bluuebunny/crossref_metadata_2025_split huggingface dataset:
Petreatment with Rituximab Reduces the Incidence of Chronic Graft-Versus-Host Disease after Allogeneic Stem Cell Transplantation in Patients with B Cell Lymphoma.
Authors: Bornhaeuser Martin , Schwerdtfeger Rainer , Kroschinsky Frank , Hanel Mathias , Treptow Uli , Theuser Catrin , Ehninger Gerhard 
Published: April 2020

Abstract
The use of anti-B cell antibodies before allogeneic hematopoietic stem cell transplantation (HSCT) induces a depletion of recipient B cells. This leads to a reduction of the number of potential antigen presenting cells and thereby might hamper the induction phase of graft-versus-host disease (GvHD). The humanized anti-CD20 antibody Rituximab has been used with increasing frequency to treat patients with B cell Non-Hodgkin’s Lymphoma. Only recently, several investigators have reported on the efficacy of Rituximab for the treatment of chronic GvHD. We therefore hypothesized, that patients who received Rituximab before being scheduled for allogeneic HSCT may have a decreased risk of GvHD.
A retrospective analysis in 72 patients after allogeneic hematopoietic stem cell transplantation for indolent (n=18), aggressive (n=45) B cell lymphoma or mantle cell lymphoma (n=9) was performed at two centres. Whereas 28 patients had received chemotherapy only before allogeneic HSCT, 44 patients had received 4–8 (median 6) cycles of 375 mg/m2 Rituximab before the start of conditioning therapy. Patient characteristics’ were comparable except for a higher proportion of unrelated donors (73% vs. 54%) and a higher median age (48 vs. 42 years) in the cohort with Rituximab pre-treatment. In addition the proportion of patients having received a previous autologous transplantation was higher (63% vs. 46%) and therefore more recipients received dose reduced conditioning before allogeneic tranplantation (70% vs. 50%) in the Rituximab group.
With a median follow-up of 24 months for all patients, no significant differences in overall survival, progression-free survival and treatment-related mortality could be detected between both groups. Although the proportion of transplants from unrelated donors was lower in the No-Rituximab group, the rate of acute GvHD grade I–IV was higher (88% vs. 63%, p=0.03). The incidence of grade II–IV and grade III–IV GvHD was not significantly different between both cohorts. Chronic GvHD occurred more often in patients who had not received Rituximab before allogeneic transplantation (52% vs. 23%, p=0.02).
Although the retrospective nature of our study does not allow to draw definitive conclusions, the rate of chronic GvHD seems to be lower when anti-CD20 treatment is performed before allogeneic HSCT. Prospective studies incorporating Rituximab for the prophylaxis and therapy of GvHD are warranted.
Improved Immune Reconstitution with Low Incidence of Severe Graft-Versus-Host Disease after Transplantation of CD34+ or CD133+ Enriched Stem Cells with Add-Back of Ten Million T-Cells Per Kg.
Authors: Greil Johann , Lang Peter , Bader Peter , Eyrich Matthias , Schlegel Paul G. , Schumm Michael , Feuchtinger Tobias , Niethammer Dietrich 
Published: April 2020

Abstract
Unrelated donors are commonly used for hematopoietic stem cell transplants, but graft-versus-host disease (GVHD) is a major problem. We investigated whether transplantation of CD34+ or CD133+ enriched stem cells with add-back of ten million T-cells per kg from unrelated donors would prevent acute GVHD in pediatric patients in combination with pharmacologic immunosuppression. Eighteen patients (1 CML in second chronic phase, 2 MDS, 4 ALL in CR1, 4 ALL in CR2, 1 JMML, 1 AML in CR1, 3 AML in CR2, 1 Wiskott-Aldrich syndrome and 1 NHL in CR2) were transplanted with G-CSF mobilized peripheral blood stem cells (PBSC) from HLA-matched unrelated donors (n = 18). Median of age was 8.9 years (0.5 to 18 years). Conditioning regimens were performed according to national therapy protocol guidelines. On the day of transplant patients received a median of 13.5 (4.5 to 30.0 x 106) CD34+ or CD133+ enriched stem cells and an aliquot of unmanipulated PBSC containing 10 x 106 T-cells per kg. GVHD prophylaxis consisted of cyclosporine A (CSA) and short course methotrexate (MTX) on day +1, +3 and +6. Engraftment was rapid with a median of 19.6 days in sixteen patients. Two patients failed to engraft at first. However, full donor chimerism and stable engraftment was achieved in both patients after cessation of CSA treatment and an additional stem cell boost without any reconditioning. One patient with ALL developed acute GVHD grade III (skin and gut) after cessation of CSA treatment, but responded well to treatment with CSA and steroids. None of the other 17 patients developed acute GVHD &gt; grade I. Thirteen are alive and well with a median follow-up of 578 days (101 to 1095 days). Three patients died of severe infectious complications and two due to relapse (JMML, AML). Compared to a historical control group of patients transplanted with highly purified CD34+ selected cells, the group with add-back of 10 x 106 T-cells per kg showed significantly higher T-cell counts (p=0.008, Wilcoxon Rank sum test) on day 90 after transplantation with a median number of six T-cells/μl in the control group and a median number of 294 T-cells/μl in the study group. We conclude that add-back of 10 x 106 T-cells per kg in combination with CSA and short course MTX improves T-cell recovery and appears to be a safe T-cell dosage regarding acute GVHD in the setting of allogeneic peripheral blood stem cell transplantation from unrelated donors.
Increased Expression of Activating Receptors on NK Cells in the Post-Transplant Period.
Authors: Boyiadzis Michael , Carson Jesse M. , Memon Sarfraz A. , Dean Robert , Castro Kate , Odom Jeanne , Bishop Michael R. , Gress Ronald E. , Hakim Frances T. 
Published: April 2020

Abstract
Following allogeneic and autologous hematopoietic stem cell transplantation (HSCT) natural killer (NK) cells are among the first lymphocyte populations to recover, returning to normal levels within one month after HSCT. We previously reported that levels of IL-15, a cytokine critical to NK cell homeostasis, are elevated during the peri-transplant period (ASH, 2003). Modulation of the relative frequency and intensity of expression of the activating NK cell receptors by IL-15 may contribute to NK-mediated cytotoxicity post transplant. We compared the expression of activating NK receptors in the donor apheresis product and in recipient peripheral blood at one and three months following reduced intensity allogeneic HSCT from HLA-matched siblings. NK cells, divided into CD56bright CD16− and CD56dimCD16+ subsets, were assessed for receptors that trigger cytotoxicity: the natural cytotoxicity receptors (NCR) NKp30 and NKp46, and the C-type lectin receptors NKG2D and NKG2C-CD94. During the first post transplant month the NK cell numbers recovered to normal levels, but the CD56brightCD16− subset increased disproportionately (p = .005). By three months the proportions of the NK cell subsets returned to pre-transplant levels. In both NK subsets, the percentage of cells expressing NCR NKp46 or NKp30 doubled by one month and was sustained at three months. The percentage of cells expressing the C-type lectin receptor CD94 similarly increased by 50% in both NK subsets. The heterodimeric activating partner of CD94, NKG2C, was also increased, but the increase in frequency of cells expressing this receptor was less than those pairing CD94 with its inhibitory NKG2A partner. The homodimeric activating receptor NKG2D increased in the NK CD56brightCD16− subset, but did not significantly change in the CD56dim CD16+ NK population. Consistent with responsiveness to elevated IL-15 during the peri-transplant period, expression of both the IL-2/IL-15 beta and common gamma receptors increased at one and three months. Furthermore, when NK cells from normal donors were cultured with IL-15, we observed both a population shift towards CD56brightCD16− and an up-regulation of the NK cell activating receptors, similar to the changes observed during the early post transplant period. These results suggest that IL-15-dependent changes in NK subset distribution and activating NK receptor repertoire occur during the early post-transplant period. Up-regulation of activating NK cell receptors may contribute to NK cell anti-tumor efficacy in the post transplant period.
Faster Immune Reconstitution after Allogeneic Hematopoietic Stem Cell Transplantation of Younger Children Compared with Older Children and Adults.
Authors: Fehse Natalja , Kabisch Hartmut , Erttmann Rudolf , Kroeger Nicolaus , Zabelina Tatjana , Freiberger Petra , Zander Axel R. 
Published: April 2020

Abstract
Allogeneic stem cell transplantation (SCT) is a potentially curative treatment of otherwise incurable malignant hematological disorders and severe immunodeficiency diseases. The post-SCT immunoincompetence can last for up to several years and might cause significant morbidity and mortality. Immune reconstitution after SCT has been studied extensively in adults, but only a few reports have been published on a paediatric population or have compared paediatric and adult population. This study was carried out in order to determine the influence of age on quantitative reconstitution of peripheral blood lymphoid subsets. A cohort of 69 adult and 109 paediatric patients received allogeneic SCT from related or unrelated fully HLA matched or 1 locus -mismatched donors for different malignant and non-malignant diseases. Analysis of lymphocyte subsets was performed at various time points: until day 30, between days 31–60, 61–90, 91–180, 181–365, between 1–2 years, between 2–5 years and more than 5 years post transplant (at the last time point only paediatric patients were analysed).
Comparing the paediatric with adult patients, we observed statistically significant faster recovery of CD19+, CD19+CD5+, CD4+CD45RA+ cells during the whole period of observation, and of total lymphocyte count (TLC) (&lt; 60d and 1-2y), CD3+ (&lt; 60d and 90d-2y), CD3+HLA-DR+ (31-60d), CD3+CD4+ (&lt; 60d and &gt; 90d), CD4+CD45R0+ (&lt; 60d), CD3+CD8+ (&lt; 60d and 90d - 2 y) and CD56+CD3−cells (&lt; 31d) in the paediatric group. Interestingly, the recovery of CD56+CD3− cells was faster from day 61-365 in the adult group (significant faster on day 91–180).
The paediatric patients were divided in two subgroups: 1) aged &lt; 10 years; 2) aged &gt; 10 years. The following lymphocyte subsets recovered significant faster in the younger group: TLC (31-60d, 91-365d and &gt; 2y), CD3+ (91-180d and &gt; 1y), CD3+HLA-DR+ (31-60d), CD3+CD4+ (31-60d and &gt; day 90), CD4+CD45RA+ (61-90d and &gt; day 180), CD4+CD45R0+ (91-180d and 2-5y), CD3+CD8+ (91-180d and &gt; 2y), CD19+ (31-60d, 91-180d and 2-5y), CD19+CD5+ (31-60d, 181-365d and 2-5y) and CD56+CD3− cells (61-180d).
The thymopoietic pathway of T helper cells is important throughout childhood; however, the thymus has a diminished capacity for the regeneration of these cells in adulthood. This can be the explanation of the faster recovery of T helper cells in the younger patients in this study. Besides, we found, that paediatric patients have a faster reconstitution of B cells and cytotoxic T cells. This might be because of the higher number of stem cells in the paediatric transplantation.
Therapeutic Plasma Levels of OKT3 Muromonab Are Effective as Second-Line and Subsequent Treatment in Recipients of Stem Cell Allografts with Steroid-Resistant Acute Graft-Versus-Host Disease.
Authors: Knop Stefan , Hebart Holger , Gscheidle Heidi , Holler Ernst , Kolb Hans J. , Niederwieser Dietger W. , Einsele Hermann 
Published: April 2020

Abstract
Acute graft-versus-host disease (aGvHD) remains a major complication in recipients of hematopoietic cell allografts and is associated with poor outcome. Salvage treatment is difficult after failure of steroids. OKT3 muromonab is a monoclonal murine IgG2a antibody directed against the CD3 antigen on T-lymphocytes that has proven effective in transplant settings both of solid organs and bone marrow. Here we report on the results of OKT3 treatment in 43 patients who had received stem-cell or bone marrow allografts for hematologic malignancies and subsequently been diagnosed with steroid-resistant aGvHD. Overall, 48 treatment cycles of OKT3 (5 mg intravenous injection once daily) were administered for acute GvHD of grades II (n = 12), III (n = 27) and IV (n = 9). Mean duration of OKT3 therapy was 9 (range, 1 –16) days. Twenty cycles of OKT3 were administered as second line and 28 as third-plus line (third line, n = 14; fourth line, n = 14) treatment regimens. Side effects were mild to moderate, comprising fever, dyspnea, tachycardia, and vomiting. Thirty of 42 evaluable patients responded to therapy for an overall response rate of 69 %. Complete remissions were seen in 6 (11.9 %) and partial remissions in 24 patients (57.1 %). Response to treatment was 88.8 % for skin GvHD; 53.3 % for gut aGvHD; and 8.7 % for liver involvement. Recurrent or progressive acute GvHD was observed in 23 patients (54.8 %) after a median of 9 (range, 1 – 58) days. Pharmacokinetic studies of monoclonal antibody OKT3 were performed in 17 patients, 16 of whom achieved at least a partial remission. Treatment duration was &gt; 5 days in 16 patients. Adequate plasma levels (&gt; 1000 ng/ml) were observed in 13 patients after a median of 6 (1 – 11) days on treatment. OKT3 concentration became undetectable at a median of merely 4 (3 – 11) days after discontinuation of therapy. Survival at 100 days from transplantation was 58.1 % and at one year, 25.5 % with the most important causes of death being infectious complications (44.7%) and GvHD (33.2%). Patients treated with OKT3 earlier in the course of aGVHD showed a trend to better survival: 35% of patients with 2nd-line therapy were alive at one year from transplantation, compared to 17.4% with 3rd- plus line treatment (p=0.21). There was also a trend to a better one-year survival for patients with grade II (33% of patients alive) when compared to grade III/IV aGvHD (16% of patients alive; p=0.24). Thus, a significant proportion of patients with corticosteroid-resistant acute GvHD showed a response to salvage treatment with OKT3. More effective antimicrobial prophylaxis and treatment are mandatory to overcome considerable treatment-related mortality in those high-risk patients.
Treatment of Steroid-Refractory Acute Graft Versus Host Disease with Low Dose Combination Monoclonal Antibody Therapy: A Pilot Study.
Authors: Moore John J. , Johnston Amanda , David Ma D.F. , Sam Milliken , Keith Fay , Tony Dodds 
Published: April 2020

Abstract
Severe (Grade III–IV) acute graft versus host disease (aGVHD) is a serious complication of allogeneic haemopoietic stem cell transplantation. Patients not responding to high dose corticosteroids have a very poor prognosis (60–90% mortality). Previous second line agents, such as anti-thymocyte globulin, have had limited success in controlling steroid-refractory aGVHD. Recently monoclonal antibodies that target cytokines (TNF and IL-2) have been trialled, with several case series reporting responses to high dose infliximab (10mg/kg, x1-9 doses) and daclizumab (1mg/kg, x5-8 doses), alone or in combination. Here we report the results of a pilot study using reduced-dose infliximab and daclizumab, along with strict antimicrobial prophylaxis, for refractory aGVHD. The aim was to maintain the efficacy of monoclonal antibody therapy whilst reducing the infectious complications seen in previous series.
Since May 2003 we have treated 10 allogeneic transplant patients with one or both of the monoclonal antibodies. All had failed a minimum of 3 days methylprednisolone 2mg/kg/day, started a median of 35 days post-transplant. The first four patients received infliximab 10mg/kg, x2-3 doses. Two of four had a complete response with one patient alive 15 months after antibody treatment. The remaining three died of opportunistic infections. Following this study, a second cohort of patients (n=6) were given low dose, combination therapy (infliximab 5mg/kg, x2 doses and daclizumab 1mg/kg, x4 doses). 5/6 patients had complete resolution of aGVHD and 5/6 are alive at a median of 130 days post-treatment. One patient died of MRSA pneumonia despite resolution of GVHD. The remaining 5 patients have had minimal and manageable infectious complications whilst on strict prophylactic regimens. In conclusion, low dose combination monoclonal antibody therapy appears to be a promising and safe treatment for refractory aGVHD.
ADAMTS13 Binds VWF Via its C-Terminal CUB2 Domain.
Authors: Zhang Weirui , Motto David , Ginsburg David 
Published: April 2020

Abstract
Thrombotic thrombocytopenic purpura (TTP) is a life threatening illness due to a deficiency of the VWF-cleaving protease, ADAMTS13. The ADAMTS13 protein is composed of a propeptide, followed by a typical zinc metalloprotease domain. The C-terminal 2/3 of the molecule contains disintegrin-like, cystine-rich, and spacer domains, as well as a total of eight TSP1 motifs and two CUB domains. The function of this C-terminal portion of the molecule and its composite motifs is unknown, though TSP1 and CUB domains of other proteins have been shown to mediate protein-protein interactions. To further explore the interaction between ADAMTS13 and VWF, we cloned full length human cDNAs for both ADAMTS13 and VWF into the mammalian expression vector pcDNA3.1. These constructs were transiently transfected into 293T cells and COS cells respectively, and conditioned media collected for analysis. Using an anti-myc antibody, myc-tagged VWF co-immunoprecipitated (co-IP) with ADAMTS13, as demonstrated by western blot analysis using antisera raised against a C-terminal peptide derived from the predicted ADAMTS13 sequence. This direct interaction required partial denaturation of VWF in 1M urea, with no co-IP observed in the absence of urea. To map the segment within ADAMTS13 responsible for VWF binding, we cloned a series of overlapping ADAMTS13 fragments into the bacterial expression vector, Pet44b. Fusion proteins were purified by binding of the included His-tag to Ni-NTA beads and incubated with recombinant myc-VWF in the presence of 1M urea. Association with VWF was analyzed by co-IP with anti-myc followed by western blot analysis using an antibody to the C-terminal HSV-tag present in each fusion protein. The CUB2 (Glu1298- Thr1427) fusion protein co-IP’d with full-length VWF and also demonstrated concentration-dependent competition with full-length ADAMTS13 for VWF binding. In summary, we have demonstrated a direct protein-protein interaction between VWF and ADAMTS13. Binding requires partial denaturation of VWF and appears to be mediated primarily through contacts with the ADAMTS13 CUB2 domain. This interaction may account for the previously observed co-purification of VWF and ADAMTS13 from human plasma. Furthermore, the requirement for 1M urea suggests that this interaction may only occur physiologically under conditions of high shear. Though others have shown that the C-terminal domains of ADAMTS13, including CUB2, are not required for VWF cleavage in vitro, our data, together with several C-terminal mutations previously reported in TTP patients, suggest that interactions between VWF and the ADAMTS13 CUB2 domain may be important in vivo.
Calpain I Null Mice Display Lymphoid Hyperplasia.
Authors: Kuchay Shafi , Nunez Rafael , Bartholomew Amelia M. , Chishti Athar H. 
Published: April 2020

Abstract
Calpains are ubiquitous calcium-regulated cysteine proteases that have been implicated in cytoskeletal organization, homeostasis, cell proliferation, cell motility, and apoptosis. The conventional calpains include mu-calpain (calpain I) that requires micromolar calcium for half maximal activation and m-calpain (calpain II) that functions at millimolar calcium concentration in vitro. To evaluate the physiological function(s) of mu-calpain, we utilized genetically altered mu-calpain null mice as the model system. Spleen biopsy of mu-calpain null adult mice showed consistent expansion of white pulp (lymphoid hyperplasia) due to an increase in number and size of follicles and PALS (periarteriolar lymphatic sheets), relative to matched wild type mice. Also, cervical lymph nodes of mu-calpain null mice showed lymphoid hyperplasia and marked plasmacytosis. The majority of mu-calpain null mice displayed multi-organ lymphoid infiltration. Using immunofluorescence labeling and multiparameter flow cytometry, the cellular composition of the hematopoietic compartment was quantified in both bone marrow and spleen of mu-calpain null mice. The mu-calpain null mice exhibited an overall increase in the number of B-cell lineage; showing ~40% increase of B220+ cells in the bone marrow and ~19% increase of B220+ cells in the spleen. Interestingly, T-cell compartment (CD3+, CD4, CD8+) and NK lineage (NK1.1+) cells were significantly reduced. Myeloid specific GR1+, CD11b+ granulocytic cell lineage, and megakaryocytic CD41+ cells were also noticeably increased in the bone marrow of mu-calpain null mice. Together, these results reveal a central role of mu-calpain in the maintenance of lymphoid homeostasis, contributing to the regulation of B-cell lineage. Ongoing experiments involve investigating quantitative effects of mu-calpain deletion on the survival and apoptotic cell death of B220+ B-cell lineage in response to various insults such as Fas ligand, cycloheximide, irradiation, 9-AC, and staurosporine. The outcome of these studies will elucidate the precise biochemical function(s) of mu-calpain by identifying key substrates that regulate apoptosis in a physiologically relevant environment.