Results of Proliferation Assays of the TT-Specific T-Cell Lines Derived From Allo-BMT Recipients and Their Donors, and Overview of the Generation of TT-Specific T-Cell Clones
| Sample . | SI5-150 ± SD . | No. Clones5-152 . | % TT Specific . | No. Identical Clones5-153 . | D/R Type5-155 . | |
|---|---|---|---|---|---|---|
| TT5-151 . | PHA . | |||||
| Donor 1 | 28 ± 9 | 84 ± 42 | 178 | ≈9%5-154 | 4/13 | D |
| Patient 1 | 7 ± 3 | 52 ± 26 | 125 | ≈7%5-154 | 3/10# | D |
| Donor 2 | 18 ± 9 | 50 ± 25 | 76 | ≈38% | 11/28 | D |
| Patient 2 | 30 ± 15 | 48 ± 24 | 43 | ≈56% | 11/22 | D |
| Sample . | SI5-150 ± SD . | No. Clones5-152 . | % TT Specific . | No. Identical Clones5-153 . | D/R Type5-155 . | |
|---|---|---|---|---|---|---|
| TT5-151 . | PHA . | |||||
| Donor 1 | 28 ± 9 | 84 ± 42 | 178 | ≈9%5-154 | 4/13 | D |
| Patient 1 | 7 ± 3 | 52 ± 26 | 125 | ≈7%5-154 | 3/10# | D |
| Donor 2 | 18 ± 9 | 50 ± 25 | 76 | ≈38% | 11/28 | D |
| Patient 2 | 30 ± 15 | 48 ± 24 | 43 | ≈56% | 11/22 | D |
Stimulation indices were calculated: SI = 3H thymidine incorporation of T cells with APC + antigen/3H thymidine incorporation of T cells with APC only ± standard deviation (SD). The background values varied between 100 and 900 cpm, the values of interest varied between 20,000 and 50,000 cpm for PHA and between 2,000 and 10,000 cpm for TT.
In the proliferation assay, TT and PHA were used as antigen.
All minicultures that were derived from 96-well plates with at least 33% negative wells/plate after limiting dilution were considered to be clonal, and used for the proliferation assays and molecular analysis.
Numbers of identical TT-specific T-cell clones (identical TCR sequence, Table 6) divided by total numbers of TT-specific T-cell clones.
Donor (D) or recipient (R) type T-cell clone as determined via FACS analysis of HLA-DR expression, after staining with anti–HLA-DR MoAbs.
% of TT specific clones of donor 1 and patient 1: mean of 3 independent cloning experiments.
#Due to the low number of clones, all clones once specific for TT were included for further molecular analysis.