Single-cell analysis of HRS cells and CD30−/EBER+ cells
| Case no. . | Cells . | PCR results* . | Cells with† . | ||||
|---|---|---|---|---|---|---|---|
| EBNA . | Ig rearrangements . | EBNA + Ig rearrangements . | HRS cell-specific clonal rearrangements . | Other clonal rearrangements . | Unique rearrangements . | ||
| 1 | HRS | 19/20 | 18/20 | 18/20 (90%) | 18 | — | — |
| Buffer | 0/8 | 0/8 | |||||
| CD30−/EBER+ | 54/86 | 33/86 | 29/86 (33.7%) | 2‡ | 4 (2, 2)2-153 | 23 | |
| Buffer | 0/36 | 0/36 | |||||
| CD3+ | 0/20 | ||||||
| Buffer | 0/8 | ||||||
| 2 | HRS | 11/20 | 10/20 | 10/20 (50%) | 10 | — | — |
| Buffer | 0/8 | 0/8 | |||||
| CD30−/EBER+ | 24/50 | 15/50 | 12/50 (24%) | — | 7 (2, 2, 3)2-153 | 5 | |
| Buffer | 0/23 | 0/23 | |||||
| CD3+ | 0/20 | ||||||
| Buffer | 0/8 | ||||||
| 3 | HRS | 9/20 | 9/20 | 9/20 (45%) | 92-154 | — | — |
| Buffer | 0/8 | 1/8 | 1 | ||||
| CD30−/EBER+ | 34/90 | 21/90 | 18/90 (20%) | — | 22-160 | 16 | |
| Buffer | 1/36 | 2/36 | 2 | ||||
| Case no. . | Cells . | PCR results* . | Cells with† . | ||||
|---|---|---|---|---|---|---|---|
| EBNA . | Ig rearrangements . | EBNA + Ig rearrangements . | HRS cell-specific clonal rearrangements . | Other clonal rearrangements . | Unique rearrangements . | ||
| 1 | HRS | 19/20 | 18/20 | 18/20 (90%) | 18 | — | — |
| Buffer | 0/8 | 0/8 | |||||
| CD30−/EBER+ | 54/86 | 33/86 | 29/86 (33.7%) | 2‡ | 4 (2, 2)2-153 | 23 | |
| Buffer | 0/36 | 0/36 | |||||
| CD3+ | 0/20 | ||||||
| Buffer | 0/8 | ||||||
| 2 | HRS | 11/20 | 10/20 | 10/20 (50%) | 10 | — | — |
| Buffer | 0/8 | 0/8 | |||||
| CD30−/EBER+ | 24/50 | 15/50 | 12/50 (24%) | — | 7 (2, 2, 3)2-153 | 5 | |
| Buffer | 0/23 | 0/23 | |||||
| CD3+ | 0/20 | ||||||
| Buffer | 0/8 | ||||||
| 3 | HRS | 9/20 | 9/20 | 9/20 (45%) | 92-154 | — | — |
| Buffer | 0/8 | 1/8 | 1 | ||||
| CD30−/EBER+ | 34/90 | 21/90 | 18/90 (20%) | — | 22-160 | 16 | |
| Buffer | 1/36 | 2/36 | 2 | ||||
Given are the numbers of cells positive for EBNA1PCR, Ig gene rearrangement PCR and both, per cells. CD3+cells were only for Ig gene rearrangements.
Sequence analysis was performed for Ig rearrangements of all EBNA+ cells. All sequences have been deposited in the EMBL database under accession numbers HSA 298442-298549. For the nature of the clonal Ig gene rearrangements see Table 3.
From one cell the clonal HRS–cell specific VH3-20 and VκA17 rearrangement, from the other cell both clonal HRS–cell specific Vκ rearrangements were amplified.
Numbers of members per clone in parentheses.
From one cell in addition to the clonal VκB3 and Vλ6a rearrangements one unmutated VH and one unmutated Vκ rearrangement and from another cell, in addition to the clonal VκB3 and Vλ6a rearrangements, one unmutated VH rearrangement was amplified. These rearrangements most likely represent cellular or other contamination.
These 2 Vκ/Jκ rearrangements with identical sequence were unmutated and rearranged without N nucleotide insertion. Thus, it can not be excluded that they derive from 2 clonally independent cells.