bluuebunny/crossref_metadata_2025_split huggingface dataset:
Age Related Changes in Hoechst 33342 Efflux Dynamics and Side Population Phenotype in Murine Bone Marrow.
Authors: Greenwood Matthew J. , Lansdorp Peter M. 
Published: October 2019

Abstract
The mechanisms underlying the aging of the hematopoietic stem cell (HSC) compartment remain poorly understood. The ATP-binding cassette cell surface transport protein, ABCG2, has been identified as the transporter responsible for Hoechst 33342 (Hst) efflux in primitive stem cells and its expression is associated with the side population (SP) phenotype in both murine and human bone marrow (BM). ABCG2 expression and Hst efflux activity is highest in those cells with the greatest repopulating potential and is progressively downregulated during differentiation. The substrate profile of ABCG2, which includes a number of antineoplastic drugs, protoporphyrin IX and the chlorophyll breakdown product pheophorbide, suggest that ABCG2 transporters may function to protect stem cells from cytotoxic insults, a function which may be of great importance in stem cell maintenance. Amongst laboratory mice, the C57BL/6 strain is the longest lived and appears to accumulate HSC’s with age as assessed by both phenotype and colony forming assays. While the phenotypic features of the SP profile have been well characterized in both humans and young mice, little is known of the Hst efflux dynamics or phenotype of the SP profile in old and very old C57BL/6 mice. In order to further characterize the SP profile in old mice, whole BM was extracted from the femurs, tibiae, pelvis and thoracolumbar vertebral bodies of young (9–13 week) and old (95–108 week) C57BL/6 (Ly5.1) mice. Cells were stained with 5μg/ml Hst followed by staining with a combination of CD45.1 FITC, Sca1 PE, c-kit APC, CD34 FITC, biotinylated CD34 and lineage markers and strep PE-Texas Red. In addition, serial sampling of Hst incubated cells was performed to assess Hst efflux activity at 20 mins incubation through to 100mins. Six-color flow analysis was performed on a FACS Vantage™ (BD) cytometer and data analyzed using FlowJo™ software. Results show a marked increase in cells with an SP phenotype in old vs young mice (mean±SD 1.85%±0.88 vs 0.15%±0.09) which were more highly enriched for CD34-Sca1+ckit+ (22.2%±8.65 vs 8.89%±6.7) cells. Subdividing the SP profile into four regions (R1 to R4) from highest to lowest Hst efflux activity revealed that in old mice, SP cells with the highest Hst efflux activity were almost exclusively of a CD34-Sca1+ckit+ phenotype (82.3%±14.0 vs 11.5%±7.8), with a decreasing proportion of these cells represented throughout the remaining SP tail, though a significant proportion of cells within R4 remain CD34-Sca1+ckit+ (15.3%±7.83 vs 4.19±3.01). Similar patterns have been observed in both whole and lineage depleted BM. In addition, BM cells from old C57BL/6 mice show prolonged Hst efflux activity with an increase in cells in the SP gate at 100 mins (1.51%±0.50 vs 0.10%±0.06). We conclude that in old C57Bl/6 mice, cells accumulate which have the capacity to efflux Hst in agreement with previous reports of an increase in HSC number with age in this mouse strain.
VEGF-VEGF Receptor 1 (Flt-1) Interactions Significantly Modulate Endothelial Progenitor Cell (EPC) Migration but Have Only Minor Effects on the Migration of Differentiated Endothelial Cells.
Authors: Li Bin , Teleron Amylynn A. , Lin Charles , Young Pampee P. 
Published: October 2019

Abstract
There is a growing body of data demonstrating that vasculogenesis, whereby bone marrow -derived circulating progenitor cells (EPCs) home to sites of neovascularization, results in significant contribution to blood vessel formation during tumor growth, ischemic injury and wound healing. Vascular Endothelial Growth Factor (VEGF) has recently been shown to augment vasculogenesis. In the current study, we examined if VEGF/VEGF Receptor (VEGFR) interactions are important for EPC recruitment. Both VEGFR1 (flt-1) and VEGFR2 (flk-1) are strongly expressed, as detected by immunofluorescent and FACS analysis, on EPCs obtained by ex vivo expansion of human peripheral blood. In a modified Boyden chamber migration assay, EPCs showed dose dependent migration to VEGF. To examine receptor specificity, EPCs were preincubated with receptor blocking anti-VEGFR1 or -VEGFR2 prior to the migration assay. Level of inhibition by VEGFR1 blocking antibody was commensurate with blocking VEGFR2. Furthermore, migration in response to a VEGFR1-specific agonist, PlGF, was comparable to that induced by VEGF and was completely ablated by preincubation with VEGFR1 blocking antibody. By contrast, differentiated endothelial cells had diminished migration in response to PlGF as compared to VEGF. Furthermore, blocking VEGFR1 only mildly disrupted VEGF-induced migration of differentiated endothelial cells in vitro. Hence, unlike differentiated endothelial cells, EPC migration in vitro was mediated by both VEGF receptors. By quantitative RT-PCR, we examined the level of VEGFR1 and VEGFR2 mRNA transcripts in EPCs versus differentiated endothelial cells. VEGFR1 transcripts in EPCs were expressed 3-fold higher than in differentiated endothelial cells. VEGFR2 and neuropilin transcript levels in EPCs, however, were lower than in differentiated endothelial cells. These results suggest that VEGF/VEGFR1 interactions are important in EPC migration in vitro. We have subcloned VEGFR1 cDNA into a retrovirus vector and have shown by western blot that we can direct increased cellular expression of VEGFR1. In further experiments, we will examine the role of VEGFR1 in human EPC recruitment using murine xenotransplant models of hindlimb ischemia and wound healing. These studies will provide valuable insight towards developing EPCs as cellular therapy to augment blood vessel formation.
Primary Nasal NK/T Cell Lymphoma: Cytology and Level of the Antiapoptotic PI9 Protein Have Prognostic Relevance.
Authors: Bossard Céline , Belhadj Karim , Reyes Felix , Martin-Garcia Nadine , Kümmer Alain , Berger Françoise , Brière Josette , Baglin Anne-Catherine , Casiraghi Odile , Lepage Eric , Gaulard Philippe ,   for the GELA
Published: October 2019

Abstract
Nasal NK/T cell lymphoma is a rare disease entity characterized by a nasopharyngeal presentation, a common origin from NK cells, an association with EBV and a predilection for Asians and South Americans contrasting with its rarity in Western countries. Several studies (Chim C.S. et al., Blood 2004; Li C.C. et al., Cancer 2004) indicate that the disease has a poor outcome following conventional combined chemoradiotherapy treatments, which might be related to resistance to therapy-induced apoptosis. It has been shown (Hermine O. et al., Blood 1996; Ten Berge R. et al., Blood 2002) that high level of expression of antiapoptotic proteins by lymphoma cells is associated with poor outcome in a variety of lymphomas.
In a retrospective analysis of 37 patients treated with first-line chemotherapy (n=34) or chemoradiotherapy (n=3) according to the LNH87, LNH93 and LNH98 trials of the GELA, we have analyzed by immunohistochemistry the expression of the granzyme B-protease inhibitor 9 (PI9) regarded as an inhibitor of apoptosis, and of the active form of caspase-3 (aC-3), an effector of apoptosis. The diagnosis was based on biopsies of the upper respiratory tract showing a proliferation of neoplastic cells with a CD3+/CD5− (100%), CD56+ (90%), TIA1/GrB+ (100%) phenotype and EBV association (100%). Among the 37 patients, 73% were < 60y, 68% were male. 76% had stage I, 5% Stage II, 19% stage IV and 68% had an IPI score of 0–1. With a median follow-up of 6.3 y, 5y-event-free and overall survivals were 38% and 47%, respectively. According to cytology, tumors were subclassified into predominatly small cell (52%) and predominatly large cell (48%) categories. PI9 was scored as positive (>10% of tumor cells) in 68% of the cases, whereas 35% were assigned a high number (>10 per field at high magnification) of aC-3 positive tumor cells. Univariate analysis showed that small cell cytology and absence of PI9 expression were associated with poor outcome, but not the level of aC-3 expression nor IPI score. By multivariate analysis, cytology and PI9 downregulation were shown to independently affect event-free (P=0.01 and 0.05, respectively) and overall survival (P=0.009 and 0.006). We conclude that nasal NK/T cell lymphomas disclose cytological heterogeneity which has prognostic relevance. We also found that lack of expression of PI9 is associated with poor outcome, an unexpected finding in view of the known antiapoptotic function of this protein. Since PI9 is constitutively expressed by normal NK cells, it might be postulated that its impaired expression in tumor cells reflects a yet unknown mechanism associated with progression.
Serum-Soluble Tumor Necrosis Factor Receptor 2 (sTNF-R2) Level Determines Clinical Outcome in Patients with Aggressive Non-Hodgkin’s Lymphoma.
Authors: Tsurumi Hisashi , Goto Naoe , Takemura Masao , Hara Takeshi , Sawada Michio , Yamada Toshiki , Kasahara Senji , Kanemura Nobuhiro , Takahashi Takeshi , Oyama Masami , Tomita Eiichi , Takami Tsuyoshi , Moriwaki Hisataka 
Published: October 2019

Abstract
The tumor necrosis factor (TNF) plays a key role in inflammatory processes, as this cytokine is one of the earliest to be produced in such a condition, and triggers the following cytokine cascade. In addition, the TNF and their receptor system are believed to play a key role in the growth, differentiation, and/or apoptosis of the malignant cells. As for TNF receptors, the two types, the 55 kDa (p55, TNFR; TNF-R1) and the 75 kDa (p75, TNFR; TNF-R2) are simultaneously expressed on many cells at different levels. The extracellular domains of these two receptors are released from the cell membrane by cleavage of TNF-Rs as soluble TNF-Rs (sTNF-R1, sTNF-R2). Reportedly the serum TNF-Rs level rise in patients with some malignancies. The aim of the present study was to assess the prognostic significance of serum sTNF-R in aggressive non-Hodgkin’s lymphoma (NHL). Consecutive 110 previously untreated patients with aggressive NHL (diffuse large B-cell lymphoma; 94, peripheral T-cell lymphoma; 16) prospectively participated in this study between 1997 and 2002. The patients were treated with 6–8 cycles of CHOP or THP-COP regimens. To evaluate serum levels of sTNF-Rs (p55; TNF-R1, p75; TNF-R2), venous blood samples were drawn from patients immediately before the initiation of treatment. Serum sTNF-R1 and sTNF-R2 were determined using a sandwich enzyme-linked immunosorbent assay (ELISA). In healthy control subjects, the median of serum sTNF-R1 and sTNF-R2 levels were 1.2 ng/ml (range 0.3–2.9) and 4.17 ng/ml (range 1.91–8.51), respectively. High serum sTNF-R level was associated with some poor prognostic factors and low complete remission (CR) rate. Patients with high sTNF-R1(4 ng/ml and over) and sTNF-R2 (15 ng/ml and over) at onset had significantly lower survival rates (5-year: 19%, 19%) than those with low sTNF-R1 (under 4 ng/ml) and sTNF-R2 (under 15 ng/ml) (62%, 69%), respectively (p<0.0005, p<0.0001). Multivariate analysis employing sTNF-R2 and some conventional prognostic factors demonstrated that sTNF-R2 and performance status for overall survival (OS) and sTNF-R2, sIL-2R, and LDH for event free survival (EFS) were significantly poor prognostic factors. As for TNFa, a serum TNFa level is not related with sTNF-R1 or sTNF-R2 level in aggressive NHL. In addition, serum TNFa level is not associated with OS and EFS. In conclusion, serum sTNF-R2 might be a significant prognostic factor for aggressive NHL and a useful tool for selecting the appropriate therapeutic strategy in the treatment of aggressive NHL. The most reliable prognostic factor and the best combination of some prognostic factors for aggressive NHL should be clarified in order to assist in selecting appropriate treatment.
Cyclophosphamide Induced CD11b+ Cells Can Inhibit Anti-CD40 Monoclonal Antibody (mAb) Therapy of B-Cell Lymphoma.
Authors: Honeychurch Jamie , Glennie Martin J. , Johnson Peter W. , Illidge Tim M. 
Published: October 2019

Abstract
Immunotherapy with mAb is emerging as an important component of treatment for B-cell lymphoma. Furthermore, anti-tumor effects may be enhanced by combining mAb with a range of conventional cancer treatments and recently impressive results have been acheived with mAb in combination with chemotherapy. Here, we have assessed the efficacy of combining an agonistic anti-CD40 mAb with cyclophosphamide (CY). We have previously shown that large repeated doses of anti-CD40 mAb alone may stimulate cytotoxic T-cell lymphocyte (CTL) responses and provide robust protection in syngeneic murine tumors, such as BCL1 lymphoma. In this study we therefore used single smaller doses of the mAb that alone fail to provide long-term clearance thus allowing us to look for additive or synergistic effects of anti-CD40 and CY. Interestingly, the timing of CY relative to mAb administration appeared critical to therapeutic outcome. Cohorts given CY 7 days (d7 CY) prior to anti-CD40 show reduced survival compared to those receiving mAb alone. In contrast treatment with CY given 1 day before, or on the same day as mAb resulted in greatly increased tumor free survival. In vivo tracking experiments revealed that there is a potent CTL response that clears tumor around 5 days after anti-CD40 treatment. However, when CY was given d7 prior to anti-CD40 mAb, CTL expansion was reduced compared with other treatments, and instead resulted in increased numbers of splenic CD11b+ cells. These CD11b+ cells have an immature myeloid precursor phenotype and were able to produce nitric oxide (NO). Co-culture of CD11b+ cells with murine splenocytes resulted in greatly reduced cytotoxic CTL proliferation, but no apoptosis, which was reversed in the presence of NO inhibitors. Similarly, splenic CTL proliferation in vivo is inhibited by this increased population of CD11b+ cells. Importantly adoptive transfer of CY-induced CD11b+ into non-CY-treated mice inhibited anti-CD40 mAb tumor protection in BCL1 lymphoma. This data indicate that combining CY with immunotherapy can enhance therapeutic responses, but the timing of CY relative to immunotherapy is critical to the therapeutic outcome.
Antagonist Anti-CD40 Monoclonal Antibody, CHIR-12.12, Inhibits Growth of a Rituximab-Resistant NHL Xenograft Model and Achieves Synergistic Activity When Combined with Ineffective Rituximab.
Authors: Long Li , Tong Xia , Patawaran Montesa , Aukerman Lea , Klinger Julie , Jallal Bahija , Luqman Mohammad 
Published: October 2019

Abstract
CD40 and CD20 are expressed in several B-cell malignancies and represent attractive targets for therapeutic intervention. The anti-CD20 monoclonal antibody, rituximab, is an approved drug for the treatment of non-Hodgkin’s lymphomas. however, the existence of patients with rituximab-resistant disease limits its clinical utility. We have previously reported that the novel, highly potent, fully human antagonistic anti-CD40 monoclonal antibody, CHIR-12.12, generated from XenoMouse® mice (Abgenix, Inc) has greater anti-tumor activity than rituximab in both rituximab-responsive and rituximab-resistant human NHL models. In this study, we evaluated the potential therapeutic application of combining CHIR-12.12 and rituximab for the treatment of NHL. Namalwa is a Burkitt’s lymphoma cell line that gives rise to aggressive rituximab-resistant tumors when implanted in nude mice. Direct treatment of these tumor cells with CHIR-12.12 or rituximab in culture does not affect cell growth when compared to treatment with an isotype control antibody. Although Namalwa cells express more CD20 than CD40 (average of 10,059 CD20 and 3,138 CD40 molecules per cell respectively, P=0.05), when tested for in vitro ADCC killing using human NK cells as effectors, CHIR-12.12 mediated stronger target cell lysis than rituximab (31.43% vs. 14.15%, P<0.0001). Adding CHIR-12.12 and rituximab together did not enhance the in vitro ADCC killing. When CHIR-12.12 and rituximab were tested in a subcutaneous Namalwa xenograft model, CHIR-12.12 alone caused 60% tumor growth inhibition (P=0.028) whereas rituximab alone at 10 and 20 mg/kg did not inhibit Namalwa tumor growth. When tumor-bearing mice were administered rituximab at 10 mg/kg plus CHIR-12.12 at 5 or 10 mg/kg, synergistic anti-tumor activity was observed in a CHIR-12.12 dose-dependent manner. The mean tumor volume reduction in combination groups is 77% with CHIR-12.12 at 5 mg/kg (P=0.0037) and 83% with CHIR-12.12 at 10 mg/kg (P=0.0018), respectively. The potential interaction between CD40 and CD20 molecules was evaluated in vitro by treating the tumor cells with CHIR-12.12 and assessing the change in CD20 expression and vice versa. The result showed no augmentation of one antigen expression by treating the tumor cells with the other antibody. The mechanism of anti-tumor synergy observed in this combination is under evaluation. Taken together, these data suggest that the combination therapy with anti-CD40 CHIR-12.12 and rituximab has the potential to improve patient outcome in B-cell malignancies co-expressing CD20 and CD40 antigens and support the further development of CHIR-12.12 antibody for treatment of B-cell malignancies.
The Geldanamycin Analogue Heat Shock Protein 90 (HSP90) Inhibitor 17-AAG in Lymphomas: Molecular Mechanisms of 17-AAG-Induced Cell Death in Hodgkin Disease and Non- Hodgkin Lymphoma Cell Lines.
Authors: Georgakis Georgios V. , Li Yang , Younes Anas 
Published: October 2019

Abstract
Heat Shock Proteins (HSP) are stress-induced proteins that act as molecular chaperones in all benign and malignant cells. HSPs are important for maintaining the function of several client proteins involved in transcriptional regulation, signal transduction, and cell-cycle control. HSP90 has been recently identified as a potential therapeutic target in several types of tumors. However, the effect HSP90 inhibition in Hodgkin disease (HD) and non-Hodgkin lymphomas (NHL) remains unknown. The geldanamycin analogue 17-allylamino-17-demethoxy-geldanamycin (17-AAG) is a small molecule inhibitor of HSP90, which interferes with ATP-dependent chaperones activity, with subsequent degredation of client proteins by the proteasome. We studied the activity of 17-AAG in a panel of cell lines of HD (HD-MyZ, HD-LM2, L-428, KM-H2) and NHL (SUDHL1, and Jurkat) origin. HSP90 was expressed in all HD cell lines but not in normal peripheral blood lymphocytes of healthy donors. 17-AAG demonstrated a significant antiproliferative effect in a dose (01 micromolar - 5 micromolar) and time (24 hours to 72 hours) dependent manner in all cells lines, but was most prominant in the HD-LM2 cells. In all HD-derived cell lines, 17-AAG had more than 50% inhibitory effect over control cells. This effect was most prominant after 72 hours of incubation. Subsequently, we examined the molecular mechanisms responsible for this antiproliferative effect. Both cell cycle arrest and induction of apoptosis were observed. In the HD-LM2 cell line, 17-AAG predominantly induced a G0/G1 cell cycle phase arrest, while in the L-428 cell line there was a G2/M phase arrest (as determined by propidium iodide staining and flow-cytometric analysis). This effect was associated with alterations in MDM2 and cyclin D1 proteins. Cell cycle arrest was followed by apoptosis as determined by Annexin-V binding assay, which was predominantly mediated by caspase 9 activation. 17-AAG also reduced the cellular content of Bcl-2, cFLIP, and AKT, but had no significant effect on XIAP or cIAP2. Furthermore, 17-AAG decreased the total ERK1/2 which has been shown to play an important role in HRS cell survival. 17-AAG-induced apoptosis was partially reversed by the pancaspase inhibitor ZVAD-FMK. Preliminary data suggested that 17-AAG did not enhance the effect of doxoorubicin or gemcitabine chemotherapy. These data suggest a first proof-of-principle that inhibition of HSP90 might be of therapeutic value in HD and NHL.
Primary Mediastinal Large B-Cell Lymphoma: Long Term Results with an Alternating Chemotherapy Regimen Based on High-Dose Methotrexate.
Authors: Fietz Thomas J. , Fiedler F. , Franke Astrid , Freund Mathias , Niederwieser Dietger W. , Thiel Eckhard , Knauf Wolfgang U. ,   East German Study Group on Hematology and Oncology
Published: October 2019

Abstract
Primary mediastinal large B-cell lymphomas (MLCL) differ from other diffuse large cell lymphomas: they convey a unique gene expression profile reminding of Hodgkin’s disease, are characterized by an occlusive infiltration pattern (e.g. with vena cava superior syndrome due to the concomitant sclerosis) and treatment with CHOP-based chemotherapy regimen has been disappointing. A dose intensification improves long-term results, but no standard therapy has been established so far. We investigated the use of a high-dose methotrexate based alternating chemotherapy regimen (B-ALL protocol of the GM-ALL) first as a single-center, later as a multicenter trial in 43 patients; depending on the extend of the disease mediastinal radiotherapy and/or high-dose chemotherapy with stem cell rescue were applied additionally.
15 patients have been included in our pilot single-center study (median age 31.5 years). 11/15 (73%) showed infiltration of either esophagus, pleura, lung or V.cava superior syndrome, 6/15 (40%) had pericardial effusion. In this cohort 14/15 (93%) achieved a remission (10/15 CR, 4/15 PR), which is long lasting up to 15 years as observed for the first CR patients.
28 patients have been included into the recent multicenter trial. Here the rate of evaluable remissions after 6 cycles of chemotherapy was identical with 95% (18/19) complete or partial remissions. However, 2 patients died after the first cycle of chemotherapy following septic complications. In 136 chemotherapy cycles the rates of a grade 3 or 4 toxicity were 21% (mucositis), 41% (neutropenia), 30% (thrombocytopenia) and 10% (neutropenic fever).
A methotrexate based alternating chemotherapy regimen leads to excellent remission rates in patients with primary mediastinal large B-cell lymphoma. With regard to the patients included in our single-center trial with a long follow-up these remissions are reliable and long-lasting. Nevertheless, early death following treatment initiation does occur and requires close monitoring and treatment in specialized institutions.