Post on 13-Jan-2016
description
CIS INTERACTION IN NK CELLS
Presented by Refael CohenPresented by Refael CohenProject Advisor: Prof. Ramit Mehr
Asya Kaplan
Bar-Ilan UniversityFaculty of Life SciencesComputational Biology
2009
Presented by Refael CohenPresented by Refael CohenProject Advisor: Prof. Ramit Mehr
Asya Kaplan
Bar-Ilan UniversityFaculty of Life SciencesComputational Biology
2009
The receptors on NK cell:The receptors on NK cell:
• Adhesion molecules – LFA-1/ICAM-1Adhesion molecules – LFA-1/ICAM-1• Activating complexes – NKG2D/MICAActivating complexes – NKG2D/MICA• Inhibitory complexes – Ly49/MHC-1Inhibitory complexes – Ly49/MHC-1• Lipid raftsLipid rafts
INTRODUCTION: NK cell Immunological Synapse (NKIS)
Active Synapse: Increase in integrated activating signal.
Our model:Each bound inhibitory receptor inhibits all
the bound activating receptors within a
Certain radius.
Lysis of target cell.
INTRODUCTION: NK cell Immunological Synapse (NKIS)
S total = S+ - Inhibited (S+)
Activating Inhibitory adhesionActivating Inhibitory adhesion
Active Synapse:
The order:The order:
1. LFA1 binds to ICAM.
2. The number of bound adhesion molecules > Cmin.
3. NKG2D and Ly49 bind their ligands.
4. Smin < integrated activating signal.
5. Directed movement of NKG2D & lipid rafts.
6. Surrounding ring of LFA1 – stress of membrane.
7. Switch point of NKG2D molecules.
8. Increase in integrated activating signal.
9. Lysis of target cell.
INTRODUCTION: NK cell Immunological Synapse (NKIS)
Active Synapse: LFA1 binds to ICAM. The number of
bound adhesion
molecules > Cmin.
INTRODUCTION: NK cell Immunological Synapse (NKIS)
Active Synapse: NKG2D and Ly49 bind their ligands. When integrated activating signal > Smin -
Directed movement of NKG2D & lipid rafts.
INTRODUCTION: NK cell Immunological Synapse (NKIS)
Active Synapse: Surrounding ring of LFA1 -Short and long complexes in adjacent membrane sites may cause a structural stress in the membrane, pushing the long
ones (LFA1 complex) out from the short ones .
INTRODUCTION: NK cell Immunological Synapse (NKIS)
Active Synapse: Switch point of NKG2D molecules.
INTRODUCTION: NK cell Immunological Synapse (NKIS)
Num
ber
of
NK
G2D
Num
ber
of
NK
G2D
Time stepTime step
• cSMACcSMAC
• pSMSCpSMSC
Cis interaction
The inhibitory Ly49A NK cell receptor not only binds to its
H-2Dd ligand expressed on potential target cells (in trans) but also is constitutively associated with H-2Dd in cis (on the same cell).
This binding probably does not transduce an inhibitory signal.
Cis interaction
Cis interaction
The long Ly49 stalk region (67-137 aa) and 3 -helices shows considerable flexibility. Cis association and trans interaction occur through the same binding site (2M) (Marie-Agnes Douceyl, Leonardo Scarpellino,, Jacques Zimmer,
Philippe Guillaume, Immanuel Luescher, Claude Bron & Werner Held, Cis association of Ly49A with MHC class I restricts natural killer cell inhibitionj, Nature Immuno. 2004)
Cis interaction
Cis interaction
Cis interaction
Cis interaction
Cis association restricts the number of Ly49 receptors available for binding of H-2Dd on target cells and reduces NK cell inhibition through Ly49.
The result: more activating integrated signal, and faster lysis.
Cis interaction
Cis interaction: Goals and Methods
My projects:
Part 1:To update the simulation of NK cell synapse (over 10,000 C++ code lines)
with cis interaction by hundreds of additions and changes, include graphic exhibit.
Cis interaction: Goals and Methods.
Part 2:
To run the update program and find when will accept positive synapse.
Also to compare the results with old program (without cis interaction).
Part 2: To run the update program and check What are the effects of cis interaction on behavior of synapse?Also to compare the results with old program (without cis interaction).To draft graphs with MATLAB programs. MOVIEMOVIE
Cis interaction: Goals and Methods.
Parameters:
cis interaction exist in mouse only, and old program was on human, therefore we need to find new data for synapse.
We find data from article on: bond length, diffusion rate, microdomain preference, dissociation rate, association rate of all percipient molecules.
Other data remain as parameters.
▲ - 0 MICA, a ■ – 1000MICA, a ♦ – 3000 MICA, and a ● – 5000 MICA. parameters investigation:
0 2000 4000 6000-500
0
500
1000
1500
2000
2500
30002000KIR2000NKG2D
0 2000 4000 6000-500
0
500
1000
1500
2000
2500
3000
2000KIR1000NKG2D
0 2000 4000 6000-500
0
500
1000
1500
2000
2500
30001000KIR2000NKG2D
0 2000 4000 6000-500
0
500
1000
1500
2000
2500
3000
1000KIR1000NKG2D
0 2000 4000 6000-500
0
500
1000
1500
2000
2500
3000
2000KIR2000NKG2D
0 2000 4000 6000-500
0
500
1000
1500
2000
2500
30002000KIR1000NKG2D
0 2000 4000 6000-500
0
500
1000
1500
2000
2500
30001000KIR2000NKG2D
0 2000 4000 6000-500
0
500
1000
1500
2000
2500
3000
1000KIR1000NKG2D
0 2000 4000 6000-500
0
500
1000
1500
2000
2500
30002000KIR2000NKG2D
0 2000 4000 6000-500
0
500
1000
1500
2000
2500
30002000KIR1000NKG2D
0 2000 4000 6000-500
0
500
1000
1500
2000
2500
3000
1000KIR2000NKG2D
0 2000 4000 6000-500
0
500
1000
1500
2000
2500
30001000KIR1000NKG2D
0 2000 4000 6000-500
0
500
1000
1500
2000
2500
30002000KIR2000NKG2D
0 2000 4000 6000-500
0
500
1000
1500
2000
2500
3000
2000KIR1000NKG2D
0 2000 4000 6000-500
0
500
1000
1500
2000
2500
30001000KIR2000NKG2D
0 2000 4000 6000-500
0
500
1000
1500
2000
2500
3000
1000KIR1000NKG2D
0 2000 4000 6000-500
0
500
1000
1500
2000
2500
30002000KIR2000NKG2D
0 2000 4000 6000-500
0
500
1000
1500
2000
2500
30002000KIR1000NKG2D
0 2000 4000 6000-500
0
500
1000
1500
2000
2500
30001000KIR2000NKG2D
0 2000 4000 6000-500
0
500
1000
1500
2000
2500
30001000KIR1000NKG2D
A. R
adiu
s 1
B. R
adiu
s 3
C
. Rad
ius
5
D. R
adiu
s 10
E
. Rad
ius
50
MHCMHC
Ste
ady-
Sta
te I
nte
gra
ted
Sig
nal
s
▲ - 0 MICA, a ■ – 1000MICA, a ♦ – 3000 MICA, and a ● – 5000 MICA. parameters investigation:
0 2000 4000 6000-500
0
500
1000
1500
2000
2500
30001000KIR2000NKG2D
MHCMHC
Inte
grat
ed s
igna
lIn
tegr
ated
sig
nal
0 2000 4000 6000-500
0
500
1000
1500
2000
2500
30002000KIR2000NKG2D
0 2000 4000 6000-500
0
500
1000
1500
2000
2500
3000
2000KIR1000NKG2D
0 2000 4000 6000-500
0
500
1000
1500
2000
2500
30001000KIR2000NKG2D
0 2000 4000 6000-500
0
500
1000
1500
2000
2500
3000
1000KIR1000NKG2D
0 2000 4000 6000-500
0
500
1000
1500
2000
2500
3000
2000KIR2000NKG2D
0 2000 4000 6000-500
0
500
1000
1500
2000
2500
30002000KIR1000NKG2D
0 2000 4000 6000-500
0
500
1000
1500
2000
2500
30001000KIR2000NKG2D
0 2000 4000 6000-500
0
500
1000
1500
2000
2500
3000
1000KIR1000NKG2D
0 2000 4000 6000-500
0
500
1000
1500
2000
2500
30002000KIR2000NKG2D
0 2000 4000 6000-500
0
500
1000
1500
2000
2500
30002000KIR1000NKG2D
0 2000 4000 6000-500
0
500
1000
1500
2000
2500
3000
1000KIR2000NKG2D
0 2000 4000 6000-500
0
500
1000
1500
2000
2500
30001000KIR1000NKG2D
0 2000 4000 6000-500
0
500
1000
1500
2000
2500
30002000KIR2000NKG2D
0 2000 4000 6000-500
0
500
1000
1500
2000
2500
3000
2000KIR1000NKG2D
0 2000 4000 6000-500
0
500
1000
1500
2000
2500
30001000KIR2000NKG2D
0 2000 4000 6000-500
0
500
1000
1500
2000
2500
3000
1000KIR1000NKG2D
0 2000 4000 6000-500
0
500
1000
1500
2000
2500
30002000KIR2000NKG2D
0 2000 4000 6000-500
0
500
1000
1500
2000
2500
30002000KIR1000NKG2D
0 2000 4000 6000-500
0
500
1000
1500
2000
2500
30001000KIR2000NKG2D
0 2000 4000 6000-500
0
500
1000
1500
2000
2500
30001000KIR1000NKG2D
A. R
adiu
s 1
B. R
adiu
s 3
C
. Rad
ius
5
D. R
adiu
s 10
E
. Rad
ius
50
MHCMHC
▲ - 0 MICA, a ■ – 1000MICA, a ♦ – 3000 MICA, and a ● – 5000 MICA. parameters investigation: S
tead
y-S
tate
In
teg
rate
d S
ign
als
parameters investigation : 1000 MHC on NK cell and target cell (30% cis)1000 MHC on NK cell and target cell (30% cis)::
Activating Integrated signal: x2Activating Integrated signal: x2Ly49-MHC trans
0
1020
30
4050
60
70
8090
100
0 5000 10000 15000 20000 25000 30000 35000
Without cis:
Ly49-MHC trans
010
20
30
40
50
60
7080
90
100
0 5000 10000 15000 20000 25000 30000 35000
With cis: integrated signal
0
50
100
150
200
250
300
0 5000 10000 15000 20000 25000 30000 35000
With cis:
integrated signal
0
50
100
150
200
250
300
0 5000 10000 15000 20000 25000 30000 35000
Without cis:
pSMAC pSMAC cSMAC cSMAC
Number of NKG2D and MICA: a bit more in centerNumber of NKG2D and MICA: a bit more in centerMICA
SP=1200
0
100
200
300
400
500
600
700
800
0 5000 10000 15000 20000 25000 30000 35000
Without cis:
NKG2DSP=2300
0
100
200
300
400
500
600
700
800
0 5000 10000 15000 20000 25000 30000 35000
With cis: MICASP=1000
0
100
200
300
400
500
600
700
800
0 5000 10000 15000 20000 25000 30000 35000
With cis:
NKG2DSP=2300
0
100
200
300
400
500
600
700
800
0 5000 10000 15000 20000 25000 30000 35000
Without cis:
parameters investigation : 1000 MHC on NK cell and target cell (30% cis)1000 MHC on NK cell and target cell (30% cis)::
pSMAC pSMAC cSMAC cSMAC
Number of Ly49 and MHC: a bit less in centerNumber of Ly49 and MHC: a bit less in centerLy49
0
100
200
300
400
500
600
700
800
0 5000 10000 15000 20000 25000 30000 35000
Without cis: MHC on target
0
100
200
300
400
500
600
700
800
0 5000 10000 15000 20000 25000 30000 35000
Without cis:
Ly49
0
100
200
300
400
500
600
700
800
0 5000 10000 15000 20000 25000 30000 35000
With cis: MHC on target
0
100
200
300
400
500
600
700
800
0 5000 10000 15000 20000 25000 30000 35000
With cis:
parameters investigation : 1000 MHC on NK cell and target cell (30% cis)1000 MHC on NK cell and target cell (30% cis)::
pSMAC pSMAC cSMAC cSMAC
Number of LFA-1 and ICAM: later switchNumber of LFA-1 and ICAM: later switch
LFA-1SP=2800
050
100150200250300350400450
0 5000 10000 15000 20000 25000 30000 35000
Without cis: ICAM
0
50
100
150
200
250
300
350
400
0 5000 10000 15000 20000 25000 30000 35000
Without cis:
LFA-1SP=3200
0
50
100
150
200
250
300
350
400
450
0 5000 10000 15000 20000 25000 30000 35000
With cis: ICAM
0
50
100
150
200
250
300
350
400
0 5000 10000 15000 20000 25000 30000 35000
With cis:
parameters investigation : 1000 MHC on NK cell and target cell (30% cis)1000 MHC on NK cell and target cell (30% cis)::
pSMAC pSMAC cSMAC cSMAC
parameters investigation :
Effects of varying the directed movement factors on Effects of varying the directed movement factors on molecule distributions.molecule distributions.
Without cis:Without cis:
0 2 4 6 8 10 120
20
40
60
80
100
0 2 4 6 8 10 120
20
40
60
80
100
0 2 4 6 8 10 120
20
40
60
80
100
0 2 4 6 8 10 120
20
40
60
80
100
0 2 4 6 8 10 120
20
40
60
80
100
0 2 4 6 8 10 120
20
40
60
80
100
120
0 2 4 6 8 10 120
20
40
60
80
100
120
0 2 4 6 8 10 120
20
40
60
80
100
120
0 2 4 6 8 10 120
20
40
60
80
100
0 2 4 6 8 10 120
20
40
60
80
100
0 2 4 6 8 10 125
10
15
20
25
30
35
0 2 4 6 8 10 125
10
15
20
25
30
35
0 2 4 6 8 10 125
10
15
20
25
30
35
0 2 4 6 8 10 120
10
20
30
40
0 2 4 6 8 10 120
10
20
30
40
1 1.5 2 5 10 .
% N
KG
2D in
ce
nte
r
% K
IR in
ce
nte
r
% L
FA
-1 in
ce
nte
r
Activated receptor and ligand directed movement factor:
Raft: ▲: 10; ■: 5; ♦: 2; ●: 1Raft: ▲: 10; ■: 5; ♦: 2; ●: 1
0 2 4 6 8 10 120
20
40
60
80
100
120
0 2 4 6 8 10 120
20
40
60
80
100
120
Inhibition receptor and ligandInhibition receptor and ligand
parameters investigation:
Effects of varying the directed movement factors on Effects of varying the directed movement factors on molecule distributions.molecule distributions.
With cis:With cis:
0 2 4 6 8 1 0 1 20
2 0
4 0
6 0
8 0
1 0 0
I n h ib it o r y D M0 2 4 6 8 1 0 1 2
0
2 0
4 0
6 0
8 0
1 0 0
0 2 4 6 8 1 0 1 20
2 0
4 0
6 0
8 0
1 0 0
0 2 4 6 8 1 0 1 20
2 0
4 0
6 0
8 0
1 0 0
0 2 4 6 8 1 0 1 20
2 0
4 0
6 0
8 0
1 0 0
1 2 0
0 2 4 6 8 1 0 1 20
2 0
4 0
6 0
8 0
1 0 0
1 2 0
0 2 4 6 8 1 0 1 20
2 0
4 0
6 0
8 0
1 0 0
1 2 0
0 2 4 6 8 1 0 1 20
2 0
4 0
6 0
8 0
1 0 0
1 2 0
0 2 4 6 8 1 0 1 20
2 0
4 0
6 0
8 0
1 0 0
0 2 4 6 8 1 0 1 20
2 0
4 0
6 0
8 0
1 0 0
0 2 4 6 8 1 0 1 25
1 0
1 5
2 0
2 5
3 0
3 5
0 2 4 6 8 1 0 1 25
1 0
1 5
2 0
2 5
3 0
3 5
0 2 4 6 8 1 0 1 25
1 0
1 5
2 0
2 5
3 0
3 5
0 2 4 6 8 1 0 1 25
1 0
1 5
2 0
2 5
3 0
3 5
0 2 4 6 8 1 0 1 20
1 0
2 0
3 0
4 0
Activated receptor and ligand directed movement factor:
% N
KG
2D
in c
ente
r
% K
IR in
ce
nter
% L
FA-1
in
cen
ter
11 1.5 2 5 10 1.5 2 5 10
. .
Activated receptor and ligand directed movement factor:
Raft: ▲: 10; ■: 5; ♦: 2; ●: 1Raft: ▲: 10; ■: 5; ♦: 2; ●: 1
0 2 4 6 8 10 120
20
40
60
80
100
120
0 2 4 6 8 10 120
20
40
60
80
100
120
Inhibition receptor and ligandInhibition receptor and ligand
conclusions:
Conclusions:Conclusions:
Very increasing in integrated signal in program with cis Very increasing in integrated signal in program with cis interaction.interaction.
The synapse not faster with cis (switch point of NKG2D The synapse not faster with cis (switch point of NKG2D not faster) because integrated signal not very increase not faster) because integrated signal not very increase in initial of the synapse.in initial of the synapse.
Less Ly49 in cSMAC with cis, maybe because Less Ly49 in cSMAC with cis, maybe because interruption of cis complexes to Ly49 to move to center interruption of cis complexes to Ly49 to move to center (no Directed Movement to Ly49).(no Directed Movement to Ly49).
As less Ly49 in cSMAC as more NKG2D in cSMAC – less As less Ly49 in cSMAC as more NKG2D in cSMAC – less crowded in center.crowded in center.
Switch point of LFA1 is slower with cis, maybe because Switch point of LFA1 is slower with cis, maybe because interruption of cis complexes to LFA1 to move to center interruption of cis complexes to LFA1 to move to center (no Directed Movement to LFA1).(no Directed Movement to LFA1).