive selection of mouse T cells developing in FP THY grafts
Mouse CD8
10
4
FL2-H -->
10
1
10
2
10
3
10
1
10
2
10
3
10
4
FL1-H -->
Mouse CD4
Peripheral repopulation of mouse T cells In athymic mice by grafting FP thymus
% of mouse CD4+ cells in PBL
The major population of mouse T cells in FP THY-grafted ATX B6 mice
ATX B6
FP THY-grafted ATX B6
NL B6
CD45RB
CD62L
CD44
Reconstitution of naïve T cells in T celldepleted ATX mice by grafting pig thymus
MEAN % OF CD4+ CELLS
100 80
* * * ** *** **
B10 (5) ATX B10 (3) FP LIV ATX B10 (11) FP THY/LIV
60 40 20 0
MEL-14+
HIGH CD45RB
LOW CD44
KLH antigen immunization
0
20
40 60 % of PCP
80
100
Summary
Xenogeneic pig thymus can efficiently support mouse T maturation.
Mouse T cells maturing in Pig thymus grafts are functional
HOST THYMUS
SELECTING I-A
b/k
10
4
NON-SELECTING I-A k
FL3-H -->
10
10
4
FL3-H -->
2
10
3.1
84.5 8.3
2.7
80.7 3.1
3
10
1
10
CD8
10
4
1
10
2
10
3
10
4
10
1
10
2
3
10
4
1
10
2
10
3
10
4
FL2-H -->
10
CD8
FL3-H -->
2
10
1
10
10
1
10
2
10
3
10
4
31.6
10
1
10.2
10
1
10
2
10
3
10
4
10
2
10
3
10
4
FL2-H -->
FL2-H -->
CD4
Mouse MHC does not participate in positive selection of mouse thymocytes in FP THY grafts
HOST THY 3A9 HOST THY 3A9 X IIKO F4 FP THY ATX 3A9 FP THY ATX 3A9 X IIKO F4
CD8
CD4
MEAN % OF THYMOCYTES
40 30 20
* * ** ** ** * * * 3A9 (6) FP THY IN ATX 3A9 (6) 3A9 X IIKO F4 (6) FP THY IN ATX 3A9 X IIKO F4 (7)
CD4/8 DP
positive selection (<5%): Thymic epithelial cells MHC restriction
CD4/8 DN
CD4 SP
CD4
Pig MHC mediates the positive selection of mouse thymocytes in FP THY grafts
异种移植
赵 勇 研究员 生物膜与膜生物工程国家重点实验室 中国科学院动物研究所，北京
异种胸腺移植
重建受者T细胞免疫力 及诱导移植免疫耐受
胸腺与T细胞的发育分化
胸 腺 是 T 细 胞 发 育 的 重 要 器 官
T细胞发育分化
基 因 重 排 ， 阳 性 / 阴 性 选 择
双阴性 Pre-TCR TCR CD4 CD8 CLP TCR- rearrangement CD4SP TCR CD8 双阳性 单阳性 CD4 TCR
MEAN CELL NUMBER (X106)
40 30 20 10 0 80 60 40 20 0
CD4 SP
DP
I-Ab
I-Ak
H-2s/b
I-Ab I-Ak H-2s/b FP THY grafts
Host thymus
ONLY PIG MHC MEDIATES THE POSITIVE SELECTION OF MOUSE THYMOCYTES IN PIG THYMUS GRAFTS
B10
>2WKS
x
ATX
>11 WKS
2 months
PCP
FP THY/LIV TISSUE
PC
Mouse T cells maturing in pig thymus grafts Make host resistant to microorganism infection
Euthymic B6 mice FP THY-grafted ATX B6 mice
10 0
4
10
1
10
2
10
89.6
2.1
3
88.4
3
7.1
10
4
10
2
10
3
10
1
10
2
10
3
10
1
10
2
10
3
10
4
FL1-Hwk.baidu.com-->
FL1-H -->
FL1-H -->
Mouse CD4
“AND” TCR-transgenic mice
CD4+ T cells express Va11V3TCR, recognizing PCC. Positive selecting MHC II: I-Ab Negative selecting MHC II: H-2s Non-selecting MHC II: I-Ak
25
Mean stimulation index
CONTROL 25-9-27 28-16-8 25-5-16 14-4-4
20 15
Anti- B6 MHC class II
Control Ab
10
*
5 0 ATX B6(3) CONTROL
*
*
* * *
ATX B6 (3) FP THY/LIV
DN
DP
TCR- rearrangement 阳性选择
CD8SP 阴性选择
Mouse (human) bone marrow cells
Pig thymus
Mouse (human) T cells
T cells maturing in xenogeneic thymus
T cell development in xenothymus Xenogeneic T cell tolerance induction Tumor adoptive immunotherapy Treatment of AIDS
EUTHYMIC B6(3) NAIVE B6 (1) CONTROL
Zhao Y, et al, J of Immunol 1997, Zhao Y, et al, J of Immunol 1998
Protocol
Thymectomy mAbs to deplete T/NK cells (days -6, and -1) mAbs on days 7 &14
T cell development in xenothymus
Thymus
Genetic athymic mice (nude mice)
BMC
Peripheral T cells
X
PROTOCOL
Thymectomy mAbs to deplete T/NK cells (days -6, and -1) 3Gy TBI mAbs on days 7 &14
MATURATION OF TCR TRANSGENIC MOUSE T CELLS IN FP THY GRAFTS IN “AND” MICE WITH SELECTING H-2
"AND" THYMUS 4.4 48.1
4
FP THY GRAFT 2.8 78.7
10
3
10
FL3-H -->
Mouse thymus
4 4
MHC II-/- mouse thymus
10
FP THY grafts in MHC II-/- mice
10
4
3.1 85.8
FL2-H -->
10
6.7
3
Mouse CD8
10
10
FL2-H -->
FL2-H -->
2
2
10
10
1
1
10
10
8.7
10
1
0.7
10
T cell development in the thymus
CD8
CD8 SP CD4/8 DP Negative selection (95%, cell death)
CD4/8 DN
CD4 SP
CD4
Development of mouse T cells in pig thymus grafts
25 20 15 10 5 0 5 10 15 20 25 FP THY-grafted, T cell-depleted ATX B6 mice (31) T cell-depleted ATX B6 mice (12) T cell-depleted euthymic B6 mice (23)
Weeks post-FP THY grafting
>2WKS
B10
x
ATX
>11 WKS
FP THY/LIV TISSUE + 1 X10 8 FLC I.P.
Zhao Y, et al Nature Medicine 1996； Zhao Y, et al, J of Immunol 1997, Zhao Y, et al, J of Immunol 1998； Zhao Y, et al, Xenotransplantation 1998 Zhao Y, et al, Transplantation 2000； Zhao Y, et al, Transplantation 2000 Zhao Y, et al, Transplantation 2003； Zhao Y, et al, Cel Mol Biol 2001
2 wks
In vitro assay: T cell proliferative responses to Ag
FP THY-grafted ATX B6 mice
Mouse T cells maturing in pig thymus respond to antigens presented by mouse MHC
Positive and negative selection of mouse thymocytes in pig thymus grafts
Positive and negative selection of thymocytes
CD8
CD8 SP
Negative selection (95%, cell death) BMC-derived APC T cell tolerance
DEX-treated B6 mice
ATX B6 mice
T cells maturing in xenogeneic thymus grafts have the ability against microorganism infection
T cell-depleted ATX B6 (4) FP THY-grafted, T cell-depleted ATX B6 (4) T cell-depleted euthymic B6 (5) Normal euthymic B6 (5) DEX-treated euthymic B6 (3)
The phenotypes of naïve and memory T cells
Thymus
Antigen
Immune response
Naïve T cells
Activated/Memory T cells
CD45RBhigh CD62Lhigh CD44low
CD45RBlow CD62Llow CD44high