bluuebunny/crossref_metadata_2025_split huggingface dataset:
Resting B Cells Suppress CD8+ T Cell Function and Prevent the Induction of Graft Versus Host Disease.
Authors: Ritchie David S. , Watt Victoria 
Published: October 2019

Abstract
B cells have been variously shown to induce direct tolerance of antigen specific CD8+ T cells, induce T cell anergy via TGF-b production, down regulate IL-12 production by dendritic cells (DC) and influence Th1/Th2 differentiation via the production of regulatory cytokines. Through these mechanisms, B cells can exert a regulatory function in in vivo models of T cell immunity including, experimental autoimmune encephalitis (EAE) and rheumatoid arthritis (RA). Recently, B cells have been shown to be essential in the prevention of effector T cell differentiation in a model of autoimmunity.
We have previously shown that resting B cells inhibited tumor protection induced by dendritic cells vaccination. Inhibition of DC immunity by B cells was independent of presentation of major histocompatibility molecule (MHC) class-I bound tumor antigen but dependent on the expression of class-II MHC. Furthermore the inhibitory effect of B cells was lost if the B cells were activated by CD40L or if CD4+/CD25+ regulatory T cells (Treg) were depleted. These studies have been further extended to examine the role of resting B cells on the induction and severity of graft versus host disease (GVHD) induced in a major MHC mismatch model. We have found that mice transplanted with B cell depleted marrow revealed more rapid CD8+ T cell engraftment, higher IL-2 and IFN-γ production, more severe GVHD and shorter survival. Conversely, those who received additional resting B cells at the time of marrow infusion were substantially protected from GVHD. These findings indicate that resting B cells may regulate T cell activation, in part via the suppressive effects of Treg, but also through their important role in T cell homeostasis. Resting B cells may therefore limit the efficacy of DC based immunotherapy or alternatively be used therapeutically to limit CD8+ T cell autoimmunity including GVHD.
Delayed Versus Initial Risk-Categorization of Pediatric Cancer Patients with Prolonged Febrile Neutropenia. A Randomized, Prospective Clinical Trial.
Authors: Ahmed Nabil M. , Mahallawy Hadir El , El Haddad Alaa M. 
Published: October 2019

Abstract
Purpose:
Hospitalization with empiric broad spectrum intravenous antibiotics has been the standard of care for treatment of high-risk febrile neutropenia. In an attempt to improved hospital utilization we attempt to identify a group of patients at a lower risk of sepsis that would be eligible for outpatient therapy. We hypothesized that patients initially considered at high risk for sepsis can be candidates for outpatient therapy if at 72 hours satisfied certain selection criteria.
Patients and methods:
A prospective, randomized clinical trial was conducted in 119 febrile neutropenic pediatric cancer patients. Patients were randomized into two groups depending on the time of risk-categorization: Initial (control) vs. delayed (study) groups. Patients in the study group were risk-assessed at 72 hours and accordingly considered for outpatient therapy if afebrile, had an absolute neutrophil count (ANC) > 100/uL, negative blood cultures, and no signs of infection among other carefully defined “Early Discharge Criteria”. Those randomized to the control group were treated in the hospital. We analyzed the clinical features, microbiological data, hospital course, duration of hospitalization and treatment cost. Intention-to-treat analysis was used. Results: We found a statistically significant difference between the duration of hospitalization in the group risk-assessed at 72 hours and control group (p<0.01). “Delayed Risk-categorization” correlated better with the duration of neutropenia, need for antibiotic change and episode outcome. Duration of hospitalization and per-episode median treatment cost were significantly less in the study group vs. control (p< 0.001). There was a significant tendency for blood cultures to cluster around the first 48 hours. Comparing the blood culture yield between patients treated in the hospital to those treated as outpatients after risk-assessment at 72 hours there was no statistically significant difference demonstrated [p=0.35].
Readmission rate after early discharge was 3.3%.
Conclusion: We conclude that pediatric febrile-neutropenic patients initially at considered at “high risk for sepsis” can be reevaluated at 72 hours for outpatient therapy. The practice of “Delayed Risk-Categoriztion” could be particularly useful to reduce the overall treatment costs, and duration of hospitalization. In addition, this practice could potenitally offer a superior quality of life.
Response to Immunizations Following Allogeneic HCT with Adjuvant Rituximab Therapy.
Authors: Pao Mary , Papadopoulos Esperanza B. , Boulad Farid , Castro-Malaspina Hugo , Jakubowski Ann , Kernan Nancy A. , Perales Miguel , Prockop Susan , Ford Ruth , Young James W. , O’Reilly Richard J. , Small Trudy N. 
Published: October 2019

Abstract
Use of anti-CD20 monoclonal antibody after allogeneic HCT for the treatment EBV lymphoma or viremia, autoimmune cytopenias, and the prevention or treatment of recurrent NHL has become more frequent. There is limited data on the effect of this therapy on specific antibody production following allogeneic HCT. We therefore examined the response to standard childhood immunizations of 30 patients, including 20 adults (>18 years of age) who received rituximab following a T cell depleted (n=26) or unmodified (n=4) HCT at our institution. T cell function was assessed in vitro by proliferative response to PHA and B cell reconstitution was confirmed by recovery of CD19+ and CD20+ circulating lymphocytes. Patients received a median (range) of 4 (1–12) doses of rituximab, with 15 patients receiving 4–6 doses. The median age of the patient population was 28.5 years with a range of 0.8–63.0 years. Children (median age: 8.5 yrs) received a transplant from an unrelated (n=7), HLA mismatched related (n=2), or HLA matched related (n=1) donor and adults (median age: 42.5 yrs), received a transplant from an unrelated (n=7), HLA mismatched related (n=2), or HLA-matched related (n=11) donor. Patients were vaccinated when their PHA response was at least 75% of the lower limit of normal and CD20+ cells were >100/ul. Vaccination was initiated at a median of 270 and 525 days following the last dose of rituximab in children and adults, respectively. A two to 3-fold rise in titers following a series of 3 tetanus and IPV vaccinations occurred in 19 of 21 and 19 of 19 evaluable patients, respectively. Although 8 of 9 children responded to the H flu conjugate vaccine, only 5 of 13 adults mounted an adequate response. Eighteen patients received the recombinant Hepatitis B vaccine. Nine responded following the initial series (5/7 children, 4/11 adults). Two of three patients who failed initial Hepatitis B vaccination, responded to a second series (1 child, 1 adult). Response to pneumococcal vaccination was the least consistent. None of 11 adults who received the unconjugated 23-valent pneumococcal vaccine developed an adequate response. In view of this observation, children given rituximab were immunized with the conjugated pneumococcal vaccine. Four of eight children responded to the initial series, and two additional children responded following a second series. These preliminary data suggest that both adult and pediatric patients given rituximab following an allogeneic HCT can mount successful antibody responses to standard T-helper cell dependent vaccines. However, B cell responses to T cell independent vaccines may be particularly impaired. Larger prospective trials, stratified by age, comparing vaccine responses in patients following allogeneic transplant in the presence or absence of post transplant rituximab therapy are warranted.
Assessment of Minimal Residual Disease (MRD) by Qualitative RT-PCR (Q-PCR) vs Real-Time Quantitative RT-PCR (RQ-PCR) in a Phase II Study of Acute Promyelocytic Leukemia (APL).
Authors: Gallagher Robert E. , Schachter-Tokarz Esther L. , Jones Dan A. , Estey Elihu H. 
Published: October 2019

Abstract
Assessment of MRD in APL by Q-PCR analysis for the presence or absence of PML-RARα mRNA after the completion of consolidation therapy in treatment regimens employing all-trans retinoic acid (ATRA) and chemotherapy (CT) has provided a reasonably accurate but not infallible prognostic test for disease recurrence (DR). This assessment has been made at detection sensitivity levels up to 1 in 104, which is considered to have optimal predictive value, since non-quantified lower levels of PML-RARα mRNA may not be relevant to clinical outcome. In the current study, we compared the results of Q-PCR (touchdown PCR for 40 cycles followed by post-PCR blot hybridization with a RARα chemiluminescent probe) to results from RQ-PCR in 15 patients on a Phase II clinical trial designed to eliminate CT (low-risk/LR patients, WBC <10K) or minimize CT (high-risk/HR patients, WBC >10K), using combined ATRA and arsenic trioxide therapy. All patients achieved complete remission (CR) and have been followed for a median of 1.2 years. DR occurred in 0/8 LR, 0/1 unclassified and 2/6 HR patients. At CR, all Q-PCR and RQ-PCR assays were positive. For RQ-PCR, the normalized quotient (NQ) value of PML-RARα, i.e., the PML-RARα copy number divided by the housekeeping gene glyceraldehyde-3′-phosphate dehydrogenase (GAPDH) copy number, ranged widely from 1.5 x 10−6 to 7.0 x 10−3. Post-CR follow-up data were:
Patient Group Number Q-PCR+ RQ-PCR+ NQ Range LR/Unclassified 9 0/24 16/24 9 x 10−9 – 1 x 10−7 HR/no DR 4 0/23 8/23 2 x 10−9 – 3 x 10−7 HR/DR 2 1/7 5/7 1 x 10−8 – 5 x 10−5
For the 2 HR/DR patients, the last pre-DR results were, respectively: Q-PCR, positive and negative; RQ-PCR, NQ = 5 x 10−5 and NQ = 2.3 x 10−7. In overall comparisons to RQ-PCR results, all Q-PCR assays were positive for NQ >10−5, 3/4 were positive in the NQ 10−6 to 10−5 range, and all were negative for NQ <10−6. Additionally, dilution-reconstruction experiments confirmed the stochastic nature of PCR assays near the detection limit and indicated that RQ-PCR was about 10-fold more sensitive than Q-PCR. Although larger studies are needed, our results suggest that HR patients who are Q-PCR-negative but RQ-PCR-positive in the >10−7 – <10−5 range at a critical post-CR checkpoint may be at increased risk of DR and, more generally, they suggest that the results of RQ-PCR testing at a critical checkpoint can be used in combination with clinical risk assessment to stratify the intensity of subsequent MRD monitoring in individual patients.
Splenectomy for Splenic Marginal B-Cell Lymphoma.
Authors: Segota Ena , Shaheen Philip , Hsi Eric , Kalaycio Matt 
Published: October 2019

Abstract
Background: Splenic marginal zone lymphoma (SMZL) is an uncommon indolent B-cell malignancy usually involving the spleen, bone marrow and blood. Splenectomy is frequently performed for both diagnostic and therapeutic purposes, but it is unclear if it affects outcome. To further explore this issue, we reviewed clinical and pathological characteristics, and treatment outcomes of SMZL patients (pts) who underwent splenectomy in our institution.
Methods: Medical records of pts who had splenectomy at the Cleveland Clinic and were diagnosed with a B-cell lymphoproliferative disorder between 1994 and 2004 were reviewed. Pathology slides were reviewed to confirm the diagnosis of SMZL by WHO criteria. Data collection included: clinical and biochemical characteristics at diagnosis including symptoms; reason for and date of splenectomy; hematological response following surgery; additional therapies other than surgery including rationale, regimen used and outcome; and follow-up data.
Results: We identified 18 pts with SMZL having splenectomy in our institution. Twelve (67%) were females and 6 (33%) males. The median age was 65 years (range 43–85). At the time of diagnosis, 9 pts had splenic and bone marrow involvement; 8 splenic, bone marrow and nodal; 1 spleen involvement only. Four pts were treated with chemotherapy prior to splenectomy for presumed CLL, hairy-cell leukemia or unspecified B-cell lymphoproliferative disorder, but none achieved CR. None of the pts received radiation. Splenectomy was performed in 9 pts for diagnostic, and in 9 pts for therapeutic purposes. The most common reason for therapeutic splenectomy was cytopenia (n=5) and abdominal discomfort (n=4). Median follow-up from splenectomy to death or last follow-up is 30 months (range 1–99). Of the 9 pts who had splenectomy for diagnostic purposes the median time to subsequent systemic therapy (n=5) was 7 months (range 0–33). Of the 9 pts treated for therapeutic purposes abdominal symptoms resolved in all (n=4) and 4/5 with cytopenias achieved hematologic response. Seven of 9 pts with only spleen and marrow involvement, achieved complete hematologic remission following surgery lasting from 8+ to 46+months. Of 8 pts presenting with spleen and lymph node involvement, 6 had cytopenias that resolved in only 3 pts following splenectomy. Six of these 8 pts received chemotherapy after splenectomy with median time from surgery to systemic treatment of 16 months (range 3–60). One pt continues to have stable lymphadenopathy observed clinically for 33 months, and 1 pt had stable lymphadenopathy for 24 months and was subsequently lost to follow-up.
Conclusion: Splenectomy, whether performed diagnostically or therapeutically, may result in long remissions in the absence of chemotherapy. Splenectomy appears to be a viable treatment alternative for SMZL.
Identification and Hierarchy of Clonogenic Factors for Human Myeloma Cells: IL-6 for CD45+ Versus IL-6 and Growth Factors for CD45-.
Authors: Amiot Martine , Bataille Regis , Collette Madeleine , Descamps Geraldine , Pellat-deceunynck Catherine 
Published: October 2019

Abstract
Multiple Myeloma (MM) is a fatal plasma-cell malignancy characterized by the accumulation of malignant plasma cells within the bone marrow. IL-6 and IGF-1 are known to be essential growth and survival factors in this malignancy. Beside these well characterized growth factors, other growth factors such as HGF, HB-EGF and FGF have been involved in this malignancy. Even though all of them seem to be involved in myeloma cell proliferation, the hierarchy of each growth factor remains to be established. In the present study, a serum-free cytokine-free and collagen-based assay which does not allow the generation of endogeneous myeloma colonies was used to identify the clonogenic factors of fourteen myeloma cell lines. We selected seven myeloma cell lines expressing CD45 on 100% of cells and seven other cell lines lacking CD45 expression. IL-6 is the only clonogenic factor able to stimulate both CD45+ and CD45- myeloma cell lines, generating myeloma colonies from 10 of 14 myeloma cell lines. For some cell lines (LP-1, L363 and XG-2) the percentage of clonogenic cells reached 40% to 50% indicating that clonogenic cells are CD138+ and do not represent a particular CD138-subpopulation. In contrast, the other growth factors (IGF-1, FGF, HGF and HB-EGF) stimulate only some CD45-myeloma cell lines and at a less extend than IL-6. Among them, IGF-1 is the most potent, generating myeloma colonies from 5 of 8 CD45- myeloma cell lines. We searched for a correlation between myeloma-colony formation and the signaling pathway induced by IGF-1 and FGF in MM cells. It appears that activation of both the PI3Kinase and ERK pathway by IGF-1 and FGF fully correlates with their capacity to stimulate the clonogenicity of CD45-myeloma cell lines. In conclusion, the CD45 phenotype of myeloma cells discriminates their signaling and proliferative response to IL-6 and growth factors. These results give a strong rational to the stratification of prognostic value discriminating CD45+ from CD45- MM. Furthermore, treatment strategies for CD45- patients should combined the disruption of signaling induced by both IL-6 and IGF-1 (or other growth factors) whereas CD45+ patients could be targeted through IL-6 only.
CD34+ Marrow Cells from MDS Patients Express Homing-Associated Cell Surface Molecules but Impaired In Vitro Migration.
Authors: Thanopoulou Eleni , Kouraklis-Symeonidis Alexandra , Symeonidis Argiris , Tzouvara Evangelia , Pherson Lorna , Zoumbos Nicholas 
Published: October 2019

Abstract
Homing and retention of hematopoietic stem cells in the marrow microenvironment are multi-step processes mainly controlled by interactions between chemokines, adhesion receptors and proteases. In the case of MDS, whether these interactions are perturbed and how they are implicated in the pathogenesis of the disease or progression to AML is not known. In this study we evaluated the cell surface expression of homing-associated adhesion molecules, CD11a, CD26, CD45, CD49d, CD49e and CXCR4 by flow cytometric analysis in BM mononuclear (MN) and CD34+ cells from 19 MDS patients [7 RA, 1RARS, 11RAEB(t)] and age-matched healthy individuals (n=9). CD34+ and MN cells from RAEB(t) samples expressed higher levels of CD11a and CD49e (2–3 fold increase) and 1,5–3 fold lower levels of CD45 compared with the cells from the healthy control group, as measured by the mean fluorescent intensity. None of the remaining molecules was found to have altered expression in the RAEB(t) subtype. A similar expression pattern of all homing-associated molecules was seen in the cells from the RA/RARS and normal control samples. However, when CD34+ cells from MDS patients were allowed to migrate in vitro toward a gradient of SDF-1α (125ng/ml), they displayed an impaired chemotactic response [mean values of % input: 9% (0,5–30%)], whereas those from healthy individuals actively migrated [mean values of % input: 20% (10–50%)]. Both migrated and non migrated CD34+ cells from MDS patients displayed little hematopoietic activity in vitro (CFC and LTC-IC). The addition of neutralizing monoclonal antibody anti-CXCR4 decreased in vitro chemotactic response of normal and MDS derived CD34+ cells by 90% and 80% respectively indicating that migration was dependent on SDF1/CXCR4 interactions. To further investigate the differences observed in response to SDF1a, immunofluorescence detection of intracellular expression of CXCR4 was performed. MDS derived CD34+ and MN cells from all the subtypes studied significantly expressed high levels of intracellular CXCR4 and were induced to cell surface CXCR4 expression after 48 hours of SCF (100ng/ml) and IL-6 (20ng/ml) stimulation, as was seen with normal cells.
In conclusion marrow CD34+ cells from MDS patients showed reduced capacity to migrate in vitro, although no significant perturbations of adhesion molecules expression were detected. We are currently investigating the signaling events and pathways, which control homing and adhesion of hematopoietic cells in the marrow.
Overcoming TNF-α Induced Suppression of Erythroid Differentiation of Human Cord Blood CD34+ Cells with Arsenic Trioxide.
Authors: Won Jong-Ho , Cheong Hee-Jeong , Kim Sook-Ja , Bae Sang-Byung , Kim Chan-Kyu , Lee Nam-Su , Lee Kyu-Taeg , Park Sung-Kyu , Hong Dae-Sik , Park Hee-Sook 
Published: October 2019

Abstract
The anemia of chronic disease-which encompasses inflammation, infection, tissue injury, and conditions associated with the release of proinflammatory cytokines (such as cancer)- is one of the most common forms of anemia seen clinically. Symptomatic anemia requires treatment. The two major forms of treatment are transfusions and erythropoietin. Arsenic trioxide (As2O3) used to treat human diseases for centuries in traditional Chinese medicine. Our recent studies suggest that low dose of As2O3 induces erythroid differentiation of K562 human leukemic cells and high dose of As2O3 induce apoptosis. In this study, we have investigated in vitro effect of As2O3 on the erythroid differentiation and it could inhibit TNF-α induced suppression of erythroid differentiation of human cord blood CD34+ cells. Expression of glycophorin A was 35.94 ± 7.94% after 7 days culture of human cord blood CD34+ cells and was decreased to 17.63 ± 7.33% when culture of human cord blood CD34+ cells with 100ng/mL of TNF-α. Expression of glycophorin A was increased in dose dependent manner after 7 days treatment with As2O3 and As2O3 increased percentage of glycophorin A in culture with TNF-α compared to TNF-α alone. The results of colony assay of CFU-MIX and BFU-E after culture with various conditions revealed similar patterns with expression of glycophorin A. These results suggest that As2O3 induces erythroid differentiation of human cord blood CD34+ cells and can reverse TNF-α induced suppression of erythroid differentiation of human cord blood CD34+ cells.
The BFU-E colony assay of the human cord blood CD34+ cells after culture with TNF-α or/and Arsenic trioxide. The BFU-E colony assay of the human cord blood CD34+ cells after culture with TNF-α or/and Arsenic trioxide.
Large Deletion of Chromosome 11 Causes Acute Intermittent Porphyria in an Italian Family.
Authors: Di Pierro Elena , Besana Valeria , Moriondo Valeria , Tavazzi Dario , Casalgrande Giovanna , Rocchi Emilio , Cappellini Maria D. 
Published: October 2019

Abstract
Acute intermittent porphyria (AIP) is an autosomal dominant disorder caused by a partial deficiency of hydroxymethylbilane synthase (HMBS), the third enzyme of the heme biosynthetic pathway. The clinical features of AIP are intermittent attacks of neurological dysfunction, including abdominal pain and neuropsychiatric symptoms. Most patients remain asymptomatic throughout their life but 10–20% have severe acute attacks. AIP is normally diagnosed on the basis of clinical symptoms and urinary overproduction of porphyrin precursors, d-aminolevulinic acid (ALA) and porphobilinogen (PBG). The measurement of erythrocyte HMBS activity can confirm the diagnosis and it is also used to detect asymptomatic relatives. So far, more than 290 different mutations in the HMBS gene have been identified among AIP patients. Most of the reported mutations have been detected only in single families. In an Italian family with typical clinical and biochemical signs of AIP, no mutation was found by direct sequencing of 5′ UTR, 3′ UTR, erythroid promoter and the entire coding region of HMBS gene. All sympomatic patients showed apparent homozigosity for eleven common polymorphisms along the HMBS gene. Further family studies established the absence of Mendelian segregation. Excluding interference of polymorphisms in the primer sites, we assumed the presence of a complete deletion of the HMBS gene. In order to verify this hypothesis we performed a semiquantitative RT-PCR analysis on fresh RNA and different long PCR semiquantitative analysis on DNA. All analyses revealed the presence of a normal length fragment which was less abundant compared to controls. In order to identify the size of the deletion, SNPs analysis was extended to flanking genes, H2A Histone Family member X (H2AX) and Dolichyl-Phosphate N-Acetylglucosamine Phosphotransferase 1 (DPAGT1), downstream and Vacuolar protein sorting 11 (VPS11), upstream. We found heterozygotic polymorphisms in VPS11 and DPAGT1 genes. We performed XL-PCR of 19317bp with primers situated in regions outside the heterozygotic polymorphisms that showed a shorter band of 5434bp. Sequence analysis of the abnormal fragment revealed a loss of 77 intergenic base pairs upstream of the HMBS and H2AX genes and 2500bp of DPAGT1 gene. Simultaneously, an inverted complementary sequence of 223bp of intron 9 of HMBS gene was incorporated. Use of ElDorado-Genomatix software revealed the presence of different Alu sequences near breakpoints, that could cause this deletion. The H2AX gene is a member of a classical gene family and its loss has no adverse effects on phenotype. Defects in DPAGT1 are the cause of a congenital disorder of glycosylation type Ij, characterized by severe mental and psychomotor retardation. Our patients had no other symptoms than AIP, supporting the suggestion that DPAGT1 inheritance is probably recessive.