bluuebunny/crossref_metadata_2025_split huggingface dataset:
Molecular and Clinical Features of Diffuse Large B-Cell Lymphoma with MYC Alterations: Inverse Correlation with p53 Pathway Disruption.
Authors: Moller Michael B. , Nielsen Ole , Christensen Bjarne E. , Pedersen Niels T. 
Published: October 2019

Abstract
Diffuse large B-cell lymphoma (DLBCL) is a clinically and genetically heterogeneous disease. Frequent genetic lesions include alterations of BCL2 and BCL6, as well as aberrations of the RB pathway and the p53 pathway resulting in deregulation of cell cycle progression and apoptosis. MYC translocations and amplifications have been detected in a subset of cases. Recently, MYC was shown to control a major regulatory network in B-cells (Nature Genet. 37; 382–90: 2005). We thus hypothesized that DLBCLs with constitutively active MYC are associated with distinctive genetic and clinical features when compared to cases with wild-type MYC. Thirty seven de novo DLBCLs were analyzed using immunohistochemistry for CD10, bcl6, MUM1, and bcl2 expression, as well as fluorescence in situ hybridization for MYC, BCL2, BCL6, IGH, IGK, and IGL breakpoints and amplifications. The lymphomas were previously extensively characterized for aberrations at the genetic, epigenetic, or protein level of components of the RB pathway (pRb, p16, cyclin D3, cdk4) and the p53 pathway (p53, p14ARF, MDM2, E2F1). Five cases (14%) had MYC alterations (4 breakpoints and 1 amplification). Of the MYC+ DLBCL cases, 2 showed GCB immunophenotype and 3 were of non-GCB type. Perturbation of the p53 pathway components p53, p14ARF, MDM2, and E2F1 were identified in 28%, 16%, 53%, and 20% of the MYC− cases, respectively, and 83% had aberrations of 1 or more p53 pathway component. In contrast, only 1 (20%) of the MYC+ cases had a p53 pathway aberration (MDM2 overexpression; p=0.013). Thus, MYC alterations and p53 pathway aberrations were inversely correlated. RB pathway aberrations were equally frequent among MYC+ (40%) and MYC− (43%) DLBCLs. However, the pattern of aberrations was different. Lack of pRb expression was the only alteration of the RB pathway in the MYC+ DLBCLs, whereas 30 of 31 (97%) MYC− lymphomas had normal pRb expression (p=0.045). p16 alterations (point mutation, deletion, hypermethylation, lack of expression) and/or cyclin D3 overexpression were identified in 12 (38%) MYC− cases but in none of the MYC+ cases (p=0.038). MYC status was also related to cytogenetic BCL2 alterations. Four of 5 (80%) MYC+ DLBCLs showed BCL2 breakpoints or amplification in contrast to 4 of 32 (13%) of MYC− DLBCLs (p=0.005). Furthermore, all MYC+ DLBCLs were bcl2 positive (p=0.05) and MUM1 negative (p=0.022), and the proliferation index was lower in the MYC+ DLBCLs (p=0.038). Clinically, MYC+ DLBCL patients were characterized by advanced stage disease (p=0.037) and high tumor burden (p=0.037) but favorable treatment response (p=0.05). Taken together, our data indicate that DLBCL with MYC alterations constitute a subgroup of DLBCL with distinct clinical and molecular features. The patterns of aberrations of the p53 pathway, the RB pathway, and BCL2 indicate profound differences in the molecular pathogenesis between MYC+ and MYC− DLBCL.
All IPI Factors Are Equal, but LDH Is More Equal Than Others.
Authors: van Imhoff Gustaaf W. , Graveland W. , Van der Holt B. , Van Glabbeke M. , Verdonck L. F. , Sonneveld P. , Kluin-Nelemans J. C. 
Published: October 2019

Abstract
The International Prognostic Risk Index (IPI) offers the most important prognostic information in patients with aggressive non-Hodgkin Lymphoma (NHL). Risk classification is based on presence or absence of age >60 yr, stage III or IV, more than one extranodal site, performance score 2–4, or elevated serum LDH above the upper limit of normal (ULN). Because each factor confers a more or less equal independent risk for treatment outcome, they are summed to generate a final IPI risk score (Shipp model). Dichotomization of the continuous variable age is practical, as treatment for patients >60 yr often differs from younger ones. However, as LDH also is a continuous variable we wondered if risk based on actual LDH, especially in patients with highly elevated levels, would have additional prognostic impact. IPI risk factors including actual serum LDH at diagnosis were retrieved from 1286 patients (28% >60 yr) with advanced aggressive NHL and treated with curative intent in 6 clinical trials conducted by HOVON (5 trials) and EORTC (1 trial). All LDH ULNs from the participating centers were verified. LDH levels were divided by the ULN of each center to generate normalized ratios. Six % of patients had LDH >5 times ULN, 8% 3–5 times, 11% 2–3 times, 34% 1–2 times, 20% 0.75–1 times and 20% had levels below 0.75 of ULN. In a multivariate Cox regression model similar independent hazard ratios (HR) ranging from 1.6 to 2.6 were found for the individual dichotomized risk factors according to the Shipp model except for the number of extranodal sites which turned out to be non-significant. This factor was with a HR of 1.4 also the least predictive factor in the Shipp model. In contrast to the dichotomized LDH variable (normal versus elevated), risk for inferior outcome increased linearly with actual LDH levels. Five year OS estimates were 71% for patients with LDH <0.75 x ULN; 53% for 0.75–1; 49% for 1–2; 37% for 2–3; 31% for 3–5; and 25% for LDH >5 x ULN. The HR for these groups were respectively 0.54, 1, 1.45, 2.46, 2.54 and 5.15. This analysis using actual LDH values gave a better discrimination as compared to the HR of 2.6 (95% CI 2.1–3.1) for the dichotomized LDH (i.e. normal versus elevated) in multivariate analysis. Interestingly, the 235 patients with very low LDH (<0.75 x ULN) had much better outcomes with a HR of 0.54 as compared with patients with an LDH-ratio between 0.75 and 1. In conclusion, highly elevated LDH levels in aggressive NHL confer a worse prognosis and suggest the application of a modified IPI risk index adding extra risk points for patients with highly elevated LDH levels.
Analysis of EBV-Specific T Cell Responses in Transplant Recipients with PTLD.
Authors: Sebelin Kathrin , Meier Antje , Papp-Vary Matthias , Oertel Stefan , Pezzutto Antonio , Doerken Bernd , Riess Hanno , Subklewe Marion 
Published: October 2019

Abstract
EBV causes a chronic infection in >95 % of the population and despite its strong growth transforming capacity the majority of EBV infected individuals remain asymptomatic. In contrary, in immunosuppressed patients (pts) the risk of EBV reactivation and development of posttransplant lymphoproliferative disease (PTLD) is high. This is assumed to be due to a defective T cell response. Here we analyzed the EBV-specific CD8 and CD4 T cell response to different EBV latent and lytic antigens in pts with newly diagnosed PTLD. A prospective study of 10 pts after solid organ transplantation at time of diagnosis of PTLD was performed. EBV-specific CD8 T cells were examined by flow cytometric analysis using HLA-A2, HLA-B7 and HLA-B8 restricted tetramers incorporating BMLF1 (lytic), EBNA3 and LMP2 (both latent)-derived peptides. Staining was done in conjunction with mAbs against CD8 and CCR7. The ability of CD8 T cells to produce IFN-γ in response to the same EBV-derived peptides was measured by cytokine secretion assay. In healthy, EBV+ donors, we previously have found a consistent CD4 T cell response to the latent EBV antigen EBNA1. Therefore, EBNA1-specific CD4 T cell responses were monitored for IFN-g / IL-4 secretion after protein stimulation. T cell analysis was combined with EBV-DNA quantiation by real time PCR. We found EBV-specific CD8 T cell responses at low frequency in most pts with PTLD (8/10). Half of the pts showed low frequency EBNA1 specific CD4+ T cell responses. All pts with an EBNA1 specific CD4 T cell response showed an EBV-specific CD8 T cell response. In 2/10 pts we found no EBV-specific CD4 and CD8 T cell responses and both pts died under initial therapy. EBV-viral load was found to inversely correlate to absolute CD4 T cell counts. In comparison to healthy normal donors, no significant differences in EBV-specific T cell response could be observed. However, pts EBV-specific T cells were decreased in comparison to pts with high EBV viral load after TX and no PTLD as well as in comparison to pts with infectious mononucleosis. These results indicate that impairment of EBV-specific T cells is not due to clonal depletion, but rather seems to be due to impaired functional activation and expansion. We therefore conclude that pts with PTLD have an inadequatly low EBV-specific T cell responses which correlates to a low absolute CD4 T cell count. We propose a combined immunomonitoring of EBV viral load, absolute CD4 T cell count and EBV-specific T cell enumeration in pts at risk for development of PTLD. Further studies are needed to evaluate the role of EBV-specific T cell monitoring in immunosuppressed pts for prediction of PTLD and the potential usefulness of T cell monitoring as a prognostic marker in PTLD.
Interluekin-10 (IL-10) Promoter Gene Polymorphism (−819*C) Associated with Poor Clinical Outcome in DLBCL Patients Treated with R (Rituximab)-CHOP Regimen as a First Line.
Authors: Kim Yeo-Kyeoung , Kim Hee-Nam , Lee Je-Jung , Yang Deok-Hwan , Lee Il-Kwon , Park Kyeoung-Soo , Kim Hyeoung-Joon 
Published: October 2019

Abstract
IL-10 is an anti-inflammatory cytokine inhibiting Th1 functions, which may facilitate development of NHL by supporting tumor escape from the immune response. Among the various IL-10 promotor genes, the IL-10-1082G, −819C and −592C alleles were reported to be associated with high serum IL-10 level and there were a few reports about their roles in treatment outcome of DLBCL patients. In the present study, we studied the frequency of these 3 alleles and their influence to the clinical outcome in DLBCL patients treated with standardized regimen; 8 cycles of R (375 mg/m2 on day 1 of each cycle)-CHOP every 3 weeks or 4 cycles of R-CHOP with involved field radiotherapy. 60 patients with newly diagnosed DLBCL were included and all of them represented CD20 positivity. At diagnosis, median age was 60 (19~95) and 30 (57.7%), 13 (25.0), 7 (13.5%) and 2 (3.8%) patients belonged to the low, low-intermediate, high-intermediate and high risk group according to IPI reports, retrospectively. Of total 60 patients, 9 (15%), 23 (38.3%) and 27 (45%) were found to carry IL-10-1082G, −819C and −592C alleles, retrospectively. As compared with -1082AA genotype, -1082G allele (GA/GG) failed to show better outcome (eg. higher CR or longer progression-free survival, PFS) in interim results after 4 cycles of treatment. On the other hands, after completion of 4 R-CHOP, patients with −819C allele (TC/CC) showed significantly lower response rate (CR or PR) than in patients carrying -819TT (82.6% vs. 100%, p<0.05). Furthermore, the progression-free survival (PFS) was significantly shorter in patients carrying −819C allele than TT genotype (18.4 ms. vs. 26.9 ms. p<0.05). Patients with −592C allele (AC/CC) also showed lower interim response rate than with AA genotype (85.2% vs. 100%, p<0.05). PFS also decreased in −592C allele compared with AA genotype (18.8 ms. vs. 26.8 ms., p<0.05). In multivariate analysis including IPI, stage, IL-10-1082G, −819C and −592C allele frequencies, −819C allele remained as an independent prognostic factor predicting shorter PFS (RR=4.3, p<0.05). Additionally, in a haplotype analysis, there was a trend of longer PFS in patients carrying ATA/ATA than in other haplotypes but there was no statistical significance (p=0.08). As is well known, after introduction of rituximab in the treatment regimen, patients with DLBCL have been shown better treatment outcome than before. However, it is still a matter of problem to find and to manage the patients with refractory or progressive disease despite of these reinforced regimens. This study suggests that the detection of IL-10 gene promoter polymorphism such as −819C allele helps us to predict the prognosis and to establish a stratified treatment plan in DLBCL patients.
Assessment of Favorable (F) Versus Unfavorable (U) Early Stage Hodgkin’s Disease (HD); the Stanford V + Radiotherapy (RT) Experience.
Authors: Advani Ranjana , Maeda Lauren , Hoppe Richard T. , Breslin Sheila , Rosenberg Saul A. , Baer David , Mason Joseph , Horning Sandra J. 
Published: October 2019

Abstract
Early stage favorable HD is defined at Stanford and most US centers as stage I/II A and a mediastinal mass ratio (MMR) < 1/3 of the intrathoracic diameter. Within these parameters, we treated 101 patients (pts) with 8 weeks of Stanford V chemotherapy followed by 30 Gy (n = 84) or 20 Gy (n = 17) RT to involved sites. Seven pts had sites >10 cm (2 mediastinal and 5 nodal). At a median followup (f/u) of 5.5 years (y) the freedom from progression (FFP) was 96% with an overall survival (OS) of 98%. Stage I/II patients with MMR >1/3 of the intrathoracic diameter (n = 54) were treated with 12 weeks of Stanford V chemotherapy and 36 Gy RT to sites ≥5 cm. In this large MMR group the FFP and OS were 92% at a median f/u of 8 y. Other groups (EORTC, GHSG, NCI-C and GELA) have stratified early stage patients as favorable (F) and unfavorable (U) based on several risk factors. In addition to MMR and symptoms, these systems include age, # of nodal sites, sedimentation rate (ESR), extranodal lesions and in the GELA, elements of the international prognostic score for advanced HD. Phase III clinical trials have employed different therapies for F and U early stage pt based on these factors. We therefore retrospectively applied these factors to our “favorable” Stanford V pt to determine if they were prognostic as shown.
Group (% U) % FFP % OS F U p value F U p value EORTC (33) 98 86 0.02 100 92 0.03 GHSG (64) 100 91 0.08 100 96 0.27 NCI-C (60) 100 91 0.06 100 96 0.2 GELA (61) 97 92 0.4 100 96 0.3
ifferences in FFP and OS were significant using EORTC criteria. Notably, the majority of our pt were U (including all from Kaiser) according to GHSG criteria. Among 5 relapsed pt, 4 had ≥ 3 nodal sites and 3 had ESR ≥50. All relapses were in the RT field, one combined with distant disease. Secondary therapy was successful in 4 pts although one transplant-related death occurred. In conclusion, stage I-IIA pts with MMR <1/3 treated with Stanford V + 20–30 Gy had excellent overall outcome but application of stratification factors used by other groups, particularly the EORTC, defined a less favorable subset. As HD therapy moves to further reductions in chemotherapy and RT or chemotherapy alone for early stages, these risk factors take on added importance in the interpretation of current trials results and design of future studies.
Phosphatidylserine (PS) Binding Sites in Kringle Modules Regulate the Domain Organization and Conformation of Bovine Prothrombin.
Authors: Sen Shalmali , Banerjee Mou , Koppaka Vishwanath , Zhou Chaoming , Lentz Barry R. 
Published: October 2019

Abstract
Phosphatidylserine (PS) specifically regulates prothrombin activation during blood coagulation by binding to specific on factor Xa (Koppaka et al. Biochemistry, 7483, 1996) and its cofactor, factor Va (Zhai et al. Biochemistry, 5675, 2002). Binding to PS-containing membranes also alters prothrombin conformation (Lentz et al., Biochemistry, 5460, 1994; Chen et al. ibid. 4701, 1997). We ask here whether specific PS binding sites on prothrombin also control these structural changes, and, if so, where these sites are located. Four methods (Trp fluorescence, circular dichroism [CD], differential scanning calorimetry [DSC], and quasi-elastic light scattering [QELS]) were used to define the structural consequences of soluble 1,2-dicaproyl-sn-glycero-3-phospho-L-serine; (C6PS) binding both to whole prothrombin and to its proteolytically generated fragments. Intrinsic fluorescence titrations suggested the existence of two linked C6PS binding sites in fragment 1.2 (F1.2) and prethrombin 1, both of which contain prothrombin’s kringle pair. The existence of two sites was supported by direct equilibrium binding measurements with F1.2. CD measurements at increasing C6PS concentrations with both F1.2 and the kringle pair (F1.2 minus the γ-carboxy-glutamic acid [GLA] domain) were consistent with formation of a structure akin to an anti-parallel β sheet. Thermal denaturation profiles of F1.2 suggested calcium-independent, C6PS-induced domain reorganization within this fragment. Denaturation profiles and Trp fluorescence of the N-terminal, membrane binding domain (F1) and fragment 2 (F2) did not reveal any C6PS-induced changes. F2 consists mainly of the second cys-rich kringle module. The hydrodynamic radius of prothrombin was also found to decrease substantially (from 3.3 nm to 2.6 nm) in the presence of saturating (1mM) C6PS. Other lipids (phosphatidylglycerol and phosphatidyl-D-serine) did bind to prothrombin but did not produce comparable structural changes. The results show that C6PS 1) binds specifically to linked, calcium-independent sites within prothrombin’s two kringle domains and 2) induces thereby a conformational reorganization in the whole molecule. Supported by USPHS grant HL072827 to BRL.
The CXCR4 Antagonist AMD3100 Mobilizes a More Primitive Subset of CD34+ Cells Than G-CSF.
Authors: Seeger Timon , Veldwijk Marlon , Bridger Gary , Calandra Gary , Goldschmidt Hartmut , Ho Anthony , Fruehauf Stefan 
Published: October 2019

Abstract
The CXCR4-antagonist AMD3100 mobilizes peripheral blood progenitor cells (PBPC) in 8–10 hours while a standard mobilization regimen with granulocyte colony-stimulating factor (G-CSF) takes 4 days before significant numbers of PBPC can be detected. Recently it was shown that AMD3100-mobilized PBPC have a higher reconstitution potential in the NOD/SCID mouse model than G-CSF mobilized PBPC. Clinically they also mediate rapid and sustained engraftment after transplantation. Prospective comparative studies are yet lacking. It can be expected that the functional differences are also reflected on the phenotypic level. We used 6-color and 3-color flow cytometry to study the co-expression of primitive and lineage-associated markers on mobilized PBPC of patients with multiple myeloma and non-Hodgkin’s lymphoma (n=5). Mobilization treatment consisted of 5 days G-CSF (10 μg/kg, s.c. AM) and a single dose of AMD3100 (240 μg/kg, s.c.) in the evening of day 4. CD34+ cells of day 4, before AMD3100-dosing (G-CSF group) and on day 5, 10-hours after AMD3100 (AMD3100+G-CSF group) were compared. Primitive CD34+/CD38- cells which solely mediate hematopoietic reconstitution in the NOD/SCID mouse model were increased after addition of AMD3100 in all patients by a median of 2,58 fold (range 1,19–6,66) while the myeloid lineage committed CD34+/CD117+ and the actively proliferating CD34+/CD71+ cell subsets were significantly downregulated (p<0,05 each). The myeloid CD34+/CD33+ cell population showed a similar trend (median 0,60-fold change, range 0,24–1,08). Adhesion molecules were significantly regulated (CD49b) or showed such a trend (CD49d). Quantification of aldehyde dehydrogenase (AlDH) positive cells in A+G vs. G-CSF-mobilized samples did not show significant differences between these groups. We conclude that AMD3100 mobilizes a more primitive subset of CD34+ progenitor cells than G-CSF alone. Whether this translates into lower CD34+ threshold values for transplantation to achieve a similarly rapid and sustained engraftment after transplantation or into a shorter reconstitution period if similar amounts of CD34+ cells are used as after standard G-CSF-mobilization needs further study.
CD34 coexpression analysis: AMD3100+G-CSF mobilized CD34+ cells/G-CSF mobilized CD34+ cells (fold-change) CD34+/ median min max P value CD38− 2.58 1.19 2.09 0.073 HLA-DR− 1.14 0.14 2.09 0.568 CD90+ 0.42 0.05 26.47 0.417 CD7+ 1.17 0.58 2.9 0.433 CD10+ 0.87 0.11 6.66 0.513 CD19+ 1.11 0.72 12.69 0.368 CD33+ 0.6 0.24 1.08 0.075 CD45RA+ 5.84 1.54 7.46 0.155 CD71+ 0.51 0.43 0.64 0.002 CD117+ 0.36 0.02 1.11 0.046 CD18+ 1.55 0.56 4.09 0.347 CD49b+ 6.91 2.74 11.75 0.08 CD49d+ 0.63 0.25 0.89 0.061 CD58+ 5.82 2.83 21.88 0.261 CD62L 2.9 1.43 4.36 0.419 CD184+ 0.99 0.25 1.79 0.955
Immunodeficient Mice Are Poor Mobilizers -Novel Evidence That Demonstrates a Pivotal Role of Complement in Triggering Mobilization of HSPC.
Authors: Reca Ryan , Wysoczynski Marcin , Hansen Richard , Kucia Magda , Janowska-Wieczorek Anna , Ratajczak Janina , Ratajczak Mariusz Z. 
Published: October 2019

Abstract
Complement (C) is activated by immunoglobulin-dependent classical and immunoglobulin-independent alternative pathways, and we recently reported that the activation of C enhances the responsiveness of hematopoietic stem/progenitor cells (HSPC) to an SDF-1 gradient cascade by releasing C3a and desArgC3a cleavage fragments (Blood2003;101:3784; Blood2004; 103: 2071). We also found that bone marrow (BM) concentration of C3a and desArgC3a increases during mobilization with G-CSF or cyclophosphamide (CY). To explain this phenomenon we envisioned that these mobilizing agents expose a neo-antigen in BM tissue by turning BM into a highly proteolytic microenvironment. As a consequence of this, the newly exposed neo-epitope becomes bound by natural IgM antibodies leading to the activation of the classical C cascade. In our studies to elucidate the role of C activation in triggering the mobilization of HSPC, we found that C is effectively activated in the BM of G-CSF- or CY- mobilized wild-type (wt) but not IgM-deficient (RAG2null) mice. More importantly we found that several immunodeficient murine strains (RAG2null, SCID and Xid) displayed severely reduced G-CSF-induced mobilization of HSPC which we believe is a result of lack of B lymphocytes and complement-activating immunoglobulins. Supporting this, we found T cell depletion in wt mice did not affect mobilization. Moreover, G-CSF-induced mobilization in RAG2null, SCID and Xid animals was restored after infusion of murine inmmunoglobulins. Furthermore, since mobilization by zymosan or sulfated glycans activates C via the alternative immunoglobulin-independent pathway, we focused on the role of C activation during zymosan-induced mobilization. As expected, zymosan-induced immunoglobulin-independent C activation and mobilization of HSPC were unaffected in immunodeficient (RAG2null, SCID and Xid) mice which have a normal serum level of C3. However, these processes were severely reduced in C3-deficient animals. Thus our data strongly support the notion that C and innate immunity is an important trigger of mobilization of HSPC. While G-CSF and cyclophosphamide activate C by a classical IgM-dependent pathway, zymosan and sulfated polyglycans activate it by employing an alternative pathway. In conclusion, mobilization of HSPC could be envisioned as part of a more global immune response that is triggered/mediated by C3 activation/cleavage.