bluuebunny/crossref_metadata_2025 huggingface dataset:
Influence of Platelet Depletion on Immunomagnetic CD34+ Cell Selection for Haploidentical Transplants.
Authors: Del Fante Claudia , Viarengo Gianluca , Bergamaschi Paola , Marchesi Andrea , Bellotti Laura , Parisi Cristina , Tinelli Carmine , Salvaneschi Laura , Perotti Cesare
Published: October 2019

Introduction: Immunomagnetic CD34+ cell selection (ICS) is a widely employed technology in auto and allotransplant settings. In particular, when an haploidentical transplant is performed an high dose of purified CD34+ cells together with an efficient T and B cells depletion are requested to minimize the risks of GVHD and EBV related lymphoma. In order to ameliorate the performances of the ICS device, we compared 73 ICS performed following the customer’s recommendations vs 20 ICS performed after 3 centrifugations in order to increase the platelet depletion of the starting leukapheresis (LKF) product.

Matherials and Methods: 93 ICS (from single or fractioned LKF product) were carried out using the Miltenyi Clinimacs device on 76 LKF collected from 37 healty donors mobilized with G-CSF alone (12μg/kg). In tab 1 are summarized the characteristics of the starting 93 pre-immunoselection products in terms of TNC, CD34+ cells, plt and T and B lymphocyte content. 73 LKF were prepared following the standard manufacturer’s protocol and the remaining 20 with an adjunctive plt depletion obtained by 3 low speed centrifugations (900 rpm).

Results: a statistical significance in terms of CD34+ cells recovery, final TNC (x106) and CD34+ cell (x106) absolute count was found in favour of the modified immunoselection procedure (see tab.1).

Conclusions: ICS demonstrated to be an highly efficient procedure when the Miltenyi Clinimacs device is employed. Moreover, the modification of the standard manufacturer’s technique simply by adjunctive centrifugations on the starting LKF product can further improve the recovery of stem cells maintaining, at the same time, a good purity and not influencing the T and B cells log of depletion, thus demonstrating the negative influence of plt contamination on ICS. These results are more remarkable in the context of haploidentical transplant where the plt contamination of LKF is particularly high. Table 1

Standard procedure(n=73) Modified procedure (n=20) LKF content Pre-ICS Post-ICS Pre-ICS Post-ICS Mann Withney test TNC(x10e6) 58150 (24150–146700) 264 (73.1–784.5) 54250 (32790–87570) 330.41 (126.84–537.9) p 0.02 CD34+(x10e6) 354.25 (115.67–941.22) 245.55 (59.39–768.81) 377.52 (149.84–604.23) 310.38 (115.87–489.49) p 0.04 PLT(x10e9/L) 3220 (555–9320) - - 3522 (744–6600) - - n.s. LymphocytesT(x10e6) 12880 (1200–36050) 0.13 (0.01–0.6) 11820 (1690–25050) 0.38 (0.08–2.34) n.s. LymphocytesB(x10e6) 3180 (70–15000) 1.06 (0.02–8.42) 2810 (1120–6100) 0.82 (0.19–2.98) n.s. CD34+(%)recovery 70.98 (9.5–115.32) 82.34 (66.37–99.94) p 0.04 Purity(%) 92 (69–99) 94 (89–98) n.s. LogT-Depletion 5.1 (3.74–7.91) 4.6 (3.4–5.2) n.s. LogB-Depletion 3.65 (2.45–6.08) 3.55 (2.58–4.44) n.s.
The Role of Hemoglobin Barts in Umbilical Cord Blood Harvest and Transplantation.
Authors: Regan Donna M. , Wofford Jonathan D. , McCormick Kimberly A. , Alonso J. Mario F. , Creer Michael H.
Published: October 2019

The abnormal hemoglobin present during fetal development in individuals with four-gene deletion alpha-thalassemia was characterized at St. Bartholomew’s Hospital in London and hence was named “hemoglobin Barts (Hb Barts).” In the homozygous state, when all four alpha-globin genes on chromosomes 16 are affected, no normal fetal or adult hemoglobin can be produced and the fetus becomes profoundly anemic in utero. This typically leads to stillbirth or death soon after birth due to severe hydrops fetalis accompanied by pulmonary hypoplasia and cardiac failure. Various heterozygous states can also produce detectable levels of Hb Barts, but with less dramatic clinical manifestations. These variants can be detected and quantified in umbilical cord blood (UCB) in proportions indicating one-, two- or three-gene deletion alpha-thalassemia, respectively representing silent carrier, thalassemia trait, and Hemoglobin H disease states. Silent carriers, the largest group with Hb Barts at birth, have a normal CBC, no clinically detectable problems and less than 5% Hb Barts present in cord blood sample. Individuals with alpha-thalassemia minor (or trait) have a very mild microcytic anemia but no other clinical problems, and express 5 to 10% Hb Barts in cord blood. One- and two-gene alpha-globin deletions are very common in African, Mediterranean and Asian populations, ethnic groups targeted for cord blood bank inventories.

Sixteen UCB products containing Hb Barts at levels consistent with either one or two alpha-gene deletions were exported for stem cell transplant from the St. Louis Cord Blood Bank. Survival of recipients receiving Hb Barts-containing units was not significantly different from those receiving cord products with normal newborn hemoglobin electrophoretic patterns (P =.5707). To further evaluate the effect of transplanting units with Hb Barts variants, we propose an Institutional Review Board-approved study to evaluate erythropoiesis and the disappearance of fetal hemoglobin variants at designated post-transplant time points in patients who have been transplanted with these products.

In deliveries where the infant exhibits symptoms of hydrops fetalis, obvious perinatal complications would preclude cord blood harvest for the intended use of allogeneic transplantation. Considering the need for these unique and diverse HLA types, criteria for banking should allow inclusion of UCB units with Hb Barts content of less than 10% based on immunoelectrophoretic patterns. These units should pose minimal additional risk in transplantation and increase the ethnic diversity of bank inventories. Additionally, since recipients’ genetic material remains intact after transplantation, they cannot pass these variant hemoglobins to their children.
Influence of Pretreatment with Amphotericin B on Treatement-Related Mortality (TRM) Following Allogeneic Stem Cell Transplantations.
Authors: Trenschel Rudolf , Ditschkowski Markus , Steckel Nina K. , Hlinka Michal , Elmaagacli Ahmet , Ottinger Helmut , Flasshove Micheal , Beelen Dietrich W.
Published: October 2019

Conventional amphotericin B (C-AMB) is one of the most frequently used antifungal first line agents in patients with hematological malignancies diagnosed to have invasive fungal infections or persistent neutropenic fever despite of its well documented adverse effect profile. To evaluate the influence of pretreatment with C-AMB on the outcome of allogeneic HSCT, a retrospective analysis including 270 consecutive allogeneic HSCT between 1/2002 and 12/2003 at out institution was performed. Median patient age was 42 years, female to male ratio 46/51 and CMV positivity 83 %. Diagnosis was AML (n=90), ALL (n=72), CML (n=45), MDS (n=21), NHL (n=14), MM (n=8) or micellaneous (n=20). Sixty percent (164/270) of patients were in early, the remainders in advanced disease stages. Conditioning therapy was TBI or chemotherapy in 237 and 33 patients, respectively. Donors were identical siblings (n=110), non-identical family members (n=3), matched unrelated (n=116) or mismatched unrelated (n=41). Pretreatment with antimycotics was reported in 74 cases (C-AMB, n=73; voriconazole, n=1) for FUO (n=23), not specified fungal pneumonia (n=23), pneumonia due to aspergillus sp.(n=18), hepatosplenic candidiasis (n=6), fungemia (n=3), and IFI by scedosporium prolificans (n=1). A Cox regression model was used to test C-AMB pretreatment as an independent risk factor on TRM. Age in decades, disease stage, preceding neutropenia, renal impairment (creatinine > 1mg/dl) was also included. Only previous treatment with C-AMB (p=0.001; RR 3), and advanced disease stage (p=0.008; RR2,5) remaind statistically significant. Subsequently, to identify patients at increased risk, we defined following strata: 1. patients without previous neutropenia; 2. patients with previous neutropenia; 3. patients with C-AMB pretreatment, creatinin < 1,0 mg%; 4. patients with C-AMB pretreatment and creatinin > 1,0mg%; 5. patients with renal impairment for other causes; 6. patients with renal impairment or C-AMB pretreatment and antimycotic therapy during transplant procedure. TRM was 3.2%, 5.43%, 19.57%, 26.67%, 29.63% and 24.14%, respectively. We conclude that patients being pretreated with conventional C-AMB have an increased independent risk for TRM (RR 4; p<0.001).
Abandoning Care in Laminar Air Flow (LAF) Units and Routine High Dose Intravenous Immunoglobulines (IVIG) in Allogeneic Hematopoietic Stem Cell Transplantation (HSCT) Does Not Increase Mortality and Rate of Infections.
Authors: Cantoni Nathan , Passweg Jakob , Weisser Maja , Droll Armin , Christen Susanne , Widmer Andreas , Flueckiger Ursula , Frei Reno , Tichelli André , Gratwohl Alois
Published: October 2019

Nursing in LAF units and routine high dose IVIG were considered essential elements and standard of care in allogeneic HSCT during the 1980s. They were abandoned by many teams in view of high costs, lack of formal proof of evidence and improved diagnostic and therapeutic interventions against viral and fungal infections. A recent prospective study showed no benefit of routine IVIG but few data have analysed the consequences of the combined change in practice. We made these changes at our institution in two stages during a 10-year period: change from care in LAF rooms to single rooms (SR) and change from routine high dose IVIG to targeted IVIG replacement according to CDC guidelines. Basic strategies in supportive care and data collection remained unchanged. This resulted in a retrospective analysis of 357 allogeneic HSCT in three cohorts, i.e. LAF + IVIG, SR + IVIG and SR − IVIG (Table). Endpoints analysed were survival, transplant related mortality (TRM), relapse rate, incidence and severity of GvHD and infectious complication rates defined as: sepsis, septic thrombosis, urinary tract infections, pneumonias, and CMV reactivation. Results were adjusted by multivariate analyses for all the changes over time (see Table *significant difference between groups). There were more patients in advanced stage of disease and with higher age in more recent years. There were significantly fewer septicemias per hospital day but not per day in neutropenia in the LAF + IVIG group without impact on survival. There were no significant differences in any of the other outcomes analysed. Abandoning LAF and routine high dose IVIG use did not impact negatively on HSCT outcome. These data support current practice.

Group LAF + IVIG SR + IVIG SR, no IVIG Total N 116 134 107 357 Male 55% 51% 62% 59% Age, years* 34 34 40 35 Median, range 4–58 2–62 1–62 35 Diagnosis* Leukemias 85% 83% 82% 84% Lymphoproliferative 7% 13% 15% 11% Others 8% 4% 3% 5% Early disease* 46% 38% 29% – Late disease 54% 72% 71% 66% % RIC* 0% 10% 32% – Days in hospital* 53 32 28 38 Days in neutropenia* 16 15 11 14 Day 100 TRM 18% 20% 22% 20%
The Gut-Mucosa-Barrier-Function during Hematopoietic Stem-Cell Transplantation Predicts Acute Graft-Versus-Host Disease Severity.
Authors: Johansson Jan-Erik , Ekman Tor
Published: October 2019

Background

Graft-versus-host disease (GVHD) is the primary complication of allogeneic, haemopoietic, stem-cell transplantation (HSCT). From murine models, it has been revealed that gut toxicity increases the translocation of inflammatory mediators through the impaired intestinal barrier, which aggravates systemic GVHD. In patients, the intestinal barrier is disrupted after myeloablative conditioning but preserved after reduced intensity conditioning (RIC). The present study investigates, in a clinical situation, whether the severity of acute GVHD depends on intestinal-barrier function.

Methods

In 38 patients (21 myeloablative, 17 RIC), intestinal permeability was assessed before transplant and during the transplantation course (day −1 to +14). Blood levels of cytokines were assessed in parallel.

Results

Intestinal barrier integrity was preserved in RIC patients but disrupted in myeloablatively conditioned on day 4 (p = 0.0091) and on day 7 (p = 0.0014) compared with RIC patients. Patients with mild, acute GVHD (grades 0–1) had a significantly better preserved intestinal-barrier function (p = 0.042 on day 4) and lower TNF-a levels (p = 0.013 and 0.025 on day −1 and day +7 respectively), compared with patients with more pronounced GVHD (grades 2–4).

Conclusion

Intestinal barrier function predicts the severity of acute GVHD in patients and gives support to the assumption that gut protective strategies may diminish acute GVHD.
The Useful Individualized Assessment of Immunosuppressive Therapy with Cyclosporine A (CsA) after Allogeneic Stem Cell Transplantation by Simultaneous Analyses of Cytokine Production among CD4+ T Cells and Blood CsA Levels.
Authors: Tsukada Nobuhiro , Furukawa Tatsuo , Ito Hiromi , Izumi Noriko , Momoi Akihito , Okazuka Kiyoshi , Honma Keiichiro , Masuko Masayoshi , Hashimoto Shigeo , Aoki Sadao , Aizawa Yoshifusa
Published: October 2019

Cyclosporin A (CsA) is the most common immunosuppressive agent used for recipients undergoing allogeneic stem cell transplantation (SCT). CsA require therapeutic drug monitoring (TDM) for both its effectiveness and toxicity, but the schedule of administration and optimal blood levels vary among institutions. To establish better assessment of CsA effect on individual immune responsiveness, we analyzed the proportion of IFN-γ+ or IL-2+ cells among CD4+ lymphocytes using 3-color flow cytometric analysis before and after CsA administration. CsA is administered as 3-hour infusion or oral medication twice daily in equally divided doses. 83 peripheral blood samples from 8 allogeneic BMT recipients are analyzed. Cells were cultured in the presence of phorbol 12-myristate 13-acetate, ionomycin, and brefeldin A at 37°C for 4hrs and then stained for surface markers and intracytoplasmic IFN-γ and IL-2. Blood CsA levels were simultaneously measured. We found that the proportion of IFN-γ+CD4+ or IL-2+CD4+ lymphocytes were inversely proportional to blood CsA levels (Figure 1). To achieve profound inhibition of IL-2 production, high concentration of CsA is required (e.g. 600–800 ng/ml for less that 5 % IL-2+CD4+ cells). In vitro experiments were also conducted using lymphocytes from healthy donors with or without activation with CD3 and CD28 to show how the sensitivity to CsA is different in between activated and resting T cells. We found that higher CsA concentration is required for inhibiting IL-2 production from CD4+ lymphocytes activated with CD3 and CD28 than from resting CD4+ lymphocytes (Figure 2). In this regard, our 3 hour intravenous CsA infusion can be superior to continuous intravenous infusion in inhibiting activated T cells. Further analyses are required, but the simultaneous assessment of cytokine production among CD4+ T cells and blood CsA levels may be a useful index to estimate the degree of immunosuppression afforded by CsA and may enable us individualized therapeutic drug monitoring of this agent. Since more and more patients are undergoing cord blood transplantation (CBT) or reduced-intensity stem cell transplantation (RIST), this assessment may also be useful for the establishment of immunosuppressive therapy for those patients.
An Experimental and Clinical Study of CMV Infection after Allogeneic Hematopoietic Stem Cell Transplantation.
Authors: Wu De Pei , Wu Xiaojin , Li Caixia , Ma Xiao , Sun Aining
Published: October 2019

Objective: To establish quick and reliable diagnostic methods for detecting the cytomegavirus (CMV) infection after allogeneic hematopoietic stem cell transplantation (allo-HSCT) and to evaluate their clinical value in early diagnosis of CMV infection

Methods: (1) From March 2001 to December 2003, Seventy-four patients undergoing allo-HSCT were enrolled in this study. After allo-HSCT, all the patients were detected for CMV gB DNA in peripheral blood by haplo-nested PCR methods and 35 of 74 patients for CMV PP65 antigen in peripheral blood by the fluorescent immunohistochemistry (FIHT) assay. The positive rate and correspondence of these two methods were evaluated.

Results: (1) In 35 of 74 patients who were tested for CMV infection by two assays at the same time, the CMV-viremia was first detected at a median of 36 days after transplantation by FIHT assay and 26 days by haplo-nested PCR assay during the follow-up time of 90~720 days. The median time of positive persistence was 19 days by FIHT assay and 28 days by haplo-nested PCR assay. The significant difference of them was observed (P<0.05). The corresponding of the two assays was 88.57%. The difference of the two methods was not significant statistically. The incidence of CMV-agtigenemia and CMV-DNAemia were 65.71% and 71.43% during the follow-up days, respectively. (2) The infection rate in the nonmyeloablative allogeneic peripheral stem cell transplantation (NST) group was 78.95%, in the group of related peripheral blood stem cell transplantation (R-PBSCT) and/or related bone marrow transplantation (R-BMT) 48.57%, and in the group of unrelated bone marrow transplantation (UR-BMT) 61.54%. The infection rate of CMV in the UR-BMT group was higher than that in R-PBSCT and/or R-BMT group, but without significant difference (P>0.05). The infection rate of CMV in the NST group was significantly higher than in R-PBSCT and/or R-BMT group (P<0.05), but only a litter higher than in UR-BMT group (P>0.05). Both CMV-antigen and CMV-DNA were detected in all seven patients undergoing haploidentical hematopoietic stem cell transplantation (Hi-HSCT).

Conclusion: (1) Both FIHT and haplo-nested PCR assays were quick and reliable diagnostic methods for detecting the CMV infection after allo-HSCT and their correspondence was good. However CMV infection could be found earlier by haplo-nested PCR assay than by FIHT assay. (2) The infection rate of CMV is high after allo-HSCT and might be different in different types of allo-HSCT.
Immune Suppressor Factor Confers Enhanced Supporting Activity for Hematopoietic Stem Cells in Bone Marrow Stroma.
Authors: Nakajima Hideaki , Shibata Fumi , Fukuchi Yumi , Goto-Koshino Yuko , Ito Miyuki , Kitamura Toshio
Published: October 2019

Immune suppressor factor (ISF) is a subunit of vacuolar ATPase proton pump that plays a critical role for proton transfer to extracellular space or subcellular organs. Physiological importance of vacuolar ATPase was evident in a variety of cellular functions such as re-absorption of bones by osteoclasts, acidification of lysosomes in macrophages, and acidification of urine in kidney. However, a role for vacuolar ATPase in hematopoiesis is still unknown. We have previously identified a short form of ISF (ShIF) as a stromal cell derived factor that supports factor-independent growth of a mutant subline of Ba/F3, an IL-3 dependent murine hematipoietic cell line. In this study, we addressed whether ISF supports self-renewal and expansion of primary hematopoietic stem cells (HSC). Co-culture of murine bone marrow cells with a stromal cell line overexpressing ISF or ShIF (MS10/ISF or MS10/ShIF) enhanced their colony-forming activity as revealed by the numbers and the size of the colonies. In addition, the numbers of long-term culture initiating cell (LTC-IC) were markedly increased by co-culture on MS10/ISF or MS10/ShIF. Moreover, competitive repopulating activity of c-Kit+Sca-1+Lin- HSC was significantly maintained without any added cytokines when they were co-cultured with MS10/ISF or MS10/ShIF compared with a mock control. In order to check whether these activities were dependent on the proton pump function of ISF, we generated a mutant ISF or ShIF whose proton pump activity was abolished by point mutation. Interestingly, all stem cell supporting activities described above were abolished in the mutant ISF or ShIF, indicating that proton transfer across cellular or endosomal membrane was critical. Analysis of gene expression profile of mock and ISF-transfected cell lines revealed that any cytokines or growth factors previously known to affect hematopoiesis are not modulated at mRNA level. However, downregulation of secreted frizzled related protein (sFRP)-1, an antagonist for Wnt, and up-regulation of matrix metalloproteinase-3 (MMP-3) were clearly noted in ISF/ ShIF-overexpressing cell line, suggesting that relative increase of Wnt activity and the modulation of extracellular matrix are the key molecular events underlying the enhanced supporting activity for HSC. These results propose a novel role for ISF in self-renewal and expansion of HSC in vivo.
Three Hundred and Fifty Cases of Pediatric Allogeneic Hematopoietic Stem Cell Transplantation: Single Center Study in Korea.
Authors: Park Young-Shil , Jang Pil-Sang , Choi Sangrhim , Chung Nak-Gyun , Cho Bin , Jeong Dae-Chul , Kim Hack-Ki
Published: October 2019

Background: Hematopoietic stem cell transplantation (HST) was first introduced to Korean children in 1983. Since then HST has been a curative method for pediatric malignant or non-malignant hematologic disorders and the number of children receiving HST’s has been increasing steadily.

Method: We analyzed three hundred and fifty cases of pediatric allogeneic hematopoietic stem cell transplantation between Nov. 1983 and Dec. 2003 in the Catholic Hematopoietic Stem Cell Transplantation Center of Korea.

Results: HLA-matched sibling bone marrow transplantations were performed in 206 cases (103 males, 103 females, median age 12 years) with median follow-up of 73 months. The 5-year event-free survival (EFS) of ALL and AML was 70.5 % and 67.22 %, respectively. The 5-year EFS of severe aplastic anemia was 90.9 %. The 5-year EFS of CML/MDS and rare hematologic diseases was 65.5 % and 90.0 %, respectively. Eighty-one children (56 males, 25 females, median age 9 years) underwent unrelated bone marrow transplantation (UBMT), 42 (27 males, 15 females, median age 5 years) cord blood transplantation (CBT) and 21 (14 males, 7 females, median age 8 years) familial haploidentical HST (FHT). Three-year EFS of UBMT, CBT and FHT was 55.6 %, 48.0 % and 38.0 %, respectively.

Conclusion: HLA-matched sibling allogeneic BMT showed better survival in children with hematopoietic stem cell disorders. Recently, transplants using alternative stem cell sources are increasing due to the lack of suitable sibling donors and continued efforts for reducing transplant-related complications are warranted for improved survival.
Nonmyeloablative Allogeneic Stem Cell Transplantation: Hacettepe University Institute of Oncology Experience.
Authors: Koc Yener , Ozdemir Evren , Oyan Basak , Kansu Emin
Published: October 2019

The purpose of this study was to determine the feasibility and efficacy of nonmyeloablative allogeneic stem cell transplantation in patients with hematologic malignancies and metastatic renal cell carcinoma (RCC). Thirty patients with various pathologies (AML/MDS, n=9; CML, n=5; lymphoid malignancy, n=6; Multiple Myeloma, n=7; RCC, n=3) received a fludarabine (F)-based nonmyeloablative regimen (F+Busulphan, n=5; F+TBI, n=18; F+Cyclophosphamide, n=3; others, n=4), followed by unmanipulated peripheral stem cells from HLA-identical sibling donors. Prophylaxis for graft-versus-host disease (GVHD) consisted of cyclosporine and mycophenolate mofetil. Median age was 43 years (range, 17–62), median time from diagnosis to transplant was 1.6 years (range, 0.4–9.1). Patients underwent a median of 2 prior chemotherapy regimens (range, 0–7). Prior stem cell transplantation failed in seven patients (23%) (autologous, n=6; ablative allogeneic, n=1). Disease phase prior to transplant was complete or partial remission in 16 patients (CR, n=12; PR, n=4) and active disease in 14 patients (Primary refractory, n=1; Relapse, n=10; Untreated, n=3). Recipients were male and donors were female in 15 transplants (50%). The median numbers of mononuclear and CD34+ cells infused were 8.8 x 108/kg (range, 4–26 x 108/kg) and 9.7 x 106/kg (5.2–32.5 x 106/kg) respectively. Neutrophil engraftment achieved after a median of 9 days (range, 6–12) and platelet engraftment occurred after a median of 11 days (range, 9–19). In twelve (40%) patients, absolute netrophil count did not fall below 500/mm3 and 14 patients (47%) required no platelet transfusions. The patients received a median of 2 donor lymphocyte infusions (range, 0–8). Nine patients (30%) had acute GVHD (Grade II-IV, n=5, 17%) and 12 patients (40%) had chronic GVHD (limited, n=4; extensive, n=8, 27%). Three patients (10%) died of nonrelapse treatment-related complications within 100 days post-transplant (Pneumonia, n=2; Bronchoalveolar hemorrhage, n=1). Two patients had graft failure at day 100 post-transplant. Twenty two patients (73.4%) are currently alive with a median follow-up of 6 months. Primary causes of deaths (n=8, 26.6%) are as follows; GVHD (n=3), pneumonia (n=2), bronchoalveolar hemorrhage (n=1), and relapse (n=2). Among 22 evaluable patients, response was obtained in 18 patients (82%) (CR, n=14; PR, n=2; stable disease, n=2) and 4 patients relapsed. Our data suggest that nonmyeloablative allogeneic transplantation is a safe and effective treatment strategy in hematological malignancies.
T Cell-Depleted Related Haploidentical Peripheral Blood Stem Cell Transplantation in a Patient with Fanconi Anemia. Cagliari Experience.
Authors: Sanna Maria Adele , Orofino Maria Grazia , Cossu Fausto , Addari Maria Carmen , Piroddi Antonio , Rizzo Giuseppa Fabiana , Badiali Manuela , Galanello Renzo , Argiolu Francesca
Published: October 2019

Stem cell transplantation is presently the best treatment for Fanconi Anaemia (FA) patients developing bone marrow failure. 70% of success is reported in patients with a HLA identical sibling whereas the outcome for HSCT in those transplanted from unrelated donors is in the range of 29–43%, graft rejection, GVHD and regimen related toxicity beeing the main causes of failure. This results limited the ability to perform marrow transplantation other than HLA identical siblings for this disease. Recently a fludarabine based cytoreductive regimen has been successfully used in T cell depleted haploidentical/mismatched transplant of FA patients. We report a case of a 7 year old boy with bone marrow failure since 1999. Androgens treatment was uneffective, no HLA identical family donor was available and the search for a suitable marrow or cord blood unrelated donor was unsuccessful. After 4 years he underwent T-cell depleted haploidentical PBSCT from his father. Conditioning regimen was: fludarabine 30 mg/mq from day −6 to day −3, cytoxan 300 mg/mq from day −6 to day −3, rabbit ATG (3.75 mg/kg) from day −5 to day −3. GvHD prophylaxis consisted of cyclosporine 1 mg/kg from day −1. The donor received G-CSF 8 ug/kg/dose twice daily for 6 days and underwent leukapheresis on day 5 and 6. Donor stem cells were depleted of T cells by positive selection of CD34+ cells using the Clinimacs device according to the suggested procedures (Milteny Biotec). On day 0, 15.3x106 x kg CD34+ cells were infused with 1.5 x 105 CD3 + cells. The clinical postransplant course was uneventful. Neutrophil engraftment ( >0.5 x 109 ) occurred on day 14, platelet count >100x109 on day 15. He was discharged on day 39 without signs of GVHD. Molecular analysis of DNA-VNTRs at 1, 3, 6, 9, 12 months showed >95% donor chimerism on peripheral blood. At 14 months after transplantation the patient is well, normal blood cell count (WBC 5.4 x 109/l, Hb 13.6 gr /dl, platelets 293x 109 /l). Count of T-cells are reported in the normal reference range ( CD3+ :1865 ug/l, CD8+ :1026ug/l, CD19+ :732ug/l, CD56+: 452). Karnofsky score is 100%.

Conclusion: the case reported shows that the fludarabine based regimen and the infusion of a high number of T-cell depleted CD34+ was successful in absence of peri-transplant complications and can be proposed for the cure of FA patients at high risk of clonal disease and without HLA-matched sibling donor.
Clinical Outcome of Cord Blood Transplantation by Age with Units Shipped from Tokyo Cord Blood Bank.
Authors: Nagamura-Inoue Tokiko , Yan Cui , Kodo Hideki , Mugishima Hideo , Sugo Michiko , Takada Kei , Shioya Mika , Hirai Masako , Takahashi Atsuko , Satomi Aya , Hoshino Mokaku , Tahara Yoshiyuki , Asano Shigetaka , Takahashi Tsuneo
Published: October 2019

Recently, cord blood transplantation (CBT) for adult is rapidly increasing in number, Especially for the patients over 50 years of age. By the end of 2003, 224 units were shipped for the patients Group I: <15 y.o. 39%, Group II: 15~50 y.o.40% and Group III :> 50 y.o.21%. We analyzed 152 patients with hematological malignancies including ALL, AML, CML, MDS, malignant lymphoma, myeloma and neuroblastoma reported from CBT center in the world by the end of 2003.

Patients and Methods:Group I included 58 patients with 13 standard risk and 45 high risk patients and showed mean±SD of age; 5.3 ±4.1y.o.,BW; 20.5±13.4kg, CB volume at collection; 91.6±27.1ml, NC; 5.5±3.3x107/kg, CFC; 8.6±6.4x104/kg, CD34; 1.5±1.1x105/kg, Group II: 64 cases with 25 standard and 39 high risk patients and age; 30.6±10.3y.o., BW; 51.9±9.5kg, CB volume at collection;116.6±27.7ml, NC;2.6±0.7x107/kg, CFC;5.2±2.6x104/kg and CD34;0.8±0.5x105/kg, Group III: 30 cases with 9 standard and 21 high risk patients and age; 54.1±3.3y.o., BW;55.9±11.0kg, CB vol.at collection;118.8±24.7ml NC;2.4±0.4x107/kg, CFC;4.8±1.9x104/kg and CD34;0.8±0.4x105/kg. The patients who underwent CBT for graft failure (GF) of prior transplant were excluded. Conditioning regimen in Group I demonstrated 54 patients with full regimen and 4 with reduced intensity regimen (RIST); in Group II, 62 patients with full regimen and 2 RIST; and in Group III, 14 cases full regimen and 15 cases RIST.

Results: Cumulative myeloid engraftment was seen 67.2% in Group I, 73.4% in Group II and 46.7% in Group III (*Group II vs. Group III: P<0.05). Overall survival /EFS on day 100 showed 73.4%/59.4% in Group I, 74.0%/58.6% in Group II and 43.3%/34.5% in Group III (*Group III vs. others: P<0.05). In Group III, the survival rate indicated 42.8% in full regimen group and 13.3% in RIST group at 1year after CBT. In Group I, four patients died of GF, 13 of relapse, 11 of Transplantation related disease (TRD); in Group II, 5 patients died of GF, 8 of TRD, 10 of relapse. In Group III, four patients died of TRD, 3 of GF and 3 of relapse in full regimen, while in RIST, six patients died of TRD, 1 of relapse and 1 of acute GVHD.

Conclusion: The application of CBT has been expanded to the elderly patients (>50 y.o.), although the conditioning regimen and the special medical care for the complications in the early pahse after UCBT has remained to be discussed.
Results of High Dose Chemotherapy and Autologous Stem Cell Transplantation in Patients with Acute Myeloid Leukemia.
Authors: Ault Kenneth A. , Case Delvyn C. , Boyd Marjorie A. , Ervin Thomas J. , Aronson Frederick R. , Ebrahim Kurt S. , Hedlund Jaquelyn A.
Published: October 2019

35 patients with acute myeloid leukemia have undergone transplantation at our institution over the past 12 years. Patient selection criteria included age less than 70 years, creatinine less than 2mg/ml, no active infection, cardiac ejection fraction >40%, DLCO > 50% of predicted and no other co-morbid conditions that would jeopardize survival. 31 patients were in first remission, 4 were in second or higher remission. 3 patients had favorable cytogenetics, 25 had intermediate or unfavorable cytogenetics. After achieving remission for at least 30 days, patients were consolidated with Etoposide and AraC, followed by G-CSF. Hematopoietic stem cells were collected when the WBC rebounded to at least 10,000/ml. The target dose of CD34 positive cells was 5x106/kg. The minimum dose given was 2.3 x 106/kg). The average age at transplantation was 41 years (range 22 to 61). Days of neutropenia (AGC<500/ml) ranged from 2 to 10 (average 5.1). The median length of follow up is 2.6 years. Currently 26 patients are alive, and 25 are free from progression. Overall survival is 63%, and progression-free survival is 47% at both 5 and ten years.

Figure Figure
Outcome after Relapse among Children with Standard Risk (SR) ALL Treated on CCG-1952.
Authors: Malempati Suman , Gaynon Paul S. , Sather Harland , La Mei K. , Stork Linda C.
Published: October 2019

The event-free survival (EFS) of children with SR-ALL is now > 80%. Nonetheless, ALL relapse remains a major problem in pediatric cancer, and optimal therapy for these children remains to be defined. We examined post relapse outcomes for children initially treated on the Children’s Cancer Group CCG-1952 study and compared stem cell transplant (SCT) with chemotherapy (CT) as salvage treatment in second remission (CR2). Between 5-96 and 1-00, 2176 eligible patients with SR-ALL (WBC < 50,000/mcl; age ≥ 1, < 10 years) were enrolled on CCG-1952, 321 of whom experienced a relapse after initially achieving M1 remission. Among the relapses, 196 are bone marrow (BM) ± extramedullary (EM) relapses (61%) and 125 are isolated EM (iEM: CNS, testicular, or ocular) (39%) relapses. The mean time (range) from first remission to BM relapse, iCNS relapse, and itesticular relapse is 34 ± 1 (2 to 79) month (mo.), 22 ± 1 (1 to 70) mo., and 37 ± 3 (4 to 74) mo., respectively. The 3-year EFS and overall survival (OS) after relapse for all patients are 41% and 53%. Three-year EFS and OS after BM ± EM relapse are 32% and 40% and after iEM relapse are 55% and 71%. Patients with early BM relapse (CR1 < 36 months) or early iEM relapse (CR1 < 18 months) are 2.3 times (p=0.002) and 2.8 times (p=0.01) more likely to suffer a subsequent adverse event than patients with later relapse by Cox regression analysis after adjustment for age, pre-relapse treatment, day 7/14 BM status, and type of treatment in CR2. The 3-year OS for early and late relapses are 31% vs 59% (p=0.001) for BM relapse and 50% vs 87% (p=0.01) for iEM relapse. We compared 73 SCT and 215 CT patients, excluding 33 patients with adverse events prior to the median time to SCT (130 days, range 56 to 1148 days) using Kaplan-Meier life table analysis and the log rank test. The cohorts are similar with respect to age (p=0.57), duration of CR1 (p=0.96), and initial Day 14 BM status (p=0.69). Thirty-seven percent (n=62) of patients with BM relapse (33 early; 29 late) and 9% (n=11) of patients with iEM relapse (6 early, 5 late) received SCT. Thus far, outcomes are similar between cohorts for patients with any BM relapse (OS, p=0.45; EFS, p=0.70), early BM relapse (OS, p=0.95; EFS, p=0.66), and iEM relapse (OS, p=0.44; EFS, p=0.96). Among patients with later BM relapse, a trend favors CT (OS, p=0.08; EFS, p= 0.14). In summary, duration of CR1 remains the most significant predictor of outcome after either BM or iEM relapse in this SR-ALL cohort. Prognosis after early BM relapse remains poor with either SCT or CT.
Clinical Report on the Treatment of Children with Advanced Neuroblastoma with Autologous Haematopoietic Stem Cell Transplantation.
Authors: Chen Jing , Gu Long-Jun , Tang Jing-Yan , Zhao Hui-Jun , Pan Ci , Xue Hui-Liang , Wang Yao-Ping
Published: October 2019

Neuroblastoma(NB) is a common solid tumors in children. Even with active chemotherapy, operation and irradiation, the survival rate is still very low in advanced patients. To improve the treatment results, autolougous haematopoietic stem cell transplantations have been performed in 11 patients with advanced NB in our hospital. The average age was 3.8 years old (2–6 years old) and the average weight was 15.3kg(11.6–20kg) in this group. All 9 patients were in stage IV with the primary mass in abdomen and extensive bone marrow involvement but 2 patients in stage III with the primary mass in thorax. Although high dose chemotherapy and active operation was given, 4 cases still have not got complete remission at the time of transplant. As 2 cases received 2 times transplant, totally 13 transplants were performed in these 11 children. 3 were collected stem cell from the anterior crista of iliac in both sides while another 10 were received the stem cell from the peripheral mononuclear cell harvested with CS-3000 cell separator after G-CSF mobilization. To reduce the risk of post transplant relapse, 4 cases were purged with CliniMACS based on the CD34 positive selection. All the patients conditioned with Etopside plus Carboplatin plus Melphalan. Results show the number of mononuclear cell collected from bone marrow or peripheral blood was equal to 5.7±0.9′108/kg and 5.7±1.0′108/kg respectively(p>0.05). All of them achieved the haematopoietic reconstitution after transplantation. The mean time for the neutrophile count recovering to 0.5 ′109/L was 10.5±5.7 days and the platelet recovering over 2.0 ′109/L. The mean transfusion independent day was 16.8±9.4 days with average 2 units of packed red blood cells and 4 units of platelet products transfused in the course of transplantation. The mean fellow up time was 26.7 months. 4/11 children died of relapse 5 months to 27 months after transplantation. Other 7 children are still in survival. None of our children died of complication associated with transplantation. It suggests autologous stem cell transplantation is a safe and effective measure in the treatment of children with advanced neuroblastoma and worth of further recommendation.
Long-Term Follow-Up of CML Patients Treated with Autografting Followed by IFN-A and Imatinib for Relapsed Patients.
Authors: Carella A.M. , Corsetti Maria T. , Beltrami Germana , Bodenizza Carlo , Cavaliere Marina
Published: October 2019

Residual diploid hematopoietic progenitor cells (HPC) represent a quantitatively useful reservoir from a therapeutic standpoint, particularly early in the course of the disease. Clinical evidence for the persistence of diploid HPC in CML has been provided by the fact that Ph-negative cells are mobilized into the blood of patients treated with chemotherapy/G-CSF, IFN-a and, more recently, with Imatinib. In this report, we update our experience in 50 patients with early chronic phase not previously treated with IFN-a. All patients completed the mobilization protocol (ICE/mini-ICE) and diploid or prevalently diplod HPC were mobilized in peripheral blood after G-CSF. High-dose therapy consisted of Busulfan (4 mg/kg/d x 4 days) (44 patients) or TBI-containing regimen (6 patients). No patient died of the procedure. After engraftment, all patients were treated with IFN-a (3–5 MU/d three times weekly). The median follow-up of 50 patients is 77 months (range, 8 to 142 months). At present (July 31, 2004), 39 patients (78%) are alive at a median follow-up of 89 months (range, 43 –142) from autografting: 15/39 patients maintain major cytogenetic remission (MCyR) under IFN-a (1 patient received MUD in MCyR;) 23 patients (22 in CP and 1 in AP), who relapsed cytogenetically received Imatinib. Sixteen out of 23 pts on Imatinib achieved MCyR (2 patients) or complete cytogenetic remission (CCyR) (14 patients). Eleven out of 50 patients died at a median of 35 months (range, 8 to 106 months): 9 patients of blastic transformation (2 in the Imatinib group), 1 patient of fulminant hepatitis and 1 of cardiac arrest under Imatinib treatment. In conclusion, intensive treatment with autografting/IFN-a ± Imatinib was able to control the disease in 39/50 patients of whom 31 patients are still in major/complete cytogenetic remission.
Risk Factors of Cytomegalovirus Infection in Autologous Stem Cell Transplantation (ASCT).
Authors: Bonferroni Margherita , Grasso Mariella , Tavera Silvia , Calvi Roberta , Mattei Daniele , Pistone Maria A. , Gallamini Andrea
Published: October 2019

Aim: to analyse the incidence and risk factors of cytomegalovirus (CMV) infection after ASTC.

Methods: 129 pts who underwent ASCT in our Institution from June 1997 to June 2004 were analysed retrospectively ( 40 NHL, 66 MM, 8 HD, 8AML, 7CLL). Conditioning regimen consisted in MitoMel in NHL, Melphalan in MM (100–200/m2) and BEAM in AML and CLL. All patients were monitored with Ag pp65 twice weekly from conditioning therapy to hematological recovery. CMV prophylaxis consisted of acyclovir (400 mg p.o. twice daily) and the use of leucocyte-depleted blood products. Following the report of a positive CMV Ag all patients were treated with ganciclovir 5 mg/kg or foscarnet 90 mg/kg twice daily for 2 weeks or until the test was negative.

Results: we observed 8 CMV Ag positive: 5 were MM in RP at the transplant and 2 out of 3 NHL were heavily pretreated. Six patients were older than 60 years. The number of CD34 infused was 3.8x106/kg (range 3.1–5.1 ). The mean time for PMN > 500/ul and platelets > 30.000/ul was respectively 11 and 21 days. CMV Ag occurred after a mean time of 21 days (4–45) following ASCT. All patients with a positive CMV Ag were febrile. CMV infection was considered the sole cause of fever in 4 pts; 2 had antibiotic-resistant fever, 1 acute enteritis and 1 pneumonia. The remaining 4 pts had concomitant sepsis: Klebsiella, Pseudomonas, Streptococcus Pneumoniae and Staphylococcus Epidermidis. Seven pts had CMV Ag levels >5/200000 cells and one pt >2/200000 at presentation. The resolution of fever and clearance of CMV Ag occurred after 5.6 days (3–9) from the start of antiviral treatment, but in 2 pts was delayed (respectively 14 and 18 days). Of note, these two pts had the highest CMV Ag levels observed (350 and 961) and one treated with foscarnet developed acute renal failure.

Conclusion:

1) CMV infection is an infrequent complication of ASCT( 6,2%) 2) CMV Ag detection was unrelated to the number of CD34 infused and the duration of neutropenia. 3) Pts heavly pretreated, febrile or with sepsis had high rates of CMV reactivation and disease. This subset of pts require more frequent diagnostic evaluation and prompt antiviral therapy.
Conventional Hospital Room Is as Safe as Sterile Unit for High-Dose Chemotherapy and Peripheral Blood Stem Cells (PBSC) Transplantation.
Authors: Ketterer Nicolas , Zaman Khalil , Luthi Francois , Rosselet Anne , Bally Frank , Schneider Philippe , Frossard Valerie , Leyvraz Serge
Published: October 2019

Infectious complications in transplanted patients (pts) motivate recommendations that autologous stem cell transplantation (ASCT) should be carried out in special rooms endowed with positive pressure and high-efficiency particulate air filtration (HEPA) filters. However growing indications for ASCT and cost restrictions prompt to lighten the procedure without compromising the safety. We report here 59 consecutive and non-selected ASCTs performed from 2/00 to 4/04 in conventional single rooms (without positive pressure air filtration) to analyze early toxicity, infectious complications and 100-days (d.) transplant related death (TRD). High-dose chemotherapy (HDC) and ASCT were administered for non-Hodgkin’s lymphoma (n=39), multiple myeloma (n=13) and Hodgkin’s lymphoma (n=2). Five pts with multiple myeloma received double ASCT. Median (med.) age was 54 years (range 22–71). Med. number (nb.) of previous lines of treatment was 1 (range 1–6). Thirty-nine pts were intensified in 1st remission (26 CR, 11 PR, 2 MR), 12 in 2nd remission (10 CR, 2 PR), 1 in 3rd CR and 2 in either SD or refractory relapse. Except for amphotericin-B mouthwashes and oral valacyclovir in case of HSV seropositivity, no systemic prophylactic antibiotic (abt) was used. The med. nb. of reinfused stem cells was 3.7x106 CD34+/kg (1.86–33.3). One pt received 0.94x106 CD34+ cells/kg combined with bone marrow. G-CSF was started at day 7 (2–15) after transplant. Med. time to achieve an absolute neutrophil count >0.5x109/L and >1.0x109/L was respectively (resp.) 10 d. (8–13) and 11 d. (8–13). Med. time to spontaneous platelet count >20x109/L and >50x109/L was resp. 11 d. (0–28) and 13 d. (9–81). The med. nb. of red cells and platelets transfusions was resp. 2 (0–12) and 2 (0–25). Except five pts, all developed fever (>38.3°C) and 56 out of the 59 pts received abt for a med. duration of 12 d. (1–46). Twenty-six pts (44%) had fever without any documented infection. Grade3/4 infections were diagnosed in 27 pts (46%). One patient presented candidemia. No case of aspergillosis was observed. Two pts (3%) needed transient transfer to the intensive care unit because of septicemia with reversible cardio-respiratory failure. The med duration of hospitalization was 20 d. (2–75), and was <28 d. for 54/59 pts (92%). With a med. follow up of 22.6 months (3–52), 44 patients are still alive: 33 (61%) in CR and 11 (20%) with relapse. 11 patients died: 8 from progressive disease, one following allotransplant 12 months after ASCT and one at d. 104 due to thrombotic thrombocytopenic purpura. The later was the unique TRD. The profile of toxicity and the low TRD rate observed in our non selected pts are comparable to whose reported usually in the literature for pts treated in sterile units with HEPA, confirming that HDC and ASCT can be safely realized in conventional single rooms.
Survival after Autologous Transplantation to Treat Diffuse Large B-Cell Lymphoma with a History of CNS Involvement, Bone Marrow Involvement, or Histologic Transformation.
Authors: Hayes-Lattin Brandon , Mehta Pritesh , Dunn Adam , Subbiah Nan , Leis Jose F. , Simic Aleksandra , Allen Bryon , Balliesen Suzanne , Curtin Peter T. , Epner Elliot , Fleming William H. , Jacoby Carol , Mauro Michael J. , Nichols Craig R. , Schubach Susan , Maziarz Richard T.
Published: October 2019

Background: Patients with diffuse large B-cell lymphoma (DLBCL) who present with extra-nodal involvement or histologic transformation from low-grade disease have lower rates of survival after conventional-dose therapy. The impact of these features on outcomes after high-dose therapy with autologous transplantation was investigated.

Methods: We retrospectively reviewed the outcomes of 64 consecutive patients receiving autologous transplantation from 1995 to 2003 for the treatment of DLBCL. Variables considered were age, gender, histologic transformation, history of bone marrow involvement, history of central nervous system (CNS) involvement, time from diagnosis to transplant, number of pre-transplant regimens, chemosensitivity prior to transplant, conditioning regimen, year of transplant, and the use of peripheral blood or bone marrow stem cells. Survivals were estimated by the Kaplan-Meier method. The Cox proportional hazards regression model was used to test the significance of factors on survival. Univariate association between variables and survival were tested by the log-rank test. Correlation between variables was tested with Spearman’s rank coefficient.

Results: The median age at transplant among 64 patients was 53.5 years (17–74). The median overall survival was 3.3 years. Among these patients, 12 had a history of CNS involvement, 10 had a history of bone marrow involvement, and 12 had transformed from low-grade lymphoma. A Cox regression model suggested age <50 (log-rank, p=0.093), <2 regimens prior to transplant (log-rank, p=0.052), and the lack of BEAM as conditioning regimen (log-rank, p=0.019) as multivariate factors for survival. However, the use of BEAM was highly associated with older age (R²= 0.42, p=0.001) and more prior chemotherapy regimens (R²=0.24, p=0.054). A history of extranodal involvement, including prior CNS involvement (p=0.842) or bone marrow involvement (p=0.518), was not associated with lower survival by univariate analysis. No significant association was found between survival and a history of transformation (p=0.484).

Conclusions: A history of CNS involvement, bone marrow involvement, or histologic transformation is not associated with lower rates of survival among patients undergoing autologous transplantation for diffuse large B-cell lymphoma at our institution. Patients who present with such histories remain candidates for autologous transplantation.