bluuebunny/crossref_metadata_2025_split huggingface dataset:
Competitive Repopulation Assay for Human Stem Cell Engraftment in NOD-scid IL-2rγnul Mice.
Authors: Giassi Lisa J. , Laning Joseph , Biber Kristen , Cuthbert Amy , Phillip Lowry A. , Shultz Leonard D. , Greiner Dale L. 
Published: April 2020

Abstract
Umbilical cord blood (UCB) is a source of stem cells for hematopoietic transplantation (Laughlin, et al. NEJM2004; 351:2265) but application in adults is limited by marginal stem cell numbers. Combining cord blood units has shown promise in achieving engraftment (Barker, et al. Blood2002; 100:41a [abs #142]) without crossed immunologic rejection. Short-term hematopoiesis showed contributions from both cords, but long-term engraftment derived predominantly from just one cord. To study this process we have developed an in vivo pre-clinical competitive repopulation assay using NOD-scid IL-2rγnull mice that support high levels of human UCB engraftment (Shultz, et al, J. Immunol.2005, 174:6477). NOD-scid IL-2rγnull mice were transplanted with T-cell depleted UCB containing 3x104 HLA-A2+ CD34+ cells, 3x104 HLA-A2− CD34+ cells, or a 1:1 mixture (6x104 CD34+ cells). Human hematopoietic chimerism, measured by human CD45 and HLA-A2 expression, was quantified in bone marrow 6 and 12 weeks after transplantation. Variable mixed engraftment within individual as well as between 6 separate experiments was observed. At both 6 and 12 weeks, mixed contribution from both cords was achieved although rarely in a 1:1 ratio despite the infusion of equivalent CD34+ cell numbers. HLA-A2 expression did not provide a selective advantage. The complete dominance of one cord observed in the clinical report and in animal studies (Yahata, et al. Molecular Therapy2004; 10: 882) suggests an important role for the immune activity of UCB T-cells in this competitive process, a hypothesis we are currently testing. This model has a rapid turnaround and can test multiple permutations, providing a preclinical vehicle for characterizing optimal combinations of cord cells and for testing conditions that allow one cord to ultimately prevail.
Legend: Engraftment of human CD45+ cells at 6 weeks. All bars represent single mice except for the HLA-A2+ group, which is the average (±SEM) of 3 mice. Legend: Engraftment of human CD45+ cells at 6 weeks. All bars represent single mice except for the HLA-A2+ group, which is the average (±SEM) of 3 mice.
Regulatory and Activated T Cells in Stem Cell Autografts Collected by Mobilization with High-Dose Cyclophosphamide and Granulocyte Colony Stimulating Factor.
Authors: Condomines Maud , Quittet Philippe , Lu Zhao-Yang , Nadal Laure , Latry Pascal , Lopez Ernesto , Baudard Marion , Rossi Jean-François , Tarte Karin , Klein Bernard 
Published: April 2020

Abstract
High-dose chemotherapy (HDC) supported by autograft of hematopoietic progenitors (HP) is a standard therapy for patients with multiple myeloma (MM). High-dose cyclophosphamide (CTX) and G-CSF are widely used to collect HP. As the number of lymphocytes in the autograft is a powerful prognostic factor in patients with MM, our purpose was to study how CTX-G-CSF treatment affects the phenotype and function of T cells, in particular regulatory T cells (Treg), in 15 patients with MM. CTX induced severe T cell immunosuppression with a slow and partial T-cell recovery (a threefold decrease) at the time of HP collection. CTX-G-CSF treatment did not affect the percentages of central memory (CD45RA−, CCR7+), effector memory (CD45RA−, CCR7−), and late effector (CD45RA+, CCR7−) CD4 or CD8 T cells but a decrease of naïve CD4 cells (CD45RA+, CCR7+) was found. The percentages of CD25+ cells increased two- to threefold in CD4 or CD8 T cells, respectively. Post-CTX treatment CD4CD25+ cells included both activated CD4CD25low cells and CD4CD25high T cells. The latter were Treg because they expressed high level of FOXP3 and membrane CTLA-4 mRNA and protein and displayed functional suppressor function. In CTX-G-CSF leukaphereses from 15 patients with MM, the mean Treg number was one fifth that of CD34 and the CD3, CD4 and CD8 numbers respectively 3 fold, 2 fold and equal that of CD34. Post-CTX-G-CSF treatment CD3 cells did not cell cycle in vivo and died in short-term culture in vitro. Adding IL-2 or IL-15 induced their survival and cell cycle, and stimulation with anti-CD3 MoAb led to efficient growth in vitro. These results suggest that following CTX-G-CSF treatment, CD3 cells are preactivated in vivo and do not cell cycle, likely due to a lack of T cell growth factors in vivo. The current data indicate that CTX-G-CSF treatment profoundly affects T cell function without eliminating Treg. The persistence of Treg could be explained by an opposite effect of CTX known to kill Treg and of G-CSF amplifying Treg. Given the major impact of lymphocyte count on patients’ survival post HDC and HP and T cell graft, the present data invite to define novel therapeutic strategies to improve T cell recovery in vivo while limiting Treg expansion.
Markers of Iron Overload Are Poor Prognostic Factors in Stem Cell Transplantation.
Authors: Connor Rebecca , Molnar Istvan , Lovato James , Grover Manisha , Hurd David , Pomper Gregory , Torti Suzy V. , Torti Frank M. 
Published: April 2020

Abstract
Transfusional iron overload is common in survivors of acute leukemia and hematopoietic stem cell transplantation and might cause long-term liver dysfunction. Routine evaluation for iron overload in such patients is recommended because excess iron can be readily removed from the body via phlebotomy or chelation. Iron overload might be associated with worse survival after stem cell transplantation in these diseases. We were interested in determining whether levels of the iron binding protein ferritin or the serum transferrin receptor (TfR) were predictive for survival. In a prospective study, we examined the correlation between iron parameters at the time of transplantation and overall survival. Serum ferritin, transferrin saturation, and TfR were measured before preparative regimen on patients who underwent hematopoietic stem cell transplantation between 1999 and 2004 for the diagnosis of aplastic anemia, MDS or acute leukemia (n=79). We used the number of transfusions before transplantation as a measure of iron load. Among these iron markers, serum ferritin correlated the most with the number of transfusions, regardless of remission status. High ferritin (&gt;1,500 ng/ml), low TfR (≤4 μg/ml) and low TfR/log ferritin ratio (≤2) were associated with shorter survival (p=0.005, 0.04, and 0.001 respectively)(Figure 1). Among acute leukemia patients in remission, there was no difference in overall survival between patients with high or low iron markers. Markers of iron excess (serum ferritin &gt;1,500 ng/ml, TfR/log ferritin ratio ≤2) at the time of stem cell transplantation is associated with shorter survival in MDS, aplastic anemia and acute leukemia with active disease. These results demonstrate that knowledge of patient ferritin and TfR levels can aid in risk stratification. The results also suggest that patients with high levels of ferritin may benefit from iron chelation before treatment.
Figure 1: Overall Survival based on iron parameters (A) Serum Ferritin (B) Serum Transferrin Receptor (C) TfR (mcg/ml) divided by log ferritin (ng/ml) Figure 1:. Overall Survival based on iron parameters (A) Serum Ferritin (B) Serum Transferrin Receptor (C) TfR (mcg/ml) divided by log ferritin (ng/ml)
Downregulation of Interleukin-13 by PUVA Treatment in a Human Model of Graft-Versus-Host Disease May Highlight the Importance of This Cytokine in the Mechanism of Extracorporeal Phototherapy Treatment.
Authors: Marshall Scott R. , Wang Xiao N. , McArthy Chris M. , Jackson Graham , Dickinson Anne M. 
Published: April 2020

Abstract
Graft-versus-Host disease (GvHD) remains the most serious complication following haemopoietic stem cell transplantation, with an incidence of 40–60% and can be fatal in up to 50% of cases. Extracorporeal phototherapy (ECP) is a novel treatment of both acute and chronic GvHD involving psoralen and UVA (PUVA) treatment of peripheral blood cells with high reported success even in those resistant to conventional immunosuppressive treatments. ECP appears to induce selective immune suppression without increased rates of infection or disease relapse. It is well tolerated by patients with minimal side effects and therefore is emerging as a highly desirable therapy. However, its mechanism of action remains poorly understood. An in vitro human skin explant model for GvHD has been used to study the role of cytokine changes induced by ECP. The model involves sensitising donor lymphocytes with recipient lymphocytes in vitro in a primary mixed lymphocyte reaction and then evaluating the secondary response on recipient skin biopsies by grading the graft-versus-host reactivity (grades I-IV) histopathologically using the Lerner grading system for GvHD. The assay has been proved to be superior to other assays used to predict clinical GvHD and has been used to investigate the pathophysiology of GvHD. Skin explant supernatants were collected and stored at −80 C and measured in batches using a multiplex Th1/Th2 cytokine assay. In 20 experiments where PUVA treatment of responder cells caused down regulation of GvHR in the skin explant model, statistically significant downregulation of IL-13, IL-5 and IFNgamma (p=&lt;0.05) was demonstrated. No changes of IL-6, IL-1b, IL-10, IL-2, IL-4 or TNFalpha levels were demonstrated. Further evidence for the importance of IL-13 was highlighted in 3 skin explant assays where no downregulation of GvHR was seen with PUVA treatment and IL-13 levels remained high. Jordon et Al (Blood Jan2004) have documented the association between IL-13 production and the incidence of acute GvHD. Our data adds further evidence to challenge the notion that acute GvHD is purely a Th1-type cytokine response and that there may be a significant link between the Th2-type cytokine IL-13 with acute GvHD. Furthermore, downregulation of IL-13 and by PUVA may be important in the control of GvHD. More skin explant assays will be performed to consolidate these provisional findings.
Comparison of Pathology Transform of Liver, Spleen, Intestinal Tract and Skin with Acute Graft-Versus-Host Disease in Mice.
Authors: Wang Chunyan , Tan Huo , Huang Zhenqian , Zhang Huanzhu , Guo Kunyuan 
Published: April 2020

Abstract
Objective: To study the pathology transform of liver, spleen, intestinal tract and skin with acute graft-versus-host disease after myeloablative MHC mismatched BMT in mice.
Methods: The murine model of MHC mismatched BMT was established by using (BALB/c H-2d mouse as the recipient, and C57BL/6H-2b mouse as the donor. 30 recipient mouse were divided into 3 groups (n=10) after irradiation (TBI 60Co 9.0Gy). simple-irradiation group, MHC mismatched GVHD group: the BALB/c H-2d mice were conditioned with 9.0Gy, after a interval of four hours followed by infusion of C57BL/6H-2b mice bone marrow cells (1×107/one) and spleen cells (3×107/one). syngeneic BMT group: C57BL/6H-2b mice were conditioned with 9.0Gy, after a interval of four hours followed by infusion of C57BL/6H-2b mice bone marrow cells (1×107/one). The effect was assessed by hemotopoietic reconstruction, survival time, body weight, histopathology in the recipients.
Rasults:
Life span: survival time was (5.23±0.78) days for the simple-irradiation group, survival time was (14.67±2.34)days for MHC mismatched GVHD group, beyond 30 days for syngeneic BMT group. Pathology results: GVHD pathology manifestation were found in MHC mismatched GVHD group. The severity of GVHD pathology transform in mice were based on the occurrence time of GVHD. The GVHD pathology transform of liver, spleen, intestinal tract and skin in the same mouse were not parallel which GVHD pathology transform of spleen was the severest, the nest were intestinal tract and skin, GVHD pathology transform of liver was the trivial, even more normal. 3 of 10 in GVHD group had skin III-IV GVHD, but normal liver.
Conclusion: The result have shown that the onset time of liver GVHD in four organs was the last in myeloablative MHC mismatched BMT in mice.
Rejection of Primary Lymphoma Cells Derived from c-myc-Transgenic Mice after Haploidentical Transplantation: Treatment of the Tumor May Confer to Resistance Against GvL-Effect.
Authors: Sukumar Madhusudhanan , Wilke Andrea , Mautner Josef , Kolb Hans-Jochem , Bornkamm Georg , Gerbitz Armin 
Published: April 2020

Abstract
The long term prognosis of the High Grade B-cell-lymphoma in adults is still very poor, especially in the case of resistance to chemotherapy. To date allogeneic stem cell transplantation as a last treatment option results in poor outcomes due to high transplant related mortality or disease progression. This dilemma raises the question of whether transplant related mortality could be reduced by earlier transplantation, and whether such tumors can be recognized by an allogeneic graft providing a graft versus lymphoma effect. The immunology behind allogeneic rejection of high grade lymphomas is poorly understood due to a dearth of good animal models. Using a transgenic mouse model in which the human c-myc oncogene is driven by the human immunoglobuline lambda locus mimicking a t(8;22) translocation of Burkitt’s lymphoma, we established primary lymphoma cell lines. C57BL/6c-myc tg mice were crossbred with DBA mice and lymph nodes from tumor bearing F1-mice were explanted and seeded on irradiated human MRC5 fibroblast feeder cells. Approximately 70% of all explanted lymphomas established long term cultures which could be maintained free of feeder cells after 4 to 6 weeks. FACS analysis of lymphoma cells displayed strong CD19 expression, reduced expression of MHC Class I and II and, in several cases, low expression of costimulatory molecules such as CD80, CD86 and CD54. The phenotype was maintained over time with a follow up of up to 4 months. As few as 100 lymphoma cells gave rise to orthotopic tumors or systemic disease within 40 days after s.c. or i.v. injection, respectively. On the contrary, when 50 Mio. F1-lymphoma cells were injected into haploidentical C57BL6 parental animals, no tumor growth could be observed after at least 100 days of follow up. Despite fast growth and low expression of MHC Class I and costimulatory molecules, lymphoma cells were easily rejected in a haploidentical setting. Ex vivo chemotherapy treatment of lymphoma cells resulted in resistance after 2 courses of 100nM Adriamycine. Increased doses up to 500nM Adriamycine further accelerated resistance and growth speed of the cells. Flow cytometric analysis indicated a reduced fraction of apoptotic cells in culture suggesting selection for cells bearing alteration in the apoptotic pathways. In further experiments altered cells will be tested in haploidentical settings to confirm clinical experience in our model.
Effect of Intravenous Busulfan Versus Total Body Irradiation on Donor Chimerism and Acute Graft-Versus-Host Disease (GVHD) after Allogeneic Stem Cell Transplantation (AlloSCT) in Mice.
Authors: Wang Jie , Zhou Xiaotao , Symons Heather , Levy Moshe Y. , Fuchs Ephraim J. 
Published: April 2020

Abstract
The goals of administering transplantation conditioning to leukemia patients undergoing alloSCT are to kill tumor cells, prevent rejection of the donor graft, create “space” for donor hematopoiesis, and provide an overall milieu that favors a donor T cell-mediated “graft-versus-leukemia” effect. However, transplantation conditioning also increases the incidence and severity of acute GVHD by damaging non-hematopoietic tissues and by inducing lymphopenia, which augments the antigen-driven expansion of alloreactive T cells. The purpose of this study was to compare the relative safety and efficacy of transplantation conditioning with intravenous busulfan (Busulfex; ESP Pharma, Edison, NJ) versus total body irradiation (TBI) by administering titrated doses of either agent to mice and comparing their effects on: 1) donor chimerism after syngeneic and semi-allogeneic (parent into F1) SCT; 2) graft rejection after MHC-matched allogeneic SCT; and 3) acute GVHD after semi-allogeneic SCT. Both agents induced titratable donor chimerism after syngeneic BMT, but TBI was more immunosuppressive, since allogeneic marrow engrafted after 700 cGy TBI but not after an equivalently myelotoxic dose of Busulfex (60 mg/kg). When given in equivalently myelotoxic doses, TBI always induced more GVHD than did IV busulfan, as measured by weight loss and the augmentation of donor T cell and myeloid chimerism. As a result, stable mixed hematopoietic chimerism was generated more readily in mice conditioned with nonmyeloablative doses of busulfan than in mice conditioned with equitoxic doses of TBI. These results demonstrate that IV busulfan promotes GVHD to a lesser extent than does an equivalently myelotoxic dose of TBI. IV busulfan may be the preferred agent if stable mixed hematopoietic chimerism is the desired outcome. The relative roles of lymphopenia versus tissue damage in the differential outcomes of TBI versus Busulfex conditioning are being investigated.
Poor Graft Function Following Allogeneic Stem Cell Transplantation: High Rate of Complete Recovery Using Boost CD34+ Selected Peripheral Blood Stem Cells without Further Conditioning.
Authors: Larocca Alessandra , Bruno Barbara , Di Grazia Carmen , Gualandi Francesca , Occhini Domenico , Raiola Anna Maria , Dominietto Alida , Bregante Stefania , Lamparelli Teresa , Van Lint Maria Teresa , Pogliani Enrico Maria , Bacigalupo Andrea 
Published: April 2020

Abstract
Background: we have shown in a previous study that approximately 30% of patients may experience poor graft function (PGF) following an allogeneic hemopoietic stem cell transplant (HSCT); it is uncertain whether a boost of donor stem cells is beneficial in terms of trilineage recovery (TLR) and transplant related mortality (TRM).
Aim of the study: To compare infusion of boost donor stem cells in patients with primary PGF (who failed to achieve sustained graft function) and secondary PGF (patients who developed PGF after complete hematologic recovery).
Methods: we have attempted to rescue PGF using stem cells from the same donor in 54 patients affected by hematologic malignancies: 20 patients received no further infusion (group A), 14 received a boost of unmanipulated marrow or blood cells from the original donor, without further conditioning (group B), and 20 received donor cells after CD34 selection without conditioning (group C).
Results: Respectively 40%, 36% and 75% of patients in groups A, B and C obtained a trilineage recovery (p=0.02). TLR was more frequent in secondary PGF (69%) vs primary PGF (36%) (p=0.09), and more frequent in recipients of PB boost donor cells (p=0.02). There was no effect of donor type on hematologic recovery. TRM was 55%, 64%, 25% respectively in groups A, B and C (p=0.06) and was highly correlated with TLR (p=0.00001). PGF was the primary cause of death in 30%, 21%, 10% in the 3 groups, graft vs host disease in 5%, 36%, 10%.
Conclusion: The study indicates that a boost of CD34 selected PB donor cells is associated with a high chance of trilineage recovery and low risk of acute GvHD. A boost of unmanipulated donor cells does not seem to offer an advantage over no infusion. TRM is lower when using PB CD34 selected cells.
Figure 1: trilineage response in the 3 groups Figure 1:. trilineage response in the 3 groups Figure 2: TRM according to hematologic recovery Figure 2:. TRM according to hematologic recovery
The Different Dynamics of Chimerism Evolution Determines the Success of Myeloablative, Reduced Intensity Conditioning and T-Cell Depleted Allogeneic Stem Cell Transplantation.
Authors: Buño Ismael , Balsalobre Pascual , Simón Ainhoa , Iglesias Gustavo , Manzano Cintia , Carrión Rafael , Serrano David , Gómez-Pineda Alfonso , Díez-Martín José L. 
Published: April 2020

Abstract
Background: The dynamics of chimerism evolution determines the success of allogeneic stem cell transplantation (SCT). Several factors such as the intensity of the conditioning regimen, the T-cell content of the graft or the GVHD prophylaxis, influence the degree of chimerism after SCT. Objective: To evaluate the dynamics of chimerism after different SCT settings (ablative, reduced intensity conditioning -RIC- and T-cell depleted -TCD-) and its influence in the success of the procedure.
Patients and Methods: The study includes 68 SCT: 32 ablative (including 7 from HLA matched unrelated donors -UD-), 19 RIC and 17 TCD (including 8 from haploidentical donors and 2 from UD with 1 HLA disparity). Chimerism analysis was performed by FISH for the sex chromosomes or STR-PCR (sensitivity 1 Samples obtained on days +30, +100, +180, +365 and once a year thereafter, included bone marrow (BM) and peripheral blood (PB). Moreover, chimerism was analyzed in PB and leukocyte lineages (T lymphocytes CD3+, B lymphocytes CD19+ and myeloid cells CD15+ isolated (purity &gt;95%) by immunomagnetic means, AutoMACS, Miltenyi Biotec), every 2 weeks, starting on day +15 (except in ablative), and until complete chimerism (CC) was achieved. Results of chimerism follow-up were censored once the diagnosis of relapse or rejection was established.
Results: The incidence (% patients) of mixed chimerism (MC) on day +30 (ablative: BM 27%, PB 15%; RIC: BM 40%, PB 41%, CD3 40%; TCD: BM 31%, PB 33%, CD3 54%), as well as its dynamics (MC on day +100: ablative BM 8%, PB 4%; RIC BM 8%, PB 15%, CD3 22%; TCD BM 20%, PB 40%, CD3 37%) were different in the three SCT settings. All except 2 CD15 samples analyzed showed CC. Moreover, the percentage of recipient cells (%R) was significantly higher after RIC and TCD than after ablative SCT, as well as in T lymphocytes than in BM or PB (7/8 cases with simultaneous studies showed MC in T lymphocytes and CC in PB). All RIC SCT evolved to CC by day +180 while TCD SCT showed persistent MC (2 patients with stable MC after one year). The incidence of rejection was greater after RIC (2/19) and TCD (4/17) than after ablative SCT (2/32). All these patients showed MC, mainly in T lymphocytes, which allowed early diagnosis and successful treatment with immunosuppression withdrawal and donor leukocyte infusion. Patients with CC in PB/T lymphocytes on day +30 had a higher incidence of GVHD&gt;I than those with MC. In the present series, however, a relationship between chimerism and relapse, disease free survival or overall survival, was not observed.
Conclusions: SCT with greater incidence of MC (RIC and TCD) favor immune tolerance between donor and recipient which reduces the risk/severity of GVHD at the expense of a higher incidence of graft rejection.