bluuebunny/crossref_metadata_2025_split huggingface dataset:
Performance of Free Light Chain Assays in Clinical Practice.
Authors: Katzmann Jerry A. , Dispenzieri Angela , Abraham Roshini S. , Kyle Robert A. 
Published: October 2019

The quantitative assay for free light chains (FLC) is a recently introduced commercial product (FREELITETM, The Binding Site, Ltd.) that has been reported to be sensitive and specific for detecting and monitoring free light chain diseases such as primary systemic amyloidosis (AL), light chain deposition disease (LCDD), non-secretory multiple myeloma (NSMM), and LCMM (light chain multiple myeloma). We have prospectively evaluated the test performance in clinical practice.
Results : In 2003, our Clinical Laboratory received samples for FLC assays from 1020 Mayo Clinic patients. The majority of these patients (88%) had plasma cell disorders (PCD). All 120 patients who did not have PCD had normal K/L FLC ratios. Among these, 52 had non-AL amyloidosis: localized amyloid (23), hereditary (16), senile (6), secondary (3), and amyloid of unknown type (4). The 68 other patients who did not have a PCD were being tested because of peripheral neuropathy, rule out AL, anemia, proteinuria, lymphoproliferative disease, and a number of other indications with small numbers of patients (n=13). Among the monoclonal gammopathy patients were 330 with MM, 269 AL, 115 MGUS, 72 SMM, 22 plasmacytomas, 20 NSMM, 9 macroglobulinemia, 7 LCDD, and a variety of other diagnoses with smaller numbers of patients. The sensitivity of the K/L FLC ratio was 100% in LCDD (7/7) and 70% in NSMM (14/20). The 6 NSMM patients with normal K/L FLC ratios had all been treated with SCT and 5 of the 6 had achieved hematologic remission by bone marrow plasma cells. Among the 110 AL patients who had not been previously treated and who had a FLC assay performed within 120 days of diagnosis, the sensitivity of the K/L FLC ratio was 92% compared to 71% for serum IFE and 84% for urine IFE. Using all 3 assays, there was 99.1% (109/110) sensitivity for detecting monoclonal light chain in AL.
Conclusion : The performance of the FLC assay in this prospective analysis matches the results from published retrospective validation studies.
Diagnostic Imaging of Multiple Myeloma - FDG PET and MRI Complementary for Tracking Short Vs. Long Term Tumor Response.
Authors: Walker Ronald , Jones-Jackson L. , Rasmussen Eric , Miceli M. , Anaissie Elias , Epstein Joshua , Tricot Guido , Shaughnessy John , Barlogie Bart 
Published: October 2019

Abstract
Multiple myeloma results in both diffuse infiltration and/or focal lesions (FL) of the marrow. MRI detects FL long before X-ray. We routinely perform MRI of skull, entire spine, pelvis, sternum and shoulders. MRI-defined number of FL (#FL) is an adverse prognostic variable for both EFS and OS for patients treated according to Total Therapy 2, 2nd in importance only to cytogenetic abnormalities (CA) (univariate and Cox multivariate models using standard prognostic factors, CA, and #FL). Of these patients who also had FDG PET (PET), baseline analysis revealed the same axial skeleton FL as on MRI (r=0.47, p<0.001). Another 268 FL in long bones and 11 sites of extramedullary tumor were seen with PET due to MRI’s restricted field of view. Serial MRI and PET in 59 patients with FL on MRI (Fig. 1) and 123 patients with hyperintense MRI signal on STIR (no FL) revealed that patients without FL normalize MRI and PET scans synchronously with clinical (M-protein and bone marrow) response (nCR/CR) (Fig. 2a). However, in patients with FL, PET normalization follows nCR/CR very closely, while regression of MRI-FL (MRI-CR) lags in proportion to MRI #FL (Fig. 2b). We conclude PET is superior to MRI for detection of extent of disease and monitoring treatment response. However, PET-negative MRI-FL usually contain viable myeloma cells on biopsy which eventually progress. Thus, long term serial MRI is superior to PET in assessing completeness of response.
Targeting Mitochondrial Factor Smac/DIABLO as Therapy for Multiple Myeloma (MM).
Authors: Chauhan Dharminder , Li Guilan , Hideshima Teru , Podar Klaus , Tassone Pierfrancesco , Neri Paola E. , Mitsiades Constantine , Mitsiades Nicholas , Raje Noopur , Kumar Shaji , Schlossman Robert , Sharma Sushil , Munshi Nikhil , Zawel Leigh , Anderson Kenneth C. 
Published: October 2019

Abstract
Mitochondrial signaling is an important component of chemotherapy-induced apoptosis in cancer cells. Stress-inducers trigger the following sequence of events: Smac (second mitochondria-derived activator of caspases)/ DIABLO is released from the mitochondrial inner membrane to cytosol; activated/cytosolic Smac binds to XIAP, a protein belonging to the family of IAPs (Inhibitors of apoptosis proteins), thereby abrogating its inhibitory effects on caspase-9 and resulting in activation of caspase-9-mediated apoptotic cascade. IAPs play a key role in regulating the apoptotic signaling, and overexpression of IAPs confer chemoresistance in various tumor types. Here we show that a low molecular weight synthetic Smac mimetic (Novartis) by virtue of its ability to bind and inhibit IAPs, block the growth of MM cell lines, including those resistant to conventional agents including dexamethasone, melphalan or doxorubicin. Examination of purified patient MM cells demonstrated similar results. In contrast, no significant toxicity of Smac mimetic was noted against normal peripheral blood mononuclear cells. Moreover, Smac mimetic also did not affect viability of MM patient-derived bone marrow stromal cells (BMSCs). Treatment of MM cells overexpressing anti-apoptotic protein Bcl2 with Smac mimetic significantly decreases their viability. Importantly, combined treatment of MM cells with Smac mimetic and Tumor necrosis factor-related apoptosis inducing ligand (TRAIL, Apo-2 ligand) trigger synergistic anti-MM activity, without significant toxicity in normal cells. Finally, Smac mimetic enhances the anti-MM activity of conventional Melphalan. Collectively, these findings provide the rationale for clinical evaluation of Smac mimetic alone and in combination with Melphalan or TRAIL, to enhance MM cell killing, overcome drug-resistance, and improve patient outcome in MM.
Identification of a Novel Toll-Like Receptor-Independent Immunoadjuvant Pathway That Depends upon Programmed Cell Death.
Authors: Hoebe Kasper , Janssen Edith , Beutler Bruce 
Published: October 2019

Abstract
Molecules of microbial origin, and synthetic derivatives of these molecules, have long been used for their immuno-adjuvant effect, and as the key sensors of microbial infection, Toll-like receptors (TLRs) are thought to be essential for adjuvanticity. To the contrary, we now demonstrate the existence of a robust, TLR-independent pathway for adjuvant effect: one that is actually far stronger than the TLR-dependent pathway. Activation of Toll-like receptors (TLRs) and the subsequent production of cytokines such as type I interferon leads to the maturation of dendritic cells (DCs) with upregulation of MHC molecules and costimulatory molecules such as CD40, CD80 and CD86, allowing for optimal interaction between DCs and T-cells. We have determined that TLR signal transduction is minimally dependent upon two adapter proteins, MyD88 and TRIF. In compound homozygous mutant (DKO) mice that lack functional MyD88 and TRIF, there is complete abrogation of all TLR signaling. Such animals therefore comprise a unique model with which to study TLR-independent immune responses. We have now used DKO mice to determine whether an adaptive immune response can be obtained in the absence of TLR signaling. As expected, adjuvanticity obtained via “classical” microbial adjuvants such as complete Freund’s adjuvant or LPS was completely absent in DKO mice. However, subcutaneous administration of syngeneic murine cells expressing ovalbumin and rendered apoptotic by exposure to ultraviolet light resulted in a strong T-cell response in vivo, with impressive production of interferon-g by CD8+ cells and efficient killing of EL-4 cells that expressed CD8-specific OVA peptides, both in wildtype and DKO mice. Adjuvanticity was observed only in the context of apoptosis, in that living cells, not exposed to ultraviolet light before injection, induced little or no response. Moreover, the mixture of the protein antigen with apoptotic cells was insufficient to induce an adaptive immune response; rather, only cells that expressed the protein prior to induction of apoptosis were stimulatory. These results indicate the existence of a specific, cell death-dependent mechanism for adjuvanticity that is TLR-independent and induced by endogenous molecules. We propose that this new adjuvant pathway is of fundamental importance to immune responses at large. We believe that it is required for initiation of the adaptive immune response witnessed in the context of allograft rejection, graft-versus-host disease, and autoimmune diseases as well.
Reduced-Intensity Conditioning (RIC) Followed by Allogeneic Transplantation in Relapsed Lymphomas: Evidence for Graft-Versus-Lymphoma Effect in Low and High Grade Histologies, but Not in Hodgkin Disease.
Authors: Corradini Paolo , Dodero Anna , Bregni Marco , Olivieri Attilio , Farina Lucia , Rizzo Elena , Milani Raffaella , Scime Rosanna , Rambaldi Alessandro , Bonifazi Francesca , Patriarca Francesca , Locasciulli Anna , Bacigalupo Andrea , Gianni Alessandro Massimo , Tarella Corrado 
Published: October 2019

Abstract
Reduced intensity conditioning (RIC) followed by allogeneic stem cell transplantation (SCT) is associated with durable engraftment, low transplant-related mortality (TRM) and clinical remissions in hematologic malignancies. In addition, RIC followed by allograft is feasible also in patients who had failed previous autologous (auto) SCT. Here, we report the outcome of 118 lymphoma pts receiving RIC and allogeneic SCT from HLA-identical sibling donors. Histologic types included in the study were: chronic lymphocytic leukemia/small lymphocytic lymphoma (CLL/SLL; n=26), low-grade non-Hodgkin lymphoma (LG-NHL; n=24, 21 follicular NHL), high-grade NHL (HG-NHL, B cell n=28, T cell n=17), Hodgkin disease (HD; n=23). Median age was 50 years (range: 20–69). 47% of pts had refractory disease (46% CLL/SLL; 33% LG-NHL; 44% HG-NHL; 56% HD) and 48% have failed an auto SCT (15% CLL/SLL; 33% LG-NHL; 55% HG-NHL; 82% HD). All pts received debulkying chemotherapy followed by the same RIC regimen with thiotepa (10 mg/kg), fludarabine (60 mg/ms) and cyclophosphamide (60 mg/kg). GVHD prophylaxis consisted of cyclosporine A and short-course methotrexate. All pts engrafted. De-novo acute GVHD developed in 43% of pts and chronic GVHD in 45%. The 2-year TRM rate was 13% and was not influenced by previous auto SCT or hystological type. At a median follow-up of 20 months (range, 6–62 months), 91 pts (77%) are alive (n=69 CR, n=8 PR, n=14 with disease) and 27 died (23%) for disease (n=17) or TRM (n=10). The 2 years PFS rates for CLL/SLL, LG-NHL, HG-NHL, HD were 63%, 78%, 64%, 16%, respectively. Chemorefractory disease at time of SCT was associated with a worse 2-year PFS compared to chemosensitive disease (41% versus 74%, p<0.001). There was a trend toward lower relapse risk for pts developing acute or chronic GVHD (37% versus 50%, p=0.2). The 2-year OS rates for CLL/SLL, LG-NHL, HG-NHL, HD were 77%, 90%, 76%, 56%, respectively. Chemosensitive disease had significant impact on OS (84% 2-year OS compared with 64% for chemorefractory patients; p< 0.007) as well the occurrence of chronic GVHD (87% versus 57%; p<0.0059). Interestingly, previous auto SCT and age > 55 years did not influence OS. Among pts in clinical CR, 24 were suitable for PCR monitoring of residual disease and 17 pts (71%) are in continuous molecular remission. Our study indicates: 1) low TRM in a multicenter phase II study; 2) low risk of relapse and good survival rates in indolent lymphomas; 3) encouraging results in HG-NHL 4) disappointing results in HD
Regulation of CD95 in EBV infected B-Cells.
Authors: Le Clorennec Christophe , Youlyouz-Marfak Ibtissam , Adriaenssens Eric , Coll Jean , Bordessoule Dominique , Bornkamm Georg W. , Feuillard Jean 
Published: October 2019

Abstract
Expression of CD95 is regulated by various agents such as TNFa, IFNg, or Daunorubicine, able to induce the transcriptional factors NF-kB, STAT1 and p53 respectively. The CD95 promoter is known to harbour binding sites for these factors. These factors are modulated both in expression and activity during EBV infection of B-cells. EBV is known to induce CD95 expression. The objective of this work was to understand the contribution of p53, STAT1 and NF-kB in the regulation of CD95 expression by EBV.
To control the activity of NF-kB, STAT1, p53 and LMP1, we have developed and used various episomal inducible vectors. One of the characteristics of these vectors is that the gene of interest is expressed from a bidirectional doxycycline regulatable promoter that allowing simultaneous expression of truncated NGF receptor, used as a surrogate marker of inducibility. We have subcloned the following cDNAs into these vectors: dominant negative IkBa, active (STAT1a) and dominant inactive natural isoform of STAT1 (STAT1b), p53, a dominant negative mutant of LMP1 (LMP1CT) and LMP1 wild type. After stable transfection of an LCL, induction of the cDNA of interest was performed with doxycycline for 24H. Positive cells for NGFR were purified using magnetic beads and were additionally treated with TNFa, IFNg or Fludarabine for 24H. To control the latency III program, we have used the EREB2-5 cell line harboring an estrogen-regulatable EBNA2 gene. The function of CD95 was studied by quantifying apoptosis induction after CD95 cross linking with a specific antibody.
We found that induction of latency III program in EREB 2-5 cells was associated with an increase of both p53 and CD95 expression.
Inhibition of LMP1 by LMP1CT decreased the expression of CD95. CD95 expression was hardly regulated by p53. Inhibition of both NF-kB and STAT1 decreased CD95 expression. Only IFNg was able to increase expression of CD95. Finally, we show that induction of CD95 expression by EBV render the infected B-cell sensitive to the induction of CD95-mediated apoptosis. Our results suggest that the latencyc III program of EBV sensitize the infected B-cells to CD95-mediated apoptosis via STAT1 and NF-kB activated by LMP1. In the view of the virus/host equilibrium, this process may render EBV-infected B-cells in latency III susceptible to elimination by the immune system.
The SCL Transcription Factor Supports Erythroid Cell Survival and Differentiation Downstream of the Erythropoietin Receptor.
Authors: Lahlil Rachid , Martin Richard , Aplan Peter D. , Begley C. Glenn , Damen Jacqueline E. , Krystal Gerald , Hoang Trang 
Published: October 2019

Abstract
Erythroid cell development critically depends on the SCL/Tal1 transcription factor and on erythropoietin signalling. In the present study, we have taken several approaches to show that the two genes operate within the same pathway to consolidate the erythroid lineage. Signaling through the erythropoietin receptor (EpoR) upregulates SCL protein levels in a clonal cell line (TF-1) in vitro, and in murine fetal liver cells in vivo, when Epor−/− cells were compared to those of wild type littermates at E12.5. In addition, we provide functional evidence for a linear pathway from EpoR to SCL that regulates erythropoiesis. Interfering with SCL induction or SCL function prevents the anti-apoptotic effect of Epo in TF-1 cells and conversely, ectopic SCL expression is sufficient to substitute for Epo to transiently maintain cell survival. In vivo, SCL gain of function complements the cellular defects in Epor−/− embryos to support cell survival and maturation during primitive and definitive erythropoiesis, as assessed by cellular and histological analyses of Epor−/− SCLtg embryos. Moreover, several erythroid specific genes that are decreased in Epor−/− embryos are rescued by the SCL transgene including glycophorinA, bH1 and bmaj globin, providing molecular confirmation of the functional and genetic interaction between Epor and SCL. Conversely, erythropoiesis becomes deficient in compound Epor+/−SCL+/− heterozygote mice, indicating that the genetic interaction between EpoR and SCL is synthetic. Finally, using EpoR mutants that harbour well defined signalling deficiencies, combined with gain and loss of function approaches for specific kinases, we identify MAPK as the major signal transduction pathway downstream of EpoR that upregulates SCL function, necessary for erythroid cell survival and differentiation. Taken together, our observations are consistent with the view that cytokines can influence cell fate by altering the dosage of lineage transcriptional regulators.
Adult T Cell Leukemia/Lymphoma (ATLL) Is a Malignant Counterpart of Regulatory T Cells (Treg).
Authors: Kohno Tomoko , Yamada Yasuaki , Akamatsu Norihiko , Kamihira Shimeru , Tomonaga Masao , Matsuyama Tosifumi 
Published: October 2019

Abstract
Adult T cell leukemia/lymphoma (ATLL) is a lymphoproliferative disorder caused by a retrovirus, human T-lymphotropic virus type 1 (HTLV-1). ATLL is subclassified into four subtypes, smoldering, chronic, acute, and lymphoma. The acute type progresses rapidly and is usually resistant to conventional chemotherapy. In contrast, the chronic type shows an indolent clinical course and the patients survive for several years, even without chemotherapy. Irrespective of the subtypes, however, ATLL patients are in a severely immune-suppressed condition and can easily acquire opportunistic infections such as Pneumocystis Carinii pneumonitis. Suppression of cell-mediated immunity has also been reported in HTLV-1 carriers. Although ATLL cells show the activated helper/inducer T-cell phenotypes, CD4+ and CD25+, they exhibit strong immune-suppressive activity in vitro. The recent notion of CD4+ CD25+ regulatory T cells (Treg) prompted us to investigate the origin of ATLL cells from the standpoint of Treg. Forkhead/winged helix transcription factor (Foxp3) is a functional marker of Treg, which plays a central role in their generation. There are other marker molecules for Treg, including glucocorticoid-induced TNFR family-related protein (GITR) and the chemokine receptors CCR4 and CCR8.
In the present study, we examined primary ATLL cells from 48 patients: 36 patients with acute type and 12 patients with chronic type. We also examined ATLL cell lines, HTLV-1-infected T-cell lines and peripheral blood mononuclear cells (PBMC) from healthy adults as control cells. We used RT-PCR for detection of Foxp3, GITR, CCR4, and CCR8 mRNA expression. Foxp3 and/or GITR mRNA were detected in over 90% of the patients, and 50% of the patients expressed both. There was no difference between subtypes. In contrast, Foxp3 and GITR mRNA were scarcely detected in the PBMC from healthy adults. Furthermore, we confirmed GITR expression at the protein level by flow cytometry. CCR4 and CCR8 mRNA were also detected in almost all ATLL samples, at significantly higher levels than in the normal PBMC. Corresponding to the results of the primary cells, ATLL cell lines and HTLV-1-infected T-cell lines also expressed GITR mRNA, although HTLV-1-negative cell lines, Jurkat and Molt4, completely lack it. Next, we examined whether GITR affects ATLL cell proliferation using a GITR- expressing IL-2-dependent ATLL cell line, KK1. We found that GITR ligand induced proliferation of KK1 cells in an IL-2-negative condition. Thus, these results indicate the Treg origin of ATLL cells and show that GITR expression is possibly involved in the development of ATLL.
Junctional Adhesion Molecule-1 Is Involved in bFGF-Induced Angiogenesis In Vivo.
Authors: Cooke Vesselina G. , Naik Meghna U. , Skarnes William , Naik Ulhas P. 
Published: October 2019

Abstract
Neovascularization is a multistep process that occurs in the body in both physiological and pathological conditions. We have recently shown that Junctional Adhesion Molecule-1 (JAM-1), a member of the Ig superfamily of molecules, is involved in endothelial cell adhesion and migration, leading to angiogenesis. In quiescent endothelial cells, JAM-1 is located at the cell-cell junctions where it forms a complex with integrin αvβ3. Upon treatment of the cells with growth factors, such as bFGF, JAM-1 dissociates from its complex with αvβ3 and redistributes to the cell surface. Blockage of the extracellular domain of JAM-1 inhibits bFGF-induced endothelial cell morphology, proliferation and angiogenesis. Additionally, functional knock-down of JAM-1 using the RNAi technique in endothelial cells showed decreased adhesion and migration of these cells, indicating a possible role for JAM-1 in angiogenesis. In this report, we show that JAM-1 has an important role in bFGF-induced angiogenesis in vivo. Here we present for the first time the generation JAM-1 knock-out mice, using the gene trap strategy. We have successfully confirmed the JAM-1 −/− genotype via Southern, Northern, and Western blot analyses. JAM-1 −/− mice are viable and do not seem to have any external abnormalities, except that they appear to be smaller in size. Retinal fluorescein angiogram revealed no evidence for morphological defects in the vasculature of JAM-1 −/− mice. To evaluate the role of JAM-1 in angiogenesis, we performed an aortic ring assay with both wild type and JAM-1−/− mice. Mouse thoracic aortas were harvested, cross-sectioned into rings of 1-mm thickness, and cultured in a three-dimensional Matrigel supplied with 50 ng/ml bFGF. Vascular sproutings were counted every other day for a period of 7 days at which time they were stained with crystal violet and photographed. Aortic rings from WT mice treated with bFGF showed a 2.8-fold increase in microvessel growth, compared to WT controls with no supplementation of bFGF. In contrast, microvessel sproutings in bFGF treated aortic rings from JAM-1 −/− mice were no more than the vessels in the WT control mice. These results suggest that JAM-1 may be important for bFGF induced angiogenesis. To further confirm the role of JAM-1 in angiogenesis, WT and JAM-1 −/− mice were injected in their flank region with Matrigel containing 80 ng/ml bFGF and 60 U/ml heparin. Two weeks after injection, Matrigel plugs were excised, embedded in paraffin, and the presence of blood vessels was visualized by H&E staining. Matrigel plugs from control WT mice that were not treated with bFGF showed no vascularization, while bFGF supplied Matrigel plugs from WT mice showed a robust vessel growth. Interestingly, bFGF-treated Matrigel plugs form JAM-1−/− mice failed to produce any blood vessels. These ex vivo and in vivo studies using JAM-1−/− mice suggest that JAM-1 has a unique and essential role in bFGF-induced angiogenesis.