Table 11 Protocol features for Prodigy systems
From: Lymphocyte expansion in bioreactors: upgrading adoptive cell therapy
Authors | Protocol features | Starting material | Culturing | Expansion | Purity | Functionality | |||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
IL-2 [IU/mL] | Stimmul. strategy | Medium | Serum | Source | Seed [cell/ml] | Vol. [mL] | Tubing Set | Shaking | Fold | Days | Compared system | Comp. Fold | |||
T cells | |||||||||||||||
Mock [187] - 2016 | Used | CD3/CD28; Transd. | TexMACS GMP | 3% HS | T cells from PBMC | 7.45–10 × 108 | 100 | TS520 | Shaker from day 4/5. More vigorous shaking depending on cell density | 16.2 ± 7.9 | 8–10 | G-rex 10 | 22.3 ± 12.2 | No difference with G-rex;98.0% max T cell purity | Exhaustion profile did not differ. Prodigy cells produced less IFN-γ. Secretion of TNF-α and IL-2 was lower but not significant specific lysis to target |
Priesner [199] - 2016 | Not used | CD3/CD28; Transd.; IL7; IL15 | TexMACS GMP | 3% HS | CD62L+ cells from PBMC | 3 × 109 total | NP | TS520 | Static culture until day 3; clusters were dispersed | 13–23 (cells); 28–42 (Tcells) | 10–13 | Not used | 83.4–98.9% CD3 + CD45+ T cells | Transduction efficiency of 83% | |
Lock [200] - 2017 | Not used | CD3/CD28(transact); Transd.; IL7; IL15 | TexMACS GMP | 3% HS | CD4 + CD8 + CD45+ from PBMC | 2–10 × 107 total | 250 | TS520 | Automated media exchange every day | 65 ± 36 | 12 | Not used | 91.3–5.0% for Healthy donor and 88.3–7.1% for Melanoma Patient sourced | Transduct.: 34.5–11.7% for HD and 36.4–17.7% for PM. Secretion of GM-CSF, IFN-γ, IL-2, and TNF-α, no IL-4, IL-5, or IL-10 detected. | |
Blaeschke [201] - 2018 | Not used | CD3/CD28(transact); Transd.; IL7; IL15 | TexMACS GMP | 3% HS | CD4+/CD8+ from PBMC | 2.86 × 107 total | NP | TS520 | NP | 14.7–102.4 | 12 | Not used | 50.3% CD4+ and 38.7% CD8+ cells; 10.2% of the cells were NKT cells. | Transduction: 26.95%; No increased expression of exhaustion markers. CD19 CAR T cells killed 80% of the target (5:1); cells were able to secrete GM-CSF, IFN-γ, IL-2, and TNF-α | |
Zhang [202] - 2018 | 200 | CD3/CD28(transact); Transd. | TexMACS GMP |  | CD4+/CD8+ from PBMC | 108 total | 250 | TS520 | Media exchanges performed without disrupting the cells | 16 (cells); 20 (T cells) |  | Not used | CD4+ and CD8+ T cells changed from 45 and 34% to 22 and 74%.The frequency of CD4+ Tregs decreased | Transduction 60% in CD4+ and 50% in CD8+ T cells. Cells produced significant amounts of Th1 cytokine, IFN-γ and IL-2 after stimulation. Cytotoxicity ~ 60% at a 3:1 effector-to-target ratio | |
Zhu [197] - 2018 | Used | CD3/CD28(transact); Transduct. | TexMACS GMP | 3% HS | CD4+/CD8+ from PBMC | 108 total | NP | TS520 | As medium is added or exchanged, it is programmed to shake | 24.5–41.0 | 13 | G-rex100 | Signific. better | CD4+ decreased; CD8+ increased. Treg decreased. Little difference between Prodigy and G-rex in regard to phenotype | Low levels of IFN-γ. IFN-γ; all CAR-T-cell products lysed Raji cells when tested; transduction: 21 to 56.6%. better transduction in Prodigy. |
 Aleksandrova [203] - 2019 | Not used | CD3/CD28(transact); Transd.; IL7; IL15 | TexMACS GMP | HS | CD4+/CD8+ from PBMC | NR | NP | TS520 | NP | 46–81 | 12 | Not used | 97% T cells (range 96–99); some impurities of NKT and NK cells: median 2.9 and 0.07%, respectively. CD4/CD8 ratio of 2.4 decreased to 1.7 (range 1.1–2.3) in the final product. | Transduction: 22% on day 5 (range 17–41%) and 23% in the final product (range 21–45%).; CAR T cells were also cytotoxic against target cells | |
Fernández [204] - 2019 | 100 | CD3/CD28(transact); Transd. | TexMACS GMP |  | CD45RA− from PBMC | 1 × 108 total | NP | TS520 | NP | 13.4–38.6 | 10–13 | Not used | Enrichment in CD4+ vs. CD8+ T cells. Effector memory (TEM) phenotype | Cytotoxicity ≥20%. The expression of NKG2D increased during the expansion of cells. NKG2D CAR expression 55–60.6% | |
MarÃn [205] - 2019 | 1000 | T reg beads (aCD3/aCD8); Rapamycin | TexMACS GMP | 5%AB | CD4 + CD25− from PBMC | Mean: 56.9 × 106 | 250 | TS510 | Shaked from day 5; media exchange was conducted. Agitation paused to facilitate bead to cell contact | Similar kinetics | G-rex | G-rex 95.6% (range 93.7–97.0; n = 4). Prodigy R 92.8% (range 89.8–94.5%, n = 4; p = 0.08). | Suppressive capacity varied between donors but appeared to be independent of the manufacturing regimen | ||
NK cells | |||||||||||||||
Granzin [198] - 2015 | 500 | Irr. EBV-LCL | TexMACS GMP | 5% AB | CD56 + CD3- from PBMC | 2 × 106 | 210 | TS730 | Shaking after day 7 | 850 ± 509 | 14 | T75 | 1344 ± 1135 | > 99% CD3e/CD56þ,T or B cells could not be detected. | No differences in the production of IFN-γ and TNF-α and similar levels of degranulation. No reduction in telomere length. T75 and Prodigy had a comparable marker profile. NKG2C, CX3CR1 and KIR2DL2/DL3 higher in Prodigy |
 Oberschmidt [188] - 2019 | 500 | IL-21, IL-15; NK MACS supplement | NK MACS | 5%AB | CD56 + CD3- from PBMC | 2–4 × 106 | 250 | TS310 | 0.17–0.3 g shaking mode was activated on day 7. | 11.31–17.94 | 3–14 | Not used | 98.8–99.2% | Total NK cells expressed initially low to moderate levels of NKG2D, NKp30, NKp44, and NKp46. The surface markers increased during the 14-day expansion. Death receptors and activation markers increased. NK cell cytotoxicity increased | |