Clonal mutations observed in the major mutation cluster region of the first BCL-6 intron in 7 cases of lymphoma with (3;14) translocation
Case no. and diagnosis . | Mutations located within the MMC or 3′ of the MMC . | Intraclonal heterogeneity . | No. of sequenced clones . |
|---|---|---|---|
| 1. DLBCL | |||
| Untranslocated allele | 119C>G; 123C>T; 265G>A; 378G>T; 423C>T; 519C>T; 709A>G | Yes | 6 |
| Translocated allele | |||
| der(3q27) | 345G>C; 347G>A; 349-350insG; 350T>G; 359A>G; 365G>C; 423C>G; 429C>G; 441G>A; 443G>A; 478T>G; 685C>T; 697G>A; 737delT; 741T>A; 780G>A | No | 9 |
| der(14q32) | Unknown* | ||
| 2. Grade 3 FL | Yes | 9 | |
| Untranslocated allele | 18T>C; 95T>A; 542T>C; 548C>T; 782A>T | ||
| Translocated allele | |||
| der(3q27) | No clonal mutation | No | 6 |
| der(14q32) | 18T>C; 375G>A; 416G>A; 423C>G | Yes | 7 |
| 3. DLBCL | |||
| Untranslocated allele | No clonal mutation | No | 18† |
| Translocated allele | |||
| der(3q27) | 51C>G; 87T>A; 90C>G; 100A>G; 107A>G; 136A>G; 142A>G; 170C>T; 196A>T; 252C>T; (387-393)del | No | 9 |
| der(14q32) | 6G>A; 7C>A; 30T>C; 36T>C; 38T>A | Yes | 6 |
| 4. DLBCL | |||
| Untranslocated allele | No clonal mutation | No | 18† |
| Translocated allele | |||
| der(3q27) | 520delT | No | 6 |
| der(14q32) | 8T>G; 16A>T; 23T>C; 44T>G; 45C>A; 71A>C; 76T>C; 88C>A; 89T>A; 97T>A; 105T>G; 107T>A; 110T>G; 123C>G; 143T>C; 145C>T; 149A>C; 151-152insT, 153T>G | No | 6 |
| 5. DLBCL | |||
| Untranslocated allele | 202A>T; 226A>G; 267T>G; 269T>A; 344A>T; 434G>T | No | 4 |
| Translocated allele | |||
| der(3q27)‡ | 1249G>C; 1260G>C; 1284C>T; 1285C>T | No | 4 |
| der(14q32) | 23T>C; 24A>G; 32A>G; 43T>C; 44T>G; 47G>C; 57A>G; 71A>G; 73G>C; 75T>A; 84G>C; 87T>A; 90A>G; 100A>G; 104A>C; 108A>C; 116A>G; 121A>C; 122G>C; 140A>G; 141A>G; 176delA; 178G>T; 181delA; 183delA; 184G>T; 188G>A; 209T>G; 211G>C; 222G>A; 234G>C; 257T>G; 298A>C; 309G>A; 312T>A; 328G>A; 335G>C; 347G>C; 349T>G; 350T>G; 356G>C; 360T>C; 377G>A; 390A>C; 393C>G; 394C>T; 402G>A; 421C>G; 431G>A; (433-442)del; 459C>G; 466T>G; 477G>A; 500T>G; 501T>C; 505A>C; 509G>A; 510C>G; 519C>T; 526G>A; 608G>A; 620G>C 623G>A; 632T>G; 659C>G; 692C>G; 694G>A; 732T>C; 734delG; 738T>C; 742G>A; 769G>A; 775A>G; 779A>G; 782A>T | No | 9 |
| 6. DLBCL | |||
| Untranslocated allele | 46C>G; 84G>A; 142A>G; 193G>A; 211G>A; 502G>A; 520delT; 573T>C; 599T>C; 677T>A; 717A>G; 737T>G; 750T>G; 752T>A | Yes | 8 |
| Translocated allele | |||
| der(3q27)‡ | 1314G>C | No | 5 |
| der(14q32) | 25C>T; 26C>T; 84G>A; 100A>G; 106A>G; 108A>C; 119C>T; 121A>C; 122G>T; 139T>C; 173C>G; 191A>T; 195A>C; 197T>C; 200T>C; 217T>C; 261T>C; 275T>A; 279T>C; 328G>C; 329C>T; 334A>C; 337A>G; 345G>A; 350-351linsG; 400G>C; 423C>G; 424T>C; 425C>G; 445A>G; 478T>A; 479G>A; 486C>T; 487C>A; 488C>T; 499C>G; 503C>G; 507C>G; 508T>A; 509G>C; 512T>A; 524T>G; 541T>G; 545C>G; 555T>A; 561C>T; 574C>A; 579C>T; 614A>G; 617A>G; 650C>G; 671T>G; 677T>A; 679C>G; 697C>T; 718T>G; 720C>G; 727A>G; 728C>T; 735G>A; 741T>C; 761 61T>A; 763A>C; 766C>G | No | 15 |
| 7. Grade 1 FL | |||
| Untranslocated allele | No clonal mutation | No | 13† |
| Translocated allele | |||
| der(3q27) | No clonal mutation | No | 4 |
| der(14q32) | 73G>C; 177G>C; 339G>C; 397G>C; 441G>A; 445A>C; 475C>T; 504A>T; 549C>T; 613A>T; 633C>T; 735G>A; 736G>A | Yes | 4 |
Case no. and diagnosis . | Mutations located within the MMC or 3′ of the MMC . | Intraclonal heterogeneity . | No. of sequenced clones . |
|---|---|---|---|
| 1. DLBCL | |||
| Untranslocated allele | 119C>G; 123C>T; 265G>A; 378G>T; 423C>T; 519C>T; 709A>G | Yes | 6 |
| Translocated allele | |||
| der(3q27) | 345G>C; 347G>A; 349-350insG; 350T>G; 359A>G; 365G>C; 423C>G; 429C>G; 441G>A; 443G>A; 478T>G; 685C>T; 697G>A; 737delT; 741T>A; 780G>A | No | 9 |
| der(14q32) | Unknown* | ||
| 2. Grade 3 FL | Yes | 9 | |
| Untranslocated allele | 18T>C; 95T>A; 542T>C; 548C>T; 782A>T | ||
| Translocated allele | |||
| der(3q27) | No clonal mutation | No | 6 |
| der(14q32) | 18T>C; 375G>A; 416G>A; 423C>G | Yes | 7 |
| 3. DLBCL | |||
| Untranslocated allele | No clonal mutation | No | 18† |
| Translocated allele | |||
| der(3q27) | 51C>G; 87T>A; 90C>G; 100A>G; 107A>G; 136A>G; 142A>G; 170C>T; 196A>T; 252C>T; (387-393)del | No | 9 |
| der(14q32) | 6G>A; 7C>A; 30T>C; 36T>C; 38T>A | Yes | 6 |
| 4. DLBCL | |||
| Untranslocated allele | No clonal mutation | No | 18† |
| Translocated allele | |||
| der(3q27) | 520delT | No | 6 |
| der(14q32) | 8T>G; 16A>T; 23T>C; 44T>G; 45C>A; 71A>C; 76T>C; 88C>A; 89T>A; 97T>A; 105T>G; 107T>A; 110T>G; 123C>G; 143T>C; 145C>T; 149A>C; 151-152insT, 153T>G | No | 6 |
| 5. DLBCL | |||
| Untranslocated allele | 202A>T; 226A>G; 267T>G; 269T>A; 344A>T; 434G>T | No | 4 |
| Translocated allele | |||
| der(3q27)‡ | 1249G>C; 1260G>C; 1284C>T; 1285C>T | No | 4 |
| der(14q32) | 23T>C; 24A>G; 32A>G; 43T>C; 44T>G; 47G>C; 57A>G; 71A>G; 73G>C; 75T>A; 84G>C; 87T>A; 90A>G; 100A>G; 104A>C; 108A>C; 116A>G; 121A>C; 122G>C; 140A>G; 141A>G; 176delA; 178G>T; 181delA; 183delA; 184G>T; 188G>A; 209T>G; 211G>C; 222G>A; 234G>C; 257T>G; 298A>C; 309G>A; 312T>A; 328G>A; 335G>C; 347G>C; 349T>G; 350T>G; 356G>C; 360T>C; 377G>A; 390A>C; 393C>G; 394C>T; 402G>A; 421C>G; 431G>A; (433-442)del; 459C>G; 466T>G; 477G>A; 500T>G; 501T>C; 505A>C; 509G>A; 510C>G; 519C>T; 526G>A; 608G>A; 620G>C 623G>A; 632T>G; 659C>G; 692C>G; 694G>A; 732T>C; 734delG; 738T>C; 742G>A; 769G>A; 775A>G; 779A>G; 782A>T | No | 9 |
| 6. DLBCL | |||
| Untranslocated allele | 46C>G; 84G>A; 142A>G; 193G>A; 211G>A; 502G>A; 520delT; 573T>C; 599T>C; 677T>A; 717A>G; 737T>G; 750T>G; 752T>A | Yes | 8 |
| Translocated allele | |||
| der(3q27)‡ | 1314G>C | No | 5 |
| der(14q32) | 25C>T; 26C>T; 84G>A; 100A>G; 106A>G; 108A>C; 119C>T; 121A>C; 122G>T; 139T>C; 173C>G; 191A>T; 195A>C; 197T>C; 200T>C; 217T>C; 261T>C; 275T>A; 279T>C; 328G>C; 329C>T; 334A>C; 337A>G; 345G>A; 350-351linsG; 400G>C; 423C>G; 424T>C; 425C>G; 445A>G; 478T>A; 479G>A; 486C>T; 487C>A; 488C>T; 499C>G; 503C>G; 507C>G; 508T>A; 509G>C; 512T>A; 524T>G; 541T>G; 545C>G; 555T>A; 561C>T; 574C>A; 579C>T; 614A>G; 617A>G; 650C>G; 671T>G; 677T>A; 679C>G; 697C>T; 718T>G; 720C>G; 727A>G; 728C>T; 735G>A; 741T>C; 761 61T>A; 763A>C; 766C>G | No | 15 |
| 7. Grade 1 FL | |||
| Untranslocated allele | No clonal mutation | No | 13† |
| Translocated allele | |||
| der(3q27) | No clonal mutation | No | 4 |
| der(14q32) | 73G>C; 177G>C; 339G>C; 397G>C; 441G>A; 445A>C; 475C>T; 504A>T; 549C>T; 613A>T; 633C>T; 735G>A; 736G>A | Yes | 4 |
To analyze the MMC, sequences were aligned with the reported BCL-6 germ line gene9 (GenBank accession nos. AF 191831 and Z79581). The first nucleotide of the amplified BCL-6 first intron corresponds to the first nucleotide of the MMCF primer, defining position +1. In each case, multiple clones were obtained from at least 2 independent PCRs using different BCL-6 primers. Clonal mutations are observed in at least 2 independent clones. Taq error rate (1 × 10-3 bp-1) was also calculated by amplification and cloning of BCL-6 using an identical amount of monocyte DNA template not targeted for somatic mutation. MMC indicates major mutation cluster region; DLBCL, diffuse large B-cell lymphoma; FL, follicular lymphoma. Polymorphisms are shown in boldface type; biallelic mutations are underlined. del indicates deletion.
In case 1, no PCR product was obtained using different BCL-6 forward primers and switch reverse primers.
In absence of clonal mutations, 1 or 2 mutations per clone may have resulted from Taq error or may represent unrelated cells.
In these cases, mutations were located 3′ of the MMC.