Real-Time Quantitative Y Chromosome-Specific PCR (QYCS-PCR) for Monitoring Hematopoietic Chimerism after Sex-Mismatched Allogeneic Stem Cell Transplantation

Fehse, Boris; Чухловин, А. Б.; Kühlcke, Klaus; Marinetz, Olga; Vorwig, Oliver; Renges, Helmut; Krüger, William; Zabelina, Tatyana; Дудина, О. В.; Finckenstein, Friedrich Graf; Kröger, Nicolaus; Kabisch, H; Hochhaus, Andreas; Zander, Axel R.

doi:10.1089/152581601750289028

Cited by 81 publications

(69 citation statements)

References 14 publications

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“…To standardize the data, a second PCR detecting the human hematopoietic cell kinase gene was carried out in parallel. 24 Table 2 lists the corresponding primers and probes. For the detection of HA-1-positive (HA-1 H ) microchimeric cells, we adapted an earlier established nested PCR protocol.…”

Section: Detection Of Circulating Y-chromosome ؉ and Ha-1 H Microchimmentioning

confidence: 99%

Naturally acquired tolerance and sensitization to minor histocompatibility antigens in healthy family members

Halteren

Jankowska‐Gan

Joosten

et al. 2009

Blood

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Bidirectional cell transfer during pregnancy frequently leads to postpartum persistence of allogeneic cells and alloimmune responses in both the mother and in her offspring. The life-long consequences of naturally acquired alloimmune reactivity are probably of importance for the outcome of allogeneic stem cell transplantation. We investigated the presence of CD8 pos minor histocompatibility (H) antigen-specific cytotoxic T lymphocytes (T CTL ) and CD8 pos minor H antigen-specific T regulator cells (T REG ) in peripheral blood cells obtained from 17 minor H antigen-disparate mother-offspring pairs. Absence of minor H antigen-specific T REG , as marked by the feasibility to expand T CTL from isolated tetramer pos populations, was observed in 6 mothers and 1 son. The presence of minor H alloantigen-specific T REG was observed in 4 mothers and 5 sons.These T REG were detected within isolated tetramer dim staining fractions and functioned in a CTLA-4-dependent fashion. Our study indicates that both T CTL and T REG mediated alloimmunity against minor H antigens may be present in healthy female and male hematopoietic stem cell donors, potentially influencing graftversus-host reactivity in different ways.

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Section: Detection Of Circulating Y-chromosome ؉ and Ha-1 H Microchimmentioning

confidence: 99%

Naturally acquired tolerance and sensitization to minor histocompatibility antigens in healthy family members

Halteren

Jankowska‐Gan

Joosten

et al. 2009

Blood

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“…Based thereon, standard curves could be obtained for either mutation using the abovedescribed duplex PCR and the DDct method as described earlier. 11 The amount of mutated and wild-type allele in every unknown sample thus might be determined by plotting the corresponding mean Dct value against the standard curve.…”

Section: Patientsmentioning

confidence: 99%

Screening and monitoring of MPL W515L mutation with real-time PCR in patients with myelofibrosis undergoing allogeneic-SCT

Alchalby

Badbaran

Böck

et al. 2010

Bone Marrow Transplant

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Monitoring of minimal residual disease (MRD) after allogeneic (allo)-SCT for myelofibrosis (MF) allows recognizing the depth of remission and thus guides application of appropriate therapeutic interventions. MPL W515L/K mutations, which are detected in 5-10% of JAK2V617F-negative patients, may be useful for this purpose. Using a highly sensitive quantitative PCR method, we tested 90 patients with MF who underwent allo-SCT for the presence of MPL W515L/K mutations. Two patients with primary MF were found to harbor MPLW515L while no patient was positive for MPLW515K mutation. Both patients were JAK2V617F negative and cleared the mutation rapidly after allo-SCT and remained negative for a median follow-up of 19 months. The results of molecular monitoring correlated well with other remission parameters such as normalization of peripheral blood counts and morphology and complete donor chimerism. We conclude that MPLW515L can be cleared after allo-SCT and hence may be used as an MRD marker in a proportion of JAK2V617F-negative MF patients.

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“…Four centers applied a multiplex polymerase chain reaction technique using short tandem repeatmarkers, while two centers used allele-specific sequences or the Y chromosome in sex-mismatched transplantations. [12][13][14] Definitions Standard risk of relapse was defined as a diagnosis of CML in the first chronic phase, AML in first complete remission. High risk was defined as more advanced disease or any other diagnosis.…”

Section: Supportive Care and Monitoringmentioning

confidence: 99%

Reduced-intensity conditioning with busulfan and fludarabine with or without antithymocyte globulin in HLA-identical sibling transplantation – a retrospective analysis

Schetelig

Bornhäuser

Kiehl

et al. 2003

Bone Marrow Transplant

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Summary:It is unknown whether the addition of antithymocyte globulin (ATG) to reduced-intensity conditioning with busulfan (BU) and fludarabine (FLU) is beneficial in HLA-identical sibling transplantation. Therefore, we analyzed retrospectively data on 83 patients, who received peripheral blood stem cells from HLA-identical siblings after conditioning with either 8 mg/kg BU and 150 mg/m 2 FLU (n ¼ 45) or 8 mg/kg BU, 180 mg/m 2 FLU and 40 mg/ kg ATG (n ¼ 38). Graft-versus-host disease (GVHD) prophylaxis consisted of CSA alone (n ¼ 32) or a combination with either MTX or MMF (n ¼ 51). The median age was 52 years. Graft failure occurred in two patients after BU/FLU and in three after BU/FLU/ATG (P ¼ 0.66). After conditioning with BU/FLU, platelet recovery was significantly faster (P ¼ 0.017), and less platelet (Po0.001) and red blood cell (P ¼ 0.002) support was needed. Incidences of acute GVHD grades II and IV were 46 and 49%, respectively. Limited chronic GVHD occurred more often after BU/FLU compared to BU/FLU/ ATG (54 vs 23%, P ¼ 0.02). The overall survival, nonrelapse and relapse mortality did not differ significantly. We conclude that in peripheral blood stem cell transplantation from HLA-identical siblings after reduced-intensity conditioning with BU and FLU, ATG has no major impact on the rate of graft rejection and acute GVHD, but it reduces the incidence of limited chronic GVHD. Reduced-intensity conditioning regimens with busulfan (BU), fludarabine (FLU) and antithymocyte globulin (ATG) are increasingly used in related and unrelated hematopoietic stem cell transplantation (HSCT). 1 While the use of ATG as an additional drug for graftversus-host disease (GVHD) prophylaxis is widely accepted in unrelated transplantation, the application in HLA-identical sibling transplantation is a matter of debate. ATG-Fresenius is a polyclonal IgG antibody raised in rabbits against the Jurkat T cell line. 2 The antibody had been incorporated into dose-reduced conditioning regimens for two reasons: first, ATG given before transplantation exerts T-cell depletion of the recipient, which facilitates donor stem cell engraftment after nonmyeloablative doses of BU and FLU and second, antibody persisting in the recipient beyond transplantation exerts in vivo depletion of donor T cells, thereby possibly reducing the risk for acute and chronic GVHD. 3,4 However, ATG has considerable short-term toxicity. 5 While the efficacy of ATG to prevent GVHD and to reduce nonrelapse mortality after myeloablative HSCT has been demonstrated in several studies, the role of ATG in reduced-intensity conditioning has not been evaluated. [6][7][8][9][10] However, reduced-intensity conditioning with BU and FLU was safe even without ATG in 24 patients as shown by our group. 11 In order to assess the impact of ATG, we retrospectively compared the engraftment, incidence and severity of GVHD and survival after BU-and FLU-based reduced-intensity conditioning with or without ATG in patients who had received peripheral blood stem cell transplantation (PBSCT...

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Real-Time Quantitative Y Chromosome-Specific PCR (QYCS-PCR) for Monitoring Hematopoietic Chimerism after Sex-Mismatched Allogeneic Stem Cell Transplantation

Cited by 81 publications

References 14 publications

Naturally acquired tolerance and sensitization to minor histocompatibility antigens in healthy family members

Naturally acquired tolerance and sensitization to minor histocompatibility antigens in healthy family members

Screening and monitoring of MPL W515L mutation with real-time PCR in patients with myelofibrosis undergoing allogeneic-SCT

Reduced-intensity conditioning with busulfan and fludarabine with or without antithymocyte globulin in HLA-identical sibling transplantation – a retrospective analysis

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