[Show abstract][Hide abstract] ABSTRACT: Hepatic veno-occlusive disease (VOD) is one of the most common and important regimen-related toxicities observed after hematopoietic stem cell transplantation (HSCT). There are no universally accepted preventative or therapeutic approaches for VOD. We prospectively evaluated the safety and efficacy of a short course of methylprednisolone (MP) in 48 patients undergoing allogeneic HSCT who were diagnosed with hepatic VOD. MP was administered at a dose of 0.5 mg/kg i.v. every 12 h for a total of 14 doses, and then discontinued without taper. Thirty (63%) patients responded with a reduction in total serum bilirubin of 50% or more after 10 days of treatment. In univariate analysis, non-responders had a higher total bilirubin at the start of MP therapy, more weight gain, evidence of fungal infection and platelet refractoriness. High SGPT and early engraftment were significant factors among responders. Twenty-five of the 30 responders survived up to day +100, whereas all but three non-responders died within 100 days post-HSCT, for a probability of survival of 58% among responders and 10% for non-responders. Prospective comparative studies are needed to confirm the observed encouraging outcome of MP therapy for VOD.
Bone Marrow Transplantation 03/2008; 41(3):287-91. · 3.54 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: The design of chemotherapy-induction regimens for acute myeloid leukaemia (AML) is directed towards the early elimination of bone marrow (BM) leukaemic blast cells (LBCs). Patients with AML after induction show LBC reduction in a hypoplastic BM and also demonstrate a varying number of residual BM plasma cells (PCs).
To relate PC number to several blood and BM parameters as well as clinical features such as infection and survival.
On the 14th day after the start of chemotherapy (D+14) BM samples were examined for residual PCs in 60 adult (>or=15 years) patients undergoing AML-induction chemotherapy, and the proportion of PCs was related to blood and BM parameters including French-American-British (FAB) subtype, other inflammatory cells, antecedent infection, attainment of complete remission and 36-month survival.
Median PC proportion of 11.3% (range 0.1-48.7%) in D+14 BM aspirates and 10.7% (0.6-41%) in trephine biopsies was observed. Their number showed a direct relationship with residual BM lymphocytes (r=0.368; p=0.025). Higher numbers of residual PCs also reflected the presence of infection before diagnosis and coincident with treatment (p=0.039). Although we could not demonstrate an association between PC numbers and 36-month survival, PC numbers were significantly higher in patients with residual leukaemia at D>14 (p=0.007).
D+14 BM PC number reflects the effectiveness of induction chemotherapy and the presence of antecedent inflammation or infection.
Journal of Clinical Pathology 05/2007; 60(5):520-3. · 2.44 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: We conducted a retrospective study with the aim of identifying risk factors and clinical characteristics associated with HBV reactivation and clinical flare after allogeneic stem cell transplantation (aSCT). We reviewed the King Faisal Specialist Hospital and Research Center International Bone Marrow Transplant Registry database from January 1998 to June 2000. Complete serological screening for HBV was available in 128 of 131 patients transplanted during that period. Fifty-four (42%) had evidence of prior infection and recovery from HBV before transplant (hepatitis B core antibody positive, B surface antigen negative). Forty-two were evaluable for HBV reactivation and clinical flare. Six (14%) reactivated with clinical flare as documented by seroconversion and/or positive HBV DNA in the serum with biochemical hepatitis at 5.5, 18, 18, 19, 21 and 23 months post-transplant. Five of fifteen patients with chronic graft-versus-host disease (cGVHD) reactivated with clinical flare in contrast to 1/27 without cGVHD (RR: 9.0, 95% CI: 1.2-70.1 P < 0.02). HBV reactivation with clinical flare occurred during immunosuppressive therapy tapering or withdrawal in all patients. In conclusion, hepatitis B core antibody positive allogeneic stem cell recipients with cGVHD are at significant risk of HBV reactivation with clinical flare.
Bone Marrow Transplantation 08/2002; 30(3):189-94. · 3.54 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Molecular monitoring of donor/recipient T-cell kinetics early post-transplant can provide clues to the immunological events that govern host-versus-graft reaction (HVGR) and graft versus-host-disease (GVHD). We have previously used fluorescence in situ hybridization (FISH) with X and Y probes to monitor recipient T (R-T) cell clearance early after myeloablative allogeneic stem cell transplantation (ASCT). We demonstrated that impaired clearance of residual host-T-cells in the early days post-transplant was associated with graft rejection, while enhanced clearance could be an indicator of increased donor anti-host alloreactivity and predictive of acute GVHD. Although FISH is the most accurate quantitative molecular tool for the determination of the exact donor/recipient-T-cell numbers at any time points post-transplant, it has the disadvantage of being limited to sex mismatched donor/recipient pairs. Our goal was to develop a molecular approach that, irrespective of gender, would be comparable to FISH in accurately determining host residual T-cell clearance after myeloablative conditioning for ASCT. We have genotyped DNA from cell lysates using polymerase chain reaction (PCR) amplification of short tandem repeats (STR) with fluorescently labeled oligonucleotide primers, and used the Genescan 672 software for accurate quantitative analysis of the amplified alleles. Here, we show that this approach allowed us to achieve in T-cells accurate quantitative analyses of amplified donor/recipient alleles in sex matched patients on days +5, +8 and +12 post-transplant, despite severe leukopenia.
Leukemia and Lymphoma 07/2002; 43(6):1281-7. · 2.61 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Extramedullary myeloid cell tumour (EMMT) localised to the mediastinum is a rare manifestation of acute myeloid leukaemia, forming less than 4% of all cases of EMMT. In contrast to other types of EMMT, cytogenetic characteristics of this rare entity are relatively unknown. This report describes a patient with EMMT who had evidence of superior vena cava syndrome and normal peripheral blood counts at diagnosis. The results from an initial biopsy specimen were consistent with a diagnosis of mediastinal large B cell lymphoma. A diagnosis of acute myeloid leukaemia was made three months after initial diagnosis by bone marrow examination. Review of the initial biopsy specimen showed strong positivity for myeloperoxidase, revealing that the patient had been initially misdiagnosed as having large B cell lymphoma. Cytogenetic studies revealed a near triploid and near tetraploid karyotype with structural abnormalities in 12 and three metaphases, respectively. Review of the literature showed that a near tetraploid or triploid karyotype is found in most of the reported cases of mediastinal EMMT. Thus, the presence of a near triploid/tetraploid karyotype and mediastinal EMMT may represent a specific subset of EMMT. The biological relevance of this observation is discussed.
Journal of Clinical Pathology 04/2002; 55(3):221-5. · 2.44 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: The use of mobilized peripheral blood (PB) stem cells for autologous transplantation initially generated much enthusiasm because of enhanced engraftment in comparison to marrow stem cells and avoidance of general anesthesia for the donor. Its application to the allogeneic setting seemed inevitable. For obvious ethical reasons, allogeneic donors are mobilized with cytokines only, mainly granulocyte colony-stimulating factor (G-CSF). Results from preliminary studies suggest that in comparison to standard bone marrow transplants, outcomes such as engraftment, host-versus-graft reaction, graft-versus-host disease, graft-versus-leukemia and immunological reconstitution may be different. Surprisingly, G-CSF, previously recognized as a late acting lineage-specific factor for neutrophil production, not only disrupts homeostasis between stem cells and their microenvironment, but also induces significant quantitative and qualitative changes in the accessory cell compartment, affecting lymphocytes, monocytes, natural killer, dendritic, and stromal cells. Furthermore, mobilization of huge numbers of non-professional antigen presenting cells (CD34+ stem cells) amplifies the tolerizing potential of PB stem cell grafts. Thus, G-CSF mobilization provides PB transplants with different immunobiologic properties in comparison to standard bone marrow grafts. Whether these immunobiologic differences will lead to better transplant outcomes remains to be shown through much awaited results of large randomized clinical trials.
Bone Marrow Transplantation 08/2000; 26(1):1-16. · 3.54 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Fusarium is a newly emerging fungal pathogen associated with significant morbidity and mortality in the immunocompromised host. We have reviewed our hospital's experience with Fusarium between 1985 and 1995. Fusarium species were isolated from 22 specimens, representing 11 patients. Cases were not clustered by time period. The median age of the patients was 36.5 years (range 17-69 years). The sources of the organism were 12 skin lesions from eight patients, seven blood cultures from two patients and one specimen each from a Hickman catheter tip, nail clippings and a bronchoalveolar lavage. Seven of the patients had chemotherapy-induced neutropenia when the Fusarium was isolated. Five of them developed invasive fusarosis during acute leukaemia induction treatment. They remained neutropenic, and none survived. The other two patients recovered from neutropenia and were treated successfully for this infection. The remaining four patients were not neutropenic or immunocompromised. Three grew Fusarium from skin or nail clippings and one from bronchial alveolar lavage (BAL). There was no evidence of invasive disease in any of the four. None of them received antifungal therapy, and they were all alive at last follow-up. We conclude that Fusarium is a newly emerging infection in neutropenic patients. A high index of suspicion, especially for skin lesions, will help in early diagnosis before systemic and visceral dissemination. Excision of the initial focus of infection and antifungal therapy, aided by speedy neutrophil recovery, are likely to protect patients threatened with these fatal infections. Fusarium isolated from non-neutropenic, non-immunosuppressed patients is not significant and does not merit systemic antifungal treatment.
British Journal of Haematology 04/2000; 108(3):544-8. · 4.94 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Bone marrow transplant (BMT) recipients are prone to bacterial, viral and fungal infections. Mycobacterium tuberculosis infection can occur in these patients, but the incidence is lower than that of other infections. This report describes four patients with Mycobacterium tuberculosis infection identified from 641 adult patients who received a BMT over a 12-year period (prevalence 0.6%). The pre-transplant diagnosis was AML in two patients and CML in the other two. Pre-transplant conditioning consisted of BU/CY in three patients and CY/TBI in one. Graft-versus-host disease (GVHD) prophylaxis was MTX/CsA in three patients and T cell depletion of the graft in one patient. Sites of infection were lung (two), spine (one) and central nervous system (one). Onset of infection ranged from 120 days to 20 months post BMT. Two patients had co-existing CMV infection. One patient had graft failure. The two patients who received anti-tuberculous (TB) therapy recovered from the infection. Although the incidence of tuberculosis in BMT patients is not as high as in patients with solid organ transplants, late diagnosis due to the slow growth of the bacterium can lead to delay in instituting anti-TB therapy. A high index of suspicion should be maintained, particularly in endemic areas.
Bone Marrow Transplantation 10/1999; 24(5):551-4. · 3.54 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Primary graft failure, secondary to either host-vs.-graft reaction or delayed engraftment, and graft-vs.-host disease (GVHD) are among the most difficult clinical problems to manage in the field of allogeneic bone marrow transplantation (BMT). Early diagnosis of both conditions would greatly improve their outcome. Using fluorescence in situ hybridization (FISH) with an X- and Y-probe mixture, we sequentially monitored chimerism of neutrophils and lymphoid cells from day 1 to 100 in 28 consecutive recipients of sex-mismatched unmanipulated bone marrow grafts. The objective was to quantitatively assess the evolution of chimerism during this crucial time interval and to determine whether chimerism patterns would be predictive of engraftment and GVHD. In recipients with primary graft failure (n=7), the presence of donor-type neutrophils and NK cells as well as the predominance of donor-type T cells distinguished patients who responded to G-CSF (n=5) from nonresponders (n=2). Furthermore, the clearance of host CD3+CD56- cells during days 5-10 posttransplantation was significantly hastened in patients who subsequently developed acute (delta=80%) or chronic (delta=81%) GVHD compared with patients without GVHD (delta=17%). Thus, our data suggest that molecular monitoring of the fate of host/donor hematopoietic cells in the early posttransplantation period could be useful in differentiating patients with delayed engraftment from those with irreversible rejection and in predicting the occurrence of GVHD as soon as day 10. This investigational approach may provide an appropriate basis on which to select adequate treatment for primary graft failure and high-risk candidates that could benefit from novel preemptive therapies for GVHD.
[Show abstract][Hide abstract] ABSTRACT: A controversy persists in autologous transplantation as to which source of progenitor cells, bone marrow (BM) or peripheral blood (PB), contains the lowest number of contaminating lymphoma cells, and how mobilization procedures affect these numbers. To accurately measure the number of non-Hodgkin's lymphoma (NHL) cells harboring the bcl-2/immunoglobulin H (IgH) rearrangement in progenitor cell grafts, we developed a nested quantitative competitive polymerase chain reaction assay (QC-PCR). DNA from lymph nodes of four patients with NHL were cloned into the pSK(+) vectors to generate four internal controls (ICs) (two with major breakpoint region [MBR] and two with minor cluster region [mcr] rearrangements). The kinetics of amplification of ICs paralleled those of bcl-2/IgH rearranged genomic DNA. When used in a QC-PCR assay, these ICs were accurate at a 0.2-log level and provided reproducible results, as shown by low intrarun and interrun variability. An excellent correlation between predicted and observed lymphoma cell content (r = .99) was observed over a range of at least 5 logs of rearranged cells. This approach was used to measure involvement by NHL cells at the time of progenitor cell harvest in 37 autologous transplant patients. The number of bcl-2/IgH rearranged cells in BM, PB, and mobilized PB (mPB) was found to vary from 1 to 1.1 x 10(5) per million cells. The number of lymphoma cells present in BM was significantly higher than in PB (P = .0001), with a median difference in lymphoma cell content between BM and PB of 0.48 log of cells (range, -0.7 to 5 logs). In contrast, we found no difference in the concentration of bcl-2/IgH rearranged cells present in BM versus PB progenitor cells mobilized with cyclophosphamide and granulocyte colony-stimulating factor (G-CSF) (mPB) (P = .57). In conclusion, the QC-PCR assay described in this study could measure accurately and reproducibly the number of bcl-2/IgH rearranged cells among normal cells. Differences in levels of contamination by lymphoma cells between BM and PB were of less than one log (10-fold), and no differences in lymphoma cell concentrations were observed between BM and mobilized PB. As more cells are usually infused with mPB than with BM grafts, mPB progenitor cell grafts may actually be associated with higher levels of contamination by lymphoma cells. Furthermore, this QC-PCR assay should provide an important tool to assess the prognostic impact of lymphoma cell burden both in progenitor cell grafts and in vivo.
[Show abstract][Hide abstract] ABSTRACT: Thymic function is severely impaired in most marrow transplant recipients. To evaluate the impact of thymic hypoplasia on T cell reconstitution following marrow transplantation, we compared the phenotype and function of T lymphocytes in thymectomized recipients with those of euthymic hosts. Irradiated C57BL/6 mice (Thy1.2+, Ly5.1+) received 10(7) T cell-depleted B6.Ly5.2 bone marrow cells (Thy1.2+, Ly5.2+), with or without 3 x 10(5) B6.PL lymph node cells (Thy1.1+, Ly5.1+) as a source of T lymphocytes. Multiparameter flow cytometry analysis showed that in euthymic mice (group 1), T cell reconstitution was carried out by donor hematopoietic stem cells that differentiated in the host's thymus, whereas the production of chimeric T cells in athymic recipients depended on the presence or absence of T cells in the graft. When T lymphocytes were present in the graft (group 2), their progeny constituted the vast majority of splenic T cells on day 100 posttransplant. When the graft did not contain T lymphocytes (group 3), T cell reconstitution resulted from extrathymic maturation of donor hematopoietic progenitors; T cells differentiating along this pathway expressed lower levels of T cell receptor and a large proportion of the CD8+ subset expressed CD8alpha alpha homodimers. The T cell receptor Vbeta profile of all chimeras was similar to that of normal C57BL/6 mice. Compared with T cells found in euthymic recipients, those in mice from groups 2 and 3 were less abundant (particularly with respect to the CD4+ subset), displayed the CD44/CD45 phenotype of activated memory cells, and expressed high levels of IL-2 receptor beta chain. These results show that both the presence or absence of the thymus and the composition of the grafted inoculum determine the source and extent of posttransplant T cell reconstitution. Because they determine the nature of the differentiation pathway taken during T cell development in the host, these two factors can exert a critical influence on the appearance of graft vs. host disease and the level of host immunocompetence.
[Show abstract][Hide abstract] ABSTRACT: Using in situ hybridization with an X and Y chromosome probe mixture, we have sequentially studied peripheral blood samples from 10 patients (four males/six females) in an HLA-matched allogeneic setting in order to monitor the kinetics of early hematopoietic reconstitution. Interphase cells from smears consisting of purified granulocytic and lymphocytic populations respectively were studied in three patients at 24, 48, 72 and 96 h post-transplant. This period was arbitrarily defined as the immediate post-transplant period. These three patients plus seven others were studied sequentially at days 5, 10, 15, 20, 25 and 50 post-transplant, defined as the intermediate post-transplant period. The X and Y probes were indirectly labelled with rhodamine and fluoresceine isothiocyanate, respectively. Donor neutrophils were detected as early as 24 h post marrow infusion followed by a significant expansion at 48 h. At 96 h post-transplant, the median percentage of donor neutrophils was > 90%. In the immediate post-transplant period, most of the lymphocytes were of recipient origin. However, we have documented a significant expansion in donor lymphocytes, starting at day 5 post-transplant in most patients. Almost complete chimerism for the myeloid and lymphoid lineages was established at days 10 and 25 post-transplant, respectively. All patients engrafted normally according to standard clinical criteria. Follow-up data for those surviving > or = 100 days (eight patients), showed persistence of this pattern of hematopoietic reconstitution in all but one patient. Molecular monitoring of early engraftment has enabled us to unravel a distinct biphasic pattern of myeloid and lymphoid engraftment.
Bone Marrow Transplantation 07/1996; 17(6):1143-8. · 3.54 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: NK cells can exert potent anti-leukemia activity after either autologous or allogeneic BMT. However, in autologous blood or marrow transplant patients, NK cell number and/or function could be reduced, and also may vary according to the sampling site. In order to evaluate the hypothesis that blood or marrow grafts from autologous transplant patients exhibit impaired NK cell activity that could contribute to disease recurrence, we evaluated the immunologic characteristics of NK cells in the bone marrow (BM) and peripheral blood (PB) from 27 patients undergoing autologous BMT, and also from 20 normal donors. We measured baseline and interleukin-2 (IL-2)-activated NK cell cytotoxicity, as well as expression of IL-2 receptors (IL-2R) (alpha-chain (p55) and beta-chain (p75)), and adhesion molecules. The cytotoxic activity of PB NK cells was significantly lower in autologous transplant patients than in normal donors (P < 0.0005) and this difference was not mitigated following IL-2 activation. In contrast, BM from autologous patients showed normal NK cell cytotoxicity, but contained higher numbers of NK cells (P < 0.025), with more intense CD56 expression (P < 0.05). Expression of p75 was lower on BM than on PB NK cells in both patients and normal donors. In addition, induction of p55 by IL-2 was abrogated in autologous PB NK cells. Therefore, depending on the site of harvest and the nature of donor cells (pre-BMT vs normal), our results show significant differences in NK cell number, function, and IL-2 receptor expression. This may affect relapse rates following autologous transplants performed with either PB or BM grafts.
Bone Marrow Transplantation 04/1996; 17(3):315-22. · 3.54 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Allogeneic bone marrow transplantation (BMT) has been shown to provide effective therapy for chronic myelogenous leukemia (CML), but previous reports have also demonstrated the persistence of bcr-abl-positive cells for months to years after BMT in the majority of patients. To evaluate the biologic significance of persistent bcr-abl-positive cells, we examined the relationship between clinical parameters known to affect the risk of relapse and the ability to detect bcr-abl-positive cells post-BMT.
We analyzed 480 samples from 92 patients at two transplant centers for the presence of bcr-abl-positive cells by polymerase chain reaction (PCR). Two different BMT preparative regimens and protocols for prevention of graft-versus-host disease (GVHD) were used. One center used cyclophosphamide plus total-body irradiation (CY/TBI) and T-cell-depleted marrow; the second center used busulfan plus cyclophosphamide (Bu/CY) and untreated marrow with cyclosporine and methotrexate (Csp/MTX) as GVHD prophylaxis.
We first determined the percent of patients at each center with > or = one PCR-positive (PCR+) result at defined intervals post-BMT. Between 0 and 6 months post-BMT, the majority of patients (80% to 83%) in both populations had PCR-detectable bcr-abl-positive cells. Between 6 and 24 months post-BMT, 80% to 88% of patients who received T-cell-depleted marrow remained PCR+, as compared with 26% to 30% of patients who received unmodified marrow. After 24 months post-BMT, the percentage of PCR+ patients was not significantly different in the two populations. This pattern of detection of bcr-abl-positive cells post-BMT followed the development of chronic GVHD in patients who received unmodified marrow. All patients were also divided into three groups based on post-BMT PCR results as follows: (1) persistent PCR+ (n = 29), (2) intermittent PCR-negative ([PCR-] n = 40), and (3) persistent PCR- (n = 23). These three groups were found to have a low, intermediate, and high probability of maintaining remission and disease-free survival, respectively (P = .0001). Intermittent or persistent PCR- results, which reflect levels of minimal residual disease < or = the limit of detection by PCR, were clearly associated with both acute (P = .004) and chronic (P = .000005) GVHD. Nevertheless, 44% of patients without GVHD also had intermittent or persistent PCR- assays.
The persistence of PCR-detectable bcr-abl-positive cells early post-BMT in more than 80% of patients suggests that neither BMT preparative regimen effectively eradicates CML cells in most patients. Subsequently, acute and/or chronic GVHD are associated with a decreased ability to detect residual bcr-abl-positive cells, which suggests that immunologic mechanisms mediated by donor cells are important for inducing long-term remissions after BMT. The demonstration that 44% of patients without GVHD had either low or undetectable levels of residual leukemia suggests the presence of mechanisms capable of suppression or eradication of CML independent of GVHD.
Journal of Clinical Oncology 08/1995; 13(7):1704-13. · 18.04 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Numerous minor histocompatibility antigens (MiHAs) show tissue-specific expression and can induce vigorous T cell responses. They therefore represent attractive targets for leukemia immunotherapy mediated by adoptive transfer of T cells. The main objective of this work was to determine whether MiHAs expressed by normal hematopoietic cells were present on leukemic cells and whether they could trigger lysis by cytotoxic T lymphocytes (CTLs). CTL assays showed that mouse leukemic cells of both lymphoid and myeloid lineages were sensitive to CTLs targeted toward some but not all MiHAs. In four out of four strain combinations in which we primed CTLs against immunodominant MiHAs, effectors killed leukemic blasts, whereas no cytotoxicity was observed when CTLs were targeted toward four immunorecessive MiHAs. Testing of HPLC fractions obtained from normal and leukemic cells provided molecular evidence that leukemic blasts expressed only some of the MiHAs found on normal mouse hematopoietic cells. Decreased density of H-2 class I molecules at the surface of leukemic cells suggests that down-regulation of genes encoding either class I molecules or proteins involved in antigen processing played a role in the aberrant expression of MiHAs. In vivo resistance to the leukemic cells by various strains of mice correlated with in vitro CTL activity. These results show that leukemic cells express only some (immunodominant) MiHAs and suggest that this subset of MiHAs represent prime targets for adoptive immunotherapy.
Journal of Clinical Investigation 05/1995; 95(4):1561-8. · 12.81 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Bone marrow is the primary site of disease in patients with acute lymphoblastic leukemia (ALL) and is frequently involved in patients with non-Hodgkin's lymphoma (NHL). At the time of autologous bone marrow transplantation, marrow grafts from patients with leukemia and lymphoma are often still contaminated by malignant cells, even when such patients achieve complete clinical remission. In this study, we evaluated the potential of anti-B4-blocked-ricin (anti-B4-bR) immunotoxin to eliminate residual ALL and NHL cells from bone marrow. Anti-B4-bR binds to the CD19 antigen, which is B-lineage specific, and, at concentrations of 5 x 10(-9) M or greater, could eliminate more than 3 logs of CD19+ Nalm-6 or Namalwa cells in a 20-fold excess of normal irradiated bone marrow after only 5 hr of incubation. This activity was abrogated by the addition of anti-B4 but not by the presence of galactose, which is the natural ligand for native ricin. Also, when used at these high concentrations, anti-B4-bR showed little nonspecific toxicity against normal hematopoietic progenitors. In conclusion, a single short exposure to anti-B4-bR is capable of inducing high levels of depletion of CD19+ leukemia and lymphoma cells without significant nonspecific toxicity against normal marrow progenitors. Therefore, anti-B4-bR offers an interesting approach to the elimination of B-lineage malignant cells prior to autologous bone marrow transplantation.
Journal of Clinical Immunology 02/1995; 15(1):51-7. · 3.38 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: 1. Background: Bone marrow engraftment is mediated by stem cells with complex self-renewal, proliferative and differentiation potentials. This process is empirically documented by the progressive rise in the absolute neutrophil count occurring 2 to 3 weeks post-marrow infusion. 2. Purpose: To investigate the kinetics of early engraftment using in situ hybridization with an X and Y chromosome probe mixture in a sex-mismatch allogeneic setting. 3. Methods: Interphase cells from peripheral blood smears consisting of 98% pure granulocytic and lymphocytic populations were studied according to the following time frame: (a) 2 HR before marrow infusion, 12, 24, 48, 72 and 96 HR post-infusion in 3 pts (b) 5, 10, 15, 20, 25 and 50 days posttransplantation in 4 pts. The X and Y chromosome probes were indirectly labeled with different colour fluorochromes and could be observed simultaneously. The X-chromosome component was a single probe that hybridizes to the centromeric region while the Y chromosome component was a collection of probes that hybridizes to most of the Y chromosome q-arm. 4. Results: Donor neutrophils were documented as early as 12 HR post-infusion with a steady rise beginning 24 HR post. By day 5, all circulating neutrophils were of donor origin. A steady rise of donor lymphoid cells could be documented only between day 10 and 15 post-marrow infusion. By day 25 most of the lymphoid cells were of donor origin. 5. Conclusion: This molecular study reveals that donor hematopoietic cells appear very early post-marrow infusion and are presumed to be the progeny of stem cells in a late stage of differentiation, responsible for a transitory phase of engraftment. The timing of permanent engraftment remains to be determined by other molecular approaches. In situ hybridization in sexmismatch transplants can be of great clinical benefit in predicting very early graft rejection especially in the setting of unrelated donors.
The American Journal of Human Genetics 01/1994; 55. · 11.20 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: We have investigated the feasibility and efficacy of administering a radiation-free preparative regimen in the setting of allogeneic bone marrow transplantation (BMT) in 40 consecutive patients with acute lymphoblastic leukaemia (ALL). Busulfan (4 mg/kg/d x 4 d) and cyclophosphamide (50 mg/kg/d x 4 d) (BuCy4) were given in 29 patients and 11 received busulfan (4 mg/kg/d x 4 d), etoposide (60 mg/kg) and cyclophosphamide (60 mg/kg/d x 2 d) (BuCy+VP - 16). Median age was 22 years (range 1-50); 11 patients were children < or = 15 years of age. All children and 20 adults were at high risk of relapse pretransplant. Nine adults and one child died from transplant-related toxicity. 11 patients relapsed at a median of 11 months post-transplant (range 2-27). The 3-year Kaplan-Meier estimated probability of relapse was 42.1% and found to be significantly lower in patients with chronic GVHD (P = 0.03). 19 patients are leukaemia-free survivors with a median follow-up of 33 months (range 7-59). The Kaplan-Meier actuarial probability of disease-free survival at 3 years was 43% for all patients. 63.6% for children versus 30.2% for adults (P = 0.24) and 51.6% for patients transplanted in first remission versus 30.2% for those transplanted in subsequent remissions (P = 0.20).
British Journal of Haematology 12/1993; 85(4):706-13. · 4.94 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Refractory anemia (RA) is the only myelodysplastic syndrome (MDS) devoid of quantitative marrow diagnostic criteria. The diagnosis rests mainly on the subjective identification of qualitative abnormalities according to the French-American-British criteria (FAB) involving one or more bone marrow hematopoietic cell lineages. The occurrence of nonrandom chromosome abnormalities remains the hallmark of the disease and the only means of investigation which confirms the disease objectively. With the purpose in mind to further characterize RA among MDS, we have undertaken a prospective high resolution banding chromosome analyses of bone marrow cells in patients with primary refractory anemia (PRA) with the aim of defining a cytogenetic phenotype and of assessing the clinical relevance of clonal abnormalities at initial diagnosis. Of 39 patients consecutively referred for chromosome analyses with a diagnosis of RA according to the FAB criteria, 27 patients had PRA and fulfilled our criteria for adequate chromosome analyses. Median age was 68 years. Fourteen of 27 patients (52%) had clonal chromosomal abnormalities at diagnosis. None of the patients showed a complex karyotype; 9/14 (64%) had a mixture of normal and abnormal cells. Interstitial or terminal deletions, involving chromosomes 5, 6, 7, 9, 11, 12, and 20, were found in 11/14 (79%) of the patients. Comparison of survival between patients with and without abnormalities showed no difference. The presence of clonal abnormalities did not predict transformation to acute myeloblastic leukemia (AML) nor was it associated with poor survival. In this study, patients with PRA were found to have a predominant pseudodiploid karyotypic pattern characterized by interstitial and/or terminal deletions as opposed to derivatives, specific and non-specific balanced translocations, or other structural and numerical abnormalities. We were unable to reveal any prognostic significance to the presence of these clonal abnormalities at initial diagnosis.
American Journal of Hematology 01/1993; 41(4):241-8. · 4.00 Impact Factor