Table 1.
Results of ELISPOT, binding assay and PREDBALB/c prediction for the 9mer peptides from the CD8-positive sequences of the yellow fever envelope protein
| 9mer position (Yellow Fever Envelope) |
Sequence |
# IFN-γ SFC (CD4-depleted splenocytes/106) (2) | Binding assay (IC50 in nmol) (PREDBALB/c score) |
||
|---|---|---|---|---|---|
| E57 to E75 | RKVCYNAVLTHVKINDKCP(1) | Dd (3) | Kd (3) | Ld (3) | |
| 57–65 | RKVCYNAVL(4) | 1 | > 70,000(5) | > 70,000 | 39,442 |
| (7.98) | (5.72) | (4.70) | |||
| 58–66 | KVCYNAVLT | 93 | > 70,000 | > 70,000 | > 70,000 |
| (2.32) | (1.84) | (0.90) | |||
| 59–67 | VCYNAVLTH | 265 | > 70,000 | 211⁎ | 55,368 |
| (7.98) | (1.60) | (4.62) | |||
| 60–68 | CYNAVLTHV | 403 | > 70,000 | 1.0⁎ | > 70,000 |
| (6.48) | (8.66⁎⁎) | (5.18) | |||
| 61–69 | YNAVLTHVK | 143 | > 70,000 | 93⁎ | > 70,000 |
| (3.46) | (1.92) | (0.90) | |||
| 62–70 | NAVLTHVKI | 7 | > 70,000 | 1542 | > 70,000 |
| (8.42⁎⁎) | (5.56) | (0.30) | |||
| 63–71 | AVLTHVKIN | 60 | > 70,000 | 54,670 | 20,708 |
| (4.10) | (1.10) | (0.88) | |||
| 64–72 | VLTHVKIND | 60 | > 70,000 | > 70,000 | > 70,000 |
| (2.94) | (5.22) | (2.22) | |||
| 65–73 | LTHVKINDK | 51 | > 70,000 | > 70,000 | > 70,000 |
| (4.02) | (1.60) | (1.50) | |||
| 66–74 | THVKINDKC | 1 | > 70,000 | > 70,000 | > 70,000 |
| (0.80) | (2.26) | (0.40) | |||
| 67–75 | HVKINDKCP | 6 | > 70,000 | > 70,000 | 48,666 |
| (1.64) | (0.58) | (2.5) | |||
| E329 to E343 | PCRIPVIVADDLTAA(6) | ||||
| 329–337 | PCRIPVIVA | 158 | > 70,000 | > 70,000 | > 70,000 |
| (4.78) | (0.50) | (0.44) | |||
| 330–338 | CRIPVIVAD | 371 | > 70,000 | 915 | 54,239 |
| (2.36) | (2.40) | (2.12) | |||
| 331–339 | RIPVIVADD | 140 | > 70,000 | > 70,000 | > 70,000 |
| (8.50⁎⁎) | (4.18) | (1.42) | |||
| 332–340 | IPVIVADDL | 63 | > 70,000 | 33,005 | 56⁎ |
| (6.72) | (6.00) | (7.50) | |||
| 333–341 | PVIVADDLT | 26 | > 70,000 | > 70,000 | 3719 |
| (3.40) | (1.80) | (0.00) | |||
| 334–342 | VIVADDLTA | 86 | > 70,000 | > 70,000 | 19,242 |
| (2.70) | (3.10) | (1.50) | |||
| 335–343 | IVADDLTAA | 46 | > 70,000 | > 70,000 | 38,032 |
| (3.46) | (1.96) | (0.90) | |||
(1) Sequence of 19 amino acids that correspond to 15mer peptides E57–71 and E61–75 that were positive for CD8 response by ELISPOT (Fig. 2B); (2) peptides were considered ELISPOT positives when > 20 SFC/106 cells; (3) molecules of H2d class I; (4) sequence of each 9mer in relation to the sequence from where it is found in the envelope protein; (5) IC50 for the binding assay for class I molecule (nmol); (6) sequence of 15 amino acids from the 15mer peptide E329–343 (Fig. 2B); ⁎values of IC50 below the cutoff of 500 nmol, suggestive of a possible binder peptide; ⁎⁎ score above 8 from PREDBALB/c software prediction.