bluuebunny/crossref_metadata_2025_split huggingface dataset:
Immunogenicity of a BCR-ABL e1a2 p190 Translocation in a Balb/c Mouse Model.
Authors: Gomez-Nunez Marta , Pinilla-Ibarz Javier , Dao Tao , Zakhaleva Victoriya , Korontsvit Tatyana , May Rena J. , Scheinberg David A. 
Published: October 2019

Abstract
Intracellular processing of the breakpoint products of the bcr-abl fusion gene may generate novel peptides that, if capable of binding to HLA class I molecules, would be potential targets for a cytotoxic T lymphocyte (CTL) response. In humans, peptides derived from the e1a2 p190 breakpoint have generated peptide-specific MHC class II proliferative responses. Short peptides have shown binding to MHC class I molecules, although processing and presentation of endogenous p190 protein has not been shown. In a Kd Balb/c mouse model, we tested the hypothesis that single, key amino acids near the breakpoint could be the reason of lack of immunogenicity of the p190 breakpoint peptides. We synthesized a native peptide from the 190 breakpoint (AFHGDAEAL) as well as a synthetic mutated peptide (AYHGDAEAL), which showed excellent predicted binding on the BIMAS algorithm (1152 and 2880 respectively), although in vivo experiments did not show any specific CTL response. In order to assess if the lack of immunogenicity in vivo was due to the absence of binding to the MHC class I molecule rather than to poor TCR interactions, we designed a series of peptides where neutral amino acid, alanine, substitutions were introduced at different potential binding sites in the synthetic peptide: at position three (AYAGDAEAL), position four (AYHADAEAL), position five (AYHGAAEAL) and position seven (AYHGDAAAL). The binding of these altered peptides to H2 class I was assessed using a MHC stabilization assay on T2-Kd cells (TAP deficient cells). In spite of the good computer prediction for binding the MHC stabilization assay did not show evidence of binding of the native and synthetic peptides (<1.9 over the background). In contrast, alanine substitution in position five resulted in the best binding (3.5x over the background). Alanine substitutions in positions three, four and seven also showed improved binding to Kd molecules (2.6x, 3.1x and 3x over the background respectively). These results were confirmed by immunization in vivo with these altered peptides. Only the peptide with the alanine substitution in position five generated a immune response in a CD8 gamma-IFN Elispot assay. Since the altered peptides with the alanine substitutions in positions three, four and seven have shown binding to the Kd molecules, but not a CTL response, change of the aspartic acid in position five near to the fusion breakpoint allows appropriate presentation and recognition of the sequence by the TCR.
Autologous Hematopoietic Stem Cell Transplantation for Refractory Crohn’s Disease, the Summary of Phase I Trial.
Authors: Oyama Yu , Craig Robert , Traynor Ann , Statkute Laivyde , Burt Richard 
Published: October 2019

Abstract
Background &Aims
Crohn’s disease (CD) is an immunologically mediated inflammatory disease of the gastrointestinal tract. In theory, immune ablation followed by autologous hematopoietic stem cell transplantation (HSCT) can induce remission by reconstituting a new immune system.
Methods
We conducted a phase I HSCT study in 17 patients with severe CD. Candidates were less than 60 years old with a Crohn’s Disease Activity Index (CDAI) of 250–400 or Craig Crohn’s Severity Index (CCSI) of >17 despite conventional therapy including infliximab. Peripheral blood stem cells were mobilized with cyclophosphamide and granulocyte colony-stimulating factor and CD34+ enriched. The immune ablative conditioning regimen consisted of 200mg/kg cyclophosphamide, and 90mg/kg equine antithymocyte globulin.
Results
There was no treatment related mortality. The procedure was well tolerated with anticipated cytopenias, neutropenic fever, and disease-related fever, diarrhea, anorexia, nausea, vomiting. The median days for neutrophil and platelet engraftment were 9 (range 7–11) and 9 (range 8–18, three patients never below 20,000), respectively. The median infused CD34+ and CD3+ cell counts were 4.83 x 106/kg (range 1.73–24.88) and 0.56 x 104/kg (range 0.0–3.09), respectively. Initial median CDAI and CCSI were 282 (range 101–358) and 24 (range 18–33). Symptoms and CDAI/CCSI improved quickly and dramatically (figure), and radiographic and colonoscopy findings improved gradually over months to years following HSCT. For patients with >6 months follow up, twelve out of fifteen patients entered sustained remission defined by a CDAI < 150 or reduction of a CCSI >10 after median follow-up of 29 months (range 11–48).
Conclusion
Autologous HSCT can be performed safely and has a marked durable salutary effect on CD activity. A randomized study {KISS study: C(K)rohn’s Immune Suppression vs. Stem Cell Transplant} comparing HSCT to standard therapy is now enrolling patients.
Figure Figure
Humanized Anti CD-40 Antibody SGN-40 Effectively Induces Cytotoxicity Against Chronic Lymphocytic Leukemia (CLL) Cells through Antibody Mediated Cytotoxicity and Demonstrates Modest Biologic Evidence of CD40 Activation.
Authors: Gowda Aruna C. , Zhao Xiaobin B. , Cheney Carolyn , Mehter Najma , Lozanski Gerard , Lin Thomas S. , Guster Sara , Drachman J. G. , Muthusamy Natarajan , Byrd John C. 
Published: October 2019

Abstract
The CD40 antigen is involved in cell survival and differentiation of B-cells and is uniformly expressed on chronic lymphocytic leukemia (CLL) cells. The CD40/CD40L interaction stimulates B-cells, dendritic cells and monocytes to proliferate, differentiate, up regulate co-stimulatory molecules and increase antigen presentation. While activation of CD40 can protect CLL cells against early fludarabine-induced apoptosis, these cells become sensitive to delayed death by extrinsic pathway apoptosis. (Blood, 105: 3193–8, 2005). SGN-40 is a humanized anti-CD40 antibody entering clinical trials and has been reported to have weak agonistic properties following CD40 ligation. To pursue rational clinical development of SGN-40, we studied the effects of this antibody in fresh, non-cryopreserved primary CLL cells. These studies included classic antibody mediated killing mechanisms and evidence of both CLL cell activation and protection against early fludarabine-mediated apoptosis. CLL cells treated with SGN-40 (10 mcg/ml) for 2 hours (hrs) in the presence of human serum promoted no complement mediated cytoxicity (CDC) in 8 pts tested. Direct SGN-40 induced apoptosis of human CLL cells with or without anti-Fc IgG cross-linking at 24, 48 and 72 hrs was not increased over that observed with the isotype control antibody trastuzumab in 8 pts studied. In contrast, SGN-40 induced antibody dependent cellular cytotoxicity (ADCC) against CLL cells an average of 12% (±11.39 SD, range 2–32%) killing at 4 hrs (effector to target cell ratio 25:1) in 6 pts tested. The SGN-40 induced ADCC against CLL cells were similar to that observed with alemtuzumab (average 19%, SD 6.9, range10–30%) or rituximab (average 18%, SD 12.48, range 8–42.5%). SGN-40 also mediated death in Raji and 697 lymphoblastic lymphoma cell lines via ADCC. Similar to reports by others with CD40 ligand, SGN-40 mediated activation was noted with modest up-regulation of CD80 and HLA-DR at 48hrs. When administered prior to fludarabine, SGN-40 also protected against death in 5 consecutive samples, although this was less than observed with CD40 ligand transfected HeLa cells, consistent with incomplete CD40 activation. Concurrent administration of SGN-40 and fludarabine did not protect from drug-mediated apoptosis. In conclusion, these findings suggest that SGN-40 has dual property of mediating cytotoxic effect by ADCC and partial CD40 activation. Development of SGN-40 as a therapeutic agent in CLL is justified and future studies by our group are focusing on enhancing SGN-40 mediated ADCC against CLL cells and potentially designing combination studies with SGN-40 to exploit this agent’s ability to engage the CD40/CD40L network.
SPC2996 - A New Bcl-2 Inhibitor for the Treatment of CLL.
Authors: Kearney Phil , Westergaard Majken , Bo Hansen Jens , Hansen Henrik F. , Thrue Charlotte A. , Frieden Miriam , Koch Troels , Ørum Henrik 
Published: October 2019

Abstract
The proapoptotic oligonucleotide agent - SPC2996, has been developed by Santaris Pharma A/S, as a specific Bcl-2 mRNA antagonist. SPC2996 was selected from our inhouse discovery research in a library of short antisense oligonucleotides with Locked Nucleic Acid (LNA). LNA brings a significant increase in the affinity towards complementary RNA and in addition remarkably increases the bio-stability.
Stability was evaluated in human and rodent plasma with SPC2996 capable of 20day half life in human serum. mRNA and protein analyses were used for showing specific downregulation of Bcl-2 in cancer cell lines and this reduction was associated with induction of apoptosis. Cellular Bcl-2 responses on mRNA and protein are seen from 1nM of SPC2996. These effects lead to apoptosis induction. Anti- tumor activity was studied in xenotransplanted athymic mice models of human cancer. SPC2996 was equally effective as Genasense in these models.
That SPC2996 specifically dowregulates Bcl-2 was demonstrated in primate studies. Side by side comparison with Oblimersen sodium in these studies showed SPC2996 effectively downregulated Bcl-2 at 3mg/kg while Oblimersen sodium showed little or no target downregulation. Microarray analyses were used to demonstrate specificity of target downregulation.
With SPC2996 we bring the first member of a new generation of safe, potent and specific mRNA antagonist drugs, exploiting the benefits of LNA. SPC2996 provides the cancer patient with the possibility of lower and less frequent dosing, which is unprecedented by any other oligonucleotide inhibitor of Bcl-2 used in clinical trials.
Liposomal Amphotericin B (L-AMB) as Initial Therapy for Invasive Filamentous Fungal Infections (IFFI): A Randomized, Prospective Trial of a High Loading Regimen vs. Standard Dosing (AmBiLoad Trial).
Authors: Cornely Oliver A. , Maertens Johan , Bresnik Mark , Herbrecht Raoul 
Published: October 2019

Abstract
Background: L-AMB exhibits non-linear pharmacokinetics, with maximal values for Cmax and AUC achieved at a dose of 10 mg/kg/d. Maximal tolerated dose is >15 mg/kg/d. In animal models improved efficacy has been demonstrated with higher doses of L-AMB, but comparative clinical data for efficacy of higher than standard doses of L-AMB are lacking. A randomized, double blind study was performed to compare the efficacy and safety of a high loading dose regimen (HD) to standard dosing (SD) as initial therapy of IFFI.
Methods: Patients with proven or probable IFFI by modified EORTC/MSG criteria were randomized to receive L-AMB 3 or 10 mg/kg/d x14d, then 3 mg/kg/d until investigator-defined end of study drug treatment (EOT). Study drug was blinded for the first 14 days of treatment. The primary endpoint was favorable overall response (FOR) assessed at EOT. FOR=complete responses + partial responses. Survival was followed up to 12 wks. An independent Data Review Board (DRB) confirmed all IFFI diagnoses and response assessments.
Results: 201 patients with DRB confirmed IFFI diagnoses comprised the MITT population. 107 received SD, 94 HD. Groups were well matched in terms of risk factors. 93% of patients in each group had underlying hematological malignancies. 16% of SD and 19% of HD patients had allogeneic SCT. Neutropenia was present at baseline in 73% of patients overall, and persisted through EOT in 42%. Invasive aspergillosis (IA) accounted for 97% of cases. Median duration of study drug treatment was SD 15d (range 1–60d) and HD 14d (range 1–57d). FOR at EOT was 50% for SD vs. 46% for HD (p= NS). No significant differences in FOR by treatment group were seen for the subsets of IA, allo-SCT, neutropenia subsets, or site of IFFI. Survival at 12 wks was 72% SD vs. 59% HD (p= NS). Nephrotoxicity (serum creatinine ≥ 2x baseline) occurred in 14% SD vs. 31% HD (p<.01). Grade 3 or greater hypokalemia (K+<3.0 mmol/L) developed in 16% SD vs. 30% HD (p<.02), but no difference was found in grade 4 hypokalemia (K+ ≤ 2.5 mmol/L) 3% SD vs. 4% HD (p= NS).
Conclusions: In a population of highly immunocompromised patients (93% with underlying hematological malignancies, 73% with neutropenia at study entry), L-AMB as initial treatment of invasive aspergillosis and other filamentous fungal infections at a standard dose of 3 mg/kg/d had an overall favorable response rate of 50% and a 12 wk survival rate of 72%. L-AMB given as a high loading regimen of 10 mg/kg/d x14d did not demonstrate any benefit in overall response or survival, and was associated with higher rates of nephrotoxicity and hypokalemia.
CD117 Expression Is a Poor Prognostic Factor (PF) for Progression Free Survival (PFS) and Freedom from Progression (FFP) in Patients with Newly Diagnosed Acute Myelogenous Leukemia (AML).
Authors: Saber Wael , Abou-Jawde Rony , Hsi Eric , Jin Tao , Kalaycio Matt , Sobecks Ronald , Sekeres Mikkael , Pohlman Brad , Tripp Barbara L. , Advani Anjali 
Published: October 2019

Abstract
Background: There are conflicting reports on the prognosis of patients with newly diagnosed c-KIT positive (CD117) AML. In addition there is controversy on the best way to define CD117 positivity by flow cytometry (FC). We retrospectively reviewed data on 95 patients with newly diagnosed AML during the years 1998–2002 to determine the prognostic value of CD117 expression. Methods: CD117 expression was determined by staining cells with antibodies to CD45 and CD117 (BD Biosciences San Jose, CA). CD45-stained cells without CD117 antibody were used as a negative control. FC was performed on FACSCalibur instruments and data acquired using CellQuest software (BD Biosciences). Using a CD45/orthogonal light scatter gate to isolate blasts, the mean fluorescence index (MFI) was calculated as the CD117 mean channel number (MCN) of the blasts/MCN auto fluorescence. Statistical analysis was performed using Cox proportional hazards (CPH) and log rank (LR) testing; Recursive Partitioning analysis was used to Identify CD117 MFI cut-point.
Results: The median age of patients was 59 years. 30% had a history of an antecedent hematological disorder. Good, normal, unfavorable, and other cytogenetics were seen in 10%, 49%, 30%, and 11% of patients, respectively. Patients were treated with anthracycline based induction therapy, followed by consolidation therapy (45% chemotherapy, 23% stem cell transplant). The complete remission (CR) rate was 69.5% (58.7% for age ≥60, 79.6% for age <60). Median Follow up was 16.5 months (m). Median PFS and median overall survival (OS) was 11.2 m (8.5 m for age ≥60, 12.7m for age <60) and 21.3m (19.9m for age ≥ 60, 37.1m for age <60), respectively. CD117 expression (MFI) did not correlate with age, cytogenetics, CR rates or OS. However, patients with CD117 MFI > 10.7 had a median PFS of 9.5m vs. 14.0m for MFI ≤ 10.7 (p=0.033, LR). Median FFP for MFI > 10.7 was 10.0m vs. 33.6m for MFI ≤ 10.7 (p=0.019, LR). On univariate CPH analysis, CD117 (MFI >10.7) was associated with worse PFS (HR: 1.86, 1.04–3.32, p=0.036) and FFP (HR: 2.38, 1.13–5.03, p=0.023). Multivariate CPH analysis showed increasing CD117 expression (per 5 unit MFI increase) was associated with worse FFP (HR: 1.19, 1.01–1.4, p=0.042).
Conclusion: CD117 expression does not correlate with age, cytogenetics, or OS. However, CD117 (MFI>10.7) is a poor PF for PFS and FFP in AML patients. Higher CD117 ratio is also an independent poor PF for FFP. Whether or not agents that inhibit CD117, such as Imatinib Mesylate, can improve prognosis is the basis of ongoing trials.
T-bet Down-Modulation in Tolerized Th1 Effector CD4 Cells Confers a TCR-Distal Signaling Defect That Selectively Impairs IFN- γ Expression.
Authors: Long Meixiao , Slaiby Aaron M. , Hagymasi Adam T. , Mihalyo Marianne A. , Lichtler Alexander C. , Reiner Steven L. , Adler Adam J. 
Published: October 2019

Abstract
When Th1 effector CD4 cells encounter tolerizing antigen in vivo their capacity to express the effector cytokines IFN-γ and TNF-α is lost more rapidly than non-effector functions such as IL-2 production and proliferation. To localize the relevant intracellular signaling defects, cytokine expression was compared following restimulation with antigen vs agents that bypass TCR-proximal signaling. IFN-γ and TNF-α expression were both partially rescued when TCR-proximal signaling was bypassed, indicating that both TCR-proximal and distal signaling defects impair the expression of these two effector cytokines. In contrast, bypassing TCR-proximal signaling fully rescued IL-2 expression. T-bet, a transcription and chromatin remodeling factor that is required to direct the differentiation of naive CD4 cells into IFN-γ-expressing Th1 effectors, was partially down-modulated in tolerized Th1 effectors. Enforcing T-bet expression during tolerization selectively rescued the ability to express IFN-γ, but not TNF-α. Conversely, expression of a dominant-negative T-bet in Th1 effectors selectively impaired the ability to express IFN-γ, but not TNF-α. Analysis of histone acetylation at the IFN-γ promoter further suggests that down-modulation of T-bet expression during Th1 effector CD4 cell tolerization does not impair IFN γ expression potential through alterations in chromatin structure.
Development of Novel CD138 Antigen-Specific Peptide Capable of Eliciting Myeloma-Specific Cytotoxic T Lymphocytes Response.
Authors: Bae Jooeun , Martinson Jeff , Yu-Tzu Tai , Neri Paola , Tassone Pierfrancesco , Li Xianfeng , Coffey Rory , Xia Zhinan , Prabhala Rao , Anderson Kenneth , Munshi Nikhil 
Published: October 2019

Abstract
Multiple myeloma (MM) is characterized by the accumulation of mature plasma cells in the bone marrow. Despite progress in treatment, the disease still remains incurable, therefore novel therapeutic approaches are required to achieve better treatment. Development of peptide-based immunotherapies against specific tumor-associated antigens offers an attractive approach that may lead to tumor-targeted cellular therapy in MM patients. CD138 (Syndecan-1) is an unique target expressed on the surface of MM cells and normal plasma cells with very limited other distribution or expression. CD138 has been reported to play an important role in myeloma cell survival and, infact, is used to purify MM cells in laboratory experiments. The purpose of this study was to develop immunogenic peptides derived from CD138 antigen for the induction of antigen-specific cytotoxic T lymphocytes (CTLs) against MM. We have identified an immunogeneic HLA-A2-specific CD138 peptide that is capable of inducing MM specific-CTLs. To induce the specific CTLs, HLA-A2-positive normal human T lymphocytes were stimulated with autologous dendritic cells or T2 cells pulsed with the CD138 peptide. The generated CTLs showed a distinct phenotype consisting of 67% of CD69+/CD45RO+ and 1% of CD45RA+/CCR7+ T cells, characteristic of effector memory cell population. A significant (p < 0.05) increase in T cell proliferation and IFN-g secretion were observed by the CTLs following restimulation with HLA-A2+/CD138+ MM cells. The CTLs displayed HLA-A2-restricted CD138-specific cytotoxic activity against MM cell, demonstrated as 76% and 86% cytotoxicity at Effector:Target ratios of 20:1 or 60:1, respectively. The MM cell-specific cytotoxicity was confirmed in “cold” target inhibition assays using HLA-A2.1+/CD138+ MM cells as “cold” inhibitor, demonstrated 25% and 39% reduction in cytotoxicity when incubated with cold inhibitors at E:T ratios of 20:1 or 60:1, respectively. A distinct CD138 peptide-specific tetramer+/CD8+ population was also confirmed. Taken together, these data demonstrate the efficacy of the immunogenic CD138 peptide in inducing a cellular immune response in MM for future clinical application.
Two Functional Mutations in cis of the Ferrochelatase Gene (FECH) Cause Erythropoietic Protoporphyria (EPP).
Authors: Di Pierro Elena , Brancaleoni Valentina , Moriondo Valeria , Cappellini Maria D. 
Published: October 2019

Abstract
Erythropoietic protoporphyria is an autosomal dominant disease with incomplete penetrance, due to reduced activity of ferrochelatase (FECH), a mitochondrial enzyme that catalyzes the final step of the heme biosynthetic pathway. The clinical phenotype of EPP results from coinheritance of a mutated allele and a wild-type low expressed allele of the FECH gene. To date, more than 100 different mutations have been identified in the FECH gene of EPP patients. Different evidence suggests that an entire haplotype (−251G, IVS1-23T, and IVS3-48C) reduces allele expression. In two Italian EPP patients carring the −250G>C mutation in the promoter region we recently demonstrated the loss of an SP1 binding site causing a strong impairment of promoter activity. In both patients we searched for the GTC haplotype on the other allele. Family segregation studies established that the GTC haplotype was in trans to the −250C mutation. Indeed, asymptomatic parents carried either the three polymorphisms or the point mutation. In both patients only the IVS3-48C polymorphism was in apparent homozygosity since family studies established the absence of Mendelian segregation. To reduce the possibility of interfering polymorphisms in the primer sites, a second set of sequencing primers was designed. Apparent homozygosity was confirmed. We assumed the presence of an intronic deletion on the same allele carrying the −250G>C mutation. In order to establish the size of the deletion we performed a semiquantitative RT-PCR analysis on fresh RNA and XL-PCR analysis on DNA. In both cases the primers were situated in regions outside the heterozygotic polymorphisms. As expected RNA analysis revealed the presence of a normal fragment of 612bp and a shorter fragment of 343bp that was less abundant compared to controls. Sequence analysis on the isolated abnormal fragment showed loss of exons 3 and 4. DNA analysis revealed the presence of two fragments of 21440bp and 15864bp respectively. We performed an internal long PCR to obtain two shorter fragments 9717bp and 4141bp respectively. Sequence analysis on the isolated abnormal fragment showed a 5576bp deletion defined by two short direct repeats of about 40bp. The deleted region includes both a first repeat and interposed sequence. These results showed the presence of two functional mutations on the same allele. Deletion contributed to create a null allele preventing formation of the correct mRNA that is already strongly impaired by the −250G>C mutation. The deletion is caused by slipped strand mispairing or more probably by unequal intragenic recombination. These two mutations in the promoter region and intron 3 respectively, are located in the same regions as the −251G and ivs3-48C polymorphisms, strongly suggesting a functional relationship between these two regions.