the Clinic The V Foundation May 2 nd 2016 Michael A Caligiuri MD Human large g ranular lymphocytes or natural k iller NK cells Human NK cell subsets NK cell receptor biology Clinical application of ID: 634214
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Slide1
Natural Killer Cells: Receptor Biology takes us to the Clinic
The V FoundationMay 2nd, 2016
Michael A. Caligiuri, MDSlide2
Human large granular lymphocytes or natural killer (NK) cells
Human NK cell subsetsNK cell receptor biologyClinical application of
NK cell receptor biology
Monocyte
NK Cell
NK Cell
T Cell
2Slide3
Emerging model for human
NK cell subsets:
innate
immunoregulator
,
CD56
bright CD56bright
CD16dim/neg CD94/NK2GA
NKRs+ KIR+++ CD94/NKG2ATNF-βGM-CSF
TNF-αIFN-gIL-10
High
CytokineProduction
C-kit
L-selectin high
IL-2R
α
IL-2/15R
β
g
c
CD56
bright
NK Cell
3Slide4
Secondary Lymphoid Tissue
Contributions of the CD56bright NK cells in
the human immune response
Early Response
Late Response
monokines
infection
APC
NK
IFN-
γ
CD56
bright
Fehniger
et al, Blood,
2003
APC
NK
T
MHC
TCR
IL-12
IL-2
IFN-
γ
IFN-
γ
IL-12R
IL-2R
CD56
bright
IL-2R
4
T.
FehnigerSlide5
Emerging model for human
NK cell subsets: innate
cytotoxic effector,
CD56
dim
Low
Cytokine
Production KIR CD56dim
CD16brightIL-2/15Rβ
gcPEN5/PSGL1(~70%)Effector Functions+++ ADcCC+++ LAK+++ Natural Cytotoxicity
CD56dim
NK Cell
NKRs
+++ KIR
+ CD94/NKG2A
5Slide6
Natural killing by an NK cell
Getting Inside The Mind Of A Killer
6Slide7
Emerging model for human
NK cell subsets: innate
cytotoxic effector,
CD56
dim
Low
Cytokine
Production KIR CD56dim
CD16brightIL-2/15Rβ
gcPEN5/PSGL1(~70%)Effector Functions+++ ADcCC+++ LAK+++ Natural Cytotoxicity
CD56dim
NK Cell
NKRs
+++ KIR
+ CD94/NKG2A
7Slide8
How do inhibitory NKR work?
KIR sees and engages MHC Class I molecules and send an inhibitory
signal
I
n
the absence of
an activating ligand KIR prevent
autologous NK killing of normal cellsKIR 2DL2Inhibitory ReceptorNKHLA-Cw3SELF
Activating
Receptor
Normal
HematopoieticStem Cell
8Slide9
How do KIR work?
KIR sees and engages MHC Class I molecules and send an inhibitory signal
The KIR inhibitory signal overrides NK killing triggered by activation signal
NO
KILL
Autologous
Leukemia
TargetActivatingLigand
NKActivatingReceptorKIR 2DL2Inhibitory
ReceptorHLA-Cw3
9Slide10
KILL
NK
How do inhibitory NKR work?
Absent or
defective MHC Class
I fails to trigger inhibitory KIR
NK cell kills tumor cell target if tumor expresses ligand to activating receptor
LysisLeukemiaTargetKIR 2DL2Inhibitory Receptor
Activating
LigandActivatingReceptor10Slide11
Human NK cells have KIR that bind to specific EPITOPES that are shared among some but not other MHC class I molecules
Binds HLA
-Cw2, 4, 5, and 6
(Group 2 HLA-C epitopes)
HLA-Cw1, 3, 7, and 8
(Group 1 HLA-C epitopes)
KIR 2DL1
KIR 2DL2
11Slide12
Human NK cells have KIR that bind to specific EPITOPES that are shared among select MHC class I molecules
KIR 2DL2
KIR 2DL2
KIR 2DL2
KIR 2DL2
Cw1
Cw3
Cw7
Cw8
12Slide13
Haplo-mismatch BMT
Donor has KIR on some NK cells that bind to Cw3That KIR is called KIR 2DL2KIR 2DL2 recognizes an epitope shared by group 1 HLA-C,that includes HLA-Cw1, 3, 7, and 8If the recipient AML blast lacks group 1 HLA-C,and expresses a ligand to an activating receptor, lysis occurs
A24 B35 Cw2
A3 B62
Cw3
A24 B35 Cw2
A1 B14
Cw2
Donor
Recipient (leukemia)13Slide14
KILL
NK
Lysis
Leukemia
Target
Activating
Receptor
ActivatingLigandKIR 2DL2Inhibitory
ReceptorHaplo-mismatch BMT
KIR 2DL2 expressed on some donor NK cells recognizes an epitope shared by HLA-Cw1, 3, 7, 8
The AML blast from
recipient
expresses
HLA-Cw2: no
engagement
of KIR
Donor
A24 B35 Cw2
A3 B62 Cw3
HLA-Cw2
(Group 2)
Recipient
A24 B35 Cw2
A24 B35
Cw2
14Slide15
Haplo-mismatch BMT
Donor has KIR on some NK cells that binds to HLA-Cw3That KIR is called KIR 2DL2KIR 2DL2 recognizes an epitope shared by group 1 HLA-C that includes HLA-Cw1, 3, 7, and 8If the recipient AML blast expresses group 1 HLA-C, NO lysis
A24 B35 Cw2
A3 B62
Cw3
A24 B35 Cw2
A1 B14
Cw8
Donor
Recipient (leukemia)15Slide16
Human NK cells have KIR that bind to specific EPITOPES that are shared among select MHC class I molecules
KIR 2DL2
KIR 2DL2
KIR 2DL2
KIR 2DL2
Cw1
Cw3
Cw7
Cw8
16Slide17
Haplo-mismatch BMT
KIR 2DL2 expressed on
donor NK cells
recognizes
an
epitope shared
by HLA-Cw1, 3, 7, 8
The AML blast from recipient expressesHLA-Cw8: engagement of KIRDonorA24 B35 Cw2A3 B62 Cw3RecipientA24 B35 Cw2A24 B35
Cw8
NO
KILL
Leukemia
TargetNo Lysis
ActivatingLigand
NK
Activating
Receptor
Inhibitory
KIR2DL2
(group
1
specific
)
HLA-Cw8
(group 1)
17Slide18
Haplo-mismatch BMT
DonorA24 B35 Cw2A3 B62 Cw3
Recipient
A24 B35 Cw2
A24 B35
Cw8
DonorA24 B35 Cw2A3 B62 Cw3RecipientA24 B35 Cw2A24 B35 Cw2
NO
KILL
Leukemia
Target
No Lysis
NK
Inhibitory
KIR2DL2
(group
1
specific
)
HLA-Cw8
(group 1)
KILL
NK
Lysis
Leukemia
Target
Activating
Receptor
Activating
Ligand
KIR
2DL2
Inhibitory
Receptor
HLA-Cw2
(Group 2)
Activating
Receptor
Activating
Ligand
18Slide19
Relevant clinical issues for haploidentical KIR epitope incompatible transplantation for AML in CR
Do DONOR stem cells, infused without T-cells, get rejected by HOST NK when KIR are incompatible in host vs graft direction?Do DONOR NK cells, upon encountering KIR incompatibility with normal HOST tissues, cause graft versus HOST disease in the absence of T-cells?Do DONOR NK cells, upon encountering KIR incompatibility with HOST leukemia, cause a graft versus LEUKEMIA effect in the absence of T-cells?
19Slide20
Role of NK cell alloreactivity in haploidentical KIR epitope incompatible transplantation for AML in CR
60 HLA haplotype-mismatched, T-cell depleted hematopoietic stem cell transplantations29 of 60 donor-recipient pairs had appropriate KIR epitope incompatibility31 of 60 donor-recipient pairs did not have appropriate KIR epitope incompatibility
No donor graft rejection in KIR mismatch patients
No graft versus host disease in KIR mismatch patients
In vitro analysis of donor NK cell lysis of host target cells was consistent with
“
rules” up to three months after the transplantL. Ruggeri et al, Blood, 2001; Science, 2002; update, 200520Slide21
60 AML patients transplanted in CR
Leukemia
Relapse
No KIR Mismatch
(
n = 31)
KIR Mismatch
(
n = 29)
p = 0.002
Years
0
2
4
6
8
10
0.0
0.2
0.4
0.6
0.8
1.0
12
Event-Free
S
urvival
No KIR Mismatch
KIR Mismatch
p = 0.03
Years
0
2
4
6
8
10
0.0
0.2
0.4
0.6
0.8
1.0
12
Ruggeri
et
al,
Science 2002, updated
2005
21Slide22
Role of NK cell alloreactivity in HLA-mismatched hematopoietic stem cell transplantation
ConclusionsIn mismatched transplants, a clinical graft-versus leukemia effect, independent of T-cell-mediated graft versus host disease, appears to positively impact on relapse rate when NK cell KIR epitope incompatibility is in the graft versus host directionAdditional study of donor versus recipient NK cell alloreactivity in mismatched hematopoietic stem cell transplants has confirmed these results
22Slide23
NK
Autologous
Leukemia/MM
Target
Activating
Receptor
Activating
LigandBlocking KIR in autologoussetting for AML and Multiple Myeloma (MM)
D. BensonLysisKILL
Benson et al, Clin Cancer Res, 201523Slide24
NK
GBM
Activating
Receptor
Activating
Ligand
Adding a Chimeric Antigen
Receptor to NK CellsRecognizing EGFR for Glioblastoma Multiforme (GBM)
LysisCARHan et al, Scientific Reports, 201524
EGFRJ. YuSlide25
NK92-CAR-EGFR Cells Reside in situ CNS GBM
25NK-92
10 × 10
2 × 10
4 × 10
Patient-Derived
GBM Tumor
M NC PC brain lung liver BL
Sp
BM
EGFR-CAR
371bp
CONFIDENTIALSlide26
Single injection of NK92-CAR-EGFR Cells Inhibit GBM Tumor Growth in vivo and Prolong Survival of Mice Implanted with U251 GBM Cells & Primary GBM
26
Primary GBM
NK92-CAR-EV
NK92-CAR
-EGFR
Vehicle
p/s/cm²/
sr
(×10
6
)
NK92-CAR
-EGFR
*
U251
NK92-CAR-EV
Vehicle
*
% Survival
Days
Days
% Survival
*
**
*
HBSS
NK92-CAR-EV
NK92-CAR-EGFR
U251
Days
U251
Vehicle
NK92-CAR-EV
NK92-CAR-EGFRSlide27
NK cells: biology to the clinic
SummaryHuman NK cell subsetsNK cell receptor biologyClinical application of NK cell receptor biology
27Slide28
caligiurilab.com
28