Table 3.
Number of samples ELISPOT patterns interpreted by B-cell phenotype in PROTCL and BFC by time
| Interpretation based on B-cell phenotype | Number of ELISPOTs showing the defined pattern |
||||||||
|---|---|---|---|---|---|---|---|---|---|
| PROTCL |
BFC |
Total |
|||||||
| Time point 1 | Time point 2 | Time point 1 | Time point 2 | Time point 1 | Time point 2 | ||||
| No evidence of B-dependent antidonor response | No response | No regulation | 3 | 1 | 5 | 11 | 8 | 12 | |
| Evidence of regulation |
Regulated antidonor response without evidence of B-dependency |
Breg: only when CD25+ cells present |
2 |
2a |
0 |
1 |
5 |
7 |
|
| Breg: when CD25+ cells present and absent |
0 |
2 |
2 |
0 |
|||||
| Breg: only when CD25+ cells absent |
0 |
1 |
1 |
1 |
|||||
| Evidence of B-dependent antidonor response | B-dependent antidonor response with evidence of regulation | Breg when CD25 present BUT Bdep when CD25 absent, |
1 |
0 |
5b |
3c |
12 | 13 | |
| Bdep: only when CD25+ cells absent |
1 |
3 |
4 |
3 |
|||||
| Bdep when CD25 present, Breg when CD25+ cells absent | 0 |
1 |
1 |
3 |
|||||
| No evidence of regulation |
Unregulated B-dependent antidonor response |
Bdep: when CD25+ cells present and absent |
3 |
0 |
7 |
3 |
13 |
11 |
|
| Bdep: only when CD25+ cells present |
2 |
4 |
1 |
4 |
|||||
| Not done / Not viable / Not interpretabled | 2 ND 1 NDSR |
1 NDSR | 5 ND 3 NDSR 3 DSR |
5 ND 3 NDSR |
14 | 9 | |||
Several other viable samples at later time points were collected and analyzed and included in Shiu et al.7 but are not considered further here.
Bdep, decrease in spot count of ≥20% when CD19+ cells depleted; BFC, for-cause biopsy; DSR, donor-specific reactivity; Breg, increase in spot count of ≥20% when CD19+ cells depleted; ELISPOT, enzyme-linked immunosorbent spot; IFN-γ, interferon-γ; NDSR, no donor-specific reactivity; PBMC, peripheral blood mononuclear cells; PROTCL, protocol.
One of these samples was DSR (i.e., IFN-γ produced by CD8-depleted PBMCs but spot count increased by >20% with B-cell depletion. Therefore, there were 7 DSR and 8 NDSR in PROTCL at time point 2.
Three of these samples were DSR (i.e., IFN-γ produced by CD8-depleted PBMCs but spot count increased by >20% with B-cell depletion. All 3 also showed increases >20% with depletion of CD25+ cells followed by reduction (>20%) in spot count when CD19+ cells additionally depleted. Therefore, there were 14 DSR and 18 NDSR in BFC at time point 1.
One of these samples was DSR (i.e., IFN-γ produced by CD8-depleted PBMCs but spot count increased by >20% with B-cell depletion. Both also showed increases >20% with depletion of CD25+ cells followed by reduction (>20%) in spot count when CD19+ cells additionally depleted. Therefore, there were 11 DSR and 21 NDSR in BFC at time point 2.
In some samples, it was not possible to perform all 4 depletion combinations to enable interpretation based on B-cell phenotype, but classification as DSR/NDSR was possible.