Quantitative analysis of chimerism after allogeneic stem cell transplantation by PCR amplification of microsatellite markers and capillary electrophoresis with fluorescence detection: the Paris–Robert Debré experience

Acquaviva, Cécile; Duval, Michel; Mirebeau, Delphine; Bertin, Raphaëlle; Cavé, Hélène

doi:10.1038/sj.leu.2402762

Cited by 38 publications

(27 citation statements)

References 17 publications

(22 reference statements)

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“…This real-time PCR has a less quantitative accuracy with a variation coefficient of only 30-50% 54,55 compared to 5% of the STR systems. 44,46,[57][58][59] This does not hamper the analysis when only very low levels of mixed chimerism are needed to be quantified. However, in the range above 5% autologous cells, a variation coefficient of approximately 50% does not yet permit accurate quantitative investigations of the dynamic of mixed chimerism post transplant.…”

Section: Methods For Chimerism Analysismentioning

confidence: 99%

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How and when should we monitor chimerism after allogeneic stem cell transplantation?

Bader

Niethammer

Willasch

et al. 2004

Bone Marrow Transplant

180

155

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Summary:Chimerism analysis has become an important tool for the peri-transplant surveillance of engraftment. It offers the possibility to realize impending graft rejection and can serve as an indicator for the recurrence of the underlying malignant or nonmalignant disease. Most recently, these investigations have become the basis for treatment intervention, for example, to avoid graft rejection, to maintain engraftment and to treat imminent relapse by pre-emptive immunotherapy. This invited review focuses on the clinical implications of characterization of hematopoietic chimerism in stem cell transplantation.

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Section: Methods For Chimerism Analysismentioning

confidence: 99%

“…Semiautomated PCR analyses using the appropriate hardware allowed moreover a high sample throughput. 41,[44][45][46][47] This made it possible to study chimerism in all patients and in short time intervals (already early) after transplantation. Reconditioning and second transplant probably necessary.…”

Section: Methods For Chimerism Analysismentioning

confidence: 99%

How and when should we monitor chimerism after allogeneic stem cell transplantation?

Bader

Niethammer

Willasch

et al. 2004

Bone Marrow Transplant

180

155

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“…Chimerism was assessed by quantitative PCR of STR by LabCorp (San Antonio, TX, USA) for San Antonio patients or using a technique we described elsewhere for Montreal patients. 29 The sensitivity of these chimerism assays was ± 5%. Complete donor chimerism was defined as 490% cells of donor origin in blood.…”

Section: Assessment Of Trial Endpointsmentioning

confidence: 99%

Engraftment of unrelated cord blood after reduced-intensity conditioning regimen in children with refractory neuroblastoma: a feasibility trial

Jubert

Wall

Grimley

et al. 2010

Bone Marrow Transplant

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Relapsed or refractory neuroblastoma is uniformly fatal. We hypothesized that allogeneic response could provide a platform for immunotherapy in neuroblastoma. We therefore undertook a pilot trial of unrelated cord blood transplantation after reduced intensity conditioning regimen (RIC) in children with relapsed neuroblastoma to assess engraftment and tolerability in this heavily pretreated population. The RIC included CY (50 mg/kg, day À6), fludarabine (40 mg/m 2 , days À6 to À2), total body irradiation (200 cGy, day À1), and rabbit antithymocyte globulin (2.5 mg/kg, days À3 to À1). Six patients were enrolled: four were in partial responsive relapse, one with a mixed response and one in refractory relapse. All patients tolerated the regimen well and had donor engraftment with full neutrophil and plt recovery (median time 12 and 35 days, respectively). One patient never experienced neutropenia and another did not need plt transfusions. All patients progressed after transplant (median time 55 days, 26-180 days). Natural killer (NK) cell counts were normal within 2 months, whereas T-cell recovery was slower. In conclusion, unrelated cord blood engrafts after RIC in children with refractory neuroblastoma. Future research should be aimed at transplanting patients with minimal residual disease, using less intensive immunosuppression and adding NK-cell based post transplant immunotherapy.

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“…Several methods for evaluating chimerism have been developed, including cytogenetic and molecular techniques. The most widely used technique for evaluating chimerism is polymerase chain reaction (PCR) amplification of short tandem repeats (STR) [4][5][6][7][8][9][10]. However, the sensitivity of this method is only approximately 1-5%, even when fluorescent-labeled primers and an automated DNA sequencer are used [4][5][6].…”

Section: Introductionmentioning

confidence: 99%

Long‐term persistence of host cells detected by X‐chromosome gene‐based assay in patients undergoing gender‐mismatched hematopoietic stem cell transplantation

et al. 2005

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Using our recently developed human androgen receptor (HUMARA) gene-based chimerism assay, long-term chimerism was investigated in female patients who underwent hematopoietic stem cell transplantation (HCT) with cells from male donors. After restriction digestion of samples, we detected a small number of female-derived cells within a large population of male-derived cells, with a sensitivity from 0.1% to 0.05%. Chimerism was examined in four patients with myeloid malignancies: two patients with acute myeloid leukemia (AML) from myelodysplastic syndrome (MDS), and one patient each with AML M3 and AML M4. All patients underwent myeloablative conditioning regimens and exhibited good clinical results during a median follow-up period of 6.6 years (range, 3.4-7.5 years). Female-derived cells were detected throughout the entire follow-up period in all bone marrow samples, but they became undetectable in the peripheral blood samples of 3 patients. Moreover, the HUMARA band pattern suggests that these residual host cells were normal cells. This study confirms the usefulness of the HUMARA gene-based assay, which showed that patients undergoing HCT frequently show mixed chimerism (MC) for a long period, especially in bone marrow, although the possibility of contamination by host stromal cells cannot be excluded.

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Quantitative analysis of chimerism after allogeneic stem cell transplantation by PCR amplification of microsatellite markers and capillary electrophoresis with fluorescence detection: the Paris–Robert Debré experience

Cited by 38 publications

References 17 publications

How and when should we monitor chimerism after allogeneic stem cell transplantation?

How and when should we monitor chimerism after allogeneic stem cell transplantation?

Engraftment of unrelated cord blood after reduced-intensity conditioning regimen in children with refractory neuroblastoma: a feasibility trial

Long‐term persistence of host cells detected by X‐chromosome gene‐based assay in patients undergoing gender‐mismatched hematopoietic stem cell transplantation

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