bluuebunny/crossref_metadata_2025_split huggingface dataset:
Decrease of FoxP3 Gene Expression Precedes and Inversely Correlates with Onset of Acute GVHD.
Authors: Tabellini Laura , Haeusermann Peter , Damodaran Anand C. , Hansen John A. 
Published: April 2020

Abstract
Background: Regulatory CD4+CD25+FoxP3+ T-cells are hypothesized to play a role in modulating the severity of acute GVHD and induction of non-responsiveness or tolerance after hematopoietic cell transplantation. Recent studies in patients post transplant demonstrated association between decrease of FoxP3 and development of acute GVHD. However sequential changes of FoxP3 expression prior to onset of clinical GVHD have not been reported.
Methods: Thirteen patients who received myeloablative-conditioning regimen for hematological malignancies and cyclosporine and methotrexate for GVHD prophylaxis have been studied on two or more occasions prior to onset of clinically significant grade II-IV acute GVHD. Six of these patients developed GVHD by day 30 (range, 29–31 days), 4 patients by day 37 (range, 35–39 days), and 3 patients showed no evidence of GHVD during the first 100 days post-transplant. Sequential measurements of FoxP3 gene expression by real time PCR were made at day 21, day 28 and day 35 post-HSCT. All samples analyzed were collected before initiation of systemic steroid therapy. The 6 patients with onset by day 30 and 1 patient without GVHD were studied at days 21 and 28. RNA was extracted from white blood cells and cDNA templates were synthesized using 1μg RNA. The standard curve method was used for relative quantitation of Foxp3.
Results: Using the paired t test, we found a significant decrease of FoxP3 gene expression between day 21 (median 0.3; range 0.01–1.9) and day 28 (median 0.13; range 0.003–0.321) (p=0.05). Additionally, a clear trend for a decreasing expression of Foxp3 was found between day 28 and day 35 (median 0.1; range 0.06–0.54). Two of the 3 patients who remained GVHD free the first 100 days showed either an increase or no change in FoxP3 expression between days 21, 28 and 35 and between days 21 and 28 respectively.
Conclusion: These preliminary results indicate a pattern of decreasing FoxP3 gene expression in patients preceding the development of acute GVHD independent of the day of onset of this disease. These studies are being extended in a new cohort of transplant patients, and monitoring of patients without GVHD is continuing to determine if there are any patterns of FoxP3 expression associated with the onset of chronic GVHD. The available preliminary data suggest that sequential measurements of FoxP3 may predict onset of clinically significant acute GVHD.
Expression of Interleukin-7 Receptor and Activation Markers on Central Memory CD8 Cells in HIV-Infected Individuals Treated with Antiretrovirals.
Authors: Routy Jean Pierre , Boulassel Med-Rachid , Mercier Francois , Grignon Chantal , Young Mavis , Legault Mario , Tremblay Cecile , Cheynier Remi , Sekaly Rafick-Pierre 
Published: April 2020

Abstract
We have recently shown that high levels of interleukin-7 (IL-7) predict poor virological response in HIV-1-infected patients. To understand the underlying mechanisms associated with increase in IL-7 levels, we assess the expression of IL-7 receptor and activation markers on different CD8 subsets in there well defined groups of patients including successfully treated (ST; n=8), untreated (UT; n=6), failing HAART due to drug resistance (FT; n=12) and healthy subjects (n=5).
The expression of IL-7/IL-2 receptors (CD127/CD25) and activation markers (CD38/HLA-DR) on CD8 naïve cells (CD45RA+CCR7+CD27+), central memory (TCM; CD45RA−CCR7+CD27+), pre-terminal effector memory (TPEM; CD45RA−CCR7−CD27−) and terminal effector memory cells (TTEM; CD45RA+CCR7−CD27−) was measured in fresh blood samples using 8-color flow cytometry. Mann Whitney U test and Spearman correlation were used.
Although there was a trend toward a decrease in naïve, TCM, TPEM and TTEM cells from ST to UT and then to FT, this did not reach significance (P=0.09). FT and UT subjects showed higher number of CD8 activated cells than ST or controls (22%, 20%, 3% and 4% respectively; P=0.07). Similarly, the percentages of TCM activated cells were significantly higher in FT and UT than ST or controls (28%, 27%, 7% and 0.07%; P=0.02), whereas naïve and TTEM activated cells were similar in all groups. Interestingly, the percentage of TCM cells expressing CD127+CD25+ was lower in FT compared to UT or ST (0.7%, 2.1 and 7.4%; P=0.02). A strong positive correlation was observed between viral load and CD8+ or TCM activated cells (r=0.46, P=0.02; r=0.65, P=0.004 respectively) but not with naïve CD8+ cells. These data suggest that a substantial proportion of TCM but not naïve, TPEM and TTEM cells expressing immune activation markers with lower IL-7 receptor expression is present in subjects with persistent antigen stimulation, thus providing an explanation for increasing circulating IL-7 levels.
Successful Allogeneic HSCT in a Patient with Chronic Granulomatous Disease and McLeod Phenotype after Sensitization to Kx- and K- Antigens.
Authors: Hoenig Manfred , Schulz Ansgar , Schuetz Catharina , Freihorst Joachim , Baumann Ulrich , Flegel Willy A. , Debatin Klaus-Michael , Friedrich Wilhelm 
Published: April 2020

Abstract
Chronic granulomatous disease (CGD) is a primary immunodeficiency characterized by the defective generation of active oxygen metabolites by phagocytic cells. CGD can be associated with a condition called McLeod phenotype if a large X-chromosomal deletion includes the genes for gp91phox and the XK-protein. The absence of XK on erythrocyte membranes concomitantly leads to a reduced expression of Kell antigens. If transfused with red cells expressing these proteins, patients with CGD and McLeod phenotype become sensitized which may limit future transfusions. We present a patient with CGD and McLeod phenotype, who had received Kx- (the antigen of XK) and K- positive red cells at the age of 10 months; 9 years later he was given granulocyte transfusions following lung surgery because of pulmonary aspergillosis. Shortly thereafter he required a transfusion of red cells, to which he responded with a severe hemolytic adverse reaction. Antibodies against Kx- and K- antigens were detected. Hematopoietic stem cell transplantation (HSCT) is potentially curative in CGD and was planned for this boy at the age of 14 years because of progressive aspergillosis. Transfusions in this situation can only be given from Kx- and K- antigen negative donors suffering from McLeod syndrome themselves. Resources for these products are extremely limited. Furthermore any transplant would contain Kx- antigen and could potentially boost immunological reactions inducing graft rejection. To cope with these potential complications the following strategy was used: before HSCT autologous red cells as well as bone marrow for an autologous rescue therapy in case of graft rejection were cryopreserved. B-cells were depleted in the patient by two infusions of Mabthera (anti-CD20 antibody). Total body irradiation (12Gy), fludarabine and ATG were given for conditioning. No blood products were necessary during conditioning and before HSCT from a HLA- matched unrelated donor. Transfusions of platelets and red cells from Kx- antigen positive and K- negative donors were well tolerated 11 days after HSCT. No GvHD occurred on prophylaxis with CSA. Hematological and immunological reconstitution with complete donor chimerism was uneventful except a transient episode of immune mediated thrombocytopenia at 5 months after HSCT which responded promptly to steroids. After 10 months of follow up the patient is currently fully engrafted, in an excellent clinical condition and back to school. Conclusion: In patients with X- linked CGD screening for McLeod phenotype needs to be performed prior to transfusion of blood products. Even if sensitization to Kx- and K- antigens has occurred, HSCT can be curative as demonstrated for the first time in this report.
Secondary Cytogenetic Abnormalities and Outcome in Children with TEL-AML1-Positive Acute Lymphoblastic Leukemia.
Authors: Abla Oussama A. , Hitzler Johann , Ye Charles , Abdelhaleem Mohamed , Grant Ronald , Nie Gloria , Kangesan Shanthini , Teshima Ikuko , Shago Mary 
Published: April 2020

Abstract
The t(12;21) translocation, which results in the fusion of the TEL (ETV6) and AML1 (RUNX1) genes, is present in ~25% of pediatric B-precusor acute lymphoblastic leukemia (ALL) patients. Although initially thought to be a favorable prognostic indicator, the t(12;21) was associated with a similar rate of relapse as t(12;21)-negative pre-B ALL in subsequent studies. While secondary cytogenetic abnormalities are often present, their prognostic significance is unknown. The objective of this study is therefore to examine the type, frequency and prognostic impact of secondary cytogenetic abnormalites in t(12;21)-positive blasts of children with ALL. We studied retrospectively 56 patients diagnosed with t(12;21)-positive ALL at the Hospital for Sick Children between 2000–2005. The mean age at diagnosis was 4.9 years (range &lt;1 to 12 years). Nine patients (16%) presented with a white blood cell count of greater than 50 × 109/L. Cytogenetic studies consisted of a combination of FISH, G-banding and spectral karyotyping. The most frequent secondary cytogenetic changes in patients with t(12;21)-positive ALL were: the deletion of the non-translocated TEL allele (38%), an extra copy of the TEL-AML1 fusion gene (14%). Additional numerical abnormalities were seen in approximately one-third of the cases, with the most common being a gain of chromosome 21(13%). Recurrent deletions of chromosomal regions 6q and 11q were observed (14%). A surprising degree of karyotypic complexity was noted. Three or more additional chromosome abnormalities, two abnormal clonal lines, and complex structural rearrangements involving TEL-AML1 were detected in 28%, 21%, and 9% of patients, respectively. Five children (9%) in this cohort have developed a relapse of ALL. The 5-year event-free survival (EFS) was 83±7%. The 5-year EFS for patients with a deleted second TEL allele or an extra TEL-AML1 fusion was not statistically different. Of note, 4 of the children who relapsed had high-risk features at presentation (NCI criteria). All were treated with chemotherapy protocols developed for lower risk ALL. The realization that TEL-AML1 does not always predict excellent prognosis indicates that an on-going study of patients and their survival, with detailed analysis of complex genetic changes, is necessary for a reliable assessment of the prognostic value of the t(12;21) in pediatric ALL.
Perforin Polymorphism A91V and Susceptibility to Childhood Acute Lymphoblastic Leukemia (ALL).
Authors: Mehta Parinda A. , Davies Stella M. , Kumar Ashish , Devidas Meenakshi , Lee Susan , Wilke Tiffany , Elliott James , Villanueva Joyce , Pullen Jeanette , Zewge Yetnayet , Filipovich Alexandra H. 
Published: April 2020

Abstract
Perforin plays a key role in the cytotoxicity of NK cells and cytotoxic T lymphocytes. Genetic mutations in perforin gene (PRF1) give rise to 30–40% of cases of familial hemophagocytic lymphohistiocytosis. We recently studied two boys who had been treated for PreB ALL and subsequently developed HLH. Both showed decreased perforin expression in NK cells and were found to be heterozygous for PRF1 A91V. The functional consequences of the PRF1 A91V (resulting from C272T transition) are controversial; between 3 and 17% of apparently normal people are heterozygous for this polymorphism depending on the population studied. Recent data suggest that A91V results in conformational change that may impair processing of perforin protein to the active form, and that the polymorphism may increase susceptibility to ALL in children (Trambas et al, Blood2005; 106:932; Santoro et al, Haematologica2005;90:697). We have genotyped 655 normal blood donors and 2,272 children with ALL treated on Pediatric Oncology Group 9900 clinical trials. Genotyping was performed using a specific PCR assay (Taqman). Genotypes in the normal US blood donors showed significant variation in allele frequency by race, with a very low frequency of the variant allele in black controls (0.7% compared with 4% in white controls). Allele frequencies did not vary by gender and genotype frequencies were in Hardy Weinberg equilibrium (whites C/C 92%; C/T 8%; T/T 0.2%). In light of the low frequency of the variant allele in blacks, analysis of leukemia cases was restricted to a comparison of white cases and white controls. Analyses were performed for all B-lineage ALL cases combined, and were also further stratified for known genetic subtypes of ALL.
Perforin genotype CC Perforin genotype CT or TT p-value (cases versus controls Controls (n=507) 464 (91.5%) 43 (8.5%) - Cases (all; n= 1321) 1198 (90.7%) 123 (9.3%) 0.58 BCR-ABL 22 (76%) 7 (24%) 0.0048 Trisomy 4,10 238 (93.7%) 16 (6.3%) 0.29 TEL-AML1 250 (90.6%) 26 (9.4%) 0.66 E2A-PBX 24 (92.3%) 2 (7.7%) 0.89 MLL abn. 40 (87%) 6 (13%) 0.29 Hyperdiploid 244 (92.4%) 20 (7.6%) 0.66 Hypodiploid 8 (100%) 0 (0%) 0.40
Taken together, these data indicate that the A91V polymorphism is not associated with an overall increased risk of childhood ALL, in contrast to the previous report. PRF1 A91V was identified with significantly increased frequency in children with BCR-ABL positive ALL. However, this observation includes a relatively small number of cases and the potential association should be explored further, perhaps in adult ALL series in which the frequency of BCR-ABL positivity is likely to be high.
Activating FLT3 Mutations in CD4+/CD8− Pediatric T-Cell Acute Lymphoblastic Leukemias.
Authors: Van Vlierberghe Pieter , Meijerink Jules P.P. , Stam Ronald W. , van der Smissen Wendy , van Wering Elisabeth R. , Beverloo H. Berna , Pieters Rob 
Published: April 2020

Abstract
Activating mutations in the FMS-like tyrosine kinase 3 gene (FLT3) are the most common genetic aberration in acute myeloid leukemia (AML). Internal tandem duplications (ITD) in the juxtamembrane (JM) domain, or point mutations (PM) in the activation loop of the tyrosine kinase domain lead to a constitutive activated state of the FLT3 tyrosine kinase. Recently, FLT3 mutations were identified in a cohort of 69 adult T-ALL patients, showing that this genetic abnormality is not only restricted to myeloid leukemias. To validate the incidence of FLT3 mutations in pediatric T-ALL and investigate its relation to outcome and other clinical and immunophenotypical parameters, we screened 72 diagnostic pediatric T-ALL samples. FLT3/ITD mutations were identified in 2/72 pediatric T-ALLs (2.7%), whereas 0/72 showed point mutations in the kinase domain. Immunophenotypic analysis revealed a similar profile for both FLT3 mutated patient samples, i.e. TdT+, CD2+, CD5+, CD7+, CD4+/CD8−, cytoplasmic CD3+, surface CD3− and CD10−. Although representing early T-cell differentiation stages for both patient samples, these cases seem to have a more advanced immunophenotype compared to the FLT3 mutated adult T-ALL cases, previously described (CD34+, CD4−/CD8−). Both FLT3 mutated patients showed high level LYL1 and LMO2 expression. In addition, both pediatric samples contained a HOX11L2 translocation, which was not present in the FLT3 mutated adult T-ALL cases. The first FLT3 mutated patient suffered a relapse 13 months after initial diagnosis, whereas the other is still in continued complete remission for 61+ months. Interestingly, the relapse material showed no FLT3/ITD mutation, indicating that the FLT3 mutated T-ALL subclone seems to be effectively eradicated by current chemotherapy. These data suggest that the application of FLT3 inhibitors for FLT3-mutated T-ALLs, as recently suggested in literature, may not further improve treatment outcome in pediatric T-ALL.
Successful In Vitro Priming of EBV-Specific CD8+ T Cells Endowed with Strong Cytotoxic Function for Prophylaxis or Treatment of PTLD in EBV Seronegative Hosts.
Authors: Comoli Patrizia , Locatelli Franco , Frasson Chiara , Basso Sabrina , Labirio Massimo , Moretta Antonia , Montagna Daniela , Ginevri Fabrizio , Maccario Rita 
Published: April 2020

Abstract
EBV-seronegative solid organ transplant recipients, or recipients of T-cell depleted hematopoietic stem cell from a EBV-seronegative donor, are cohorts at high risk for the development of EBV-associated posttransplant lymphoproliferative disease (PTLD), and would maximally benefit from an EBV-directed T-cell therapy for prevention or treatment of PTLD. Reported experiences have shown that CTL obtained from EBV-seronegative individuals are almost exclusively HLA-class II-restricted CD4+ cytotoxic T cells, and efficiency of this CTL subset in the control of PTLD has not yet been demonstrated in vivo. Conversely, the central role of CD8+ CTL in the cell-mediated containment of EBV infection is well established. Therefore, we endeavoured to develop a protocol of EBV-CTL priming that would allow optimal expansion and high specific cytotoxic activity of CD8+ CTL. We employed naïve lymphocytes obtained from 5 EBV seronegative patients with end-stage renal failure receiving dialysis while listed for a kidney transplantation (4 children, 1 adult). In particular, we compared the CD8+ CTL priming efficiency of three different modified activation protocols, based on LCL stimulation potentially enhanced by either LCL presentation through DC, or selection of IFNg+ cultured cells, or culture in the presence of rhIL-12 and rhIL-7, to the standard protocol for reactivation of EBV-specific CTL from EBV-seropositive individuals. We found that while all protocols employed, with the exception of IFN-g+ T cell selection, allowed for the amplification of CD8+ and CD4+ mediated EBV-specific cytotoxic T cell responses in the EBV-seronegative adult, only specific EBV-transformed lymphoblastoid cell lines (LCL) stimulation in the presence of rhIL-12 and rhIL-7 was able to reproducibly generate and expand EBV-specific CD8+ CTL endowed with strong cytotoxic activity from truly EBV-seronegative children (mean lysis at a E:T ratio of 10:1: 14±10% with the standard protocol vs. 66±30% with the addition of rhIL-7 and rhIL-12). The lines thus activated showed a higher percentage CD8+ T cells, with less than 10% naïve CD8+/CCR7+/CD45RA+ cells. Overall, the total number of CD8+ central memory cells, and of CCR7- T cell effectors was comparable to that observed in healthy EBV-seropositive controls. In conclusion, stimulation with LCL in the presence of IL-12 and IL-7 activates EBV-specific, highly cytotoxic CD8+ CTL from EBV-seronegative subjects, this opening new perspectives for prevention/treatment of PTLD.
In Vitro Expansion of Autologous Follicular Lymphoma-Specific T Cells for Adoptive Transfer.
Authors: Lee Seung-Tae , Jiang Yun Fang , Cha Soung-Chul , Qin Hong , Kwak Larry W. , Neelapu Sattva S. 
Published: April 2020

Abstract
Advanced stage follicular lymphoma remains an incurable disease with a median survival of 8 to 10 years that has not significantly changed over the last four decades. Therefore, novel treatment options are necessary to improve the clinical outcome in these patients. The observation of spontaneous regressions in a small percentage of patients suggested that augmenting the host immune response could potentially control this malignancy. Strategies using active specific immunotherapy with idiotype vaccines led to induction of clinical and molecular responses in a few patients but have met with only limited success possibly due to the low frequency of antigen-specific T cells induced in the patients. In contrast to active immunization, T cells of a given specificity and function may be selected and expanded in vitro to the desired number for adoptive cell transfer. Towards this goal, we stimulated tumor infiltrating lymphocytes (TILs) or peripheral blood mononuclear cells (PBMCs) from five follicular lymphoma patients with CD40 ligand-activated autologous tumor cells at approximately ten-day intervals in the presence of IL-2 and IL-15. After four rounds of stimulations, T cell lines generated from 3/5 patients recognized autologous unmodified tumor cells by producing significant amounts of TNF-α, GM-CSF and/or IFN-γ. By phenotypic analysis, the T cell lines were predominantly CD4+ T cells (&gt; 70%), and intracellular cytokine assay showed that up to 40% of the CD4+ T cells were tumor-reactive. The inhibition of cytokine production by anti-HLA class II but not class I blocking antibodies confirmed that the CD4+ T cells were tumor-reactive. Further characterization revealed that the T cells from one patient recognized autologous tumor but not autologous normal B cells suggesting that they were tumor-specific. While in a second patient CD4+ T cell clones generated from the T cell line by limiting dilution recognized autologous tumor and autologous normal B cells but not autologous monocytes suggesting that they were B cell lineage-specific. We conclude that follicular lymphoma-specific T cells exist and can be efficiently expanded in vitro from both TILs and PBMCs using CD40 ligand-activated autologous tumor cells for adoptive T cell therapy. Additionally, identification of antigens recognized by these T cells could lead to development of novel immunotherapeutic strategies for lymphomas.
Chemo-Immunotherapy with Hyper-CVAD Plus Ritixumab for Adult Burkitt’s and Burkitt’s Type Lymphoma (BL) or Acute Lymphoblastic Leukemia (B-ALL).
Authors: Thomas Deborah A. , Faderl Stefan , O’Brien Susan , Bueso-Ramos Carlos , Cortes Jorge , Garcia-Manero Guillermo , Giles Francis , Verstovsek Srdan , Wierda William , Pierce Sherry , Shan Jiaquin , Brandt Mark , Hagemeister Fredrick , Cabanillas Fernando , Keating Michael J. , Kantarjian Hagop 
Published: April 2020

Abstract
Prognosis has improved for BL/B-ALL, particularly with short, intensive, multi-agent chemotherapy. Outcome of 26 non-HIV BL/B-ALL patients (pts) treated with hyper-CVAD (fractionated cyclophosphamide, vincristine, doxorubicin, dexamethasone) showed CR rate of 81% with induction mortality 19% owing to systemic fungal infections in pts aged 60 years (yrs) and older [Thomas et al, JCO 17:2461, 1999]. Older age was also an adverse feature predicting relapse with shorter 3 yr survival compared with younger pts (17% vs 77%). To evaluate the addition of rituximab, a CD20 monoclonal antibody, to hyper-CVAD, 31 adults with newly diagnosed HIV negative BL (n=15) or B-ALL (n=16) received hyper-CVAD with rituximab. Their median age was 46 years; 29% were 60 years or older. Course 1 was given in laminar air flow room for pts aged 60 years or older. Rituximab 375 mg/m2 was given intravenously days 1 and 11 of hyper-CVAD courses and days 1 and 8 of methotrexate and cytarabine courses with 16 intrathecal treatments (methotrexate day 2 and cytarabine day 7 of each course). No maintenance chemotherapy was given. No allogeneic or autologous stem cell transplant was planned in first CR. Complete remission (CR) was achieved in 24 of 28 evaluable patients (86%); 3 had a partial response and 1 had resistant disease. There were no induction deaths. The 3-year overall survival (OS), event-free survival, and disease-free survival rates were 89%, 80% and 88% respectively. All nine elderly pts aged ≥ 60 yrs achieved CR with all but one in continuous CR (death in CR from infection); 3-year OS rate was 89%. Nineteen pts (61%) were older than 40 yrs; OS rate was 90% compared with 35% for historical group (48 pts treated with hyper-CVAD alone). Reduction in the relapse rate with rituximab was observed compared with hyper-CVAD alone (7% versus 34% overall, p=0.008; 0% versus 50% for elderly, p=0.02; 11% versus 29% for age &lt; 60 years, p=0.11). Toxicity profile was similar to hyper-CVAD alone. Multivariate analysis of the current cohort and historical group identified age and treatment with rituximab as factors predicting cure. The addition of rituximab to hyper-CVAD appears to improve outcome in adult BL or B-ALL, particularly in elderly patients.