Interferon-γ ELISpot responses before and after allogeneic hematopoietic cell transplantation
| Virus . | Patient 1 . | Patient 2 . | ||||||||
|---|---|---|---|---|---|---|---|---|---|---|
| HLA restriction . | Protein and region . | Peptide sequence . | IFN-γ SFC* . | HLA restriction . | Protein and region† . | Peptide sequence . | IFN-γ SFC* . | |||
| Pre-HCT . | Day +81 . | Pre-HCT . | Day +81 . | |||||||
| CMV† | Pool | 678 | 487 | Pool | 0 | ND | ||||
| EBV† | Pool 1 | 435 | 1180 | Pool 1 | 3017 | 980 | ||||
| Pool 2 | 1870 | 137 | Pool 2 | 150 | 110 | |||||
| Pool 3 | 1013 | 620 | Pool 3 | 171 | 130 | |||||
| A2 | BMLFI (259-267) | GLCTLVAML | 120 | 120 | B8 | EBNA3A (158– 166) | QAKWRLQTL | 150 | ND | |
| A24 | BRLFI (28-37) | DYCNVLNKEF | 955 | 503 | BZLF1 (190–197) | RAKFKQLL | 3492 | 1110 | ||
| B27 | BRLFI (28-37) | RRIYDLIEL | 1455 | 237 | B35 | EBNA3A (458– 466) | YPLHEQHGM | 158 | 150 | |
| FluA† | A2 | Matrix 1 (58-66) | GILGFVFTL | 205 | 77 | B8 | NP (380–388) | ELRSRYWAI | 117 | − |
| B27 | NP (383-391) | SRYWAIRTR | 53 | 350 | ||||||
| HIV | A2 | Gag p17 (77-85) | SLYNTVATL | − | 67 | A26 | Gag p24 (35-43) | EVIPMFSAL | 142 | − |
| Env gp160 (311-320) | RGPGRAFVTI | 120 | 90 | B8 | Gag p17 (24-32) | GGKKKYKL | 129 | 50 | ||
| RT(33-41) | ALVEICTEM | − | 120 | Gag p24 (128-135) | EIYKRWII | 54 | − | |||
| Pol (22-31) | MASDFNLPPV | 195 | 63 | Nef (90-97) | FLKEKGGL | 158 | 50 | |||
| Vpr (59-67) | AIIRILQQL | 155 | − | RT (118-127) | VPLDEDFRKY | − | 100 | |||
| Nef (136-145) | PLTFGWCYKL | 170 | − | RT (18-26) | GPKVKQWPL | − | 90 | |||
| A24 | Gag p17 (28-36) | KYKLKHIVW | − | 53 | Env gp41 (95-103) | TAVPWNASW | − | 50 | ||
| B27 | Gag p24 (131-140) | KRWIILGLNK | 805 | − | ||||||
| Env gp160 (786-795) | GRRGWEALKY | − | 257 | |||||||
| Nef (105-114) | RRQDILDLWI | − | 77 | |||||||
| Cw12 | Tat (32-37) | CCFHCQVC | 100 | 67 | ||||||
| Virus . | Patient 1 . | Patient 2 . | ||||||||
|---|---|---|---|---|---|---|---|---|---|---|
| HLA restriction . | Protein and region . | Peptide sequence . | IFN-γ SFC* . | HLA restriction . | Protein and region† . | Peptide sequence . | IFN-γ SFC* . | |||
| Pre-HCT . | Day +81 . | Pre-HCT . | Day +81 . | |||||||
| CMV† | Pool | 678 | 487 | Pool | 0 | ND | ||||
| EBV† | Pool 1 | 435 | 1180 | Pool 1 | 3017 | 980 | ||||
| Pool 2 | 1870 | 137 | Pool 2 | 150 | 110 | |||||
| Pool 3 | 1013 | 620 | Pool 3 | 171 | 130 | |||||
| A2 | BMLFI (259-267) | GLCTLVAML | 120 | 120 | B8 | EBNA3A (158– 166) | QAKWRLQTL | 150 | ND | |
| A24 | BRLFI (28-37) | DYCNVLNKEF | 955 | 503 | BZLF1 (190–197) | RAKFKQLL | 3492 | 1110 | ||
| B27 | BRLFI (28-37) | RRIYDLIEL | 1455 | 237 | B35 | EBNA3A (458– 466) | YPLHEQHGM | 158 | 150 | |
| FluA† | A2 | Matrix 1 (58-66) | GILGFVFTL | 205 | 77 | B8 | NP (380–388) | ELRSRYWAI | 117 | − |
| B27 | NP (383-391) | SRYWAIRTR | 53 | 350 | ||||||
| HIV | A2 | Gag p17 (77-85) | SLYNTVATL | − | 67 | A26 | Gag p24 (35-43) | EVIPMFSAL | 142 | − |
| Env gp160 (311-320) | RGPGRAFVTI | 120 | 90 | B8 | Gag p17 (24-32) | GGKKKYKL | 129 | 50 | ||
| RT(33-41) | ALVEICTEM | − | 120 | Gag p24 (128-135) | EIYKRWII | 54 | − | |||
| Pol (22-31) | MASDFNLPPV | 195 | 63 | Nef (90-97) | FLKEKGGL | 158 | 50 | |||
| Vpr (59-67) | AIIRILQQL | 155 | − | RT (118-127) | VPLDEDFRKY | − | 100 | |||
| Nef (136-145) | PLTFGWCYKL | 170 | − | RT (18-26) | GPKVKQWPL | − | 90 | |||
| A24 | Gag p17 (28-36) | KYKLKHIVW | − | 53 | Env gp41 (95-103) | TAVPWNASW | − | 50 | ||
| B27 | Gag p24 (131-140) | KRWIILGLNK | 805 | − | ||||||
| Env gp160 (786-795) | GRRGWEALKY | − | 257 | |||||||
| Nef (105-114) | RRQDILDLWI | − | 77 | |||||||
| Cw12 | Tat (32-37) | CCFHCQVC | 100 | 67 | ||||||
CMV indicates cytomegalovirus; EBV, Epstein-Barr virus; HIV, human immunodeficiency virus; IFN, interferon; PBMC, peripheral blood mononuclear cell; and SFC, spot-forming cell.
The value of SFC/106 PBMC is shown for all positive responses. Negative responses are indicated by −. Patient 1 had no response in either the pre- or posttransplant samples to the following epitopes: HLA-A2–restricted: Env gp160 (813-822) SLLNATDIAV, RT (179-187) VIYQYMDDL, RT (309-317) ILKEPVHGV, Nef (180-189) VLEWRFDSRL; HLA-A2, A24 restricted: Nef (190-198) AFHHVAREL; HLA-A24–restricted: Gag p24 (162-172) RDYVDRFFKTL, Env gp160 (52-61) LFCASDAKAY, Env gp160 (585-593) RYLKDQQLL, Gag p17 (19-27) IRLRPGGKK; HLA-B39–restricted: Gag p24 (61-69) GHQAAMQML, HLA-Cw1–restricted: Gag p24 (36-43) VIPMFSAL, Gag_Pol_TF (24-31) NSPTRREL, Env gp160 (217-226) YCAPAGFAIL, Env gp160 (712-720) YSPLSLQTL, Vpu (74-82) HAPWDVNDL. Patient 2 had no response in either the pre or posttransplant samples to the following epitopes: HLA-B8–restricted: Gag p17 (74-81) ELRSLYNTV, Gag p24 (197-205) DCKTILKAL, Env gp120 (2-10) RVKEKYQHL, Env gp41 (75-82) YLKDQQLL, Nef (13-20) WPTVRERM, Env gp41 (337-345) LQGLERALL; HLA-B35–restricted: Gag p17 (36-44) WASRELERF, Gag p17 (124-132) NSSKVSQNY, Gag p24 (122-130) PPIPVGDIY, RT (107-115) TVLDVGDAY, RT (175-183) NPDIVIYQY, RT (175-183) HPDIVIYQY, Env gp120 (42-52) VPVWKEATTTL, Env gp120 (78-86) DPNPQEVVL, Nef (74-81) VPLRPMT.
T-cell responses to EBV, CMV, and FluA epitopes were used as controls. The CMV and EBV peptide pools consisted of published HLA class I–restricted CD8+ T-cell epitopes.