KIR-ligand, KIR, NK alloreactive subsets, and lytic capability in the various donor/recipient pairs
| Case . | KIR-ligands (donor)* . | KIR-ligands (patient)† . | Permissive inhibitory KIR (donor)‡ . | Activating KIR (donor)§ . | Alloreactive subset, % (donor)¶ . | Alloreactive subset, % (recipient post-HSCT)¶ . | Lysis vs B-EBV, % (donor)# . | Lysis vs B-EBV, % (recipient post-HSCT)# . |
|---|---|---|---|---|---|---|---|---|
| 1 | Bw4, C1, C2 | Bw4, C2 | 2DL2/3 3DL2 | 2DS1 2DS2 3DS1 | 26 | 54 (7 mo) | 31 (60) | 57 (68) |
| 2 | Bw4, C1, C2 | Bw4, C2 | 2DL2/3 3DL2 | 2DS2 | 6 | 8 (3 mo) | 1 (57) | 1 (71) |
| 3 | Bw4, C1, C2 | Bw4, C2 | 2DL2/3 3DL2 | 2DS2 | 36 | 23 (2 mo) | 57 (83) | 27 (76) |
| 4 | A3, Bw4, C1, C2 | Bw4, C2 | 2DL2/3 3DL2 | 2DS2 3DS1 | 15 | 3 (2 mo) | 2 (36) | Nd |
| 5 | Bw4, C1, C2 | Bw4, C2 | 2DL2/3 3DL2 | 2DS1 2DS2 3DS1 | 36 | 34 (2 mo) | 52 (42) | 45 (55) |
| 6 | Bw4, C1, C2 | A11, Bw4, C2 | 2DL2/3 | 2DS1 2DS2 3DS1 | 9 | 69 (3 mo) | 19 (51) | 7 (40) |
| 7 | Bw4, C1, C2 | Bw4, C2 | 2DL3 3DL2 | NO | 2 | 3 (2 mo) | 6 (53) | 0 (59) |
| 8 | Bw4, C1, C2 | Bw4, C2 | 2DL3 3DL2 | 2DS1 | 5 | 17 (3 mo) | 12 (74) | 39 (83) |
| 9 | Bw4, C1, C2 | Bw4, C1 | 2DL1 3DL2 | 2DS2 | (92) | (2) (3 mo) | 80 (82) | 32 (90) |
| 10 | Bw4, C1, C2 | Bw4, C1 | 2DL1 3DL2 | 2DS2 | (3) | (0.5) (3 mo) | 11 (76) | 27 (80) |
| 11 | C1, C2 | Bw4, C1 | 2DL1 3DL2 | NO | 9 | 10 (5 mo) | 20 (83) | 26 (91) |
| 12 | Bw4, C1, C2 | Bw4, C1 | 2DL1 3DL2 | 2DS1 3DS1 | 11 | 8 (3 mo) | 58 (100) | 42 (97) |
| 13 | Bw4, C1, C2 | Bw4, C1 | 2DL1 3DL2 | 2DS2 | (1) | (0.5) (3 mo) | 25 (96) | 10 (62) |
| 14 | Bw4, C1, C2 | Bw4, C1 | 2DL1 3DL2 | 2DS1 3DS1 | 6 | 3 (7 mo) | 43 (71) | 26 (84) |
| 15 | A11, Bw4, C1, C2 | Bw4, C1 | 2DL1 3DL2 | 2DS1 3DS1 | 13 | 14 (6 mo) | 27 (83) | 14 (88) |
| 16 | Bw4, C1 | C1, C2 | 3DL1 3DL2 | NO | 3 | 3 (2 mo) | 15 (61) | 13 (66) |
| 17 | Bw4, C1, C2 | C1, C2 | 3DL1 3DL2 | NO | 5 | 5 (3 mo) | 26 (83) | 43 (81) |
| 18 | Bw4, C1, C2 | C1, C2 | 3DL1 3DL2 | NO | 8 | 4 (6 mo) | 34 (95) | 23 (66) |
| 19 | Bw4, C1, C2 | A3, C1, C2 | NO | 2DS1 3DS1 | 2 | 2 (3 mo) | n.d. | 10 (56) |
| 20 | Bw4, C1 | C1, C2 | NO 3DL2 | 2DS1 3DS1 | 2 | 21 (6 mo) | 26 (59) | 66 (78) |
| 21 | A3, Bw4, C2 | Bw4, C1, C2 | 3DL2 | NO | 0 | 7 (13 mo) | 14 (43) | 27 (75) |
| Case . | KIR-ligands (donor)* . | KIR-ligands (patient)† . | Permissive inhibitory KIR (donor)‡ . | Activating KIR (donor)§ . | Alloreactive subset, % (donor)¶ . | Alloreactive subset, % (recipient post-HSCT)¶ . | Lysis vs B-EBV, % (donor)# . | Lysis vs B-EBV, % (recipient post-HSCT)# . |
|---|---|---|---|---|---|---|---|---|
| 1 | Bw4, C1, C2 | Bw4, C2 | 2DL2/3 3DL2 | 2DS1 2DS2 3DS1 | 26 | 54 (7 mo) | 31 (60) | 57 (68) |
| 2 | Bw4, C1, C2 | Bw4, C2 | 2DL2/3 3DL2 | 2DS2 | 6 | 8 (3 mo) | 1 (57) | 1 (71) |
| 3 | Bw4, C1, C2 | Bw4, C2 | 2DL2/3 3DL2 | 2DS2 | 36 | 23 (2 mo) | 57 (83) | 27 (76) |
| 4 | A3, Bw4, C1, C2 | Bw4, C2 | 2DL2/3 3DL2 | 2DS2 3DS1 | 15 | 3 (2 mo) | 2 (36) | Nd |
| 5 | Bw4, C1, C2 | Bw4, C2 | 2DL2/3 3DL2 | 2DS1 2DS2 3DS1 | 36 | 34 (2 mo) | 52 (42) | 45 (55) |
| 6 | Bw4, C1, C2 | A11, Bw4, C2 | 2DL2/3 | 2DS1 2DS2 3DS1 | 9 | 69 (3 mo) | 19 (51) | 7 (40) |
| 7 | Bw4, C1, C2 | Bw4, C2 | 2DL3 3DL2 | NO | 2 | 3 (2 mo) | 6 (53) | 0 (59) |
| 8 | Bw4, C1, C2 | Bw4, C2 | 2DL3 3DL2 | 2DS1 | 5 | 17 (3 mo) | 12 (74) | 39 (83) |
| 9 | Bw4, C1, C2 | Bw4, C1 | 2DL1 3DL2 | 2DS2 | (92) | (2) (3 mo) | 80 (82) | 32 (90) |
| 10 | Bw4, C1, C2 | Bw4, C1 | 2DL1 3DL2 | 2DS2 | (3) | (0.5) (3 mo) | 11 (76) | 27 (80) |
| 11 | C1, C2 | Bw4, C1 | 2DL1 3DL2 | NO | 9 | 10 (5 mo) | 20 (83) | 26 (91) |
| 12 | Bw4, C1, C2 | Bw4, C1 | 2DL1 3DL2 | 2DS1 3DS1 | 11 | 8 (3 mo) | 58 (100) | 42 (97) |
| 13 | Bw4, C1, C2 | Bw4, C1 | 2DL1 3DL2 | 2DS2 | (1) | (0.5) (3 mo) | 25 (96) | 10 (62) |
| 14 | Bw4, C1, C2 | Bw4, C1 | 2DL1 3DL2 | 2DS1 3DS1 | 6 | 3 (7 mo) | 43 (71) | 26 (84) |
| 15 | A11, Bw4, C1, C2 | Bw4, C1 | 2DL1 3DL2 | 2DS1 3DS1 | 13 | 14 (6 mo) | 27 (83) | 14 (88) |
| 16 | Bw4, C1 | C1, C2 | 3DL1 3DL2 | NO | 3 | 3 (2 mo) | 15 (61) | 13 (66) |
| 17 | Bw4, C1, C2 | C1, C2 | 3DL1 3DL2 | NO | 5 | 5 (3 mo) | 26 (83) | 43 (81) |
| 18 | Bw4, C1, C2 | C1, C2 | 3DL1 3DL2 | NO | 8 | 4 (6 mo) | 34 (95) | 23 (66) |
| 19 | Bw4, C1, C2 | A3, C1, C2 | NO | 2DS1 3DS1 | 2 | 2 (3 mo) | n.d. | 10 (56) |
| 20 | Bw4, C1 | C1, C2 | NO 3DL2 | 2DS1 3DS1 | 2 | 21 (6 mo) | 26 (59) | 66 (78) |
| 21 | A3, Bw4, C2 | Bw4, C1, C2 | 3DL2 | NO | 0 | 7 (13 mo) | 14 (43) | 27 (75) |
KIR-ligands expressed in donor HLA-class I genotype; the KIR ligand-mismatch in GVH direction is in bold.
KIR-ligands expressed in patient HLA-class I genotype.
Expressed donor KIR permissive for alloreactivity (which do not recognize patient's HLA-class I alleles); in bold is the KIR specific for KIR ligand-mismatch. Cases no. 19 and 20 were missing KIR3DL1, which is the KIR specific for Bw4-mismatch.
KIR2DS1, 2DS2 and 3DS1 are indicated when expressed in donor KIR genotype. For expression of the other activating KIR (KIR2DS4, 2DS3 and 2DS5) see Table S1.
The percentage was evaluated in polyclonal NK-cell populations by staining with an appropriate combination of anti-KIR and anti-NKG2A mAb followed by appropriate fluorochrome-conjugated second reagents using the methodology described in Figure 2A. Percentages that could not be precisely calculated due to the presence of activating KIR (undistinguishable from the inhibitory form, such as KIR2DS2 and KIR2DL2) are indicated in parentheses. The percentage of alloreactive subset in the recipient (patient after HSCT) was evaluated at the indicated time point (months after HSCT).
Percent-specific lysis, either in the absence or in the presence (in parentheses) of anti-HLA class I mAb, was evaluated in polyclonal NK-cell populations derived from the donor or the recipient. Their cytolytic assays were performed in parallel (ie, at the same time) to phenotypic evaluations. Target cells were represented by appropriate B-EBV cell lines expressing the same KIR-ligands of the patient (E:T of 20:1).