Characteristics of study participants
| CyTOF study cohort . | Patients who underwent HSCT and demonstrated aGVHD . | No aGVHD . | No HSCT . | ||
|---|---|---|---|---|---|
| Steroid-refractory aGVHD responsive to MSC-based second-line therapy (n = 11) . | Steroid-refractory aGVHD nonresponsive to MSC-based second-line therapy group (n = 6) . | Steroid-responsive aGVHD (n = 7) . | HSCT control participants (n = 11) . | Healthy control participants (n = 7) . | |
| Age (y)∗ | 12.5 (1.3-18.1) | 12.6 (1.3-16.9) | 9.9 (2.4-15) | 11.1 (0.3-17.8) | 12.1 (8-18.3) |
| Transplantation period | 2005-2013 | 2005-2013 | 2005-2010 | 2005-2012 | N/A |
| Male sex | 6 (55%) | 4 (67%) | 5 (71%) | 7 (64%) | 5 (71%) |
| HSCT indication | |||||
| Malignant disease | 8 (73%) | 3 (50%) | 5 (71%) | 10 (91%) | N/A |
| Nonmalignant disease | 3 (27%) | 3 (50%) | 2 (29%) | 1 (9%) | |
| Graft type† | |||||
| BM | 8 (73%) | 5 (83%) | 5 (71%) | 9 (82%) | N/A |
| Other | 3 (27%) | 1 (17%) | 2 (29%) | 2 (18%) | |
| Donor type‡ | |||||
| IRD/ORD | 6 (55%) | 2 (33%) | 0 | 6 (54%) | N/A |
| MUD | 5 (45%) | 4 (67%) | 7 (100%) | 5 (45%) | |
| Conditioning regimen§ | |||||
| Bu-Cy based | 5 (45%) | 1 (17%) | 3 (43%) | 6 (55%) | N/A |
| Bu-Flu based | 2 (18%) | 1 (17%) | 1 (14%) | 0 | |
| Other | 4 (36%) | 4 (67%) | 3 (43%) | 5 (45%) | |
| Serotherapy‖ | |||||
| ATG | 3 (27%) | 3 (50%) | 7 (100%) | 4 (36%) | N/A |
| Alemtuzumab | 1 (9%) | 0 | 0 | 1 (9%) | |
| None | 7 (64%) | 3 (50%) | 0 | 6 (55%) | |
| GVHD prophylaxis¶ | |||||
| CsA ± MTX | 8 (73%) | 1 (17%) | 4 (57%) | 9 (82%) | N/A |
| CsA ± MTX + other | 2 (18%) | 2 (33%) | 3 (43%) | 2 (18%) | |
| Other | 0 | 3 (50%) | 0 | 0 | |
| None | 1 (9%) | 0 | 0 | 0 | |
| GVHD onset (day + range) | 57 (12-86, 195)∗∗ | 28 (14-62) | 30 (15-89) | N/A | N/A |
| GVHD grade# | |||||
| I | 0 | 0 | 2 (29%) | N/A | N/A |
| II | 0 | 0 | 3 (43%) | ||
| III | 7 (64%) | 2 (33%) | 2 (29%) | ||
| IV | 4 (36%) | 4 (67%) | 0 | ||
| Organ involvement | N/A | N/A | |||
| Skin | |||||
| 0 | 5 (46%) | 0 | 2 (29%) | ||
| 1-2 | 3 (27%) | 2 (33%) | 3 (43%) | ||
| 3-4 | 3 (27%) | 4 (67%) | 2 (23%) | ||
| Liver | |||||
| 0 | 6 (55%) | 4 (67%) | 6 (85%) | ||
| 1-2 | 2 (18%) | 0 | 1 (14%) | ||
| 3-4 | 3 (27%) | 2 (33%) | 0 | ||
| GI tract | |||||
| 0 | 0 | 0 | 5 (71%) | ||
| 1-2 | 3 (27%) | 1 (17%) | 0 | ||
| 3-4 | 8 (73%) | 5 (83%) | 2 (29%) | ||
| GVHD treatment†† | |||||
| Steroids | 4 (36%) | 0 | 2 (29%) | N/A | N/A |
| Steroids + other | 7 (64%) | 6 (100%) | 5 (71%) | ||
| No. of MSC infusions | |||||
| 1 | 8 (73%) | 2 (33%) | N/A | N/A | N/A |
| 2 | 3 (27%) | 4 (67%) | |||
| Viral reactivations‡‡ | |||||
| Yes | 8 (73%) | 5 (83%) | 6 (86%) | 6 (55%) | N/A |
| Adenovirus | 1 (9%) | 0 | 0 | 0 | |
| CMV | 2 (18%) | 1 (17%) | 0 | 2 (18%) | |
| EBV | 4 (36%) | 1 (17%) | 3 (43%) | 3 (27%) | |
| Combination | 1 (9%) | 3 (50%) | 3 (43%) | 1 (9%) | |
| 100% chimerism§§ | 11 (100%) | 4 (67%) | 7 (100%) | 9 (82%) | N/A |
| Alive at d + 365‖‖ | 9 (82%) | 1 (17%) | 7 (100%) | 10 (91%) | N/A |
| CyTOF study cohort . | Patients who underwent HSCT and demonstrated aGVHD . | No aGVHD . | No HSCT . | ||
|---|---|---|---|---|---|
| Steroid-refractory aGVHD responsive to MSC-based second-line therapy (n = 11) . | Steroid-refractory aGVHD nonresponsive to MSC-based second-line therapy group (n = 6) . | Steroid-responsive aGVHD (n = 7) . | HSCT control participants (n = 11) . | Healthy control participants (n = 7) . | |
| Age (y)∗ | 12.5 (1.3-18.1) | 12.6 (1.3-16.9) | 9.9 (2.4-15) | 11.1 (0.3-17.8) | 12.1 (8-18.3) |
| Transplantation period | 2005-2013 | 2005-2013 | 2005-2010 | 2005-2012 | N/A |
| Male sex | 6 (55%) | 4 (67%) | 5 (71%) | 7 (64%) | 5 (71%) |
| HSCT indication | |||||
| Malignant disease | 8 (73%) | 3 (50%) | 5 (71%) | 10 (91%) | N/A |
| Nonmalignant disease | 3 (27%) | 3 (50%) | 2 (29%) | 1 (9%) | |
| Graft type† | |||||
| BM | 8 (73%) | 5 (83%) | 5 (71%) | 9 (82%) | N/A |
| Other | 3 (27%) | 1 (17%) | 2 (29%) | 2 (18%) | |
| Donor type‡ | |||||
| IRD/ORD | 6 (55%) | 2 (33%) | 0 | 6 (54%) | N/A |
| MUD | 5 (45%) | 4 (67%) | 7 (100%) | 5 (45%) | |
| Conditioning regimen§ | |||||
| Bu-Cy based | 5 (45%) | 1 (17%) | 3 (43%) | 6 (55%) | N/A |
| Bu-Flu based | 2 (18%) | 1 (17%) | 1 (14%) | 0 | |
| Other | 4 (36%) | 4 (67%) | 3 (43%) | 5 (45%) | |
| Serotherapy‖ | |||||
| ATG | 3 (27%) | 3 (50%) | 7 (100%) | 4 (36%) | N/A |
| Alemtuzumab | 1 (9%) | 0 | 0 | 1 (9%) | |
| None | 7 (64%) | 3 (50%) | 0 | 6 (55%) | |
| GVHD prophylaxis¶ | |||||
| CsA ± MTX | 8 (73%) | 1 (17%) | 4 (57%) | 9 (82%) | N/A |
| CsA ± MTX + other | 2 (18%) | 2 (33%) | 3 (43%) | 2 (18%) | |
| Other | 0 | 3 (50%) | 0 | 0 | |
| None | 1 (9%) | 0 | 0 | 0 | |
| GVHD onset (day + range) | 57 (12-86, 195)∗∗ | 28 (14-62) | 30 (15-89) | N/A | N/A |
| GVHD grade# | |||||
| I | 0 | 0 | 2 (29%) | N/A | N/A |
| II | 0 | 0 | 3 (43%) | ||
| III | 7 (64%) | 2 (33%) | 2 (29%) | ||
| IV | 4 (36%) | 4 (67%) | 0 | ||
| Organ involvement | N/A | N/A | |||
| Skin | |||||
| 0 | 5 (46%) | 0 | 2 (29%) | ||
| 1-2 | 3 (27%) | 2 (33%) | 3 (43%) | ||
| 3-4 | 3 (27%) | 4 (67%) | 2 (23%) | ||
| Liver | |||||
| 0 | 6 (55%) | 4 (67%) | 6 (85%) | ||
| 1-2 | 2 (18%) | 0 | 1 (14%) | ||
| 3-4 | 3 (27%) | 2 (33%) | 0 | ||
| GI tract | |||||
| 0 | 0 | 0 | 5 (71%) | ||
| 1-2 | 3 (27%) | 1 (17%) | 0 | ||
| 3-4 | 8 (73%) | 5 (83%) | 2 (29%) | ||
| GVHD treatment†† | |||||
| Steroids | 4 (36%) | 0 | 2 (29%) | N/A | N/A |
| Steroids + other | 7 (64%) | 6 (100%) | 5 (71%) | ||
| No. of MSC infusions | |||||
| 1 | 8 (73%) | 2 (33%) | N/A | N/A | N/A |
| 2 | 3 (27%) | 4 (67%) | |||
| Viral reactivations‡‡ | |||||
| Yes | 8 (73%) | 5 (83%) | 6 (86%) | 6 (55%) | N/A |
| Adenovirus | 1 (9%) | 0 | 0 | 0 | |
| CMV | 2 (18%) | 1 (17%) | 0 | 2 (18%) | |
| EBV | 4 (36%) | 1 (17%) | 3 (43%) | 3 (27%) | |
| Combination | 1 (9%) | 3 (50%) | 3 (43%) | 1 (9%) | |
| 100% chimerism§§ | 11 (100%) | 4 (67%) | 7 (100%) | 9 (82%) | N/A |
| Alive at d + 365‖‖ | 9 (82%) | 1 (17%) | 7 (100%) | 10 (91%) | N/A |
aGVHD, acute graft-versus-host disease; ATG, antithymocyte globulin; BM, bone marrow; Bu, busulfan; CB, cord blood; CMV, cytomegalovirus; CsA, ciclosporin; Cy, cyclophosphamide; CyTOF, mass cytometry; EBV, Epstein-Barr virus; Flu, fludarabine; GI, gastrointestinal; GVHD, graft-versus-host disease; HSCT, hematopoietic stem cell transplantation; IRD, identical related donor; MSC, mesenchymal stromal cell; MTX, methotrexate; MUD, matched unrelated donor; ORD, other related donor; N/A, not applicable.
Median age (range) in years at the day of hematopoietic stem cell donation or infusion.
Graft types include BM, CB, or granulocyte colony stimulating factor (G-CSF)-mobilized peripheral blood stem cells.
Donor types include MUD (minimal 10/12 matched), IRD, and ORD.
Other agents used are listed in supplemental Table 1.
T-cell-depleting agents include ATG or Campath (anti-CD52).
GVHD prophylactic medication included CsA and MTX. Other agents applied are listed in supplemental Table 1.
GVHD diagnoses were based on both clinical symptoms and histologic evaluation of biopsy samples obtained from affected sites, as reported earlier.1,68 Clinical symptoms of GVHD were scored and included degree of skin rash, total bilirubin levels, and output per day of diarrhea with or without abdominal pain.68 Steroid-refractory aGVHD was defined as either no response at least 7 d after starting a minimum daily dose of 2 mg/kg prednisone, or aGVHD progression of at least 1 grade within 72 h after prednisone-based treatment was initiated.
One patient demonstrated aGVHD at day d + 195 after receiving unselected donor lymphocytes administered at d + 156 for the conversion of a declined percentage of donor chimerism.
Other immune-suppressive drugs applied for GVHD treatment are listed in supplemental Table 1.
Viral reactivations were detected by routine polymerase chain reaction-based monitoring of blood samples collected up to the last PBMC sample (t = 2 or t = 3) included in the study.
Number of patients per group in whom 100% of PBMC (collected at or shortly before t = 1) displayed donor-specific DNA sequence as assessed by short tandem repeat assay.
Causes of death include GVHD, infectious complications, or relapse of the original malignancy, as detailed in supplemental Table 1.