TABLE 3.
Summary of On-target and Near Off-target Activity of HPV E629–38 TCR and Examples of Light-chain and Heavy-chain Optimized CARs in Jurkat:T2 and Acute Primary T-Cell Assays
| Jurkat:T2 Assay | Primary T-Cell Assay | ||||||||
|---|---|---|---|---|---|---|---|---|---|
| Constructs | Construct Format/Source | Test Peptide Name | Test Peptide Sequence | On/Off Target | EC50 (nM) | ECminimum (nM) | Emaximum (RLU) | Selectivity | Primary T-Cell EC 50 (nM) |
| C882 | TCR | HPV E711–19 | YMLDLQPET | On | 0.01 | 0.007 | 11,000 | NA | 0.05 |
| C881 | TCR | HPV E629–38 | TIHDIILECV | On | 0.6 | 0.05 | 18,000 | NA | 0.4 |
| PRR5 | NLVDQILESV | Off | >100,000 | NA | NA | >167 | ND | ||
| PTPRD | FIHDALLEAV | Off | >100,000 | NA | NA | >167 | ND | ||
| C1044 | Primary CAR | HPV E629–38 | TIHDIILECV | On | 2200 | NA | 17,315 | NA | NA |
| C1053 | Primary CAR | HPV E629–38 | TIHDIILECV | On | 1500 | NA | 30,000 | NA | NA |
| C1068 | Primary CAR | HPV E629–38 | TIHDIILECV | On | 930 | NA | 34,000 | NA | NA |
| C1072 | Primary CAR | HPV E629–38 | TIHDIILECV | On | 1100 | NA | 7500 | NA | NA |
| C1073 | Primary CAR | HPV E629–38 | TIHDIILECV | On | 3400 | NA | 9200 | NA | NA |
| C1078 | Primary CAR | HPV E629–38 | TIHDIILECV | On | 550 | NA | 12,500 | NA | NA |
| C1987 | Light-chain optimized CAR (parent of CT503) | HPV E629–38 | TIHDIILECV | On | 28 | 0.56 | 50,000 | NA | 5235 |
| PRR5 | NLVDQILESV | Off | >250 | NA | NA | >9 | ND | ||
| PTPRD | FIHDAILEA | Off | 800 | ND | 38,000 | 29 | ND | ||
| C2388 | Light-chain optimized CAR | HPV E629–38 | TIHDIILECV | On | 7.7 | 0.28 | 71,000 | NA | 600 |
| PRR5 | NLVDQILESV | Off | 88 | ND | 57,000 | 11 | ND | ||
| PTPRD | FIHDAILEA | Off | >100,000 | NA | NA | >12,000 | ND | ||
| C1992 | Light-chain optimized CAR (parent of CT512) | HPV E629–38 | TIHDIILECV | On | 5 | 0.02 | 114,000 | NA | 108 |
| PRR5 | NLVDQILESV | Off | 930 | ND | 55,000 | 186 | ND | ||
| PTPRD | FIHDALLEAV | Off | >100,000 | NA | NA | >20,000 | ND | ||
| CT503 | Light-chain and heavy-chain optimized CAR | HPV E629–38 | TIHDIILECV | On | 0.5 | 0.02 | 30,000 | NA | 6.5 |
| PRR5 | NLVDQILESV | Off | 121 | ND | 28,000 | 242 | ND | ||
| PTPRD | FIHDALLEAV | Off | 34 | ND | 30,000 | 68 | ND | ||
| CT512 | Light-chain and heavy-chain optimized CAR | HPV E629–38 | TIHDIILECV | On | 0.2 | 0.06 | 30,000 | NA | 5.2 |
| PRR5 | NLVDQILESV | Off | 3.5 | ND | 27,000 | 18 | ND | ||
| PTPRD | FIHDALLEAV | Off | 280 | ND | 5200 | 1400 | ND | ||
CAR indicates chimeric antigen receptor; HPV, human papillomavirus; NA, not applicable; ND, not determined; RLU, relative luminescence unit; TCR, T-cell receptor.