Allogeneic Transplant

Bone Marrow Transplantation (2003) 31, 551–557. doi:10.1038/sj.bmt.1703898

Reduced-intensity rituximab-BEAM-CAMPATH allogeneic haematopoietic stem cell transplantation for follicular lymphoma is feasible and induces durable molecular remissions

A Y L Ho1, S Devereux1, G J Mufti1 and A Pagliuca1

1Department of Haematological Medicine, Guy's, King's and St Thomas' School of Medicine, King's College Hospital, Denmark Hill, London SE5 9RS, UK

Correspondence: Dr A Ho, Department of Haematological Medicine, King's College Hospital, Denmark Hill, London SE5 9RS, UK

Received 10 July 2002; Accepted 1 October 2002.

Top

Abstract

The indolent non-Hodgkin's lymphomas are theoretically curable through allogeneic haematopoietic stem cell transplantation (allo-HSCT). The applicability of standard conditioning allo-HSCT is, however, restricted by its toxicity. Recently, reduced-intensity conditioning regimens have demonstrated efficacy with significantly reduced early morbidity and mortality. We examined the safety and efficacy of rituximab-BEAM-CAMPATH as reduced-intensity conditioning for allo-HSCT in follicular lymphomas. Minimal residual disease was assessed by polymerase chain reaction (PCR) for bcl-2/IgH translocations, and chimerism by X,Y-FISH or PCR amplification of short tandem repeat sequences. At a median follow-up of 521 days (371–719), four of five patients were alive and three were in complete molecular remission. Three patients required pre-emptive treatment for CMV reactivation. One succumbed to a perforated viscus and one had slowly progressive disease. A graft-versus-lymphoma effect was demonstrable and quantitative PCR for bcl-2/IgH may allow better accuracy in scheduling post-allograft rituximab and/or donor lymphocyte infusions. Although follow-up is short, reduced-intensity allo-HSCT with BEAM-CAMPATH conditioning appears to be safe, effective in inducing a molecular remission and is potentially curative. Further recruitment and much longer follow-up will be necessary to determine if this impacts favourably upon disease-free and overall survival.

Keywords:

reduced-intensity HSCT, follicular NHL, qPCR

Follicle centre cell lymphoma usually presents at an advanced median age of approximately 60 years and in an advanced stage. However, a significant proportion of patients present at a younger age. Although it has an indolent natural history with a median survival of 7–9 years and complete clinical remission may be achieved with standard chemotherapy, a chronic relapsing course with progressively shorter periods of remission is typical, with most patients eventually succumbing to their disease.1

The t(14;18)(q32;q21) is the karyotypic hallmark of follicular lymphomas. This results in an overexpression of the bcl-2 gene as it comes under the influence of the IgH promoter and immortalisation of the cell. In the majority (approximately 70%) of patients, the translocation breakpoint on chromosome 18 occurs within the major breakpoint region (MBR) within exon III of the bcl-2 gene.2 Polymerase chain reaction (PCR) methods, both qualitative and quantitative, targeted at these breakpoints have been developed, which enable the detection of as few as one cell harbouring the bcl-2/IgH translocation in 105–106 normal cells. However, the threshold above which t(14;18) positivity becomes significant is unknown3 as the translocation can also be detected in normal healthy individuals.4,5,6

High-dose therapy and autologous transplantation have been undertaken to improve disease-free survival. However, autologous transplantation of unmanipulated haematopoietic stem cells (HSCT) is associated with a high relapse rate.7,8 Relapse is strongly associated with the re-emergence or persistence of molecular marker positive cells detected by PCR.9,10,11,12 In vivo or in vitro purging of the graft with monoclonal antibodies has demonstrated an improvement in disease-free survival, most marked in patients whose infused cells were negative for molecular markers.11,13,14,15 Persistent marrow negativity is strongly predictive of continued complete remission,9 with peripheral blood being less predictive.16 Unfortunately, the risk of developing treatment-related myelodysplastic syndrome is substantial, at up to 19.8% at 10 years with no evidence of a plateau.17

At present, the only treatment modality that may be delivered with curative intent is allogeneic HSCT (allo-HSCT), with its attendant graft-versus-lymphoma (GvL) effects.18,19 However, the transplant-related mortality (TRM) associated with conventional transplant conditioning is high at 25–40%,20,21 resulting in no apparent improvement in overall survival although disease-free survival is significantly improved.8,21,22,23 The high toxicity and TRM of conventional transplant conditioning restricts its applicability in an 'indolent' disease in a group of advanced age and often with significant comorbidity.

Within the last few years, 'reduced-intensity' conditioning regimens have been shown to be feasible and applicable to a wide range of malignant and nonmalignant disorders. The improvements in toxicity and accompanying TRM are impressive, with rapid engraftment and demonstrable graft-versus-malignancy effects.24,25,26,27,28

Rituximab is a chimeric monoclonal anti-CD20 antibody with proven activity against follicular lymphoma.29,30 In combination with CHOP, molecular remissions have been demonstrable,31 which translates to significantly improved relapse-free survival.32 Its use as an in vivo purging agent in the autologous transplant scenario can lead to PCR negative harvests, and there is suggestion of a synergistic effect with cytotoxic drugs.14

We therefore established an allogeneic transplant protocol in follicular lymphoma utilising conditioning with 'BEAM', a proven conditioning regimen for autologous33 and allogeneic34 transplantation, together with CAMPATH-1H (anti-CD52) to minimise the risk of rejection, graft-versus-host disease35 and to maximise tumour bulk reduction. Previous series have demonstrated its feasibility.34,36 Rituximab delivered within 120 days preconditioning was used as in vivo purging. Molecular monitoring of MRD was achieved through qualitative and quantitative PCR for the bcl-2/IgH translocation, and chimerism by XY-FISH or PCR of short tandem repeat sequences (STR).

Top

Materials and methods

Qualitative t(14;18) PCR

Genomic DNA was extracted from the mononuclear cell fraction of clinical blood and bone marrow samples following density-gradient centrifugation on Ficoll-Histopaque®-1077 (Sigma-Aldrich, Dorset, UK) using the Wizard® Genomic DNA Purification Kit (Promega UK Ltd, Southampton). Precipitated DNA was rehydrated in 10 mM Tris-HCl (pH 7.4) and 1 mM EDTA (pH 8.0) and quantitated by measuring the optical density at 260 nm. Positive control DNA was extracted from the DOHH-2 cell line (DSMZ, Berlin, Germany) and negative control DNA from the K562 cell line (DSMZ, Berlin, Germany) by an identical methodology.

DNA integrity was confirmed by amplification with beta-actin primers (forward AAA TCT GGC ACC ACA CCT TC) (reverse AAC GGC AGA AGA GAG AAC CA) (MWG Biotech, Milton Keynes), 0.6 U Taq DNA polymerase (Promega UK Ltd), 1 mM dNTPs (Roche Molecular Diagnostics, UK) and 1.5 mM magnesium chloride in PCR buffer (Sigma-Aldrich, Dorset, UK) in a total volume of 25 mul. Reactions were denatured at 95°C for 3 min, subject to 40 cycles of 95°C/60 s; 60°C/60 s; 72°C/60 s, and a final elongation step of 72°C for 7 min in a GeneAmp PCR System 2700 thermal cycler (Applied Biosystems, Cheshire, UK).

Four replicates of 0.1 mug of DNA were amplified with 400 nM forward primer MBR1 (TAT GGT GGT TTG ACC TTT AG), 400 nM consensus JH primer (ACC TGA GGA GAC GGT GAC) (MWG Biotech, Milton Keynes),37 0.6 U Taq DNA polymerase (Promega UK Ltd), 1 mM dNTPs (Roche Molecular Diagnostics, UK) and 1.5 mM magnesium chloride in PCR buffer (Sigma-Aldrich, Dorset, UK) in a total volume of 25 mul. Reactions were denatured at 95°C for 3 min, subject to 40 cycles of 95°C/60 s, 60°C/60 s, 72°C/60 s, and a final elongation phase of 72°C for 7 min. The products were visualised after electrophoresis through an ethidium bromide stained 2% agarose gel. The reproducible sensitivity of this assay was approximately 1:1000 cells.

A 1 mul aliquot of this first round product was subjected to a second round reaction using 250 nM internal primer MBR2 (GAG TTG CTT TAC GTG GCC TG), 250 nM consensus JH (ACC TGA GGA GAC GGT GAC) and 125 nM dual labelled internal hydrolysis probe (6FAM-TCA ACA CAG ACC CAC CCA GAG CCC(TAMRA)Tp) (TIBMolbiol, Berlin, Germany), with 1 mM dNTPs, 0.6 U Taq DNA polymerase, 1.5 mM magnesium chloride in PCR buffer (Sigma-Aldrich, Dorset, UK) in a total volume of 20 mul. The reaction mixture was subjected to 1 min denaturation at 95°C, then 20 cycles of 95°C/15 s and 58°C/30 s in glass capillary tubes within a LightCycler® (Roche Diagnostics, Lewes) instrument. The combined reproducible sensitivity was approximately 1:10 000 cells.

Quantitative t(14;18) PCR

Two replicates of 0.1 mug of DNA were amplified with primer MBR2 (GAG TTG CTT TAC GTG GCC TG), 250 nM consensus JH (ACC TGA GGA GAC GGT GAC) and 125 nM dual labelled internal hydrolysis probe (6FAM-TCA ACA CAG ACC CAC CCA GAG CCC (TAMRA)Tp), with 1 mM dNTPs, 0.6 U Taq DNA polymerase, 1.5 mM magnesium chloride in PCR buffer (Sigma-Aldrich, Dorset, UK) in a total volume of 20 mul. The reaction mixture was subjected to 1 min denaturation at 95°C, then 60 cycles of 95°C/15 s and 58°C/30 s in glass capillary tubes within a LightCycler® (Roche Diagnostics, Lewes). Replicate 0.1 mug aliquots were amplified in parallel with 250 nM of each beta-actin primer (forward ACC CAC ACT GTG CCC ATC TA) (reverse CGG AAC CGC TCA TTG CC) and 125 nM probe (6FAM-ATG CCC XT CCC CCA TGC CAT CCT GCG Tp) utilising identical reaction mixes and conditions. A semiquantitative result was derived by utilising the ratio of the calculated t(14;18) to the calculated beta-actin. The sensitivity of the quantitative assay was approximately 1:2000 cells.

Chimerism analysis

DNA was extracted from whole blood using the QIAamp DNA blood mini-kit (Qiagen UK Ltd). The resulting DNA was processed using GenePrint® Fluorescent STR Multiplex-CSF1PO,TPOX, TH01, vWA and GenePrint® Fluorescent STR Multiplex-GammaSTRTM systems (Promega UK Ltd) following the manufacturer's instructions, and analysed on an ABI Prism 377 DNA sequencer (Applied Biosystems, Cheshire, UK). Interphase FISH analysis was performed on mononuclear cells separated from whole blood using a direct labelled Chromosome X/Y probe cocktail (Qbiogene, UK) following the manufacturer's instructions.

Patients

Patients with CD20 positive follicular lymphoma, refractory, in 1st partial or 2nd complete remission were eligible. The study protocol was approved by the local ethics committee and informed consent was obtained from each patient. Rituximab (Mabthera®) was delivered at a dose of 375 mg/m2 weekly for 4 weeks within 12 weeks prior to conditioning. Reduced-intensity conditioning comprised 'BEAM-CAMPATH', scheduled as follows: CAMPATH-1H 20 mg once daily i.v. on Days –5 to –1; BCNU 300 mg/m2 i.v. on D-6; cytarabine 200 mg/m2 12 hourly i.v. on Days –5 to –2; etoposide 200 mg/m2 once daily i.v. on Days –5 to –2; melphalan 140 mg/m2 i.v. Day –1; HSC infusion D0. G-CSF (filgrastim, Neupogen®) 300 mug daily was administered from D+7 to neutrophil recovery. Immunosuppression was with cyclosporin, titrated to achieve plasma levels of 150–200 ng/l and tapered from Day +58 in the absence of clinical GvHD. Toxicity was assessed utilising the abbreviated Common Toxicity Criteria of the NCIC38 and response assessment was made using the guidelines of the International workshop.39 GvHD was graded according to the Consensus Conference on GvHD grading.40

All patients received standard nursing and supportive care protocols for neutropenic patients. Infection prophylaxis comprised itraconazole suspension 200 mg p.o. twice daily, high-dose acyclovir 500 mg/m2 8 hourly i.v. from Day –5 in high-risk patients (patient and/or donor CMV positive), otherwise 200 mg 8 hourly p.o. and aerosolised pentamidine 300 mg on Day –10. Blood products were universally leucodepleted and irradiated to 25 cGy. CMV seronegative recipients received CMV seronegative blood products. Prior to August 2000, CMV monitoring was performed using a fluorescent antigen detection methodology (CENFA). Subsequently, a PCR-based DNA assay several-fold more sensitive than the CENFA was utilised. All patients were monitored weekly until Day +100 or until immunosuppression was completely withdrawn and no graft-versus-host disease (GvHD) was present. Two positive PCR assays or a single positive CENFA were indications for pre-emptive therapy with intravenous ganciclovir.

All data were censored on 30 June 2002.

Top

Results

At the time of reporting, a total of five patients (two male; three female), with a median age at transplant of 50.9 years (42.6–55.8), had been recruited, over a period of approximately 6 months. The median age at diagnosis was 49.3 (39.4–52.7) years and at transplant 50.9 (42.6–55.8) years. At a median follow-up of 521 days (371–719), four of five patients were alive and three were in complete radiological and molecular remission. Patient and disease characteristics are shown in Table 1.


At transplant, all were in complete or partial remission after a median of three previous courses of chemotherapy.2,3,4,5,6,7 All initial stage IV patients had continued marrow infiltration morphologically. Residual clinical/radiological evidence of pathological lymphadenopathy was present in two patients and three had detectable bcl-2/IgH translocations. Four received peripheral blood stem cells from a sibling donor with a median CD34 cell count of 5.21 times 106/kg (2.9 times 106–6.95 times 106) and one bone marrow from a volunteer-unrelated donor with 3.45 times 106/kg CD34+ cells.

The immediate post-transplant toxicities were limited to NCIC grade 1–2 stomatitis, nausea, vomiting and grade 3 febrile neutropenia. The median time to neutrophil (>0.5 times 109/l) and platelet (>20 times 109/l) recovery was 12 days (Table 2). There was no delayed or secondary neutropenia following regeneration.


One patient (Patient no. 1) pancytopenic with secondary myelodysplasia, did not receive pre-transplant rituximab, as it was felt that this would result in an unnecessary delay to allografting. Following demonstration of a persistent marrow infiltrate, predominantly recipient chimerism and rapidly progressive nodal disease at D+80, cyclosporin was completely withdrawn and rituximab delivered on D+82. Although he then achieved morphological clearance of his bone marrow and bcl-2/IgH negativity, he failed to attain MRD because of the persistence of pathological lymphadenopathy. DLI was delivered on D+204 with 5 times 106 CD3+/kg resulting in complete remission and conversion to predominantly donor chimerism but with grade III gastrointestinal and liver GvHD. He succumbed to a perforated viscus on D+371 (Table 3).


Patient no. 2 failed to achieve either a clinical or molecular response to the transplant or to post-transplant rituximab delivered from D+199, despite morphological clearance of marrow, with stable disease on imaging. Late graft failure with autologous regeneration was demonstrated by chimerism studies on D+409. Quantitative PCR demonstrated a progressive relative increase in bcl-2/IgH positivity, preceding the initial clinical evidence of progressive disease on D+497 (Figure 1). At the time of reporting, he had slowly progressive disease and is awaiting a second reduced-intensity allograft.

Figure 1.
Figure 1 - Unfortunately we are unable to provide accessible alternative text for this. If you require assistance to access this image, please contact help@nature.com or the author

Patient no. 2 quantitative PCR post-allograft, y-axis shows the ratio of bcl-2lgH positive cells to beta-actin positive cells.

Full figure and legend (17K)

Two patients (Patient nos. 4 and 5) achieved clinical and molecular remissions at first assessment following allograft. Patient no. 3 achieved a clinical and morphological remission and full donor chimerism, but demonstrated persistent bcl-2/IgH positivity on D+110. Chronic limited cutaneous GvHD developed at D+129 following complete withdrawal of cyclosporin, with subsequent bcl-2/IgH negativity. These three patients remain in complete clinical and molecular remission.

Donor lymphocyte infusions were delivered to three patients: Patient no. 1 for disease progression and declining donor chimerism, Patient no. 4 for the emergence of a small t(8;14) clone on D+482 and Patient no. 5 for declining donor chimerism. DLI was effective against the indications for administration.

CMV reactivation was demonstrated in three patients: Patient no. 4 at D+249, Patient no. 5 at D+38, Patient no. 3 at D+54, D+242 and D+345. All responded well to intravenous gancyclovir. In addition, Patient no. 3 required aerosolised ribavirin therapy for community-acquired RSV upper respiratory tract infection at D+53 and D+74.

Top

Discussion

At present, allo-HSCT represents the only therapeutic modality for the indolent lymphomas, which may be delivered with curative intent. Its applicability, however, is limited by the availability of an allogeneic HSC donor and the toxicity of the conditioning regimen.

Despite the dramatic improvements achieved to date in reducing transplant-related complications and mortality, it may be difficult to justify allo-HSCT as 'up-front' therapy for an 'indolent' disease with a long median survival. The optimal timing of allogeneic HSCT is unresolved. For this study, we therefore selected patients with relatively resistant disease beyond 1st partial remission, or patients beyond 2nd complete remission, with an available molecular marker for minimal residual disease and with a suitable sibling or volunteer-unrelated donor. These criteria have undoubtedly limited accrual into the study.

Nevertheless, reduced-intensity conditioning with rituximab-BEAM-CAMPATH appears to be safe and efficacious in permitting durable engraftment. Although effective against marrow disease, conditioning appears to have a limited effect against nodal disease present at transplant as evidenced by an absence of response in the two patients with residual lymphadenopathy. This is in keeping with the findings of other studies that chemosensitive disease and minimal disease state at the time of allografting are significantly associated with survival.8,13

A graft-versus-lymphoma effect has been demonstrated, most marked following DLI and withdrawal of immunosuppression, with regression of nodal disease in one patient and the achievement of PCR negativity in a second. Rituximab delivered post-transplant, although safe and complication-free, did not impact significantly upon residual lymphoma, again in the two patients with residual nodal disease. A more formal purging schedule with rituximab immediately prior to HSC infusion and closely there after may improve disease control41 when used in conjunction with specific conditioning protocols. While CAMPATH-1H was administered for the sole purpose of immunosuppression to minimise GvHD and graft rejection,35,42 the antilymphoma properties of CAMPATH-1H in this setting cannot be discounted.6

There appears to be a high incidence of CMV reactivation, with three of five patients requiring pre-emptive therapy. While the numbers in this study are far too small to draw any conclusions, delays in immune reconstitution following CAMPATH-1H may play a role.43,44 This may be due to delayed T-cell reconstitution or effects on antigen-presenting cells.45

Although follow-up is short, reduced-intensity allo-HSCT with rituximab-BEAM-CAMPATH conditioning appears to be safe, effective in inducing durable donor engraftment and molecular remissions and is potentially curative. Quantitative PCR for bcl-2/IgH may allow more accurate targeted scheduling of rituximab and/or DLI in the post-HSCT setting, although the 'threshold' for instituting therapy has yet to be determined. The majority of reduced-intensity conditioning protocols to date have included fludarabine because of its excellent immuno-suppressive and antilymphoma properties. However, with the inclusion of fludarabine as first line or salvage chemotherapy in lymphoma, the adoption of a nonfludarabine-containing transplant conditioning regimen may be advantageous and worthy of further investigation. It is far too early to determine if a reduced-intensity allograft approach will affect the overall or disease-free survival in this indolent lymphoma, but these early results appear encouraging. Further follow-up with a larger cohort of patients is essential, as is investigation into the role of rituximab in the pre-and post-transplant setting as well as into any potential antilymphoma properties of CAMPATH.

Top

References

  1. Johnson PW, Rohatiner AZ, Whelan JS et al. Patterns of survival in patients with recurrent follicular lymphoma: a 20-year study from a single center. J Clin Oncol 1995; 13: 140–147. | PubMed | ISI | ChemPort |
  2. Cleary ML, Sklar J. Nucleotide sequence of a t(14;18) chromosomal breakpoint in follicular lymphoma and demonstration of a breakpoint-cluster region near a transcriptionally active locus on chromosome 18. Proc Natl Acad Sci USA 1985; 82: 7439–7443. | Article | PubMed | ChemPort |
  3. Hirt C, Dolken G. Quantitative detection of t(14;18)-positive cells in patients with follicular lymphoma before and after autologous bone marrow transplantation. Bone Marrow Transplant 2000; 25: 419–426. | Article | PubMed | ISI | ChemPort |
  4. Summers KE, Goff LK, Wilson AG et al. Frequency of the Bcl-2/IgH rearrangement in normal individuals: implications for the monitoring of disease in patients with follicular lymphoma. J Clin Oncol 2001; 19: 420–424. | PubMed | ISI | ChemPort |
  5. Limpens J, Stad R, Vos C et al. Lymphoma-associated translocation t(14;18) in blood B cells of normal individuals. Blood 1995; 85: 2528–2536. | PubMed | ISI | ChemPort |
  6. Dolken G, Illerhaus G, Hirt C et al. BCL-2/JH rearrangements in circulating B cells of healthy blood donors and patients with nonmalignant diseases. J Clin Oncol 1996; 14: 1333–1344. | PubMed | ISI | ChemPort |
  7. Mijovic A, Parker J, Salisbury J et al. Autologous blood or bone marrow transplantation for relapsed and partially responding low-grade non-Hodgkin's lymphoma. Cancer Res Ther Control 1998; 5: 253–260. | ISI |
  8. Stein RS, Greer JP, Goodman S et al. High-dose therapy with autologous or allogeneic transplantation as salvage therapy for small cleaved cell lymphoma of follicular center cell origin. Bone Marrow Transplant 1999; 23: 227–233. | Article | PubMed | ISI | ChemPort |
  9. Freedman AS, Gribben JG, Neuberg D et al. High-dose therapy and autologous bone marrow transplantation in patients with follicular lymphoma during first remission. Blood 1996; 88: 2780–2786. | PubMed | ISI | ChemPort |
  10. Hardingham JE, Kotasek D, Sage RE et al. Molecular detection of residual lymphoma cells in peripheral blood stem cell harvests and following autologous transplantation. Bone Marrow Transplant 1993; 11: 15–20. | PubMed | ISI | ChemPort |
  11. Freedman AS, Neuberg D, Mauch P et al. Long-term follow-up of autologous bone marrow transplantation in patients with relapsed follicular lymphoma. Blood 1999; 94: 3325–3333. | PubMed | ISI | ChemPort |
  12. Corradini P, Astolfi M, Cherasco C et al. Molecular monitoring of minimal residual disease in follicular and mantle cell non-Hodgkin's lymphomas treated with high-dose chemotherapy and peripheral blood progenitor cell autografting. Blood 1997; 89: 724–731. | PubMed | ISI | ChemPort |
  13. Horning SJ, Negrin RS, Hoppe RT et al. High-dose therapy and autologous bone marrow transplantation for follicular lymphoma in first complete or partial remission: results of a phase II clinical trial. Blood 2001; 97: 404–409. | Article | PubMed | ISI | ChemPort |
  14. Magni M, Di Nicola M, Devizzi L et al. Successful in vivo purging of CD34-containing peripheral blood harvests in mantle cell and indolent lymphoma: evidence for a role of both chemotherapy and rituximab infusion. Blood 2000; 96: 864–869. | PubMed | ISI | ChemPort |
  15. Pappa VI, Wilkes S, Salam A et al. Use of the polymerase chain reaction and direct sequencing analysis to detect cells with the t(14;18) in autologous bone marrow from patients with follicular lymphoma, before and after in vitro treatment. Bone Marrow Transplant 1998; 22: 553–558. | Article | PubMed | ISI | ChemPort |
  16. Gribben JG, Neuberg D, Barber M et al. Detection of residual lymphoma cells by polymerase chain reaction in peripheral blood is significantly less predictive for relapse than detection in bone marrow. Blood 1994; 83: 3800–3807. | PubMed | ISI | ChemPort |
  17. Friedberg JW, Neuberg D, Stone RM et al. Outcome in patients with myelodysplastic syndrome after autologous bone marrow transplantation for non-Hodgkin's lymphoma. J Clinl Oncol 1999; 17: 3128–3135. | ChemPort |
  18. Verdonck LF. Allogeneic versus autologous bone marrow transplantation for refractory and recurrent low-grade non-Hodgkin's lymphoma: updated results of the Utrecht experience. Leukemia Lymphoma 1999; 34: 129–136. | PubMed | ISI | ChemPort |
  19. Verdonck LF, Dekker AW, Lokhorst HM et al. Allogeneic versus autologous bone marrow transplantation for refractory and recurrent low-grade non-Hodgkin's lymphoma. Blood 1997; 90: 4201–4205. | PubMed | ISI | ChemPort |
  20. Chopra R, Goldstone AH, Pearce R et al. Autologous versus allogeneic bone marrow transplantation for non-Hodgkin's lymphoma: a case-controlled analysis of the European Bone Marrow Transplant Group Registry data. J Clin Oncol 1992; 10: 1690–1695. | PubMed | ISI | ChemPort |
  21. van Besien K, Sobocinski KA, Rowlings PA et al. Allogeneic bone marrow transplantation for low-grade lymphoma. Blood 1998; 92: 1832–1836. | PubMed | ISI | ChemPort |
  22. Toze CL, Shepherd JD, Connors JM et al. Allogeneic bone marrow transplantation for low-grade lymphoma and chronic lymphocytic leukemia. Bone Marrow Transplant 2000; 25: 605–612. | Article | PubMed | ISI | ChemPort |
  23. Dann EJ, Daugherty CK, Larson RA. Allogeneic bone marrow transplantation for relapsed and refractory Hodgkin's disease and non-Hodgkin's lymphoma. Bone Marrow Transplant 1997; 20: 369–374. | Article | PubMed | ISI | ChemPort |
  24. Carella AM, Champlin R, Slavin S et al. Mini-allografts: ongoing trials in humans. Bone Marrow Transplant 2000; 25: 345–350. | Article | PubMed | ISI | ChemPort |
  25. Nagler A, Slavin S, Varadi G et al. Allogeneic peripheral blood stem cell transplantation using a fludarabine-based low intensity conditioning regimen for malignant lymphoma. Bone Marrow Transplant 2000; 25: 1021–1028. | Article | PubMed | ISI | ChemPort |
  26. Slavin S, Nagler A, Naparstek E et al. Nonmyeloablative stem cell transplantation and cell therapy as an alternative to conventional bone marrow transplantation with lethal cytoreduction for the treatment of malignant and nonmalignant hematologic diseases. Blood 1998; 91:756–763. | PubMed | ISI | ChemPort |
  27. Sykes M, Preffer F, McAfee S et al. Mixed lymphohaemopoietic chimerism and graft-versus-lymphoma effects after non-myeloablative therapy and HLA-mismatched bone-marrow transplantation. Lancet 1999; 353: 1755–1759. | Article | PubMed | ISI | ChemPort |
  28. McSweeney PA, Niederwieser D, Shizuru JA et al. Hematopoietic cell transplantation in older patients with hematologic malignancies: replacing high-dose cytotoxic therapy with graft-versus-tumor effects. Blood 2001; 97: 3390–3400. | Article | PubMed | ISI | ChemPort |
  29. Hainsworth JD, Burris HA, 3rd, Morrissey LH et al. Rituximab monoclonal antibody as initial systemic therapy for patients with low-grade non-Hodgkin lymphoma. Blood 2000; 95: 3052–3056. | PubMed | ISI | ChemPort |
  30. Colombat P, Salles G, Brousse N et al. Rituximab (anti-CD20 monoclonal antibody) as single first-line therapy for patients with follicular lymphoma with a low tumor burden: clinical and molecular evaluation. Blood 2001; 97: 101–106. | Article | PubMed | ISI | ChemPort |
  31. Czuczman MS, Grillo-Lopez AJ, McLaughlin P et al. Clearing of cells bearing the bcl-2 [t(14;18)] translocation from blood and marrow of patients treated with rituximab alone or in combination with CHOP chemotherapy. Ann Oncol 2001; 12: 109–114. | Article | PubMed | ISI | ChemPort |
  32. Coiffier B, Lepage E, Briere J et al. CHOP chemotherapy plus rituximab compared with CHOP alone in elderly patients with diffuse large-B-cell lymphoma. N Engl J Med 2002; 346: 235–242. | Article | PubMed | ISI | ChemPort |
  33. Bastion Y, Brice P, Haioun C et al. Intensive therapy with peripheral blood progenitor cell transplantation in 60 patients with poor-prognosis follicular lymphoma. Blood 1995; 86: 3257–3262. | PubMed | ISI | ChemPort |
  34. Przepiorka D, van Besien K, Khouri I et al. Carmustinem etoposide, cytarabine and melphalan as a preparative regimen for allogeneic transplantation for high-risk malignant lymphoma. Ann Oncol 1999; 10: 527–532. | PubMed | ISI | ChemPort |
  35. Hale G, Jacobs P, Wood L et al. CD52 antibodies for prevention of graft-versus-host disease and graft rejection following transplantation of allogeneic peripheral blood stem cells. Bone Marrow Transplant 2000; 26: 69–76. | Article | PubMed | ISI | ChemPort |
  36. Cull GM, Haynes AP, Byrne JL et al. Preliminary experience of allogeneic stem cell transplantation for lymphoproliferative disorders using BEAM-CAMPATH conditioning: an effective regimen with low procedure-related toxicity. Br J Haematol 2000; 108: 754–760. | Article | PubMed | ISI | ChemPort |
  37. Hosler GA, Bash RO, Bai X et al. Development and validation of a quantitative polymerase chain reaction assay to evaluate minimal residual disease for T-cell acute lymphoblastic leukemia and follicular lymphoma. Am J Pathol 1999; 154: 1023–1035. | PubMed | ISI | ChemPort |
  38. Therapy Evaluation Programme, Common Toxicity Criteria, Version 2.0. In: National Cancer Institute, 1999.
  39. Cheson BD, Horning SJ, Coiffier B et al. Report of an International Workshop to Standardize Response Criteria for Non-Hodgkin's Lymphomas. J Clin Oncol 1999; 17: 1244–1253. | PubMed | ISI | ChemPort |
  40. Przepiorka D, Weisdorf D, Martin P et al. 1994 Consensus Conference on Acute GVHD Grading. Bone Marrow Transplant 1995; 15: 825–828. | PubMed | ISI | ChemPort |
  41. Khouri IF, Saliba RM, Giralt SA et al. Nonablative allogeneic hematopoietic transplantation as adoptive immunotherapy for indolent lymphoma: low incidence of toxicity, acute graft-versus-host disease, and treatment-related mortality. Blood 2001; 98: 3595–3599. | Article | PubMed | ISI | ChemPort |
  42. Kottaridis PD, Milligan DW, Chopra R et al. In vivo CAMPATH-1H prevents graft-versus-host disease following nonmyeloablative stem cell transplantation. Blood 2000; 96: 2419–2425. | PubMed | ISI | ChemPort |
  43. Chakrabarti S, Mackinnon S, Chopra R et al. High incidence of cytomegalovirus infection after nonmyeloablative stem cell transplantation: potential role of Campath-1H in delaying immune reconstitution. Blood 2002; 99: 4357–4363. | Article | PubMed | ISI | ChemPort |
  44. Qamruddin AO, Oppenheim BA, Guiver M et al. Screening for cytomegalovirus (CMV) infection in allogeneic bone marrow transplantation using a quantitative whole blood polymerase chain reaction (PCR) method: analysis of potential risk factors for CMV infection. Bone Marrow Transplant 2001; 27: 301–306. | Article | PubMed | ISI | ChemPort |
  45. Buggins AGS, Mufti GJ, Salisbury J et al. Peripheral blood but not tissue dendritic cells express CD52 and are depleted by treatment with alemtuzumab. Blood 2002; 100: 1715–1720. | PubMed | ISI | ChemPort |
Top

Acknowledgements

We acknowledge Dr O Landt, TIB MOLBIOL, Berlin for the design and synthesis of MBR2 and bcl-2 probe.

Dr A Ho was supported by an educational grant from Roche Pharmaceuticals.

Extra navigation

.

naturejobs

More science jobs
Post a job for free
ADVERTISEMENT