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Cross-talk among MCross-talk among M, NK and cancer cells:, NK and cancer cells:
MM cells help NK cells to attack tumor by stimulatory RAE-1 but cells help NK cells to attack tumor by stimulatory RAE-1 but escape from NK killing by inhibitory Qa-1escape from NK killing by inhibitory Qa-1
Zhigang Tian, Zhixia Zhou, Cai Zhang
([email protected])([email protected])
August 19, 2010, ShenzhenAugust 19, 2010, Shenzhen
Institute of Immunology/School of Life SciencesInstitute of Immunology/School of Life SciencesUniversity of Science & Technology of China University of Science & Technology of China
Chinese Academy of SciencesChinese Academy of SciencesHefei, Anhui, ChinaHefei, Anhui, China
Figure 2-49NK cell: a professional killer of tumorNK cell: a professional killer of tumor
Nature review immunology, 2008;8:713
1.initiation 2.effector3.termination
Three main stages of cell lytic synapse
SMAC: supramolecular activation cluster
SMIC: supramolecular inhibitory cluster
Two kinds of immunological synapse
NK Cell Immunological Synapse
NK Cell Immunological SynapseNature review immunology, 2008;8:713
Annu. Rev. Immunol. 2005. 23:225–74
Natural killer cell recognition of “missing self”
KIR : killer immunoglobulin-like receptors
KLR: killer lectin-like receptors
M
NK
Death
Tumor cells
Poly I:C
1. Macrophages increase cytolysis of NK cells against cancer cells
++
% L
ysis
of
NK
cel
ls t
o M
0
1
2
3
4
5
6
50:1 25:1 5:1E/T ratio
Untreated peritoneal Mpoly I:C treated peritoneal M
0102030405060708090
5:1 25:1 50:1
**
% L
ysis
of
NK
cel
ls t
o tu
mor
NK aloneNK/M (medium)NK/M (poly I:C)
*
*
E/T ratio
M
NK
Death
Tumor cells
Poly I:C
NKG2D㈩㈩
2. Poly I:C-treated macrophages increase NKG2D expression of NK cells
Fol
d I
nd
uct
ion
of
N
KG
2D g
ene
0
1
2
3
4
5
6
NK alone NK+
RAW264.7
NK+
RAW264.7(poly I :C)
*
36.48% 32.74% 50.71%
NK onlyNK+
RAW264.7
NK+
RAW264.7 (poly I:C)
NKG2D
Ev e
nts
㈩㈩
M
Death
Tumor cells
Poly I:C
3. Increased NKG2D expression correlates to activation of NK cells
NK onlyNK+
M
NK+
M (poly I:C)
NK+
M (poly I:C)+
Isotype-control
NK+
M (poly I:C)+
anti-NKG2D
Eve
nts
25.71% 25.13% 62.52% 55.03% 38.41%
CD69
NK aloneNK+MØNK+MØ (poly I:C) NK+MØ (poly I:C)+Isotype-contyolNK+MØ (poly I:C)+ anti-NKG2D
0
5
10
15
20
25
*
*
*
Fol
d E
xpre
ssio
n
FasL TRAILPerforin
NK
NKG2D㈩㈩
㈩㈩
M
NK
Death
Tumor cells
Poly I:C
NKG2D㈩㈩
4. Increased NKG2D expression correlates to function of NK cells
IFN-
IFN-
NK+MØ (poly I:C)+ anti-NKG2D
MØ alone
NK+MØNK alone
NK+MØ (poly I:C) NK+MØ (poly I:C)+Isotype-contyol
MØ (poly I:C)
100
200
300
400
500
600
700
IFN
- (
pg/
ml)
*
E:T ratio
0
20
40
60
80
5:1 25:1 50:1
M(Medium)M(poly I:C)M(poly I:C)+Isotype-controlM(poly I:C)+Anti-NKG2D
% L
ysis
to
YA
C-1
cel
ls*
*
*
㈩㈩
M
NK
Death
Tumor cells
Poly I:C
NKG2D㈩㈩
IFN-
IFN-
IL-15 IFN-IL-12IL-18
5. Macrophage-derived cytokines play critical roles in NK cell activation
050
100150200250300350400450500
IL-15 IL-12 IL-18 IFN- IFN-
**
**
*
pg/
ml
Untreated Mpoly I:C treated M
NK NK NK
Ev e
nts
32.03% 40.24% 54.79% 47.80% 31.05% 22.08%
NK alone NK+MNK
M (poly I:C) Isotype anti-IFN- anti-IL-15M (poly I:C) M (poly I:C) M (poly I:C)
NKG2D
㈩㈩
6. Macrophage-derived cytokines play critical roles in NK cell function
M aloneNK aloneNK+ MNK+ M (poly I:C) NK+ M (poly I:C)+Isotype-contyol
NK+ M (poly I:C)+ anti-Il-15NK+ M (poly I:C)+ anti-IFN-
050
100150200250300350
IFN
- (
pg/
ml)
*
NK+ MNK+ M (poly I:C) NK+ M (poly I:C)+Isotype-control
NK+ M (poly I:C)+ anti-Il-15NK+ M (poly I:C)+ anti-IFN-
/
0
10
20
30
40
50
60
70
50:1 25:1 5:1% L
ysis
to
YA
C-1
cel
ls
****
E/T ratio
M
NK
Death
Tumor cells
Poly I:C
NKG2D㈩㈩
IFN-
IFN-
IL-15 IFN-IL-12IL-18
㈩㈩
M
NK
Death
Tumor cells
Poly I:C
NKG2D㈩㈩
RAE-1
IFN-
IFN-
7. Up-regulation of NKG2D ligands on poly I:C-treated macrophages
0 µg10 g/ml(polyI:C)
100 g/ml(polyI:C)
0
2
4
6
8
10
12
RAE-1 H60 MULT-1
20 g/ml(polyI:C)
**
*
****
**
**
Fol
d I
nd
uct
ion
Untreated peritoneal Mpoly I:C treated peritoneal M
02468
10121416
RAE-1 H60 MULT-1
**
**
**
Fol
d I
nd
uct
ion
0 g/ml 10 g/ml 20 g/ml 100 g/ml
27.99% 35.98 % 53.35% 63.76%
Eve
nts
C57BL/6
BALB/c
RAE-1
0.89% 16.78% 33.85% 74.76%
1.87% 10.69% 35.85% 63.77%E
ven
tsE
ven
ts
RAW
IL-15 IFN-IL-12IL-18
㈩㈩
M
NK
Death
Tumor cells
Poly I:C
NKG2D㈩㈩
RAE-1
IFN-
IFN-
8. TLR3 mediates the up-regulation of NKG2D ligands by macrophages
0 µg100 µg poly I:C100 µg poly I:C+si-Control100 µg poly I:C+si-TLR3
Fol
d E
xpre
ssio
n
0
1
2
3
4
5
6
RAE-1 H60 MULT-1
* * *
0
4
8
12
16
20
TLR3 TLR4
Fol
d I
nd
uct
ion **
Untreated MØpoly I:C treated M
IL-15 IFN-IL-12IL-18
㈩㈩
M
NK
Death
Tumor cells
Poly I:C
NKG2D㈩㈩
RAE-1
IFN-
IFN-
IL-15 IFN-IL-12IL-18
Qa-1b
NKG2A
㈠㈠
9. Qa-1 contributes to protect macrophages from NK cell-mediated lysisYAC-1
BALB/c MBALB/c M +poly I:C RAW264.7RAW264.7+poly I:C
Rel
ativ
e E
xpre
ssio
n
00.010.020.030.040.050.060.070.080.09
Qa-1a Qa-1b
**
****
0
2
4
18
14**
Fol
d I
nd
uct
ion
** ** ** ** **RAE-1 H60 MULT-1
YAC-1BALB/c MBALB/c M +poly I:C
RAW264.7RAW264.7+poly I:C
NK onlyNK+
RAW264.7
NK+
RAW264.7 (poly I:C)
Eve
nts 44.13% 37.43% 36.56%
NKG2A
㈩㈩
10. Qa-1 knock-down cause macrophages sensitive to NK cell killing
No treatmentpoly I:Cpoly I:C+si-Controlpoly I:C+si-Qa-1
E/T ratio
% L
ysis
to
RA
W26
4.7
0
10
20
30
40
50
5:1 25:1 50:1
*
*
*
0
10
20
30
40
50
60
50:1 25:1
*
poly I:Cpoly I:C+Isotype-controlpoly I:C+anti-Qa-1b
untreated*
% L
ysis
to
RA
W26
4.7
E/T ratio
M
NK
Death
Tumor cells
Poly I:C
NKG2D㈩㈩
RAE-1
IFN-
IFN-
IL-15 IFN-IL-12IL-18
Qa-1b
NKG2A
㈠㈠
㈩㈩
M
NKG2D
RAE-1
NK
IFN-
IL-15 IFN-IL-12IL-18
Death
Qa-1b
NKG2A
IFN-
Tumor cells
Poly I:C
㈩㈩
㈩㈩ ㈠㈠
MM cells help NK cells to cells help NK cells to attack tumor by RAE-1 but attack tumor by RAE-1 but escape from NK killing by escape from NK killing by
Qa-1Qa-1
Conclusion
Macrophages may activate NK cells to attack tumor by activating RAE-l-NKG2D recognition but protect themselves from cytolysis of NK cells via preferential inhibitory Qa-1-NKG2A recognition, by which the NK cells will constitutively be activated by macrophages to keep strong innate immunity against tumor.
Cross-talk among MCross-talk among M, NK and cancer cells, NK and cancer cells
“ STONE MONKEY” : WELCOME YOU TO
HUANG-SHAN MOUNTAIN
20NK receptor complex-ligand interactions
21
Klas Kärre
in the laboratory at the Karolinska Institute in 1983.
Nature immunology, 2008;9:477
The idea that NK cells can
distinguish aberrant cells
by recognizing ‘absence
of the expected’, rather
than ‘presence of the
unexpected’ emerged
more than 25 years ago.
Klas Kärre recapitulates
how the idea took shape,
and the first five years of
experimental work to test
its general predictions.
Natural killer cell recognition of missing self
