- Email: [email protected]
Prediction of clinical relapse after bone-marrow transplantation by PCR for Philadelphia-positive acute lymphoblastic leukaemia
890
metaboliser phenotype, may be difficult. Among extensive metabolisers we have observed a subgroup with the 44/29 genotype that shows metabolic ratios of less than 03 and lacks the 29B allele. The group includes 2 brothers of North African origin,1 Turk, and 1 British subject. We therefore believe that the 44 kb gene product may still have enzyme activity in some individuals, as has been suggestedand the presence of a 44 kb allele may not always indicate a poor metaboliser or a heterozygous extensive metaboliser. The point may be especially important in epidemiological studies on the multiracial groups of Europe and North America. Heim and Meyer suggest that the 16 + 9 kb allele (15 + 9 by our sizing) might account for the 5% of poor metaboliser alleles not detected by the genotyping method they describe. We believe this is unlikely since among our poor metaboliser group there is an association between the presence of the 15 +9 kb allele and the 29B-like allele. All 4 poor metabolisers with a single 15 + 9 kb allele were homozygous for the 29B-like allele and 1 extensive metaboliser with a 29/15 + 9 genotype also had one 29B-like allele. It may be that at least one additional PM-associated allele still has to be identified. Clearly the role of the 44 kb allele should also be elucidated. Pharmacogenetics Research Unit, Department of Pharmacological Sciences,
ANN K. DALY MARTIN ARMSTRONG JEFFREY R. IDLE
Medical School, University of Newcastle upon Tyne, Newcastle upon Tyne NE2 4HH, UK
1. Skoda RB, Gonzalez FJ, Demierre A, Meyer UA. Two mutant alleles of the human cytochrome P450dbl gene (P450IID1) associated with the genetically deficient metabolism of debrisoquine and other drugs. Proc Natl Acad Sci USA 1988; 85: 5240-43. 2. Hanioka N, Kimura S, Meyer UA, Gonzalez FJ. The human CYP2D locus associated with a common genetic defect in drug oxidation: a G1934 to A base change in introm 3 of a mutant CYP2D6 allele results m an aberrant 3’ splice recognition site. Am J Hum Genet (in press). 3. Yue QY, Bertilsson L, Dahl-Puustinen MH, et al. Dissociation between debrisoquine hydroxylation phenotype and genotype among Chinese. Lancet 1989; ii: 870.
were: primer abl-1 (abl exon 1), primer abl-2 (abl exon 2), primer BCR (5’bcr exon), and junction probe BCR-abl.’,’ After the generation of cDNAs from RNA samples of bone-marrow cells, PCR was done.8 All experiments were done in duplicate, with positive and negative controls. Care was taken to avoid crosscontamination of amplified material. All samples demonstrated correct amplification of abl transcripts with primer abl-1, confirming the presence of intact RNA. Residual bcr-abl mRNA characterised by a 128 bp fragment was detected for dilution down
used
1 leukaemic cell in IOS normal cells. Seven patients proved to be in remission by cytogenetic analysis just before BMT; however, bcr-abl mRNA was detected in all 6 patients tested. The table shows the results of cytogenetic and PCR analysis at various times after BMT. 4 patients who did not show bcr-abl mRNA continued haematological and cytogenetic remission 3-6 months after analysis. In contrast, all 4 patients who were positive for bcr-abl mRNA relapsed haematologically after a median of 1 month. Although the number of patients is small these findings suggest that Phl-ALL clone has highly proliferative potential, and, in contrast with chronic myeloid leukaemia, the existence of minimal residual Phl clone may prove to correlate with haematological
to
relapse. We thank Dr Y. Onozawa, Dr M. Yazaki, Dr K. Horibe, Dr N. Hirabayashi, Dr K. Kawashima, and Dr Y. Morishita for their help.
Department of Internal Medicine, Nagoya University School of Medicine, Showa-ku, Nagoya 466, Japan; Department of Internal Medicine and Paediatrics, Japanese Red Cross Nagoya First Hospital, Nagoya, and Department of Virology, First
National Children’s Medical Research Center, Tokyo
1.
Prediction of clinical relapse after bone-marrow transplantation by PCR for
Philadelphia-positive acute lymphoblastic leukaemia SIR,—The clinical significance of minimum residual chimeric 85 kilobase (kb) bcr-abl mRNA in patients with chronic myelogenous leukaemia after bone-marrow transplantation’ (BMT) is controversial. 1-3 We have investigated remission status in eight patients with Philadelphia-positive acute lymphoblastic leukaemia (Phl-ALL) by the polymerase chain reaction (PCR) for ALLspecific 7 kb bcr-abl mRNA before and after BMT. Phl-ALL was diagnosed clinically (acute onset of leukaemia, leukaemia cells with lymphoblastic features, or no cytogenetic abnormality in marrow during remission) and by molecular characteristics (expression of 7 kb bcr-abl mRNA).4 Preparative regimens and propyhlaxis of graft-versus-host disease are described elsewhere.5,6 In allogeneic BMT, all patients received marrow without T-cell depletion from an HLA-identical sibling. Fresh bone-marrow samples were obtained with informed consent and used for analysis just before and after BMT. The oligonucleotides CLINICAL DETAILS AND RESULTS OF PCR ANALYSIS OF Ph1-ALL PATIENTS
K. MIYAMURA Y. MORISHIMA M. TANIMOTO H. SAITO S. KOJIMA Y. KODERA S. MIZUTANI
Morgan GJ, Hughes T, Janssen JWG, et al. Polymerase chain reaction for detection of
residual leukaemia. Lancet 1989; i: 928-29. 2. Bartram CR, Janssen JWG, Schmidberger M, Lyons J, Arnold R. Minimal residual leukaemia in chronic myeloid leukaemia patients after T-cell depleted bonemarrow transplantation. Lancet 1989; i: 1260. 3. Gabert J, Thuret I, Lafage M, Carcassonne Y, Maraninchi D, Mannoni P. Detection of residual bcr/abl translocation by polymerase chain reaction in chronic myeloid leukaemia patients after bone-marrow transplantation. Lancet 1989; ii: 1125-28. 4. Fainstein E, Marcelle C, Rosner A, et al. A new fused transcript in Philadelphia chromosome positive acute lymphocytic leukaemia. Nature 1987; 330: 386-88. 5. Monshima Y, Sao H, Ueda R, et al. Preliminary clinical trial of autologous bone marrow transplantation after in-vitro monoclonal antibody and complement treatment in null-cell type
acute lymphocytic leukaemia.
Jpn J Cancer Res 1985, 76:
1222-29. 6. Morishima Y, Morishita Y, Tanimoto M, et al. Low incidence of acute graft-versushost disease by the administration of methotrexate and cyclosporine in Japanese leukemia patients after bone marrow transplantation from human leukocyte antigen compatible siblings. possible role of genetic homogeneity. Blood 1989, 74: 2252-56. 7. Shtivelman E, Lifshitz B, Gale RP, Roe BA, Canaani E. Alternative splicing of RNAs transcribed from the human abl gene and from the bcr-abl fused gene. Cell 1986, 47: 277-84. 8. Hermans A, Gow J, Selleri L, et al. bcr-abl oncogene activation in Philadelphia chromosome-positive acute lymphoblastic leukemia. Leukemia 1988; 2: 628-32
Migrating stethoscopes SIR,-Ishare the concern of your peripatetic correspondent (Sept 1, p 561) about the migration of the stethoscope from pocket to neck (first dangling, later draped) to shoulder. Le cou Laennec In fractured French, they hang together. The neck, its true, May shun a stethoscope tether. ...
...
But who in heck Would choose to hang it on the shoulder? A feckless crewAt least to this épaul’d beholder! allogeneic grafts, 5-8 autologous CR, complete remission; RL, relapse. *Karyotypes of more than 20 cells were normal Patients 1-4
grafts
Department of Neurology, University of Rochester, School of Medicine and Dentistry, Rochester, New York 14642, USA
DAVID GOLDBLATT
metaboliser phenotype, may be difficult. Among extensive metabolisers we have observed a subgroup with the 44/29 genotype that shows metabolic ratios of less than 03 and lacks the 29B allele. The group includes 2 brothers of North African origin,1 Turk, and 1 British subject. We therefore believe that the 44 kb gene product may still have enzyme activity in some individuals, as has been suggestedand the presence of a 44 kb allele may not always indicate a poor metaboliser or a heterozygous extensive metaboliser. The point may be especially important in epidemiological studies on the multiracial groups of Europe and North America. Heim and Meyer suggest that the 16 + 9 kb allele (15 + 9 by our sizing) might account for the 5% of poor metaboliser alleles not detected by the genotyping method they describe. We believe this is unlikely since among our poor metaboliser group there is an association between the presence of the 15 +9 kb allele and the 29B-like allele. All 4 poor metabolisers with a single 15 + 9 kb allele were homozygous for the 29B-like allele and 1 extensive metaboliser with a 29/15 + 9 genotype also had one 29B-like allele. It may be that at least one additional PM-associated allele still has to be identified. Clearly the role of the 44 kb allele should also be elucidated. Pharmacogenetics Research Unit, Department of Pharmacological Sciences,
ANN K. DALY MARTIN ARMSTRONG JEFFREY R. IDLE
Medical School, University of Newcastle upon Tyne, Newcastle upon Tyne NE2 4HH, UK
1. Skoda RB, Gonzalez FJ, Demierre A, Meyer UA. Two mutant alleles of the human cytochrome P450dbl gene (P450IID1) associated with the genetically deficient metabolism of debrisoquine and other drugs. Proc Natl Acad Sci USA 1988; 85: 5240-43. 2. Hanioka N, Kimura S, Meyer UA, Gonzalez FJ. The human CYP2D locus associated with a common genetic defect in drug oxidation: a G1934 to A base change in introm 3 of a mutant CYP2D6 allele results m an aberrant 3’ splice recognition site. Am J Hum Genet (in press). 3. Yue QY, Bertilsson L, Dahl-Puustinen MH, et al. Dissociation between debrisoquine hydroxylation phenotype and genotype among Chinese. Lancet 1989; ii: 870.
were: primer abl-1 (abl exon 1), primer abl-2 (abl exon 2), primer BCR (5’bcr exon), and junction probe BCR-abl.’,’ After the generation of cDNAs from RNA samples of bone-marrow cells, PCR was done.8 All experiments were done in duplicate, with positive and negative controls. Care was taken to avoid crosscontamination of amplified material. All samples demonstrated correct amplification of abl transcripts with primer abl-1, confirming the presence of intact RNA. Residual bcr-abl mRNA characterised by a 128 bp fragment was detected for dilution down
used
1 leukaemic cell in IOS normal cells. Seven patients proved to be in remission by cytogenetic analysis just before BMT; however, bcr-abl mRNA was detected in all 6 patients tested. The table shows the results of cytogenetic and PCR analysis at various times after BMT. 4 patients who did not show bcr-abl mRNA continued haematological and cytogenetic remission 3-6 months after analysis. In contrast, all 4 patients who were positive for bcr-abl mRNA relapsed haematologically after a median of 1 month. Although the number of patients is small these findings suggest that Phl-ALL clone has highly proliferative potential, and, in contrast with chronic myeloid leukaemia, the existence of minimal residual Phl clone may prove to correlate with haematological
to
relapse. We thank Dr Y. Onozawa, Dr M. Yazaki, Dr K. Horibe, Dr N. Hirabayashi, Dr K. Kawashima, and Dr Y. Morishita for their help.
Department of Internal Medicine, Nagoya University School of Medicine, Showa-ku, Nagoya 466, Japan; Department of Internal Medicine and Paediatrics, Japanese Red Cross Nagoya First Hospital, Nagoya, and Department of Virology, First
National Children’s Medical Research Center, Tokyo
1.
Prediction of clinical relapse after bone-marrow transplantation by PCR for
Philadelphia-positive acute lymphoblastic leukaemia SIR,—The clinical significance of minimum residual chimeric 85 kilobase (kb) bcr-abl mRNA in patients with chronic myelogenous leukaemia after bone-marrow transplantation’ (BMT) is controversial. 1-3 We have investigated remission status in eight patients with Philadelphia-positive acute lymphoblastic leukaemia (Phl-ALL) by the polymerase chain reaction (PCR) for ALLspecific 7 kb bcr-abl mRNA before and after BMT. Phl-ALL was diagnosed clinically (acute onset of leukaemia, leukaemia cells with lymphoblastic features, or no cytogenetic abnormality in marrow during remission) and by molecular characteristics (expression of 7 kb bcr-abl mRNA).4 Preparative regimens and propyhlaxis of graft-versus-host disease are described elsewhere.5,6 In allogeneic BMT, all patients received marrow without T-cell depletion from an HLA-identical sibling. Fresh bone-marrow samples were obtained with informed consent and used for analysis just before and after BMT. The oligonucleotides CLINICAL DETAILS AND RESULTS OF PCR ANALYSIS OF Ph1-ALL PATIENTS
K. MIYAMURA Y. MORISHIMA M. TANIMOTO H. SAITO S. KOJIMA Y. KODERA S. MIZUTANI
Morgan GJ, Hughes T, Janssen JWG, et al. Polymerase chain reaction for detection of
residual leukaemia. Lancet 1989; i: 928-29. 2. Bartram CR, Janssen JWG, Schmidberger M, Lyons J, Arnold R. Minimal residual leukaemia in chronic myeloid leukaemia patients after T-cell depleted bonemarrow transplantation. Lancet 1989; i: 1260. 3. Gabert J, Thuret I, Lafage M, Carcassonne Y, Maraninchi D, Mannoni P. Detection of residual bcr/abl translocation by polymerase chain reaction in chronic myeloid leukaemia patients after bone-marrow transplantation. Lancet 1989; ii: 1125-28. 4. Fainstein E, Marcelle C, Rosner A, et al. A new fused transcript in Philadelphia chromosome positive acute lymphocytic leukaemia. Nature 1987; 330: 386-88. 5. Monshima Y, Sao H, Ueda R, et al. Preliminary clinical trial of autologous bone marrow transplantation after in-vitro monoclonal antibody and complement treatment in null-cell type
acute lymphocytic leukaemia.
Jpn J Cancer Res 1985, 76:
1222-29. 6. Morishima Y, Morishita Y, Tanimoto M, et al. Low incidence of acute graft-versushost disease by the administration of methotrexate and cyclosporine in Japanese leukemia patients after bone marrow transplantation from human leukocyte antigen compatible siblings. possible role of genetic homogeneity. Blood 1989, 74: 2252-56. 7. Shtivelman E, Lifshitz B, Gale RP, Roe BA, Canaani E. Alternative splicing of RNAs transcribed from the human abl gene and from the bcr-abl fused gene. Cell 1986, 47: 277-84. 8. Hermans A, Gow J, Selleri L, et al. bcr-abl oncogene activation in Philadelphia chromosome-positive acute lymphoblastic leukemia. Leukemia 1988; 2: 628-32
Migrating stethoscopes SIR,-Ishare the concern of your peripatetic correspondent (Sept 1, p 561) about the migration of the stethoscope from pocket to neck (first dangling, later draped) to shoulder. Le cou Laennec In fractured French, they hang together. The neck, its true, May shun a stethoscope tether. ...
...
But who in heck Would choose to hang it on the shoulder? A feckless crewAt least to this épaul’d beholder! allogeneic grafts, 5-8 autologous CR, complete remission; RL, relapse. *Karyotypes of more than 20 cells were normal Patients 1-4
grafts
Department of Neurology, University of Rochester, School of Medicine and Dentistry, Rochester, New York 14642, USA
DAVID GOLDBLATT