Page 285 - Haematologica March 2020

Long-term eradication of ENKL by rejuvenated CTL
expression. In ENKL cells, lack of LMP1 expression was significantly correlated with PD-L1 expression.
These analyses revealed that distinct PD-L1 expression by ENKL was strongly associated with poor prognosis. CTL therapy directed at LMP1/2 and combined with PD- 1/PD-L1 axis blockade is therefore suggested as useful for the treatment of refractory ENKL.
Both LMP1-specific and LMP2-specific rejT showed strong cytotoxicity against ENKL in vitro
To examine the cytotoxicity of EBV-specific rejT against ENKL, we first determined the HLA type of the ENKL cell line NK-YS (HLA-A*2402) and SNK6 (HLA-A*02). A*2402- and A*0201-restricted LMP1/2-specific CTL clones were generated from healthy donors expressing A*2402 and an ENKL patient (Pt) (Pt 6) who expresses HLA-A*0201. T-iPSC were established from these clones and were redifferentiated into rejT with antigen specifici-
Table 1. Extranodal NK/T-cell lymphoma, nasal type: patient characteristics.
ties that matched those of the respective clones (Figure 2 A-B). We assayed the cytotoxicity of A*2402-restricted LMP2-rejT (TYGPVFMSL), A*0201-restricted LMP1-rejT (YLQQNWWTL), and A*0201-restricted LMP2-rejT (FLY- ALAL) against HLA-matched ENKL by 51Cr release. Strong killing by A*2402-restricted LMP2-rejT was shown against HLA-A*2402+ NK-YS cells (85.3%, 84.7%, 85.2%, and 83.7%; effector : target [E:T] ratios of 40:1, 20:1, 10:1 and 5:1), but not against HLA-A*2402– tumors (18.4%, 4.8%, 1.9%, and 2.4%; E:T ratios of 40:1, 20:1, 10:1 and 5:1). A*0201-restricted LMP1-rejTs also demonstrated strong cytotoxic activity against HLA-A*02+ SNK6 cells (72.0%, 69.9%, 70.4%, and 65.1%; E:T ratios of 40:1, 20:1, 10:1 and 5:1), but not against HLA-A*02– tumors (-2.3%, - 1.7%, 1.3%, and 4.8%; E:T ratios of 40:1, 20:1, 10:1 and 5:1). HLA-A*0201-restricted LMP2-rejT exhibited 76.6% and 24.7% killing at E:T ratios of 40:1 and 20:1 against HLA-A*02+ SNK6 cells, with 14.9% and 14.6% killing for
1 41
F IVB
1.9x103
8.1x102
Notreatment 1 + ++
- - -
- -- - -- - N.D. - - N.D. N.D.
Age Sex Stage EBV-DNA (Years) (copies/mL)
Treatment
Notreatment
Response to Survival treatment duration (months)
PD-1 LMP1 EBNA2
PD-L1 PD-L1 lymphoma macro- TILs
cells phages
M IVB M IVA F IVA M IVB M IIA M IVA M IVA
1% N.D. N.D. 24 +- +- -+- +- +- ---
2.0x106 2.3x102 2.4x101 not detected not detected N.D.
SMILE refractory DEX, VP-16 refractory RT-DeVIC, SMILE CR SMILE, Allo BMT CR RT-DeVIC CR RT-DeVIC, ICE refractory MILD, Allo BMT, DLI refractory
4+ +
1+ +- >63 +- +- >24 +- + >10 +- ++
2 29
3 74
4 73
5 32
6 74
7 53
8
932MIIAN.D.RT-DeVIC CR
>60+-+-N.D.N.D.
10 32
11 71
12 31
13 57
14 29
15 41
16
17
18 78 19
20 84
21 46
22 32
23 33
24 28 25 73
26 62
27 71
28 65
M IVB F IB M IIA F IIA M IVA M IVA
not detected N.D. 2.5x101 1.7x102 N.D. 5.3x105
SMILE, Auto PBSCT CR RT-DeVIC CR SMILE, RT CR RT-DeVIC CR SMILE, Auto PBSCT CR RT-DeVIC, MILD refractory
MILD unknown
RT-DeVIC
SMILE, Allo BMT RT-DeVIC
SMILE, Auto PBSCT RT-DeVIC RT-DeVIC RT-DeVIC, HD-MTX ESHAP, CHOP
RT-DeVIC
>110 +- >124 -+ >44 -
>61 - 4 - 6 -
-
- unknown - - - - - - - - - - -
+ - +- - +- - +- - +- - +- - +- - +- -
- - + - + -
+- - +- - + -
+ N.D. N.D. N.D. + + + - N.D. N.D. N.D. N.D. N.D. N.D. N.D. N.D. N.D. N.D. N.D. N.D.
+ N.D. N.D. N.D. N.D. N.D. N.D. -
M
M 9.9x102
M IVA M IVA F IVB M IVB F IA M IVA M IA M IA F IVA F IVB
M IA
1.1x104
4.5x103 1.3x104 2.3x101 N.D. N.D. N.D. N.D. N.D.
N.D.
unknown 1 CR >108 CR >66 CR 24 CR >40 CR >88 refractory 12 refractory 4 CR >120
+ - +- + - +-
-+ 1%
+- -
++ -
N.D. N.D.
- -
N.D. N.D.
ENKL: extranodal NK/T cell lymphoma, nasal type; EBV: Epstein-Barr virus; PD-1: programmed cell death 1; PD-L1: programmed death-ligand 1; LMP: latent membrane protein; EBNA: Epstein-Barr nuclear antigen; SMILE: dexamethasone, methotrexate, ifosfamide, L-asparaginase and etoposide; DEX: dexamethasone;VP-16, etoposide; RT: radiation therapy; DeVIC: dexamethasone, etoposide, ifosfamide and carboplatin; Allo BMT: allogenetic bone marrow transplantation; ICE: ifosfamide, carboplatin and etoposide; Auto PBSCT: autologous peripheral blood stem cell transplantation; MILD: methotrexate, ifosfamide, L-asparaginase and dexamethasone; HD-MTX: high dose methotrexate; ESHAP: etoposide, methylprednisolone, high dose cytarabine and cisplatin; CHOP: cyclophosphamide, doxorubicin, vincristine and prednisone; CR: complete remission; N.D., not done.
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