Characterization of immunoglobulin H or T-cell receptor β loci of single Hodgkin and Reed-Sternberg cells
| Case . | Locus . | HRS cells . | Sorted T cells* germline positive cells . | |||||
|---|---|---|---|---|---|---|---|---|
| Positive cells . | First allele . | Second allele . | Additional allele in germline positive cells . | |||||
| Positive cells . | Rearrangement . | Positive cells . | Rearrangement . | |||||
| 1 | IgH | 24/65 | 20/65 | VH2-5/DH3-10/JH5 | 9/14 | DH3-9/JH6 | 0/14 | 9/10 |
| 2 | IgH | 11/14 | 11/14 | VH3-20/DH5-5/JH4 | 1/14† | VH3-11/DH6-13/JH4/5i | 0/14 | 9/10 |
| 3 | IgH | 10/20 | 6/20 | VH4-31/DH2-8/JH5 | 9/20 | VH4-34/JH6‡ | 1/20 | 10/10 |
| 4 | IgH | 11/30 | 8/30 | VH1-8/DH3-22/JH6 | 10/30 | VH3-53/DH2-2/JH6 | 0/12 | 3/4 |
| 5 | IgH | 10/20 | 5/20 | VH3-30/DH2-15/JH6 | 9/20 | VH3-11/DH2-21/JH6 | 1/20 | 6/7 |
| 6 | TCRβ | 20/30 | 14/30 | Vβ7.1/Dβ1/Jβ1.6 | 14/30 | Dβ1/Jβ1.4 | 1/30 | 9/9 |
| Case . | Locus . | HRS cells . | Sorted T cells* germline positive cells . | |||||
|---|---|---|---|---|---|---|---|---|
| Positive cells . | First allele . | Second allele . | Additional allele in germline positive cells . | |||||
| Positive cells . | Rearrangement . | Positive cells . | Rearrangement . | |||||
| 1 | IgH | 24/65 | 20/65 | VH2-5/DH3-10/JH5 | 9/14 | DH3-9/JH6 | 0/14 | 9/10 |
| 2 | IgH | 11/14 | 11/14 | VH3-20/DH5-5/JH4 | 1/14† | VH3-11/DH6-13/JH4/5i | 0/14 | 9/10 |
| 3 | IgH | 10/20 | 6/20 | VH4-31/DH2-8/JH5 | 9/20 | VH4-34/JH6‡ | 1/20 | 10/10 |
| 4 | IgH | 11/30 | 8/30 | VH1-8/DH3-22/JH6 | 10/30 | VH3-53/DH2-2/JH6 | 0/12 | 3/4 |
| 5 | IgH | 10/20 | 5/20 | VH3-30/DH2-15/JH6 | 9/20 | VH3-11/DH2-21/JH6 | 1/20 | 6/7 |
| 6 | TCRβ | 20/30 | 14/30 | Vβ7.1/Dβ1/Jβ1.6 | 14/30 | Dβ1/Jβ1.4 | 1/30 | 9/9 |
HRS indicates Hodgkin and Reed-Sternberg; IgH, immunoglobulin H; TCRβ, T-cell receptor β; PCR, polymerase chain reaction; EMBL, European Molecular Biology Laboratory.
The 6 cases of this study have previously been described: Case 1 is case I from Müschen et al3; cases 2 and 3: cases 1 and 2 from Spieker et al20; case 4: MC case from Küppers et al14 and case II from Müschen et al3; case 5: case 8 from Kanzler et al2; and case 6: case III from Müschen et al.3 The results of the buffer controls for the germline-specific PCR are as follows: case 1: 0/4; case 2: 0/7; case 3: 0/8; case 4: 0/5; case 5: 0/8; case 6: 1/10. The results of the buffer controls for the VDJ PCR are as follows: case 1: 0/15; case 2: 0/7; case 3: 0/8; case 4: 0/6; case 5: 0/8; case 6: 0/10. Sequences are available from GenBank/EMBL data libraries under accession numbersX77417, X77416, Z77321, Z77322, AJ243643, AJ243645, AJ243647, AJ251000,AJ298441, AJ298442, AJ298443, and AJ298444. Twelve, 10, and 12 HRS cells of cases 2, 3, and 4, respectively, were also analysed for DHJH joints, but no products were obtained. From 2 HRS cells of case 1, unique VH region genes were amplified, likely representing cellular or other contamination.
B cells for case 6.
The low efficiency is likely due to the fact that this amplificate is about 600 base pairs long; such long fragments are difficult to amplify from aliquots of whole-genome PCR. The same rearrangement was amplified from 3 of 20 HRS cells previously analyzed,20 confirming that this joint represents the second clonal IgH rearrangement of this HRS cell clone.
Rearrangement has likely a deletion of FR3 and CDR3, because codon 81 is immediately followed by the JH6 primer sequence.