bluuebunny/crossref_metadata_2025_split huggingface dataset:
Endosomal Targeting Sequences from Non-Classical Antigen Presenting Molecules Can Direct Antigens into the MIIC and Other Antigen Processing Compartments.
Authors: Gansert Jennifer L. , Niazi Kayvon R. , Ochoa Maria T. , Sieling Peter A. , Modlin Robert L. 
Published: April 2020

Abstract
The ultimate goal of these studies is to enhance the development of MHC class II-restricted helper T cell responses that are required for effective and durable immunotherapy. In order to optimize the processing and presentation of MHC class II-restricted epitopes, we are using a differential targeting strategy that relies upon the unique endosomal targeting sequences from the individual isoforms of the CD1 family of non-classical antigen presenting molecules. Distinct endosomal compartments differ with respect to pH and resident proteases. MHC class II molecules can load antigen in each of the different endosomal compartments. This is in addition to the traditional antigen loading compartment, MIIC. Therefore, by forcing the expression of a tumor antigen across different endosomal compartments, we hope to maximize the number of successfully processed epitopes. To achieve this, we have developed a PCR-based strategy to link the leader, transmembrane, and endosomal targeting sequences from the four different human CD1 isoforms, CD1a-d, to DNA encoding for a surrogate antigen, green fluorescent protein (GFP). Using this strategy, we have demonstrated that fusion of individual CD1 tails to the sequence for GFP resulted in four distinct expression patterns in HELA cells transfected with these constructs. Using immunofluorescent staining and confocal laser microscopy, we determined the pattern of antigen expression in various endosomal compartments. GFP variably colocalized with recycling endosomes (ARF-6), early endosomes (CD71, transferrin receptor), late endosomes (CD63), and late endosomes/lysosmes/MIIC (CD107a, LAMP-1). Three of the four CD1 tails also resulted in co-expression of GFP with the MHC class II molecule, HLA-DR. The expression pattern for each CD1 tail is shown in Table 1. These experiments validate the feasibility of the CD1 targeting approach. Current studies are underway to apply this strategy to the leukemia antigen, WT1, using recently completed and sequenced WT1/CD1 fusion constructs.
ARF-6 CD71 CD63 CD107a CD1a −/+ − − − CD1b −/+ − + ++ CD1c + −/+ + ++ CD1d + −/+ − +
Table 1. Pattern of GFP/CD1 fusion expression in different endosomal compartments. Fusion of the endosomal targeting sequences from human CD1 molecules to the model antigen, GFP, results in different patterns of antigen expression within distinct endosomal compartments. The degree of colocalization between GFP and the indicated markers of endosomal compartments is represented as follows: (−), no colocalization; (−/+), infrequent colocalization; (+), frequent colocalization; (++), extensive colocalization.
Glutathione-S-Transferase π Expression Helps for Differential Diagnosis between Mantle Cell Lymphoma and Marginal Zone Lymphoma with t(11;14).
Authors: Rolland D. , Gazzo S. , Felman P. , Callet-Bauchu E. , Traverse-Glehen A. , Baseggio L. , Paret A. , Chapuis F. , Salles G. , Coiffier B. , Berger F. , Thieblemont C. 
Published: April 2020

Abstract
Glutathione-S-transferase π (GSTπ), whose gene is located in 11q13, is belonging to a complex multigene family of enzymes that may detoxify electrophilic xenobiotics. Recent gene expression profiling analysis showed that GSTπ transcript is belonging to the mantle cell lymphoma (MCL) signature and constitutes one of the most expressed mRNA in MCL*. Furthermore it has been reported by immunohistochemistry that overexpression of cyclin D1 in MCL was associated with a high level of GSTπ protein expression. We then looked for the specificity of a high level of GSTπ protein expression in MCL, and particularly when comparing to marginal zone lymphoma (MZL) with t(11;14). Material and Methods: Immunochemistry analysis of GSTπ and cyclinD1 expression was performed 111 non Hodgkin’s lymphoma including MCL, MZL, small lymphocytic lymphoma (SLL), follicular lymphoma (FL), and diffuse large B-cell lymphoma (DLBCL) cases with or without t(11;14). Results: A strong GSTπ expression in more than 50% of the cells was observed in 98% of the MCL. None of the MZL cases with t(11;14) expressed GSTπ, except one case that expressed a weak signal in less than 20% of the cells. None of the MZL without t(11;14) expressed GSTpi. Only two (13%) FL graded 3 according to OMS classification expressed GSTπ in less than 50% of the cells. A high GSTπ expression (&gt;50% cells stained) was detected in 29% of the DLBCL and 43% of the SLL.
* C. Thieblemont et al. Non-germinal B-cell lymphoma exhibit distinct genomic profiles allowing molecular diagnosis. Blood2004;103:2727–2737
n GST π expression &gt;50% GST π expression 5–50% π GST expression &lt;5% MCL with t(11;14) 25 23 0 2 MZL with t(11;14) 5 0 1(weak) 4 MZL without t(11;14) 31 0 1 (weak) 30 FL without t(11;14) 15 0 2 13 DLBCL without t(11;14) 14 4 1 9 SLL without t(11;14) 21 6 3 12
In conclusion, 98% of the MCL with t(11;14) expressed high level of GSTπ whereas none of our MZL with t(11;14) and high transcriptional cyclinD1 expression expressed GSTπ. This results may help for differential diagnosis between MCL and MZL with t(11;14) and suggest different transcriptional regulation of GSTπ in lymphoma subtypes with t(11;14), contributing possibly to the chemoresistant phenotype of MCL. Finally, this confirmed our previous work on MZL with t(11;14).
Campylobacter jejuni Infection in Intestinal Lymphoma: A Strong Association with Immunoproliferative Small Intestinal Disease.
Authors: Baginsky Lysa A. , Diss Tim C. , Ye Hongtao , Wren Brendan , Du Ming Qing , Isaacson Peter G. , Dogan Ahmet 
Published: April 2020

Abstract
Immunoproliferative small intestinal disease (IPSID) is a lymphoma of the mucosa-associated lymphoid tissue (MALT) that produces a characteristic truncated immunoglobulin α-heavy chain. Bacterial involvement is suspected in this disease, as some tumours resolve after antibiotic treatment and other MALT lymphomas have proven associations with infectious agents. A recent report implicated Campylobacter jejuni in the development of IPSID. In this study, we used a PCR based method to identify C. jejuni in paraffin embedded archival material of intestinal lymphomas. The cases studied included IPSID, other non-IPSID MALT lymphomas, mantle cell lymphomas (MCL), diffuse large B-cell lymphomas (DLBCL), follicular lymphomas (FL) and enteropathy-type T-cell lymphomas (ETTL) as well as inflammatory diseases such as Crohn’s disease and coeliac disease. Campylobacter specific 16S ribosomal DNA (16S rDNA) was amplified by PCR and the products were sequenced to confirm Campylobacter sequences and to classify Campylobacter species. The results are summarised in Table 1. Campylobacter 16S rDNA was amplified from 11/22 IPSID samples and from 12/121 other samples including 6/11 DLBCL. The sequence analysis showed that, in IPSID, the sequences were mostly specific for C. Jejuni whereas generic sequences consistent with several other Campylobacter species were obtained in other lymphomas. The IPSID cases originated from a very wide range of geographical locations suggesting that the association with C.jejuni was disease specific rather than environmental. Our results confirm a close association between C.jejuni infection and IPSID but also suggest that Campylobacter sequences can be found in other intestinal lymphomas, in particular DLBCL.
Table 1
Diagnosis 16S rDNA (VC6) C jejuni C jejuni or related Not sequenced IPSID 11/22 7 2 2 MCL 2/8 0 2 0 DLBCL 6/11 0 4 2 MALT 1/15 0 0 1 FL 0/13 0 0 0 ETTL 0/5 0 0 0 Crohn’s 0/12 0 0 0 Coeliac 1/13 0 0 1 Others 2/44 2 0 0     TOTAL 23/143 9 8 6
The Role of Sprouty Family Members in Hematopiesis in Zebrafish and Mammals.
Authors: Mendenhall Eric M. , Eckfeldt Craig E. , Ekker Stephen C. , Verfaillie Catherine M. 
Published: April 2020

Abstract
In an effort to identify novel regulators of hematopoiesis, we identified a Sprouty family member, SPRY1, significantly enriched (&gt;1.5 fold, p-value &lt; 0.03) in human hematopoietic stem cells (HSC) compared to more committed hematopoietic progenitor cells (HPC) using Affymetrix-based global gene expression profiling (see Eckfeldt et al. abstract). Predominantly FGF, but also VEGF, and EGF signaling induces expression of Sprouty family members (SPRY1-4). SPRY proteins form heterodimers with other Sprouty members along with homodimers. They and are intracellular antagonists and agonists of receptor tyrosine kinase signaling via the RAS/MAPK pathway. SPRY proteins are best characterized as antagonists of the FGF, EGF, and VEGF pathways. As part of a high-throughput functional analysis of the genes identified by human gene array in zebrafish, we evaluated the role of the only identified zebrafish homologue for Sprouty, SPRY4, in zebrafish hematopoiesis. Using morpholino (MO) antisense technology we knocked-down expression of zebrafish SPRY4 during early zebrafish development in a double transgenic GATA1:dsRed/Fli1:eGFP zebrafish line. The SPRY4 morpholino injected fish (SPRY4[superscript]MO[/superscript]) showed an almost complete loss of blood with normal vasculature, which was confirmed by injection of a second unique morpholino designed against SPRY4. [italics] In situ [/italics]hybridization of the hematopoietic markers SCL, GATA1 and Ikaros were all significantly reduced or absent at multiple developmental stages in the SPRY4[superscript]MO[/superscript] embryo. Together with the normal development of the vasculature, this data indicates SPRY4 acts early in hematopoiesis and after the specification of the hemangioblast. [italics] In situ[/italics] hybridization for SPRY4 demonstrated expression in the otic vesicle, tail bud and lateral plate mesoderm, the first site of hematopoietic stem cells. Coinjection of SPRY morpholino and low doses of VEGF morpholino did not rescue the SPRY blood phenotype, indicating that the blood phenotype is not likely through the VEGF pathway. Previous gain-of-function experiments injecting SPRY4 mRNA showed an expansion of hematopoietic progenitors (Furthauer et al. Development 2001), a phenotype we confirmed with overexpression of SPRY4 cDNA. Experiments are ongoing to overexpress human SPRY1 or zebrafish SPRY4 cDNAs in mammalian cells to determine if the hematopoietic phenotype correlates with the zebrafish data. Additional genes differentially expressed in human HSC and HPC evaluated via the high throughput zebrafish screen will also be presented. This is the first description of functional validation of a gene from a human microarray in zebrafish, an essential step in obtaining meaningful in vivo data from global gene profiling studies.
Orbital Non Hodgkin’s Lymphoma (NHL): A Large Monocentric Study of Initial Characteristics, Natural History and Prognostic Factors.
Authors: Jerome Meunier , Livia Lumbroso , Remy Dendale , Anne Vincent-Salomon , Bernard Asselain , Fariba Nemati , Driss Chaoui , Alain Fourquet , Laurence Desjardins , Corine Plancher , Didier Decaudin 
Published: April 2020

Abstract
Orbital NHL (ocular and/or adnexal involvement) represents less than 1% of all lymphomas but about half of malignant tumors of the eye and/or ocular adnexae. We therefore reviewed all patients treated at the Institut Curie between 1970 and 2003 for a NHL exhibiting initial orbital localization to define their initial characteristics, natural history and prognostic factors. Among 172 patients, 145 cases with completed datas were selected for the study. Pathological review according to the WHO classification showed 52 cases of MALT-type lymphoma (36%), 32 lymphoplasmocytic lymphomas (22%), 14 patients with lymphocytic lymphoma, 7 cases of follicular lymphoma, 13 unspecified low grade lymphomas [namely, 118 cases (82%) of low grade NHL], 22 patients with diffuse large B-cell lymphoma (15%), 2 cases of mantle cell lymphoma, 2 Burkitt’s lymphomas and one T-lymphoblastic lymphoma [namely, 27 cases (18%) of high-grade NHL]. Initial characteristics were: median age 66 years (range 3–96), sex ratio M/F 0.6, B symptoms 6%, PS≥2 in 4% of patients, stages III–IV 31.7%, bone marrow involvement 12%, elevated LDH in 18% and IPI 0-1/2/3/4-5 in 92/28/13 and 1 cases, respectively. Anatomic localizations were intra-orbital in 39 patients (27%), conjunctival in 38 (26%), eyelid in 9 cases, lachrymal in 8 and other in 8 cases. Treatment of selected patients consisted of abstention in 2 cases, surgical complete resection in 5 cases, mono or polychemotherapy alone (CT) in 4 cases, and radiotherapy alone in 98 cases (68%) or with CT in 36 cases (25%). With a median follow-up of 90 months (range 3–314), the 5-year relapse-free (RFS) and overall (OS) survivals were 64% and 79% for the low-grade NHL, and 43% and 50% for the high-grade NHL. Prognostic factors were determined for the 118 low-grade patients. In univariate analysis, age greater than 59 years, elevated IPI score and elevated LDH level were prognostic for lower RFS and OS. In multivariate analysis, age greater than 59 years was the only prognostic factor for both lower RFS and OS (Figure 1). In conclusion, with a median follow-up of 7.5 years, our large monocentric cohort of patients represents one of the most important series that defines the initial characteristics, natural history and prognostic factors of NHL with initial orbital localization.
Figure Figure
Long Term Results of the GELA Study, R-CHOP vs. CHOP in Elderly Patients with Diffuse Large B-Cell Lymphoma.
Authors: Coiffier Bertrand , Feugier Pierre , Sebban Catherine , Bouabdallah Reda , Delwail Vincent , Tilly Hervé , Gisselbrecht Christian , Bosly André , Salles Gilles , Reyes Felix 
Published: April 2020

Abstract
In 1998, the GELA (Groupe d’Etude des Lymphomes de l’Adulte) ran a randomized study to evaluate the benefit of the addition of rituximab to CHOP chemotherapy in elderly patients (60 to 80 years old) with diffuse large B-cell lymphoma. Early results were presented at ASH 2000 and published in NEJM (2002;346:235) with a median follow-up of 1 and 2 years, respectively. An update of these results is presented with a median follow-up of 5 years. 399 patients were included, 197 in the CHOP group and 202 in the R-CHOP group, 60% had a high risk disease according to the IPI score. Characteristics of the patients did not differ between both groups. Rituximab was given at 375 mg/m² the same day of CHOP for 8 cycles. Primary endpoint was event-free survival with events defined as progressive disease (PD) during or after treatment, relapse, institution of a new treatment, and death whatever the cause. Secondary endpoints were overall survival (OS) and progression-free survival (PFS) with progression defined as PD, relapse, or death from lymphoma or lymphoma treatment. Response rates for R-CHOP and CHOP were: CR/CRu 75% and 63%, PD 9% and 22%, and death during treatment 6% and 6%, respectively (P=.005), as previously published. With a median follow-up of 5 years, 106 events (52.5% of the patients) were observed in R-CHOP arm and 142 (72%) in CHOP arm: 19% and 31% PD or death during treatment, 5.5% and 4.5% institution of a new treatment, 20% and 34% relapses, 8% and 2.5% death in CR for R-CHOP and CHOP, respectively. Survival outcomes are presented in the table.
Table 1. 5-year survivals
R-CHOP CHOP P value Median event-free survival 3.8 y 1.1 y =0.00002 5-year event-free survival 47% 29% Median progression-free survival Not reached 1 y &lt;0.00001 5-year progression-free survival 54% 30% Median overall survival Not reached 3.1 y =0.0073 5-year overall survival 58% 45%
Progression-free survival is shown in the figure; PFS does not included 19 patients who died in CR from causes not related to lymphoma or its treatment, 5 in CHOP arm and 14 in R-CHOP arm. It better reflects the long term effect of treatment on the disease. No severe late toxicity was observed in R-CHOP treated patients and deaths not related to lymphoma did not show any pattern.
In conclusion, long term results continue to show a major benefit for the addition of rituximab to CHOP in the treatment of patients with DLBCL. This improvement increases with time.
Rituximab/Chemotherapy Induction Treatment Followed by High-Dose Therapy and Autologous Transplantation in Patients with Mantle Cell Lymphoma.
Authors: Thieblemont C. , Antal D. , Lacotte-Thierry L. , Delwail V. , Bouafia F. , Tartas S. , Michallet A.S. , Espinouse D. , Giraud C. , Salles G. , Guilhot F. , Coiffier B. 
Published: April 2020

Abstract
Between September 1998 and September 2003, 25 patients with Mantle Cell Lymphoma (MCL) were treated in our centres with a treatment combining rituximab + chemotherapy at induction with subsequent consolidation high-dose chemo/radiotherapy (HDT) and autologous PBSC-transplantation. Twenty-two (80%) patients received this treatment in first line therapy. Median age of patients was 53 (range: 41–65). All patients had a disseminated disease with a bone marrow (BM) involvement in 23 (92%) patients. Elevated LDH was present in 11 patients. All patients except one had a good performance status. IPI score was low in 5 patients, low-intermediate in 10 patients, high-intermediate in 7 patients and high in 3 patients. Chemotherapy induction regimens were: R-CHOP (n=7 pts), R-ACVB (n=4 pts), R-ESHAP (n=1 pts), R-DHAP (n=1pts), or sequential CHOP+DHAP/ESHAP+Rituximab (n=12pts) regimens. Rituximab was giving simultaneously with the chemotherapy in 13 patients or just before harvest in 12 patients. Number of rituximab injections was four in all patients except one that received 5 injections. PBSC harvest was done after cyclophosphamide or cyclophosphmide + etoposide followed by G-CSF and was successful in all patients with a median number of CD34+ cells at 6.44 106/kg. HDT included TBI in 21 patients.
After induction therapy, all patients were in response: 16 (64%) reached a CR and 9 (36%) a PR because of persistence of a weak (&lt;5%) BM involvement. Evaluation three months after transplant showed a CR in 17 (68%) patients and a PR in 7 (24%) patients because of weak BM involvement. At 3 years, 80% (20/25) of the patients are still alive (Figure). With a median follow-up at 4 years, median time to progression was 3.3 years. Four patients died from progressive disease. Transplantation toxicities were similar to those observed with regular induction treatment without rituximab with a median time of achievement of a PN count &gt; 0.5GI/L at 10 days. One patient died before neutrophil recovery because of undocumented pulmonary infection. No patient presented late neutropenia. In conclusion, combined induction chemotherapy with rituximab followed by HDT dramatically improved the overall survival and the progression free survival in MCL patients, without adding hematological toxicities and infectious complications.
Figure Figure
Myelodysplasia and Acute Myeloid Leukemia Following Therapy for Indolent Lymphoma with Fludarabine, Mitoxantrone, Dexamethasone (FND) Plus Rituximab and Interferon Alfa.
Authors: McLaughlin Peter , Estey Elihu , Glassman Armand , Romaguera Jorge , Samaniego Felipe , Ayala Ana , Hayes Kimberly , Maddox Anne Marie , Preti H. Alejandro , Hagemeister Fredrick B. 
Published: April 2020

Abstract
Treatment-related myelodysplasia (t-MDS) can occur following chemotherapy, especially with DNA-damaging agents such as alkylators or topoisomerase II inhibitors. Antimetabolites such as the nucleoside analogs have only infrequently been associated with post therapy MDS (t-MDS). In a prospective chemo-immunotherapy trial between 1997–2003 in patients with stage IV indolent lymphoma, 202 patients were treated and 8 have developed MDS between 2–5 years after therapy, including 4 who received only fludarabine, mitoxantrone, and dexamethasone (FND) for 6–8 courses, with or without rituximab. The other 4 MDS patients received other agents including alkylators, either as salvage chemotherapy or as part of the initial regimen. Complex cytogentic abnormalities were present in all patients. Abnormalities of chromosome 7 were present in six of the eight, three of whom received FND ± rituximab therapy only. The abnormalities of chromosome 7 were monosomy 7 in four (one of which had add 7p in the remaining chromosome), one del 7q, and one der 7. MDS with features classically associated with DNA damaging agents can occur following therapy with the FND combination, with or without rituximab.
A Small-Molecule Bcl-2 Inhibitor Amplifies NKT-Cell Killing of U937 Lymphoma Cells.
Authors: Lickliter Jason , Cox Joanne , Nieda Mie , Lin Henry , Nicol Andrew 
Published: April 2020

Abstract
Invariant Vα24+ Vβ11+ NKT cells regulate immune responses and are capable of direct cytotoxic activity against tumour-cell targets mediated via the perforin/granzyme-B, Fas ligand and TRAIL pathways. A potential mechanism for tumours to escape immune killing is to block downstream apoptotic pathways by overexpression of the Bcl-2 oncoprotein. We therefore evaluated whether the Bcl-2-expressing U937 lymphoma line could be sensitized to NKT-cell cytotoxicity by coexposure to the HA14-1 small-molecule Bcl-2 inhibitor. Vα24+ NKT cells were purified from donor blood samples using magnetic beads and activated by exposure to KRN7000 and IL-2. U937 cells were labelled with the PKH26 membrane dye and then cocultured with activated NKT cells in a 10:1 or 20:1 effector: target ratio in U-bottomed 96-well plates. Cultures were performed in the presence or absence of HA14-1. After 4 hours, the cells were stained with annexin V-FITC and analysed by flow cytometry. Apoptotic and viable target cells were quantified as annexin-V positive and negative cells, respectively, in the PKH26-positive gate. Results of experiments on NKT cells derived from 3 donors are shown in the figure. Bars represent the mean percentage of viable U937 cells in triplicate assays and error bars indicate the SD. At the concentration of HA14-1 used, there was only minor single-agent cytotoxicity. However, a significant increase in U937 apoptosis was observed with the combination of HA14-1 and NKT cells compared to NKT cells alone. We conclude that the addition of the HA14-1 Bcl-2 inhibitor significantly increases the cytotoxicity of NKT cells against U937 lymphoma targets. This provides preliminary data supporting the possibility that the use of a combination of a small molecule inhibitor with immune therapy may increase the clinical efficacy of immune therapies that work via cytotoxic killing of malignant targets.
Figure Figure