bluuebunny/crossref_metadata_2025_split huggingface dataset:
Pharmacovigilance of a Rituximab Biosimilar (Novex) in Argentina
Authors: Milone Gustavo , Fernandez Francisco , Penna Maria Ines , Gomez Roberto 
Published: October 2019

Abstract
INTRODUCTION
Novex is a rituximab biosimilar approved in Argentina. From January 2015 to June 2016 as part of the Risk Management Plan approved by ANMAT, every treatment detected was recorded and a follow up was implemented in order to obtain safety information.
The aim of this communication is to present the information gathered during this pharmacovigilance program.
MATERIALS AND METHODS
All treatments with at least 1 follow up point were selected and reviewed.
Treatment duration until last follow up checkpoint or date of treatment finalization was recorded.
We summarized demographic data (age, gender), indication distribution, treatment exposition (duration expressed in days and number of cycles received), adverse event frequency (cumulative incidence and incidence density) and distribution.
RESULTS
We collected 345 treatment notifications with at least 1 follow up point.
Gender distribution: 53.2 % of patients were male patients. Age range was 10 to 90 years old, with a mean of 63.9 years.
The indications distribution is summarized in table 1. More than 90% of them were hematological.
Average treatment duration was of 235 days with a median duration of 210 days (IQR = 152 - 311).
Mean number of cycles 5.5; range (1-12).During the reported period a total of 12 Individual Case Safety Reports (ICSR) were received. This represents a cumulative incidence of 3.5 %, an incidence density of 0.015 ICSR notifications per 100 days of treatment and 0.99 per 100 administered cycles.
These 12 ICSR totalized 14 different diagnoses. Of them, the most frequent was Acute Infusion Related Reaction (AIRR), as expected.
All AE but 1 were classified as serious (SAE) because they had at least 1 manifestation that prolonged hospitalization or endangered life.
As expected, the most frequent AE notified was AIRR (5 cases), followed by cardiovascular manifestations (2 arrhythmia, 1 cardiac failure and 1 ischemic stroke), infections (1 pneumonia, 1 progressive multifocal leukoencephalopathy), neurologic (1 paresthesia), cytopenias (1 pancytopenia) and cutaneous (1 bullous dermatitis).
Most of the acute manifestations occurred within the 2 initial infusions of Rituximab, as described.
CONCLUSION
The observed AE frequency and distribution was consistent with product specifications.
No AE notified were unlisted or were notified with greater frequency than expected.
Treatments registry resulted in a suitable way to detect AE that complements spontaneous reporting.

Disclosures
Fernandez: Laboratorio ELEA S.A.C.I.F y A: Employment. Penna:Laboratorio ELEA S.A.C.I.F y A: Employment. Gomez:Laboratorio ELEA S.A.C.I.F y A: Employment.
Identifying Possible Candidate Factors Influencing the Penetrance of Heterozygous NFKB1 Loss of Function Mutations By Whole Exome Sequencing
Authors: Schipp Cyrill , Borkhardt Arndt , Stepensky Polina , Fischer Ute 
Published: October 2019

Abstract
Introduction
The NFκB signaling pathway is a master regulator of immune and inflammatory responses. Recently we and other groups reported heterozygous NFKB1 loss-of-function mutations in patients with combined variable immunodeficiency (CVID) characterized by recurrent infections, autoimmunity and immunoglobulin deficiency. Pedigree analysis revealed incomplete penetrance of the disease causing mutation in 5 of the 6 analyzed families. While patients showed a severe phenotype including hypogammaglobulinemia, chronic infections and cytopenias, other carriers of the same mutation were unaffected except for slightly perturbed immunoglobulin levels indicating the existence of other factors influencing the penetrance of these mutations.
Methods
To identify genetic factors associated with complete penetrance of dominant NFKB1 mutations, whole exome sequencing was carried out using DNA extracted from blood samples derived from two patients and their families. Sequencing data of two patients and X unaffected carriers of the same NFKB1 mutations (p.R157X and p.I47fsX2) were then screened in silico for single nucleotide variations, small insertions and deletions present in modulators of immune responses in general and the NFκB pathway in particular, employing lists generated based on publicly available data on gene interactions (including e.g. data of the KEGG, and STRING databases).
Results
We detected no deleterious mutations in known modifier genes such as IL10, IL1B, IL6, CCR5, CCL5, RANTES, TGFB1 and others. But strikingly both patients harbored two polymorphisms (g.797C>A, Gly54Asp, Gly57Glu) in the Mannose Binding Lectin 2 (MBL2) gene that were previously reported as disease causing mutations in patients with primary immunodeficiency. These polymorphisms lead to reduced MBL2 expression and are linked with high susceptibility to infections. We hypothesize that low MBL2 expression in an NFKB1 haploinsufficient background may promote disease penetrance or increase the predisposition to infections.
Conclusion
Our combined next-generation sequencing and bioinformatics analyses approach identified MBL2 as an interesting candidate factor whose deficient expression may influence the penetrance of NFKB1 loss-of-function mutations. Further analysis of greater cohorts is needed to reinforce the role of MBL2 in the pathogenesis of NFKB1 haploinsufficiency.

Disclosures
No relevant conflicts of interest to declare.
Loss of NFAT2 Leads to an Acceleration of Clonal Evolution in CLL
Authors: Müller David J , Märklin Melanie , Kanz Lothar , Mueller Martin R , Wirths Stefan 
Published: October 2019

Abstract
Chronic lymphocytic leukemia (CLL) may be defined as a clonal expansion of mature B cells with stereotypic B cell receptors (BCR). Somatic hypermutation of the BCR heavy chains (IGVH) defines subgroups with different prognosis. In up to 10% of patients Richter's transformation to a high grade lymphoma with a mostly dismal prognosis is observed. The TCL1 transgenic mouse is a well accepted model of human CLL. Upon B cell-specific knock out of NFAT2, TCL1 mice develop a disease resembling Richter's syndrome with a significantly more aggressive disease phenotype. While TCL1 B cells exhibit tonic anergic BCR signaling characteristic of human CLL, loss of NFAT2 leads to readily activated BCRs indicating different BCR usage with altered downstream signaling.
Here, we analyzed BCR usage in C57BL/6 wildtype, TCL1, and TCL1-NFAT2-/-mice. Splenocyte cDNA of the respective animals at an age of 7 months was amplified by multiplex PCR covering known heavy chain VDJ alleles. The PCR products were subsequently cloned into standard bacterial plasmids and subjected to conventional DNA sequencing. In addition, PCR products were analyzed by next-generation-sequencing (NGS). For data analysis, the IMGT/HightV-QUEST online tool was applied.
A very diverse and polyclonal BCR usage was found in wild type mice with more than 4000 different clones identified. Although TCL1 mice at 7 months of age exhibit all features of CLL, their BCR usage as analyzed by unambiguously identified VDJ recombination was still polyclonal. Only with respect to VH usage, TCL1 mice were found to use a more limited set of V alleles compared to wild type mice. Loss of NFAT2 by conditional knockout on the contrary, lead to an oligoclonal usage of VDJ recombination, to a further limitation of V alleles, and also to the usage of identical VDJ recombination in different mice with high frequency - indicating BCR selection either by antigen or self-signaling in these mice. Further analyses by sequencing of hypervariable regions showed preferential usage of mutated BCRs in TCL1 mice and of unmutated BCRs in NFAT2 ko mice reflecting benign and aggressive disease in humans.
In summary, the loss of NFAT2 signaling in CLL prefers the selection of unmutated BCRs as well as the preferential usage of certain VDJ recombinations which results in the accelerated development of true oligoclonal disease.

Disclosures
No relevant conflicts of interest to declare.
Anti-FcRH5/CD3 T Cell Dependent Bispecific Antibody (TDB) for the Treatment of Multiple Myeloma
Authors: Li Ji , Stagg Nicola , Johnston Jennifer , Harris Michael , Menzies Sam , DiCara Danielle , Clark Vanessa , Cook Ryan , Slaga Dionysos , Nakamura Rin , Luke McCarty , Sukumaran Siddharth , Luis Elizabeth , Ye Zhengmao , Sumiyoshi Teiko , Danilenko Dimitry , Lee Genee , Totpal Klara , Ellerman Diego , Hötzel Isidro , James John , Junttila Teemu T 
Published: October 2019

Abstract
Bispecific antibodies that retarget cytotoxic T cell activity to kill cancer cells are currently under clinical evaluation. However, the molecular mechanism for how CD3-bispecific antibodies 'trigger' intracellular T cell signaling is not known. We demonstrate that bispecific antibodies invoke an equivalent biophysical mechanism of TCR triggering as that observed for the TCR/pMHC interaction, including target clustering and exclusion of CD45 phosphatase from the synapse. The dimensions of the target molecule play a key role in the efficiency of the synapse formation. However, we demonstrate that rational epitope selection can overcome the spatial inhibition caused by target molecules with a large extracellular domain and result in efficient synapse formation and highly potent T cell triggering. With this insight, we developed a novel T-cell dependent bispecific (TDB) antibody, anti-FcRH5/CD3 TDB, targeting the B cell lineage marker FcRH5 for multiple myeloma. Anti-FcRH5/CD3 TDB demonstrated cytotoxicity against human plasma cells and patient derived myeloma tumor cells at picomolar doses. Very low target expression level is sufficient to induce anti-FcRH5/CD3 TDB mediated killing, indicating broad activity in multiple myeloma where the prevalence of FcRH5 expression is 100%. In primates, anti-FcRH5/CD3 treatment resulted in complete depletion of tissue B cells and bone marrow plasma cells. Anti-FcRH5/CD3 TDB induces immunosuppressive feedback signaling, including PD1 up-regulation, which can be overcome by PD-L1 antibodies. These data demonstrate the potential for the anti-FcRH5/CD3 TDB, alone or in combination with inhibition of PD1/PDL1 signaling in the treatment of multiple myeloma and other B-cell malignancies.

Disclosures
Li: Genentech: Employment. Stagg:Genentech: Employment. Johnston:Genentech: Employment. DiCara:Genentech: Employment. Clark:Genentech: Employment. Cook:Genentech: Employment. Slaga:Genentech: Employment. Nakamura:Genentech: Employment. Luke:Genentech: Employment. Sukumaran:Genentech: Employment. Luis:Genentech: Employment. Ye:Genentech: Employment. Sumiyoshi:Genentech: Employment. Danilenko:Genentech: Employment. Lee:Genentech: Employment. Totpal:Genentech: Employment. Ellerman:Genentech: Employment. Hötzel:Genentech: Employment. Junttila:Genentech: Employment.
LIGHT (TNFSF14) Increases Survival and Proliferation of Human Bone Marrow-Derived Mesenchymal Stem Cells Via LTβR
Authors: Kim Hawk , Heo Sook-Kyoung , Noh Eui-Kyu , Gwon Gi-Dong , Jo Jae-Cheol , Choi Yunsuk , Park Jinny 
Published: October 2019

Abstract
Background: LIGHT (Homologous to lymphotoxins, shows inducible expression, and competes with herpes simplex virus glycoprotein D for herpes virus entry mediator (HVEM)/tumor necrosis factor (TNF)-related 2, HVEM-L, TNFSF14, or CD258) is a member of TNF superfamily. It is expressed as a homotrimer on activated T cells and also on immune dendritic cells, and has three receptors such as HVEM, LT-¥â receptor (LT-¥âR) and decoy receptor 3 (DcR3). So far, three receptors with distinct cellular expression patterns are described to interact with LIGHT. Follicular DCs and stromal cells bind LIGHT through LT-¥âR. We monitored the effects of LIGHT on human bone marrow-derived mesenchymal stem cells (BM-MSCs).
Methods: At first, we checked negative and positive differentiation markers of BM-MSCs. After rhLIGHT treatment, we monitored cell count, viability, proliferation, and cell cycle distribution. Also, PDGF and TGF¥â production by rhLIGHT were examined by ELISA, and biological mechanism were checked by immunoblotting through the rhLIGHT treatment.
Results: FACS analysis result showed that LT-¥âR receptor is expressed in human BM-MSCs, but not HVEM indicating that LIGHT binds only LT-¥âR in human BM-MSCs. (Fig. 1A/B). rhLIGHT and LT-¥âR interaction increased cell numbers in BM-MSCs using an inverted microscope for cell number changes. Cell numbers by rhLIGHT enhanced dose-dependently and time-dependently (Fig. 2 A/B). Cell viability and the expression of p-AKT, Bcl-2 and Bcl-xL by rhLIGHT were significantly increased in BM-MSCs and rhLIGHT-induced IkB-¥á degradation activated NF-kB signal (Fig. 3A/B). rhLIGHT increased cell proliferation by increasing S/G2/M phase in BM-MSCs (Fig. 3C and 4D). And cell cycle regulatory proteins were enhanced by rhLIGHT in BM-MSC including cyclin B1, D1, D3, E, and cyclin dependent kinase (CDK) 1 and 2 while CDK inhibitor, p27 was decreased by rhLIGHT treatment (Fig. 3E). Moreover, rhLIGHT-induced PDGF and TGF¥â production by STAT3 and smad3 activation accelerated BM-MSCs proliferation. And we also confirmed differentiation potential of rhLIGHT on BM-MSCs by the staining for adipogenesis (Oil Red O staining), chondrogenesis (Alcian blue staining) and Osteogenesis (Alizarin red Staining).
Conclusion: LIGHT and LT-¥âR interaction increases survival and proliferation of human BM-MSCs by activation of survival proteins, anti-apoptotic proteins, CDKs and cyclins. Moreover, LIGHT-induced STAT-3 and smad-3 activation causes PDGF and TGF-¥â production, and they enhance LIGHT signals in human BM-MSCs. We proposed the pathway of LIGHT and LT-¥âR interaction in human BM-MSCs. Therefore, LIGHT may play an important role for therapy of stem cells, and contribute to modification of MSCs.
Figure 1 Figure 1. Figure 2 Figure 2. Figure 3 Figure 3.

Disclosures
No relevant conflicts of interest to declare.
Lower Incidence of Acute Gvhd Is Associated with the Rapid Recovery of CD4+CD25+CD45RA+ Regulatory T Cells in Patients Who Received Haploidentical Allografts from NIMA-Mismatched Donors: A Retrospective (development) and Prospective (validation) Cohort-Based Study
Authors: Wang Yu , Zhao Xiang-Yu , Xu Lan-Ping , Han Wei , Chen Huan , Wang Feng-Rong , Mo Xiao-Dong , Zhang Yuan-Yuan , Zhao Xiaosu , Kong Yuan , Liu Kaiyan , Huang Xiao-Jun , Chang Ying-Jun 
Published: October 2019

Abstract
To investigate the effects of non-inherited maternal antigen (NIMA) on clinical outcomes and immune recovery, especially of regulatory T cells (Tregs), in patients who underwent unmanipulated haploidentical transplantation.A retrospective cohort (n=57) and a prospective cohort (n=88) were included. Reconstitution of immune subsets, including Tregs, was determined using multicolor flow cytometry. In the retrospective cohort, the cumulative incidence of grades II-IV acute GVHD in patients with NIMA-mismatched donors was significantly lower than that of cases with NIPA-mismatched donors (14.8% vs. 43.30%, P=0.018). Patients with higher percentages of CD4+CD25+CD45RA+ T cells (naive Tregs) within CD4+ T cells recovered on day 30 (≥1.55%) experienced a significantly lower incidence of grades II-IV acute GVHD than that of cases with lower percentages of naive Tregs (<1.55%) (13.8% vs. 46.4%, P=0.010). Multivariate analysis showed that NIMA mismatch and the percentages of naive Tregs were associated with the incidence of grades II-IV acute GVHD [P=0.050, and 0.031, respectively]. In the prospective cohort, the association of NIMA mismatch [HR=0.365, 95%CI, 0.169-0.786, P=0.010] or higher percentages of naive Tregs recovered on day 30 (≥1.55%) [HR=0.114, 95%CI, 0.027-0.479, P=0.003] with a lower cumulative incidence of grades II-IV acute GVHD was further demonstrated. No effects of NIMA mismatch on chronic GVHD, transplant-related mortality, relapse, disease-free survival, or overall survival were found. Our results confirmed the role of NIMA mismatch in acute GVHD and provided the first demonstration, based on clinical data, that recovered Tregs may be involved in the effects of NIMA on acute GVHD in a haploidentical transplant setting.

Disclosures
No relevant conflicts of interest to declare.
Characteristics of Early Recovery of Double Negative T-Cell Subpopulations after Haploidentical Hematopoietic Stem Cell Transplantation
Authors: Liu Jiangying , Bian Zhilei , Fu Qiang , Chang Ying-Jun , Huang Xiao-Jun 
Published: October 2019

Abstract
Background: Immune recovery is recognized to be crucial for decreasing morbidity of the subsequent complications in patients who underwent alloHSCT. We recently compared the recoveries of T-cell subpopulations between the haploHSCT recipients with and without EBV-DNAemia and found that lower CD4/CD8 double negative T-cell (DNT) counts were significantly correlated with the increased cumulative incidence of EBV reactivation after transplantation. Multivariate analysis further showed that hampered recovery of DNT cells was an independent risk factor to predict transplant-related mortality. Based on the expression of αβ or γδ T-cell receptors, or NK cell markers, several subpopulations of DNT cells have been identified, e.g. TCR-γ/δ+DNT, TCR-α/β+DNT, NKT cells, and other cells that are incompletely characterized. Early recovery levels of DNT subsets may correlate to the occurrence of HSCT complications. Notably, reconstitution kinetics of these special T-cell subpopulations post-HSCT remains unknown.
Method: A prospective study was conducted to observe the comprehensive characteristics of DNT recovery. Fifteen healthy donors and 60 consecutive patients who were diagnosed as hematological malignancy and underwent haploHSCT at our institute were included. Immnophenotyping analysis was performed to determine the recovery levels of DNT and its subpopulations including α/β+DNT, γ/δ+DNT (Vδ1+ and Vδ2+), and CD56+DNT at 30, 60, and 90 days after transplantation.
Result: Percentages of total DNT, α/β+DNT, and γ/δ+DNT in recipients on day 30 were comparable with those of health donors (defined as normal range). While the percentage of total DNT significantly decreased on day 60 and day 90, percentages of α/β+DNT and γ/δ+DNT were quite stable within the 3 months after haploHSCT. Interestingly, ratio of Vδ1+/ Vδ2+ on day 30 was similar with the normal control, which was significantly reversed on day 60 and day 90. In contrast, the percentage of CD56+DNT on day 30 was 3-fold higher than the normal range, which dropped down to the normal level at 60 and 90 days. In term of the absolute numbers, all detected DNT subpopulations were hampered at 30 days post-transplantation. Only the counts of Vδ1+DNT achieved the normal level at 60 and 90 days whereas numbers of other DNT subpopulations were continuously lower during the evaluation period. The pace of Vδ2+DNT recovery was the slowest compared with others.
Conclusion: Our observations give a picture of the dynamic recovery of DNT subpopulations and contribute to comprehensively understand the characteristics of immune recovery after HaploHSCT. The clinical impact of DNT-subpopulations recovery will be evaluated in the ongoing prospective study. These findings may facilitate the intervention strategies to improve the general outcome of haploHSCT recipients.

Disclosures
No relevant conflicts of interest to declare.
A Novel BRD4 Inhibitor CA2 Suppresses MM Cell Proliferation in an Orthotopic Myeloma Mouse Model
Authors: Imayoshi Natsuki , Yoshioka Makoto , Nakata Susumu , Chauhan Jay , Kado Yoko , Toda Yuki , Fletcher Steven , Strovel Jeffrey , Takata Kazuyuki , Ashihara Eishi 
Published: October 2019

Abstract
We previously demonstrated that a novel BRD4 inhibitor, CA2, suppressed MM cell proliferation in vitro in the 57th ASH Annual Meeting. In the present study, we investigated the inhibitory mechanisms of CA2 on myeloma cell proliferation in vitro and in vivo. CA2 has a unique quinolinone core and inhibited the proliferation of MM cell lines, MM.1s, AMO-1, NCI-H929, OPM-2, and IM-9. As expected, CA2 suppressed the expression of c-MYC mRNA and protein in a dose- and time-dependent manner. In addition, CA2 decreased BRD4 binding to c-MYC promoters and c-MYC enhancers in a ChIP assay using IM-9 cells and an anti-BRD4 antibody (Figure 1). Flow cytometric Annexin-V/propodium iodide staining analyses demonstrated that CA2 induced apoptosis in MM cells. Taken together, CA2 may induce apoptosis in MM cells by inhibiting the binding of BRD4 to promoters and enhancers of c-MYC gene thereby suppressing c-MYC expression.
We next performed a pharmacokinetic study of CA2 in mice. Half-lives (t1/2) of CA2 following oral administration (50 mg/kg) and subcutaneous administration (10 mg/kg) were approximately 61 min and 139.8 min, respectively. The values of area under the curve for oral and subcutaneous administrations were 179,000 hr·ng/mL and 31,000 hr·ng/mL, respectively. Lastly, we assessed the in vivo effects of CA2 using orthotopic MM mouse model, where IM-9Luc cells were inoculated intravenously through the tail veins of SCID mice that had received 2 Gy of irradiation. The mice in the vehicle-control arm died of MM by approximately 30 days after transplantation; IM-9Luc cells engrafted in many organs including bone marrow of spine and femurs, kidneys, ovaries, and stomach. CA2 treatment consisting of 20 cycles of 100 mg/kg/day b.i.d. oral dosing resulted in reduced luminescence intensity of IM-9Luc cells and prolonged the survival of mice (mean 33.7 days) compared to the vehicle-control arm (mean 27.1 days; p=0.029, Figure 2). In conclusion, CA2 is a BRD4 inhibitor with a novel structure and could be a promising molecular targeting-agent against MM.

Disclosures
Yoshioka: ConverGene LLC: Employment. Kado:Japan Community Health Care Organization Kyoto Kuramaguchi Medical Center: Employment. Strovel:ConverGene LLC: Employment.