bluuebunny/crossref_metadata_2025_split huggingface dataset:
The Frequency and Clonality of γδ+T Cells, distribution of TRGV and TRDV Repertoire in Cord Blood T Cells
Authors: Li Yang Qiu , Chen Shaohua , Yang Lijian , Li Bo , Cai Dongqing 
Published: April 2020

Abstract
Our previous study has showed the feature of distribution and clonality of T-cell receptor alpha variable region (TRAV) and TRBV subfamily T cells. Recently, data indicated the γδ+T cells may play an important role in mediated graft versus leukemia effect after stem cells transplantation and anti-cancer response. In order to farther characterize the repertoire of CB T-cells, the frequency of αβ+ and γδ+T cells were detected in cord blood (CB) by FACS, the CDR3 size of 4 TRGV and 8 TRDV subfamily genes were analyzed in mononuclear cells (MCs) from 16 cord blood samples, using RT-PCR and genescan technique. Ten healthy individuals served as control. To determine the expression level of TRGV subfamily genes, quantitative analysis of TRGV I–III subfamilies was performed by real-time PCR. Low percentage of CD3+TCRγδ+ cells (1.33±0.58%) was observed in CB. The frequency expression was found in TRGVI (93.75%) and TRGVII (81.25%) in CBMCs, whereas the TRGVIII was detected in 9 out of 16 samples (56.25%). In TRDV subfamilies, the mean value of the number of expressed subfamilies in CBMCs is higher than that from adult peripheral blood (PB) group. The most frequently expressed in CB were TRDV1 (100%), TRDV2(93.75), TRDV8 (93.75%) and TRDV3 (81.25%). The frequency of TRDV5 and TRDV8 in CBMCs were significant higher that that from PBMC. The most PCR products of TRGV and TRDV subfamilies from 10 cord blood displayed polyclonal rearrangement pattern, whereas one or two PCR products from 6 CB samples displayed oligoclonality or biclonality. In contrast, PCR products from 9 of 10 adult healthy controls contained at least an oligoclonal peak in different TRGV or TRDV subfamilies respectively. In the present study, all PCR products form TRGVIV were showed monoclonality, the direct sequencing results showed that the V region of TRGVIV joint direct to C region, indicating that there is no rearrangement in TRGVIV and it is a pseudogene. The sequence data have been submitted to genebank and the Accession Number is EU395807. The pattern of TRGV subfamily expression level in CBMCs was TRGVI&gt;TRGVIII&gt;TRGVII, in contrast, TRGVII&gt;TRGVI&gt;TRGVII was found in PBMCs. In conclusion, our results indicate polyclonal and more diverse TRDV segment usage in CB γδ+ T-cells. The pattern of TRGV expression levels was different between T cells from PB and CB.
Characterization of MHC Class-I Restricted TCRαβ+CD4−CD8− Double-Negative T Cells Recognizing the gp100 Antigen from a Melanoma Patient after gp100 Vaccination
Authors: Voelkl Simon , Moore Tamson , Rehli Michael , Nishimura Michael , Fischer Karin , Mackensen Andreas 
Published: April 2020

Abstract
The immune attack against malignant tumors requires the concerted action of CD8+ cytotoxic T lymphocytes (CTL) as well as CD4+ T helper cells. The contribution of T cell receptor (TCR)αβ+ CD4− CD8− double-negative (DN) T cells to anti-tumor immune responses is widely unknown. In previous studies, we have demonstrated that DN T cells with a broad TCR repertoire are present in humans in the peripheral blood and the lymph nodes of healthy individuals. Here we characterize a human DN T cell clone (T4H2) recognizing an HLA-A2-restricted melanoma-associated antigenic gp100-peptide isolated from the peripheral blood of a melanoma patient. Antigen recognition by the T4H2 DN clone resulted in specific secretion of IFN-γ and TNF. Although lacking the CD8 molecule the gp100-specifc DN T cell clone was able to confer antigen-specific cytotoxicity against gp100-loaded target cells as well as HLA-A2+ gp100 expressing melanoma cells. The cytotoxic capacity was found to be perforin/granzymeB-dependent. Together, these data indicate that functionally active antigen-specific DN T cells recognizing MHC class I-restricted tumor-associated antigen (TAA) may contribute to anti-tumor immunity in vivo.
Allergen-Specific B Cell Quantity Is Similar in Allergic and Nonallergic Individuals, Whereas Allergen-Specific T Cells May Be Increased in Allergic Individuals
Authors: Khan Faisal , Ueno-Yamanouchi Aito , Serushago Bazir , Bowen Tom , Lu Cathy , Luider Joanne , Storek Jan 
Published: April 2020

Abstract
Allergen-specific IgE production is a hallmark of allergic asthma, rhinitis or eczema. Theoretically this could be due to a high number of allergen-specific B cells, a high number of allergen-specific T cells helping allergen-specific B cells to differentiate into IgE plasma cells, or other mechanisms. In this study, we compared the number of allergenspecific B cells and T helper (Th) cells in 41 patients with allergic asthma/rhinitis/eczema (allergic individuals) and 34 nonallergic individuals. Allergen-specific B and Th cells were enumerated by culturing CFSE-loaded blood mononuclear cells for 7 days with allergen (cat, dog, D.pteronyssimus, Timothy or birch), and determining by flow cytometry the number of B or Th cells that had proliferated (diluted CFSE). The quantities of B cells specific for each of the 5 allergens were similar in individuals allergic to the allergen (per skin prick test result) compared to nonallergic individuals. The quantity of Timothy-specific Th cells was 3-times higher in Timothy-allergic individuals compared to nonallergic individuals (p=0.023). In contrast, the quantity of cat-specific Th cells was similar in cat-allergic and nonallergic individuals. The quantities of dog, D.pteronyssimus and birch-specific Th cells were slightly higher in dog, D.pteronyssiumus and birch-allergic compared to nonallergic individuals. No significant change in the number of allergen specific B and Th cells was found when blood drawn from the same individual at different time points was compared for 3 allergic and 3 non-allergic individuals. We conclude that for some allergens (eg, Timothy), a high number of allergen-specific Th cells, but not B cells, may play a role in the pathogenesis of allergic asthma/rhinitis/eczema. For other allergens (eg, cat), the pathogenesis may be different.
CD16/FcãRIII Antigen Expression Determines Oxidative Sensitivity in Cytotoxic Lymphocyte Subsets
Authors: Aurelius Johan , Askarieh Galia , Romero Ana I. , Thorén Fredrik B. , Brune Mats L , Hellstrand Kristoffer 
Published: April 2020

Abstract
Oxidative stress, referring to cellular toxicity induced by reactive oxygen metabolites (ROM), has been proposed as a mechanism of immunosuppression in hematological malignant diseases. ROM produced by myeloid cells such as normal or malignant phagocytes have been shown to compromise functions of lymphocytes including natural killer (NK) cells and CD4+ or CD8+ T cells. Phagocyte-derived ROM induce downmodulation of the CD3/CD16 zeta; transduction antigen in these lymphocyte subsets with ensuing loss of function and lack of responsiveness to activating cytokines such as IL- 2. In addition, ROM exposure triggers poly(ADP-ribose) polymerase 1- and apoptosisinducing factor-dependent apoptosis in NK and T cells, but the molecular details of the ROM/lymphocyte interaction are not fully understood. The present study sought to determine the role of the CD16/FcγRIII (CD16) antigen, a signal transduction molecule expressed by NK and T cells and a receptor for the Fc portion of IgG, for oxidative stress in lymphocytes. Human NK (CD56+) or T cells (CD4+ or CD8+) were sorted into pure (&gt;99%) CD16+ or CD16− populations, and incubated overnight with ROM-producing autologous mononuclear phagocytes or exogenous hydrogen peroxide. For all subsets of lymphocytes, the expression of CD16 was a strong determinant of the propensity of cells to acquire features of apoptosis after ROM exposure, as reflected by a pronounced difference of ROM sensitivity between CD16+ and CD16− NK or T cell populations. The higher degree of ROM-induced apoptosis in the CD16+ subsets was observed regardless of whether lymphocytes were incubated with autologous mononuclear phagocytes or exposed to exogenous hydrogen peroxide (p=0.04–0.001 for CD4+/16+, CD8+/16+ and CD16+/56+ lymphocytes as compared with corresponding CD16− subsets). Similar experiments using unsorted T and NK cells suggested that the differential sensitivity to oxidative stress in CD16+ and CD16− lymphocyte subsets was not explained by anti- CD16/CD16 interaction. In conclusion, our findings imply that oxidant-induced apoptosis is preferentially induced in lymphocytes expressing the CD16/FcγRIII antigen.
Clonal CD27+ CD19+ B-Cell Expansion through Inhibition of FCgIIR in HCV+ Lymphorpoliferative Disorders
Authors: De Re Valli , Pavan Alessandro , Racanelli Vito , Sansonno Silvia , Toffoli Giuseppe 
Published: April 2020

Abstract
HCV infection may or may not be associated with extra-hepatic manifestations such as type-II mixed cryoglobulinemia (MC), a clonal B cell proliferative disorder. In persistent HCV infection without MC a increase in serum immunoglobulins (Ig) is commonly observed. We found this increase is polyclonal and is determined primarily by increased levels of both HCV-specific and nonspecific IgG. Despite this hypergammaglobulinemia, memory CD27+ do not accumulate, depending on a heightened sensitivity of memory B cells to BCR-independent noncognate T cell help which speeds up their terminal differentiation into antibody secreting cells and make them more prone to activation induced cell death. In persistent HCV infection with MC, elevation of Ig is a general occurrence too. However, it is attributable to IgG and IgM. The latter include antibodies with rheumatoid factor (RF) activity, which are essential for the development of circulating, cryoprecipitable immune complexes. We found Hypergammaglobulinemia is sustained by a peripheral expansion of IgMk+restriction indicating that a limited number of antigens drives their proliferation through BCR interaction. We shown that IgM RF and their counterparts on the surface of bone marrow-resident monoclonal B cells react against a Fc-epitope of IgG and the HCV-NS3 protein. Based on the above findings, we propose a model whereby BCR, by binding the Fc of IgM/IgG/HCVNS3 immune complexes deprives FcgIIR of its natural ligand. This takes the brake off RF+CD27+ B cell proliferation and promotes their selective accumulation which is otherwise prevented by increased apoptosis susceptibility in persistent HCV infection without MC. By confocal microscopy analysis we found that both Fc and NS3 peptides are able to clusterize BCR. Cytofluorimetric analysis found activation of both SYK/ERK phosphorilation pathways. Both these observations are in agreement with the hypothesis of BCR activation in these samples. Data regarding FcR are in course.
HLA Influence on NK Cell Expansion
Authors: Schellekens Jennifer , Stserbakova Anna , Tõns Madis , Everaus Hele , Tilanus Marcel GJ , Aints Alar 
Published: April 2020

Abstract
Natural Killer (NK) cells are effector cells in the innate immune system. The anti-leukaemic capacities of NK cells in haematopoietic stem cell transplantation make these cells a potential treatment modality to improve clinical outcome. Immunotherapy with NK cells requires transfusion of large quantities, which obviates the need for an in vitro culture system for NK cells. The killer cell immunoglobulin-like receptors (KIR) on NK cells recognise defined groups of HLA class I alleles. To elucidate the influence of these interactions on proliferation, the peripheral blood mononuclear cells (PBMCs) of 29 patients and donors were cultured in CellGro SCGM with IL-2 and OKT3 antibody to expand the NK cell fraction. The killer cell immunoglobulin-like receptor (KIR) and HLA repertoire were determined by sequence specific priming and sequence based typing respectively. The percentage of NK cell expansion from the total PBMC fraction varied between 5.4% and 71.6%. A significantly better NK cell expansion was observed for individuals homozygous for HLA-C epitope group 2 (p&lt;0.05). For evaluation of cytolytic competence of the cultured NK cells, specific killing of an HLA class I expression deficient LCL 721.221 cell line and three 721.221 cell lines transfected with different HLA-C alleles was determined. A significantly better NK cell-induced specific cytotoxicity was observed towards the untransfected 721.221 cells compared to the HLA-C transfected 721.221 cells. No significant differences were observed between killing of the three HLA-C transfected 721.221 cell lines. We have shown that cytolytic capacities of the cultured NK cells are maintained and in vitro expansion of NK cells is dependant on the presence of HLA-C alleles.
KIR Gene and HLA-Cw Mismatches Affect on NK Cell Killer Activity
Authors: Cao Xiangshan , Li Jianyong 
Published: April 2020

Abstract
The KIRs were knew as natural killer (NK) cell inhibitory receptors with specificity for HLA molecules on their cellular targets. We investigated NK cell activation on the number of matches between cell killer immunoglobulin-like receptor (KIR) gene and HLA-Cw, and the level of inhibitory KIRs expressed on NK cell surface and the cytotoxicity of NK cell against AML leukemic cells in vitro. NK cell were isolated and purified from 27 healthy donors by isolation kit, Target cells were blasts derived from bone marrow of 30 patients with AML.Inhibitory KIRs expression knew as CD158a, CD158b was analyzed by flow cytometry to estimate the percentage of NK cells that could be inhibited by the HLA-Cw ligands..KIR and HLA gene typing were performed by PCR –SSP. from donors and patients respectively. NK cytotoxicity against AML leukemic cells demonstrated by MTT which showed the correlation between NK cytotoxicity and the number of KIR/HLA matches. the NK-susceptible K562 cell line which lacks HLA class I expression, was used as a positive control target in all cytotoxicity assays, autologous non-NK cell was used as negative control target cell. the cytotoxicity assays was performed in E:T 50:1 20:1 10:1 5:1 2.5:1. Results demonstrated the less number of KIR/HLA-Cw matches, the more NK cells are activated..0 match of NK cell/target cell KIR/HLA-Cw, cytotoxicity was (50.66±8.40)%,1 match and 2 matches were (38.28±6.71)%, (19.74±4.15)%, F=20.226, P&lt;0.001. NK cells expressed KIRs also had relationship with cytotoxicity, inhibitory KIRs expressed &gt;50%, the cytotoxicity is 10%, inhibitory KIRs expressed 20%–50%, the cytotoxicity is 20%, inhibitory KIRs &lt;25%, the cytotoxicity is 55%, F=16.276,p&lt;0.001. Therefore these data indicate NK cell kill AML leukemic cells mechanism follow KIR/HLA-Cw mismatch theory, the level of inhibitory KIRs expressed on NK cell surface showed the percentage of NK cells that could be inhibited by the HLA-Cw ligands. Key words: KIR NK cell CD158 HLA-Cw
A Comparative Oncogenomic Analysis Identifies Genetic Events Associated with Myeloma Progression and Drug Resistance
Authors: Chesi Marta , Braggio Esteban , Keats Jonathan J , Bergsagel P. Leif 
Published: April 2020

Abstract
We have successfully generated the first immuno-competent transgenic mouse model of multiple myeloma (MM), Vk*MYC, in which sporadic MYC activation in the germinal center is AID dependent and mediated by the somatic hypermutation process. All of Vk*MYC mice develop monoclonal PC expansion by 50 weeks of age, that is manifested by high levels of serum IgGs and major M-spikes by SPEP. The PCs in Vk*MYC mice are fully differentiated, somatically mutated, have a very low proliferation index and are found exclusively in the BM. Associated with the long lifespan (medium survival = 661d), Vk*MYC mice develop anemia and diffuse bone disease (osteoporosis with low bone mineral density), with sporadic occurrence of lytic bone lesions and hind limb paralysis. The phenotypic fidelity of the Vk*MYC mice translates in a therapeutic fidelity, as Vk*MYC mice show response to known clinically active anti-myeloma drugs (bortezomib, melphalane, dexamethasone), but not to drugs that have shown no clinical activity as single agents (vincristine, hydroxyurea, fludarabine). Furthermore, acquired bortezomib resistance could efficiently be induced in Vk*MYC mice after only four cycles of suboptimal treatment, and studies are in progress to identify genetic events associated with bortezomib resistance. Preliminary aCGH experiments on a high resolution Agilent 244K array, using genomic DNA from six Vk*MYC CD138+ selected PCs, showed that the genomic complexity of Vk*MYC tumors is quite low, with only few unbalanced rearrangements, such as trisomy of chromosome 15 detected in two samples or trisomy 9 present in another tumor. The most common abnormalities found are small focal monoallelic deletions, six of which are present in more than one tumor. A focal homozygous deletion of several exons of a gene was identified in two tumors samples. Remarkably, a comparative analysis of the syntenic chromosomal region in human MM cell lines and primary patient samples identified similar recurrent homozygous deletion in this putative novel tumor suppressor gene. Overall, these data suggest that the genomic complexity in this inbred strain of transgenic mice is significantly less then in patients with MM, simplifying the search for critical loci involved in MM tumorigenesis and drug resistance and further validate the Vk*MYC mice as a particularly faithful model of human MM.
Primary Central Nervous System Lymphoma of T-Cell Phenotype: A Rare Variant of Non-Hodgkin’s Lymphoma Treated with CNS Directed Therapy
Authors: Doolittle Nancy D , Neuwelt Edward A 
Published: April 2020

Abstract
Primary central nervous system lymphoma (PCNSL) is a non-Hodgkin’s lymphoma confined to the craniospinal axis and/or eyes, without evidence of systemic spread. Almost all PCNSL’s are classified as large B-cell lymphoma. A rare variant of PCNSL includes lymphomas of the T-cell phenotype. Until the publication of a large retrospective series of patients (pts) (n = 45) with T-cell PCNSL (Shenkier TN, et al. J Clin Oncol2005;23:2233–2239),only case reports of T-PCNSL and literature reviews were available. The large retrospective series reported that ECOG Performance Status (PS) (0 or 1 versus 2, 3 or 4) at diagnosis was significantly associated with better outcome (Log rank P &lt;.0001), as was primary treatment with high-dose methotrexate (MTX) (yes versus no) (Log rank P = .002). Median survival in the large series was 25 months (95% CI: 11 to 38 months). We reviewed our brain tumor database for pts seen between January 1980 and July 2008. Of 159 pts diagnosed with PCNSL, 3 (2M/1F) had histopathologically confirmed large T-cell phenotype. At diagnosis, the pts’ ages were 10, 23, and 64 years; in each case ECOG PS at diagnosis was 0. Brain parenchyma disease location was periventricular (1 pt), parietal (1 pt), and corpus callosum (1 pt); one pt had both brain and ocular lymphoma. First-line treatment was IA MTX-based chemotherapy with osmotic blood-brain barrier disruption (BBBD), without brain irradiation. The dose of MTX was 2.5 grams/day for 2 consecutive days, every 4 weeks for up to one year. A comprehensive neuropsychological test battery was administered at baseline prior to MTX-based BBBD treatment. Followup neuropsychological testing was conducted within 6 months after the final BBBD treatment. All patients attained CR and are alive 15.8, 8.9 and 1.2 years after diagnosis, with excellent PS and no evidence of disease progression to date. The 3 pts showed stable or improved neuropsychological function in all cognitive domains at follow-up testing. Side effects during BBBD treatment included focal seizures (1 pt) and atrial fibrillation (1 pt). In these 3 cases of T-cell PCNSL, encouraging tumor responses and survival were seen, with stable or improved neurocognitive functioning to date, warranting further investigation of CNS directed therapy in this rare variant of non-Hodgkin’s lymphoma.
A Prospective Study of Cardiotoxicity In Non-Hodgkin Lymphoma Patients with RAPID-Infusion Rituximab
Authors: Provencio Mariano , Sanchez Antonio , Mendez Miriam , Maximiano Constanza , Cantos Blanca , Vargas Juan A , Tutor Pablo , Mellor Susana , Canales Miguel A. , España Pilar 
Published: April 2020

Abstract
Background. Prospective study based on measuring the left ventricular ejection function (LVEF) of lymphoma patients treated with rapid infusion Rituximab (one hour).
Patients and Methods. 42 patients with non-Hodgkin lymphoma were treated with rapid infusion Rituximab-based chemotherapy (mostly CHOP-R). Basal LVEF was measured before chemotherapy and every six months after treatment.
Results. An average of 6.5 infusions per patient were administered (total of 273 infusions). Twenty-four (57%) patients had at least one cardiovascular risk factor. In 13 patients (31%) we observed a decreased in the post-treatment LVEF of over 10%. Patients (3) with drop &gt; 10% recovered normally, but when LVEF decreased by over 15% (6 patients), no one recovered until normal level. None of them developed clinical congestive failure.
Conclusion. Rapid infusion Rituximab added to chemotherapy does not cause clinical cardiac toxicity. Patients recover LVEF when it decreased less than 15%.
Dual PI3K and mTOR Inhibition in Waldenstrom Macroglobulinemia
Authors: Husu Emanuel N. , Roccaro Aldo M. , Sacco Antonio , Melhem Molly R. , Azab Abdel kareem , Jia Xiaoying , Ngo Hai T. , Azab Feda , Runnels Judith , Quang Phong , Leleu Xavier , Anderson Kenneth C. , Ghobrial Irene M. 
Published: April 2020

Abstract
Background. We have previously showed that PI3K/Akt is constitutively active in Waldenström Macroglobulinemia (WM) malignancies, mediating growth, survival, cell cycle regulation, and migration in primary tumor cells. Once activated, Akt phosphorylates downstream targets, including mammalian target of rapamycin (mTOR). Both PI3K/Akt and mTOR represent valid targets for antitumor therapeutic strategies. We therefore evaluated the antitumor activity of NVP-BEZ235 (Novartis, MA) in WM.
Methods: WM cell lines (BCWM.1) and IgM secreting cell lines (MEK1, Namalwa) were used. Bone marrow primary CD19+ malignant cells and bone marrow stromal cells (BMSC) were obtained from WM patients. Cytotoxicity, DNA synthesis, and cell cycle were measured using MTT assay, [3H]-thymidine uptake, PI staining/flow cytometry, respectively. Effects of NVP-BEZ235 on cell signaling cascades were determined using immunoblotting and immunofluorescence. Adhesion on fibronectin has been evaluated in WM cells in the presence of NVP-BEZ235.
Results: NVP-BEZ235 induced cytotoxicity and inhibited DNA synthesis with an IC50 of 20–25nM in BCWM.1 at 48 hours. Similar effects were demonstrated in all IgM secreting cell lines and in primary CD19+ WM cells, with an IC50 between 20nM and 50nM. No cytotoxicity was observed on peripheral blood mononuclear cells, indicating selective toxicity of the compound on the malignant lymphoplasmacytic clone. We observed that NVP-BEZ235 inhibited Akt (but not ERK phosphorylation) in a dose-dependent manner in BCWM.1 cells at 6 hours. Phosphorylation of GSK3α/β and ribosomal protein-S6, downstream target proteins of Akt, were also markedly inhibited. NVP-BEZ235-inhibited Akt phosphorylation was further confirmed by immunofluorescence. NVP-BEZ235 induced caspase-9, PARP cleavage and increased the release of Smac/DIABLO from the mitochondria to the cytosol, suggesting an induction of apoptosis in a caspase-dependent and –independent manner. We showed that NVP-BEZ235 inhibited adhesion of BCWM.1 cells to fibronectin in a dose-dependent fashion. Lastly, adherence to BMSCs did not confer protection to WM cells against NVP-BEZ235- induced cytotoxicity.
Conclusions. These data indicate that NVP-BEZ235 has significant antitumor activity in WM, thus providing the framework for clinical trials in this disease.