Table 2.

Main clinical studies with graft manipulation and Treg and Tcon add backs for prevention of GVHD

StudyHCT typePatientsTreg isolationTreg number/kgImmune suppressionGVHD outcomeRelapse incidence
Brunstein et al. (2011)66  Cord blood 23 Magnetic separation
In vitro expansion 
1 × 105 to 3 × 106  MMF, CsA aGVHD II-IV (43%)
aGVHD III-IV (17%)
cGVHD (14%) 
34.8% 
Di Ianni et al. (2011)65  Haploidentical 28 Magnetic separation 2 × 106 to 4 × 106  None aGVHD (2/26)
No cGVHD 
3.6% 
Martelli et al. (2014)67  Haploidentical 43 Magnetic separation 2 × 106  None aGVHD (6/41)
cGVHD (1/41) 
4.9% 
Brunstein et al. (2016)75  Cord blood 11 Magnetic separation
In vitro expansion 
3 × 106 to 100 × 106  Sirolimus, MMF aGVHD II-IV (9%), cGVHD (0%) 33% 
Meyer et al. (2019)69  HLA-matched Magnetic separation and cell sorting 1 × 106 to 3 × 106  Tacrolimus or sirolimus aGVHD (0%), cGVHD (0%) 42.9% 
Pierini et al. (2021)68  Haploidentical 50 Magnetic separation 2 × 106  None aGVHD (33%), cGVHD (2%) 4% 
Bader et al. (2024)70  HLA-matched 12 Magnetic separation 2.5 × 106  None aGVHD (58%), cGVHD (28%) 8.3% 
12 2.5 × 106  Tacrolimus aGVHD (8%), cGVHD (0%) 33% 
StudyHCT typePatientsTreg isolationTreg number/kgImmune suppressionGVHD outcomeRelapse incidence
Brunstein et al. (2011)66  Cord blood 23 Magnetic separation
In vitro expansion 
1 × 105 to 3 × 106  MMF, CsA aGVHD II-IV (43%)
aGVHD III-IV (17%)
cGVHD (14%) 
34.8% 
Di Ianni et al. (2011)65  Haploidentical 28 Magnetic separation 2 × 106 to 4 × 106  None aGVHD (2/26)
No cGVHD 
3.6% 
Martelli et al. (2014)67  Haploidentical 43 Magnetic separation 2 × 106  None aGVHD (6/41)
cGVHD (1/41) 
4.9% 
Brunstein et al. (2016)75  Cord blood 11 Magnetic separation
In vitro expansion 
3 × 106 to 100 × 106  Sirolimus, MMF aGVHD II-IV (9%), cGVHD (0%) 33% 
Meyer et al. (2019)69  HLA-matched Magnetic separation and cell sorting 1 × 106 to 3 × 106  Tacrolimus or sirolimus aGVHD (0%), cGVHD (0%) 42.9% 
Pierini et al. (2021)68  Haploidentical 50 Magnetic separation 2 × 106  None aGVHD (33%), cGVHD (2%) 4% 
Bader et al. (2024)70  HLA-matched 12 Magnetic separation 2.5 × 106  None aGVHD (58%), cGVHD (28%) 8.3% 
12 2.5 × 106  Tacrolimus aGVHD (8%), cGVHD (0%) 33% 

Some of the most relevant clinical studies that employed Treg and Tcon add backs to improve GVHD prevention and allow for strong antileukemic activity.

aGvHD, acute GvHD; cGvHD, chronic GvHD; CsA, cyclosporine A; MMF, mycophenolate mofetil.

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