bluuebunny/crossref_metadata_2025_split huggingface dataset:
Lack of Alloantibody Response to Red Blood Cell Antigens in Juvenile Mice Following Transfusion: Non-Immunogenic or Tolerogenic Response?.
Authors: Hendrickson Jeanne E. , Josephson Cassandra D. , Chadwick Traci E. , Zimring James C. 
Published: April 2020

Abstract
Introduction: Alloimmunization to red blood cell (RBC) antigens occurs in approximately 6% of chronically transfused adults, with some patient populations having significantly higher rates of alloimmunization. Despite these numbers, multiple studies have failed to detect alloantibodies to RBC antigens in neonates following transfusion, even after repeat exposures from the same donor. This is consistent with two possible scenarios:
RBC antigens are non-immunogenic in neonates, or RBC antigens are tolerogenic.
If early exposure to RBC antigens is a tolerogenic stimulus, then the potential clinical implications are considerable. To allow a controlled testing of these hypotheses, we have constructed a murine model of alloimmunization to RBC antigens in pediatric populations.
Methods: Weight-adjusted volumes of leukoreduced blood from mHEL donors, expressing membrane bound hen-egg lysozyme as a unique RBC antigen, were transfused into B10.BR recipients at 3 weeks (juveniles) or 3 months (adults) of age. To assess the effects of inflammation, which we have recently reported augments RBC alloimmunization in adult mice, half of the recipients were treated with Poly (I:C), a double-stranded RNA that mimics viral inflammation. Two weeks post-transfusion, RBC alloimmunization was determined by measuring serum anti-HEL IgG levels by ELISA.
Results: In 2 out of 2 experiments, as part of an ongoing study, whereas all 10 adult mice mounted a low level alloantibody response by ELISA, only one out of 10 juvenile mice made a detectable alloantibody response. The non-responsiveness of juvenile mice was not significantly altered by inflammation, as only 2 out of 10 juvenile mice treated with Poly (I:C) mounted an alloantibody response.
Conclusion: Our data demonstrate that transfused RBCs are weakly immunogenic in adult mice, but only rarely result in alloimmunization in juvenile mice. This is consistent with the pattern of alloimmunization seen in adult versus neonatal humans. Since our model system detects alloantibodies by ELISA, which is 2000 fold more sensitive than agglutination assays using mHEL RBCs, these data support the notion that the majority of juvenile mice do not make alloantibodies to the mHEL RBC antigen. Additionally, inflammation enhances alloimmunization in juvenile mice to a significantly lesser extent than in adult mice. Ongoing studies will utilize the ability of this model to assess if juvenile transfusion recipients are tolerized, by challenging with an immunogenic stimulus (transfusion in the presence of inflammation or subcutaneous injection of HEL/CFA ). If these studies demonstrate that tolerance to RBC antigens occurs after exposure in the neonatal or juvenile period, then the potential clinical implications are significant.
Determinants of PF4/Heparin Immunogenicity in a Murine Model of HIT.
Authors: Arepally Gowthami M. , Qi Rui , Hollingsworth John , Suvarna Shayela 
Published: April 2020

Abstract
Heparin-Induced Thrombocytopenia (HIT) is a life-threatening immune-mediated reaction caused by platelet factor 4 (PF4)/heparin complexes. In recent in vitro studies, we have shown that human and monoclonal HIT antibodies recognize and optimally bind to ultra-large complexes (ULC; MW&gt;670kDa) formed at equimolar ratios of PF4 and heparin. We undertook the following studies to determine if the in vivo immunogenicity of PF4/heparin complexes showed similar requirements for stoichiometric molar ratios of PF4:heparin (PHR). Using a previously described murine immunization model, mice were immunized with antigen by retro-orbital injection daily for 5 days without adjuvant. Blood was collected at baseline and at weekly intervals for 4–6 weeks. Plasma was tested for antibody development by a murine PF4/heparin (mP+H) ELISA. To determine the effects of PHR on mP+H antibody production in vivo, PF4 was mixed with unfractionated heparin (UFH) at molar ratios of 2:1, 1:1 or 1:2 and injected into mice (n=5/cohort). Antibody levels (mean A450nm ± SD) of mice injected with PHR 2:1 (0.79 ± 0.77) were higher than mice injected with PHR 1:1 (0.23± 0. 2), or PHR 1:2 (0.09 ± 0.02, p&lt;0.05 for PHR 2:1 v. PHR 1:2). Because in vitro studies showed that UFH was far more capable of generating ULCs when mixed with PF4 than low-molecular weight heparin (LMWH) or fondaparinux, mice (n=5/cohort) were injected with buffer, or mPF4 mixed with UFH, or enoxaparin (a LMWH) or the pentasaccharide, fondaparinux at doses that are therapeutically equivalent to doses given to humans. Antibody levels (mean A450nm ± SD) were significantly higher in mice injected with UFH (3.340 + 0.15) than in mice injected with buffer (0.0451 ± 0.013, p&lt;0.001 for buffer v UFH), LMWH (0.8526 ± 0.44, p&lt;0.001 for LMWH v UFH) or the pentasaccharide (0.80 ± 0.0.7, p&lt;0.001 for pentasaccharide v UFH). Because PF4/heparin ULCs are composed of repeating structural antigenic units, we next investigated the role of toll-like receptors (TLRs), a family of pattern recognition receptors (PRRs) that recognize repeating structural determinants on microbes and foreign antigens. To determine if mP+H complexes engage TLRs, we injected mP+H into wild-type (WT, n=5) or mice deficient in MyD88 (n=7), an adaptor molecule critical for intracellular signaling by most TLRs. Antibody levels (mean A450nm ± SD) did not significantly differ between WT (1.5 ± 0.6) and MyD88 null mice (0.9 ± 0.5). In summary, these studies indicate that in vivo antibody responses to mP+H complexes are critically dependent on stoichiometric ratios favoring ULC formation and that immune activation by PF4/heparin is independent of MyD88 signaling pathways.
Notch 4 Plymorphism Associated with Risk of Aplastic Anemia.
Authors: Kim Hee Nam , Choi Jeong-Hwa , Yun Ju-Hyun , Kim Yeo-Kyeoung , Lee Il-Kwon , Yang Deok-Hwan , Shin Myung-Geun , Kook Hoon , Hwang Tai Ju , Park Moo-Rim , Jo Deog-Yeon , Won Jong Ho , Kim HyeoungJoon 
Published: April 2020

Abstract
Aplastic anemia (AA) is a heterogeneous disease characterized by bone marrow aplasia and peripheral blood pancytopenia, a profound deficit of hematopoietic stem and progenitor cells. Although the causes for aplastic anemia are multiple, the pathogenic mechanism leading to the disease might be limited to two main pathways: environmental factors and genetic susceptibility. The mammalian Notch family has four homologs, Notch1 to 4. Notch4 signaling plays a role in lineage commitment in hematopoiesis. The human Notch4 gene is located at the MHC on chromosome 6p21.3. Many single nucleotide polymorphisms (SNP) have been mapped to the Notch4 locus. Therefore, we hypothesized that some of these variants could potentially influence the expression level of Notch4 protein. To investigate the role of the two Notch4 rs2071282 (+2763 C&gt;T, Pro204Leu) and 422951(+3323 A&gt;G, Thr320Ala) polymorphisms in susceptibility to AA, a case-control study was conducted in Chonnam National University Hwasun Hospital between March 2002 and June 2006. We genotyped in 151 AA patients and 552 healthy control subjects using PCR-RFLP. Here, we report that rs2071282 CT genotype is significantly associated with increased risk for AA. Using subjects with the rs2071282 CC as a reference group, the odds ratio (OR) of CT is 1.77 (95% CI 1.05–2.98, p=0.032). The haplotype CG is significantly associated with the risk of AA. Using subjects with the haplotype CA, the OR of haplotype CG is 0.70 (95% CI 0.50–0.98, p= 0.037).
In conclusion, we were able to identify two polymorphisms of Notch4, therefore, theses results suggest that Notch polymorphism may play an important role in the susceptibility to AA.
Distribution of Notch polymophisms in aplastic anemia patients and controls Control (%) AA(%) OR (95% CI) p Rs 422951 AA 302(54.8) 94(62.3) 1 AG 216(39.2) 49(32.5) 0.72(0.478–1.098) 0.128 GG 33(6.0) 8(5.3) 0.54(0.23–1.279) 0.163 Total 551 151 Rs 2071282 CC 481(87.1) 120(81.1) 1 CT 68(12.3) 28(18.9) 1.769(1.049–2.982) 0.032 TT 3(0.5) 0 - - Total Haplotype frequency for AA and controls Haplotype Control (%) AA(%) P (Chi-square) OR (95% CI) p *Haplotype frequencies were estimated from genotype data using Gibbs- algorithm.*OR and 95%Cis were estimated using based logistc regression and adjusted for sex and age C-A 743(67.7) 206(70.0) 0.062 1 C-G 281(25.6) 60(20.4) 0.70(0.50–0.98) 0.037 T-A 72(6.5) 28(9.5) 1.49(0.90–2.47) 0.126
ULBP and MICA/B as Novel Indicators for Both Diagnosis of Immune-Mediated Marrow Injury and Favorable Response to Immunosuppressive Therapy in Bone Marrow Failure Syndromes.
Authors: Hanaoka Nobuyoshi , Kawaguchi Tatsuya , Horikawa Kentaro , Nagakura Shoichi , Tsuzuki Yasuchika , Yonemura Yuji , Nakakuma Hideki 
Published: April 2020

Abstract
Bone marrow failure syndromes (BFS) including aplastic anemia (AA), myelodysplastic syndromes (MDS), and paroxysmal nocturnal hemoglobinuria (PNH) are considered to harbor immune-mediated marrow injury. Indeed, immunosuppressive therapy (IST) with antithymocyte globulin and cyclosporine ameliorates the hematopoiesis in BFS patients, despite its limit: infrequent relapse of marrow failure after IST, resistance of some patients to IST, and untoward effects like infection. To overcome the difficulties, molecular targeted therapy is alternative. However, neither incitement of marrow injury nor target molecules on marrow cells recognized by cytotoxic lymphocytes has been identified. We have currently suggested that NKG2D ligands such as ULBP and MICA/B serve as triggers for immune-mediated marrow injury in PNH (Hanaoka, Blood2006;107:1184). ULBP and MICA/B are stress-inducible membrane proteins that appear in infection and transformation. The ligands share NKG2D receptor on lymphocytes such as NK, CD8+ T, and γδ T cells and promote activation of the lymphocytes. Cells expressing the ligands are then deadly injured by NKG2D+ cells (Groh, PNAS USA 1996; Cosman, Immunity 2001). In this background, we attempted to confirm the clinical significance of the expression of NKG2D ligands in BFS. The ligands were detected by flow cytometry on the granulocytes and marrow cells in 47 (53%) of 89 patients with BFS: 28 (56%) of 50 patients with AA; 11 (65%) of 17 patients with PNH; 8 (36%) of 22 patients with MDS; and none of 17 healthy individuals. The membrane expression of the ligands was supported by their increase in plasma. It is then conceivable that blood cells were exposed to a certain stress to induce NKG2D ligands in the patients, leading to NKG2D-mediated marrow injury. There was a close association of the expression of NKG2D ligands with both progression of marrow failure and favorable response to IST. Thus, we propose that the NKG2D ligands not only are feasible predictors for both immune-mediated marrow injury and IST effects, but also serve for potential targeted therapy in BFS.
Predictive Markers of Myelodysplastic Syndrome in the Bone Marrow of Patients with Fanconi Anemia.
Authors: Al-Rahawan Mohamad M. , Alter Blanche P. , Bryant Barbara J. , Elghetany M. Tarek 
Published: April 2020

Abstract
Fanconi Anemia (FA) patients are at increased risk of myelodysplastic syndrome (MDS) and acute myeloid leukemia (AML). Impaired cell cycle regulation may have a role in this predisposition. Over-expression of p53, Ki-67 and survivin proteins has been reported in association with MDS. We analyzed bone marrow samples from 138 individuals: 31 FA, 17 acquired aplastic anemia (AA), 40 sporadic low-grade MDS (MDS-1: 35 refractory cytopenia with multilineage dysplasia [RCMD], 5 RCMD with ring sideroblasts [RCMD-RS]), 11 high-grade MDS (MDS-2: refractory anemia with excess blasts [RAEB]), 12 AML, and 27 normal. To identify cell cycle regulation abnormalities that predict risk of MDS or AML in FA patients, we analyzed bone marrow morphology, cellularity, and immunohistochemical expression of cell cycle regulation proteins: p53 (cycle arrest and apoptosis), Ki-67 (cell proliferation), caspase-3 (apoptosis), and survivin (anti-apoptosis). Protein expression was classified as over-expressed if it was detected in a percentage of cells exceeding the highest values observed in normal marrow. Between group comparisons were made using the Fisher exact test. More FA marrows were hypocellular versus sporadic MDS-1, MDS-2 and AML (p &lt;0.0001 in all). p53 was over-expressed at similar rates in FA, MDS-1, MDS-2 and AML patients (12/25 vs. 22/35, 4/11 and 8/12; p = 0.3, 0.7 and 0.3, respectively); p53 was not over-expressed in AA (0/13, p = 0.003). Ki-67 was also over-expressed at similar rates in FA, MDS-2 and AML patients (9/18 vs. 3/11 and 7/12, p = 0.3 and 0.7), in whom over-expression was higher than in AA or MDS-1 (0/12 and 4/35). Survivin expression was more common in FA than in MDS-1, MDS-2, AML or AA (12/16 vs. 2/32, 0/10, 2/12 or 1/13; p &lt;0.01 for all comparisons). In contrast, the caspase-3 over-expression was higher in MDS-2 than in FA (4/11 vs. 0/18, p = 0.014). 7/31 FA patients had RCMD marrow changes; however, there was no correlation between morphologic features of myelodysplasia and the over-expression of cell cycle markers in these patients. p53 over-expression was frequent in FA, MDS-1, MDS-2, and AML; this distinguishes these disorders from AA. Previous studies found no p53 gene mutations in FA; thus, p53 over-expression observed here may represent upregulation of p53, or diminished p53 protein catabolism. Ki-67 over-expression may be a marker for higher grade MDS. The prevalence of over-expression of survivin was unique to FA, distinguishing it clearly from any of the acquired disorders (AA, MDS or AML), and may be a marker for any inherited bone marrow failure syndrome. Our cell cycle marker results suggest that patients in general in whom there is over-expression of Ki-67 and/or p53 may be at higher risk of evolution to MDS/AML; a longitudinal study is needed to address this prediction.
Long-Term Follow up of Children with Apalstic Anemia Treated with Immunosuppressive Therapy.
Authors: Pongtanakul Bunchoo , Das Prabodh K. , Dror Yigal 
Published: April 2020

Abstract
Background: Immunosuppressive therapy (IST) is the alternative first line treatment in children with aplastic anemia (AA) who have no HLA match siblings available. The long-term outcome of patients with AA who survive after IST is unknown. We evaluated outcomes of children with AA treated with IST at long-term follow up.
Methods: we retrospectively reviewed the hospital records of children with AA from 1984 to 2004, treated at our institution with horse-derived antithymocyte globulin (hATG) 160 mg/Kg over 4 days, short course of prednisone and cyclosporine (CS).
Results: Forty two patients were treated with IST (25 boys, 18 girls). The median age at diagnosis was 8.5 years. Twenty nine (69%), eight (19%) and five (12%) patients were diagnosed with severe, very severe, and moderate AA, respectively. Nine patients (21%) had hepatitis associated AA. Twenty seven patients (64%) received one course of ATG and fifteen (36%) received 2 courses (8 received 2 courses of hATG and 7 received 1 course hATG and 1 course rabbit derived ATG). Eleven patients (26%) required G-CSF. Median follow up time was 53.3 months (range 3–244 months). Twenty six patients (62%) had a complete response (CR), eight (19%) had a partial response (PR) and eight (19%) had no response (NR). Two patients relapsed after one course of IST and needed a second course of IST and both of them had a partial response. Median time to discontinuation of CS was 13 months. Nine patients (21%) died (7 with NR and 2 with PR) and 33 patients (79%) are alive. Two patients developed myelodysplastic syndrome (MDS) 21 and 19 months post IST; both received long-term G-CSF (18 and 14 months) and had PR after 2 courses of IST. Five of 33 patients (15%) who survived had significant hypertension after CS was discontinued and one required continuous antihypertensive medication.
Conclusion: The results of this study shows promising response in children with AA treated with IST. Hypertension and MDS are late complication. Longer follow up in these patients is warranted to definite the accurate rates of the late complications and risk factors.
Monitoring Immune Function in the Allogeneic Transplant Recipient: Predicting Graft vs Host Disease (GvHD) and Infection.
Authors: Anaissie Elias J. , Diaz Jose A. , Grazziutti Monica L. , Miceli Marisa H. , Fox-Cottler Michele , Barlogie Bart , van Rhee Frits 
Published: April 2020

Abstract
Background: GvHD, intensity of immunosuppression (ISP) and infection are major causes of toxicity post allogeneic stem cell transplantation (allo-SCT). Yet, no method reliably predicts these complications. We explored the use of a novel FDA-approved immune function assay to determine whether an immunologic response to changes in immunosuppressive therapy for GvHD could predict risk of GvHD and infection and allows tailoring ISP and infection prophylaxis. The ImmuKnow is an assay of global T-cell function that measures increased ATP production after stimulating CD4+ T-helper cell and has been shown useful for managing ISP therapies in solid-organ transplantation.1 We hypothesized that ImmuKnow could assist in predicting risk of GvHD and infection.
Patients and Methods: Between March 2004 and March 2006, 24 adult allo-SCT patients (pts) (mean age 48 years; range 23–70; 50% females) underwent blood sampling for determining ImmuKnow at various times post-HSCT (267 assays, mean of 11/pt). Results were correlated with clinical course (infection, GvHD or event-free). Underlying diseases included acute myelogenous leukemia (n=9), chronic myelogenous leukemia (n=4), myeloma (n=3) and others (n=8).
Results: during the testing period, GvHD was documented in 10 pts [(acute, n=3; chronic, n=7] and 20 infections occurred [bacteremia (n=9); CMV viremia (n=4); others (n=7)]. The average immune function (ng/mL ATP) of GvHD pts was significantly higher than that of event-free pts (385 +/− 102 vs. 189 ng/mL +/−78 respectively), p&lt;0.001. Pts with infections (13 pts, 71 events) had a mean 86 ng/mL ATP (+/−87), also statistically different (p&lt;0.001) from event-free. ROC analysis showed that the positive predictive value (PPV) for GvHD with ImmuKnow &gt; 279 ng/mL was 74 with a sensitivity of 90% (95% CI, 76.9–97.2) and specificity of 83% (95% CI 73 - 91) and an odds ratio (OR) for GVHD of 42 (95% CI, 13 - 136, p&lt;0.001). For infection, ImmuKnow values &lt; 94 ng/mL were 70% sensitive (95% CI, 59 - 80) and 87% specific (95% CI, 77 - 94) with a PPV of 85 and OR of 17 (95% CI, 8–39, p&lt;0.001).
Conclusions: these preliminary data suggest that the ImmuKnow assay is an objective marker of risk for infection and GvHD post-HSCT. Tailoring intensity and duration of ISP to reduce risks of GvHD and infection in an individual patient may be possible and may improve outcomes. Validation of these preliminary data is ongoing.
Influence of Chromosomal Aberrations on Response to First Line Therapy in B-Cell Chronic Lymphocytic Leukemia: Interim Analysis of PALG-CLL3 Randomized Study Comparing Cladribine Plus Cyclophosphamide vs Fludarabine Plus Cyclophosphamide.
Authors: Robak Tadeusz , Blonski Jerzy Z. , Wawrzyniak Ewa , Palacz Aleksandra , Gora-Tybor Joanna , Jamroziak Krzysztof , Piszcz Jaroslaw , Kloczko Janusz S. , Koczkodaj Dorota , Kowal Malgorzata , Dmoszynska Anna , Mital Andrzej , Hellmann Andrzej 
Published: April 2020

Abstract
Impact of cytogenetic abnormalities on treatment with different purine nucleoside analogs in patients (pts) with B-cell chronic lymphocytic leukemia (B-CLL) is largely unknown. One of objectives of PALG-CLL3 trial, comparing cladribine plus cyclophosphamide (CC) with fludarabine plus cyclophosphamide (FC) in previously untreated progressive B-CLL, was to verify the response to treatment in subsets of pts characterized by common cytogenetic abberrations. Chromosomal abnormalities were assessed using fluorescence in situ hybridization (FISH) on interphase nuclei of lymphocytes on whole blood smears prior to the start of the study treatment. Pts were screened for trisomy 12, deletions (del) 11q, del 13q and del 17p using DNA probes: CEP12, LSI: ATM, D13S319 and p53 (Vysis), respectively. For the purpose of the present interim analysis complete cytogenetic results were available in 133 pts out of 423 pts included to the study. In this group the chromosomal aberrations were detected in 102 pts (77%) including single abnormalities observed in 69 pts (52%) and two or more aberrations in 33 pts (25%). Thirty-one pts (23%) exhibited a normal interphase FISH pattern. The most frequent single abnormality was del 13q found in 38 pts (29%), while del 17p, trisomy 12 and del 11q were identified in 14 pts (11%), 11 pts (8%), and 6 pts, (5%), respectively. The most frequently observed associations of chromosomal aberrations were: del 13q with del 11q (11 pts, 8%) and del 13q with del 17p (10 pts, 8%). Four pts (3%) revealed three chromosomal abnormalities including association of trisomy 12/del 11q/del 13q in two pts, trisomy 12/del 11q/del 17p in one pt and del 11q/del 13q/del 17p in one pt. Overall, treatment was completed and response assessed in 113 out of 133 pts with known FISH pattern. In this group of pts del 17p was the only chromosomal abnormality that correlated significantly with treatment outcome. Pts with del 17p (21, 19%) had lower probability to achieve a complete response (CR) (0.044). Interestingly, in independent analyses of both treatment arms, the negative impact of 17p was seen in pts treated with FC (p=0.002), but not in pts treated with CC (p=0.6). Moreover, comparing response rates between treatment arms we found that CC was superior to FC in terms of complete response in pts with del 17p (57% CR in CC v 14% CR in FC arm, p=0.04). In conclusion, chromosomal abnormalities can be detected in majority B-CLL pts requiring treatment. Our preliminary results suggest that CC combination may have some advantage in terms of CR achievement in B-CLL pts harboring del 17p.