A Fluorescent Microsphere Method for the Investigation of
Erythrocyte Chimaerism after Allogeneic Bone Marrow
Transplantation Using Antigenic Differences

Man, A.J.M. de; Foolen, W.J.G.; Dijk, B.A. van; Kunst, V.A.J.M.; Witte, T.M. de

doi:10.1159/000461828

Cited by 10 publications

(16 citation statements)

References 4 publications

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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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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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show abstract

“…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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“…In a study carried out by using antibody-coated fluorescent microspheres, donor-derived EA was seen in 86% of the patients on the 15th day. 3 In another study, donor-derived antigens were observed at the end of 8th week in 71% of patients. 1 It is concluded that the tube agglutination technique is as efficient as antibody-coated fluorescent microspheres or gel tests in order to detect EA engraftment.…”

Section: Discussionmentioning

confidence: 93%

“…Later, anti-human IgG-coated fluorescence microspheres were used to detect chimerism. 3 Subsequently, gel test was used to differentiate donor and recipient RBCs. 4 Flow cytometric analysis using monoclonal antibodies directed against blood group antigens enabled sensitive evaluation of dual RBC populations.…”

mentioning

confidence: 99%

Erythrocyte antigen and reticulocyte engraftment after allogeneic hematopoietic stem cell transplantation

Yildirim

Ozer

Yüksel

et al. 2004

Bone Marrow Transplant

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Summary:The aim of this study was to study the usefulness of erythrocyte antigen (EA) measurement to study engraftment after allogeneic HSCT. In all, 31 consecutive patients receiving HLA-identical bone marrow (BM) (n ¼ 13) or peripheral blood stem cells (n ¼ 18) were investigated. Apart from the ABO group, 15 EAs representing six minor blood groups were followed by the simple tube agglutination technique. A total of 20 (64.5%) patients received ABOidentical, eight (25.8%) received ABO minor and three (9.7%) received ABO major mismatched grafts. In all, 29 patients were followed for a median of 12 (6-16) months; 65% of the patients expressed donor type EA 1 month and almost all did so 6 months after transplant. Reticulocyte engraftment was significantly shorter than EA engraftment (median 18 vs 35 days) (P ¼ 0.001). Patients who received PB stem cells showed significantly faster EA and reticulocyte engraftment than patients who received BM stem cells (P ¼ 0.038 and 0.025). ABO compatibility did not have an impact on reticulocyte and EA engraftment (P ¼ 0.4 and 0.55). The earliest donor type EA detected was from the Rh and Kidd system. These data suggest that EA and reticulocyte assays are useful in monitoring engraftment. 4 Flow cytometric analysis using monoclonal antibodies directed against blood group antigens enabled sensitive evaluation of dual RBC populations. 5 Our aim was to study a simple tube hemagglutination method for monitoring RBC repopulation following HSCT. In addition, we evaluated the disappearance of recipient-derived RBC antigens and appearance of donorderived antigens after transplantation, and correlated RBC antigen engraftment with reticulocyte counts after AHSCT. Patients and methods PatientsA total of 31 consecutive HSCT patients were studied. They consisted of 11 females and 20 males with a median age of 36 years (range 18-51). The underlying diseases were chronic myeloid leukemia in 18, acute leukemia in 11, severe aplastic anemia in one and renal cell carcinoma in one patient. In all, 13 of the patients received bone marrow (BM) and 18 received peripheral blood stem cells (PBSC) from their HLA-identical siblings. Eight of the patients received minor and three received major ABO-incompatible grafts. Table 1 shows the patient and donor characteristics. Transplantation procedureThe patients were conditioned by ablative cyclophosphamidebased regimens (81%) or fludarabine-based reduced intensity regimens. 6 All the patients received short-course methotrexate and cyclosporine for GVHD prophylaxis. All the donorrecipient pairs were CMV-seropositive. Recombinant human G-CSF was used until neutrophil engraftment after day þ 1 in BM group, but not in the PBSC. All the blood products were leucofiltered and irradiated. The pretransplant evaluation of the mismatched patient-donor pairs, the decision for erythrocyte and plasma depletion of the harvest and posttransplant transfusion policy were designed by our transplant center's transfusion medicine consultant (Dr Arslan), according to recently pu...

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A Fluorescent Microsphere Method for the Investigation of Erythrocyte Chimaerism after Allogeneic Bone Marrow Transplantation Using Antigenic Differences

Cited by 10 publications

References 4 publications

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

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

Erythrocyte antigen and reticulocyte engraftment after allogeneic hematopoietic stem cell transplantation

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