TABLE 3.
Ebola virus protein sequences recognized by murine CD8+ T cellsa
EBOV protein | Epitope sequence | Amino acid position | Restriction | Result of indicated assay
|
Survivale | ||
---|---|---|---|---|---|---|---|
IFN-γ ICCb | IFN-γ ELISPOTc | 51Crd | |||||
GP | VSTGTGPGAGDFAFHK | 141-155 | H-2d | 0.24/0.02 | ND | 47 | 10/10 |
WIPYFGPAAEGIYTE | 531-545 | H-2b | 0.4/0.08 | 715 | 3 | 0/50 | |
NP | VYQVNNLEEIC | 44-52 | H-2b | 1.06/0.11 | 410 | 73 | 10/10 |
GQFLSFASL | 148-156 | H-2b | 0.88/0.11 | 295 | 45 | 19/20 | |
SFKAALSSL | 279-287 | H-2b | 0.63/0.04 | ND | 43 | 17/18 | |
DAVLYYHMM | 663-671 | H-2b | 0.99/0.11 | 910 | 57 | 17/20 | |
VP24 | KFINKLDALH | 159-168 | H-2d | 0.52/0.09 | 535 | 46 | 20/20 |
NYNGLLSSI | 171-179 | H-2d | 0.38/0.09 | 135 | 51 | 20/20 | |
PGPAKFSLL | 214-222 | H-2d | 3.34/0.09 | 957 | 43 | 18/20 | |
VP30 | KFSKSQLSLLCETHLR | 181-196 | H-2d | 0.12/0.3 | ND | 55 | 18/20 |
181-196 | H-2b | 0.45/0.15 | 190 | 47 | 19/20 | ||
DLQSLIMFITAFLNI | 231-245 | H-2b | 0.7/0.15 | 165 | 32 | 15/20 | |
231-245 | H-2d | 0.42/0.17 | ND | 38 | 16/20 | ||
VP35 | CDIENNPGL | 45-53 | H-2b | 1.99/0.15 | 40 | 80 | 19/20 |
MVAKYDHL | 138-145 | H-2b | 1.63/0.15 | 249 | 79 | 20/20 | |
TVPQSVREAFNNL | 190-202 | H-2d | 0.62/0.23 | ND | 43 | 18/20 | |
RNIMYDHL | 225-323 | H-2b | 1.53/0.15 | 202 | 87 | 19/20 | |
PGFGTAFHQLVQVICK | 233-248 | H-2d | 0.63/0.23 | ND | 47 | 17/20 | |
VP40 | LRIGNQAFLQEFVLPP | 150-165 | H-2b | 0.7/0.05 | 101 | 53 | 19/20 |
AFLQEFVLPPVQLPQ | 160-175 | H-2d | 0.45/0.22 | ND | 47 | 17/20 | |
YFTFDLTALK | 171-180 | H-2d | 0.41/0.22 | ND | 38 | 16/20 | |
TESPEKIQAI | 232-141 | H-2d | 0.6/0.22 | ND | 37 | 18/20 | |
Lassa N | RPLSAGVYMGNLSSQ | H-2b | 1.5/0.14 | 425 | 0/20 | ||
H-2d | 0.7/0.18 | 325 | 0/10 | ||||
MLR | H-2b | 0/40 | |||||
H-2d | 0/20 | ||||||
PBS | H-2b | 0/20 | |||||
H-2d | 0/20 |
Mice were vaccinated with 2.0 × 106 IU of VRP expressing Ebola virus GP, NP, VP24, VP30, VP35, or VP40. On day 28, mice were given a booster, and 7 days later, splenocytes were collected. In some cases (GP, VP35, VP30, and VP40 in BALB/c mice [H-2d]) mice received multiple booster vaccinations or larger doses of vaccine before sequences containing potential epitopes were identified.
Splenocytes were used ex vivo for identification of peptides that induced IFN-γ-expressing CD8+ T cells. Data shown represent intracellular IFN-γ data after 5 h of restimulation with peptide. The data are expressed as follows: percentage of peptide-induced IFN-γ-positive/percentage of background IFN-γ-positive cells.
Data are expressed as numbers of spot-forming cells per 106 peripheral blood mononuclear cells in the IFN-γ ELISPOT assay. Representative data are shown for one experiment. ND, not detected in assay.
Data are expressed as percent specific lysis in a chromium release assay with haplotype-matched, peptide-pulsed targets at a 25:1 effector:target ratio, except for the GP107 peptide, which is the specific lysis at a 33:1 ratio.
Data are expressed as numbers of mice surviving challenge/number tested. Peptide-specific restimulated lymphocytes were adoptively transferred to filovirus-naïve mice approximately 4 h before mice were challenged with 1,000 PFU of mouse-adapted Ebola virus. Control mice received PBS, or 5 × 106 cells from a mixed-lymphocyte reaction culture or 5 × 106 Lassa N-specific CTLs. No clinical symptoms were observed in any surviving mice.