Flow cytometric method for the routine follow‐up of red cell populations after bone marrow transplantation

Hendriks, E. C. M.; Man, A.J.M. de; Berkel, Y. C. M. Van; Stienstra, Stef; Witte, Théo de

doi:10.1046/j.1365-2141.1997.d01-2138.x

Cited by 19 publications

(19 citation statements)

References 8 publications

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“…The advantages of RCP are clear: it can be performed on blood samples; the assays are simple, accurate and very sensitive; and the results can be obtained within 24 h. Recently developed flow cytometric methods have increased the speed and efficacy of RCP. 40 The methodology of RCP is very simple: complete red cell phenotyping of patient and donor is performed using different antigens: A, B, C, c, E, D, K, Fy a , Jk a , Jk b , M, N, S and s. RCP of patients should be performed before transfusion is given. Afterwards, discriminating markers are used to detect MC with the help of the fluoro microsphere method.…”

Section: Erythrocyte Phenotypingmentioning

confidence: 99%

Significance of chimerism in hematopoietic stem cell transplantation: new variations on an old theme

Khan

Ágarwal

Agrawal

2004

Bone Marrow Transplant

146

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Summary:The main goal of post-transplantation monitoring in hematopoietic stem cell transplantation (HSCT) is to predict negative events, such as disease relapse, graft rejection and graft-versus-host disease, in order to intervene with appropriate therapy. In this context, chimerism analysis is an important method in monitoring post HSCT outcome. Mixed chimerism (MC) is mainly evaluated to define engraftment and relapse. Detection of MC is a prerequisite in both myeloablative and nonmyeloablative HSCT, in order to assess the graft status and decide later therapeutic strategies such as donor lymphocyte infusion. In this review, we discuss various techniques including erythrocyte phenotyping, cytogenetic analysis, fluorescent in situ hybridization, restriction fragment length polymorphism, STR/VNTR analysis and real-time quantitative PCR, along with the various methods used to detect minimal residual disease (MRD) in different diseases such as chronic myeloid leukemia, acute myelomonocytic leukemia or acute lymphoblastic leukemia. The review mainly highlights the optimal methodological approach, which needs to be informative, sensitive and quantitatively accurate for MC detection. Future of post HSCT graft monitoring lies in the selection of the most accurate and sensitive technique to determine both MC and MRD. Such an approach would be helpful in not only determining relapse or rejection, but also in ascertaining various responses to different treatment modalities.

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Section: Erythrocyte Phenotypingmentioning

confidence: 99%

Significance of chimerism in hematopoietic stem cell transplantation: new variations on an old theme

Khan

Ágarwal

Agrawal

2004

Bone Marrow Transplant

146

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“…Red blood cell phenotyping was performed using a flow cytometric and/or fluorescent microscopic microsphere method as described previously. [27][28][29] The sensitivity level is 0.1-0.3% for the flow cytometric method and 0.01% for the fluorescent microscopic technique. At the level of 0% patient or donor red cells, the flow cytometric results were always controlled by the fluorescent microscopic method in order to increase the sensitivity to a level of 0.01%.…”

Section: Chimerism Analyzed By Cytogenetic Analysis and Red Blood Celmentioning

confidence: 99%

Long-term follow-up of persisting mixed chimerism after partially T cell-depleted allogeneic stem cell transplantation

Schaap

Schattenberg²,

Mensink

et al. 2002

Leukemia

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Using red cell phenotyping (RCP) and/or cytogenetics (CYT) we identified 19 patients with persisting mixed chimerism (MC) among 231 patients transplanted with partially T cell-depleted stem cell grafts from HLA-identical siblings. Persisting MC is defined as MC for more than 2 years in patients without any evidence of relapse. Median leukemia-free survival in these patients was 150 (range, 50-218) months. Diagnoses were ALL (n ‫؍‬ 10); AML (n ‫؍‬ 2); CML (n ‫؍‬ 2); NHL (n ‫؍‬ 2); MDS (n ‫؍‬ 1); MM (n ‫؍‬ 1) and SAA (n ‫؍‬ 1). Purpose of this study was the longterm follow-up of MC and definition of patterns of chimerism in the various subsets of PBMCs and granulocytes. Using a PCR-STR technique CD3 + /CD4 + (T4 lymphocytes), CD3 + /CD8 + (T8 lymphocytes), CD45 + /CD19 + (B lymphocytes), CD45 + /CD14 + (monocytes), CD45 + /CD15 + (granulocytes) and CD3 − /CD56 + (NKcells) were analyzed. The majority of patients with persisting MC were conditioned with a less intensive conditioning regimen and had little GVHD. Sequential monitoring of the chimerism resulted in a group of patients (n ‫؍‬ 7) with very slow transient mixed chimerism that resulted in complete DC after median 7 years. Another nine patients had a relatively high percentage of persisting autologous cells for a median of 12 years and in three patients we observed a stable low percentage of autologous cells. Only two out of 19 patients (AML-CR1, CML-CP1) relapsed during follow-up. Both patients had a relatively high percentage of autologous cells. Chimerism in granulocytes and PBMC subsets was analyzed at a median of 8 years after SCT in nine patients. In five patients mixed chimerism simultaneously detected by RCP and CYT was associated with MC in all subsets. Within each individual patient the percentages of donor and recipient cells were very different between the different subsets. Two CML-CP1 patients were mixed chimera in only two subsets and in one patient these subsets represented pending relapse. In another two patients mixed chimerism with a very low number of autologous red cells was not found in the PBMCs because of the different sensitivity level of the RCP and the PCR-STR technique. We conclude that in patients with persisting mixed chimerism after partially T cell-depleted SCT a remarkable number of patients had lymphoid malignancies, the majority of the patients were conditioned with less intensive conditioning regimens and the mixed chimerism was not correlated with relapse. Chimerism in granulocytes and PBMC subsets did show great intra-individual differences in the subsets and these data correlated well with RCP and CYT data with the exception of the NK cells. Leukemia (2002) 16, 13-21.

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“…Red blood cell phenotyping was performed using a flow cytometric and/or fluorescent microscopic microsphere method as described previously. [37][38][39] The sensitivity level is 0.1-0.3% for the flow cytometric method and 0.01% for the fluorescent microscopic technique. At the level of 0% patient or donor red cells, the flow cytometric results were always controlled by the fluorescent microscopic method in order to increase the sensitivity of this test to a level of 0.01% in this analysis.…”

Section: Assessment Of Gvhd Relapse and Chimerismmentioning

confidence: 99%

Induction of graft-versus-leukemia to prevent relapse after partially lymphocyte-depleted allogeneic bone marrow transplantation by pre-emptive donor leukocyte infusions

Schaap¹,

Schattenberg²,

Bär³

et al. 2001

Leukemia

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In this prospective study we analyzed pre-emptive donor leukocyte infusions (DLI) in 82 consecutive patients transplanted with partially T cell-depleted grafts for acute myeloid leukemia, acute lymphoid leukemia, chronic myeloid leukemia, refractory anemia with excess of blasts, refractory anemia with excess of blasts in transformation and multiple myeloma. Donors were HLA-identical siblings. Patients without significant acute (Ͼgrade 1) and/or chronic GVHD were scheduled to be treated with DLI (35 patients) and 31 actually received DLI. Patients who developed acute GVHD Ͼgrade 1 and/or chronic GVHD were not scheduled to receive DLI and served as a comparison group (47 patients). The median interval between BMT and DLI was 22 weeks. The first six patients received 0.7 × 10 8 CD3 + cells/kg body weight (b.w.). Five out of these six patients developed acute GVHD (grade 1: n = 2, grade 3: n = 2 and grade 4: n = 1) which was more frequent and more severe than we had anticipated. In the next 25 patients the number of T lymphocytes was diminished to 0.1 × 10 8 CD3 + cells/kg b.w. which resulted in less frequent and less severe GVHD. Eight patients in this group developed acute GVHD (grade 1: n = 4, grade 2: n = 4) and three patients had limited chronic GVHD. Patients in the DLI group needed more time to establish complete donor chimerism confirmed by a higher number of mixed chimeras at 6 months after BMT. The projected 3-year probability of disease-free survival was 77% for the 35 patients intended to treat with DLI and 45% for the patients of the comparison group (P = 0.024). Relapse rate at 36 months after transplantation was 18% in the patients who were intended to treat with DLI and 44% in the comparison group (P = 0.026). We conclude that preemptive DLI is feasible and generates favorable relapse rates in patients who are at high risk for relapse. Furthermore, the incidence and severity of GVHD disease after DLI is dependent on the number of CD3 + cells infused. Leukemia (2001Leukemia ( ) 15, 1339Leukemia ( -1346

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Flow cytometric method for the routine follow‐up of red cell populations after bone marrow transplantation

Cited by 19 publications

References 8 publications

Significance of chimerism in hematopoietic stem cell transplantation: new variations on an old theme

Significance of chimerism in hematopoietic stem cell transplantation: new variations on an old theme

Long-term follow-up of persisting mixed chimerism after partially T cell-depleted allogeneic stem cell transplantation

Induction of graft-versus-leukemia to prevent relapse after partially lymphocyte-depleted allogeneic bone marrow transplantation by pre-emptive donor leukocyte infusions

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