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Shaping T cell functions by tumor
extracellular matrix.
Bei-Chang Yang (楊倍昌)
•
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Institute of Basic Medical Sciences (基醫所),
Department of Microbiology and Immunology (微免所), College of
Medicine, National Cheng Kung University, Tainan 70428, Taiwan.
鄭宇容，林育萍，黃俊淵，蘇重禎
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Fight cancer,
But, sometimes we lose
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Why T cells infiltrate and accumulate here?
NPC (CD3 stain)
Nature Med. 1999, 5:874
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This is not a single case.

Tumor structure and
extracellular matrix as a possible
barrier for therapeutic
approaches using immune cells or
adenoviruses in colorectal cancer.
Peter J.K. Kuppen et al,
Brest cancer
(invasive)
Histochem Cell Biol (2001) 115:67–72
Immune cells were abundantly
present in tumors from colorectal
origin. These cells were, however,
not found in direct contact with
tumor cells, but mainly in the
stromal part of the tumor.
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Leukocytes migration through endothelia
cells
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Molecules involve in cell-cell interaction
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Questions:



Why are T cells trapped in peripheral area of
tumor mass?
Are those T cells alive ?

Su CC, et al (2007) J Immunol. 179:4589-4597.

Hor WS, et al. (2003) J Leukocyte Biol. 73: 363-368.
Can those cells modulate consequent immune
reaction?


Do they affect tumor growth or metastasis?



IL-10 & TGF-b; Yang BC, et al. (2003) J Immunol. 171:3947-3954.
(depletion study) Chen YL, et al (2002) Br J Cancer 87:359-365.
Chen YL, et al. (2003) J Immunol. 171:1183-1192.
etc.
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黃俊淵
Q: Why do immune cells appear only in peripheral area
of tumor nest?
Possibilities:

Receptor-mediated

Secreted proteins

Extracellular matrix
A: Tumor extracellular matrix may play a role.
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
Consist of different combinations of fibrous collagen
proteins, hyaluronic acid, fibronectin, etc.

Bind to cell-surface receptor: integrin, laminin receptors, etc.

Are required for cell proliferation, differentiation,
morphogenesis, etc.
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Rational:
Why extracellular matrix?

Tumor cells secrete distinctive ECM that form a basal
membrane surrounding the tumor mass.

During the course of tumor infiltration, immune cells
have to migrate out of blood vessels and travel some
distance to get into the tumor site. There, they
encounter a distinct environment composed of various
ECM components, which are very different from those
of the bloodstream.
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Transmigration of T
cells through the
tumor monolayers.
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
Extracellular matrix recaptures the effect of tumor
monolayers.

It is integrin-dependant.
ECM was extracted by
0.5% Triton X-100.
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Jurkat cells spread poorly on glioma ECM.
(F-actin accumulated at front of lamellipodia)
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
The transcripts of collagen IV (COL IV), fibronectin (FN),
laminin-g1 (LAM) and tenascin-C (TN-C) were detected
by RT-PCR.

Glioma cells express high amount of tenascin-C.
Immunohistochemical stain
for tenascin-C on a glioma
tumor sample.
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Tenascin-C is a disulfide-bonded hexamer composed of subunits
with molecular weights in the range of 120–300 kD
Tenascin-C can interact with perlecan and
fibronectin, and cell surface receptors including
integrins a2b1, avb3 and a9b1, and annexin I.
Nature Reviews Cancer 5, 436-446 (2005)
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
Tenascin-C gene of glioma cells is knocked down by shRNA
method.

Tenascin-C-low transfectant cells allow better transmigration
of Jurkat cells.
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A better spreading of Jurkat cells on monolayer
and ECM of U-118MG(TNCshRNA) cells.
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Question: How does tenascin-C work?
Tumor contact activates ERK activities in
Jurkat cells, which are required for the
transmigration of Jurkat cells.
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
Tumor contact activates ERK activity in Jurkat cells, which
is required for the transmigration of Jurkat cells.
ERK and actin colocalize at the front of lamellipodia of Jurkat cells.
Further study on migration machinery (focal adhesion complex) is on going.
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CD3+T cells are trapped in tenascin-C-rich area.
Tenascin-C
CD3+ cells
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Conclusion:
Tenascin-C inhibits the polarization and
transmigration of T cells, that consequently
prevents a direct contact of glioma tumor and T
cells.
Are those T cells alive ?
蘇重禎
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No death occurs in Jurkat cells during coculture with
gliomas.
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Nature 407, 789 5(2000)
Caspase 3
Death substrates
Apaf-1
Caspase 9
Apoptosis
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FasLribozyme/EGFP plasmid
Fas-ligand
a-tubulin
Br J Cancer 85 (2002):1185-1192
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BC Yang
No death occurs in Jurkat cells during coculture with
gliomas.
PI-staining
A
B
A: Jurkat cells alone
B: CH-11 (1ng/ml)
C: with U-118MG(V)
D: with U118MG (R)
E: ZB4 (100ng/ml)/CH-11
C
D
E
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Coculture with tumor cells inhibits the
Fas-mediated apoptosis.
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Apoptosis program is inhibited.
Caspase 3
Death substrates
Apoptosis
Apaf-1
Caspase 9
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Direct cell contact is required.
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Integrin signal is required
Coculture with U118
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How Fas signal-mediated death is
suppressed in T cells?
25 M LY294002 for 1 h
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Fas-mediated death is suppressed by inhibitor of PI3K
but not by inhibitors of MAPK, NFkB, PKA, HSP70.
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Su CC, et al. 2007 Phosphatidylinositol 3-Kinase/Akt activation by integrin-tumor matrix
interaction suppresses Fas-mediated apoptosis in T cells. J Immunol 179:4589-4597.
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Stepwise hijacking of infiltrating T cells by tumor:
Paralyzing → Keeping alive → modifying (IL-10/TGF)

Why are T cells trapped in peripheral area of tumor mass?
Tenascin-C of glioma.

Are those T cells alive?
ECM signal suppresses apoptosis!
(IL-6/IL-8 for neutrophils)
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Glioma-associated IL-10
induction in T cell lines
Jurkat
1
2
Molt-4
3
1
2
3
IL-10
b-actin
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Direct cell-to-cell contact is required
U118(V)
Jurkat cells
1
2
3
4
IL-10
b-actin
U373(V/R) or
U118(V/R)
1. Jurkat alone
2. Jurkat separate chamber
3. Jurkat in upper chamber
/coculture in low chamber
4. Jurkat in coculture
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Fas/Fas-L signal-associated
IL-10 induction
Jurkat
U-373MG
-
V
R
U-118MG
V
R
IL-10
b-actin
Molt-4
U-373MG
-
V
R
U-118MG
V
R
IL-10
b-actin
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Thank you for listening
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