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https://www.readbyqxmd.com/read/28910385/improving-the-characterization-of-endothelial-progenitor-cell-subsets-by-an-optimized-facs-protocol
#1
Karin Huizer, Dana A M Mustafa, J Clarissa Spelt, Johan M Kros, Andrea Sacchetti
The characterization of circulating endothelial progenitor cells (EPCs) is fundamental to any study related to angiogenesis. Unfortunately, current literature lacks consistency in the definition of EPC subsets due to variations in isolation strategies and inconsistencies in the use of lineage markers. Here we address critical points in the identification of hematopoietic progenitor cells (HPCs), circulating endothelial cells (CECs), and culture-generated outgrowth endothelial cells (OECs) from blood samples of healthy adults (AB) and umbilical cord (UCB)...
2017: PloS One
https://www.readbyqxmd.com/read/28769101/allostimulatory-capacity-of-conditionally-immortalized-proximal-tubule-cell-lines-for-bioartificial-kidney-application
#2
Milos Mihajlovic, Lambertus P van den Heuvel, Joost G Hoenderop, Jitske Jansen, Martijn J Wilmer, Annemarie J F Westheim, Wil A Allebes, Dimitrios Stamatialis, Luuk B Hilbrands, Rosalinde Masereeuw
Novel renal replacement therapies, such as a bioartificial kidney (BAK), are needed to improve current hemodialysis treatment of end-stage renal disease (ESRD) patients. As BAK applications may reveal safety concerns, we assessed the alloimmunization and related safety aspects of readily available conditionally immortalized human proximal tubule epithelial cell (ciPTEC) lines to be used in BAK. Two ciPTEC lines, originally derived from urine and kidney tissue, were characterized for the expression and secretion of relevant molecules involved in alloimmunization and inflammatory responses, such as HLA class-I, HLA-DR, CD40, CD80, CD86, as wells as soluble HLA class I and proinflammatory cytokines (IL-6, IL-8 and TNF-α)...
August 2, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28758931/tumor-stroma-crosstalk-in-bone-tissue-the-osteoclastogenic-potential-of-a-breast-cancer-cell-line-in-a-co-culture-system-and-the-role-of-egfr-inhibition
#3
Laura Mercatali, Federico La Manna, Giacomo Miserocchi, Chiara Liverani, Alessandro De Vita, Chiara Spadazzi, Alberto Bongiovanni, Federica Recine, Dino Amadori, Martina Ghetti, Toni Ibrahim
Although bone metastases represent a major challenge in the natural history of breast cancer (BC), the complex interactions involved have hindered the development of robust in vitro models. The aim of this work is the development of a preclinical model of cancer and bone stromal cells to mimic the bone microenvironment. We studied the effects on osteoclastogenesis of BC cells and Mesenchymal stem cells (MSC) cultured alone or in combination. We also analyzed: (a) whether the blockade of the Epithelial Growth Factor Receptor (EGFR) pathway modified their influence on monocytes towards differentiation, and (b) the efficacy of bone-targeted therapy on osteoclasts...
July 29, 2017: International Journal of Molecular Sciences
https://www.readbyqxmd.com/read/28754882/a-regulatory-role-for-the-co-chaperone-fkbp51s-in-pd-l1-expression-in-glioma
#4
Paolo D'Arrigo, Michele Russo, Anna Rea, Martina Tufano, Elia Guadagno, Maria Laura Del Basso De Caro, Roberto Pacelli, Felix Hausch, Stefania Staibano, Gennaro Ilardi, Silvia Parisi, Maria Fiammetta Romano, Simona Romano
BACKGROUND: FKBP51 is a co-chaperone with isomerase activity, abundantly expressed in glioma. We previously identified a spliced isoform (FKBP51s) and highlighted a role for this protein in the upregulation of Programmed Death Ligand 1 (PD-L1) expression in melanoma. Because gliomas can express PD-L1 causing a defective host anti-tumoral immunity, we investigated whether FKBP51s was expressed in glioma and played a role in PD-L1 regulation in this tumour. METHODS: We used D54 and U251 glioblastoma cell lines that constitutively expressed PD-L1...
July 17, 2017: Oncotarget
https://www.readbyqxmd.com/read/28682474/autologous-serum-supplement-favours-in-vitro-regenerative-paracrine-factors-synthesis
#5
Nazmul Haque, Noor Hayaty Abu Kasim, Noor Lide Abu Kassim, Mohammad Tariqur Rahman
OBJECTIVES: Foetal bovine serum (FBS) is often the serum supplement of choice for in vitro human cell culture. This study compares the effect of FBS and autologous human serum (AuHS) supplement in human peripheral blood mononuclear cell (PBMC) culture to prepare secretome. MATERIALS AND METHODS: The PBMC (n = 7) were cultured either in RPMI-1640 containing L-glutamine and 50 units/ml Penicillin-Streptomycin (BM) or in BM with either AuHS or FBS. Viability, proliferation and differentiation of PBMC were evaluated...
August 2017: Cell Proliferation
https://www.readbyqxmd.com/read/28680763/the-influence-of-stromal-cells-and-tumor-microenvironment-derived-cytokines-and-chemokines-on-cd3-cd8-tumor-infiltrating-lymphocyte-subpopulations
#6
Stefan Koeck, Johan Kern, Marit Zwierzina, Gabriele Gamerith, Edith Lorenz, Sieghart Sopper, Heinz Zwierzina, Arno Amann
The tumor microenvironment has been identified as a major mediator of immunological processes in solid tumors. In particular, tumor-associated fibroblasts are known to interact with tumor infiltrating immune cells. We describe the influence of fibroblasts and tumor-microenvironment-derived cytokines on the infiltration capacity of CD3(+)CD8(+) cytotoxic T lymphocyte subpopulations using a multicellular 3D co-culture system. 3D tumor microtissues were cultivated using a hanging drop system. Human A549 and Calu-6 cancer cell lines were incubated alone or together with the human fibroblast cell line SV80 for 10 d to form microtissues...
2017: Oncoimmunology
https://www.readbyqxmd.com/read/28656222/activation-of-the-toll-like-receptor-8-pathway-increases-the-immunogenicity-of-mesenchymal-stem-cells-from-umbilical-cord
#7
Yu Yang, Yanwen Wang, Li Li, Fei Chen, Peng Zhang
Mesenchymal stem cells (MSCs) are now widely used in clinical cell‑based therapy due to their characteristics of low immunogenicity, multiple differentiation potency and the capability to modulate immune responses. However, accumulated research has indicated the absence of engrafted MSCs because of the increased immunogenicity of MSCs. Toll‑like receptors (TLRs) are essential for the innate immune response and regulating the biological function of MSCs. The present study used human umbilical cord‑derived MSCs (UCMSCs) and activated the TLR8 pathway of UCMSCs to study the role of TLR8 in mediating the immune status of UCMSCs...
August 2017: Molecular Medicine Reports
https://www.readbyqxmd.com/read/28637474/arsenic-trioxide-inhibits-ebv-reactivation-and-promotes-cell-death-in-ebv-positive-lymphoma-cells
#8
Qinyan Yin, Mark Sides, Christopher H Parsons, Erik K Flemington, Joseph A Lasky
BACKGROUND: Epstein-Barr Virus (EBV) is associated with hematopoietic malignancies, such as Burkitt's lymphoma, post-transplantation lymphoproliferative disorder, and diffuse large B-cell lymphoma. The current approach for EBV-associated lymphoma involves chemotherapy to eradicate cancer cells, however, normal cells may be injured and organ dysfunction may occur with currently employed regimens. This research is focused on employing arsenic trioxide (ATO) as EBV-specific cancer therapy takes advantage of the fact the EBV resides within the malignant cells...
June 21, 2017: Virology Journal
https://www.readbyqxmd.com/read/28594941/low-level-laser-lll-attenuate-lps-induced-inflammatory-responses-in-mesenchymal-stem-cells-via-the-suppression-of-nf-%C3%AE%C2%BAb-signaling-pathway-in-vitro
#9
Kan Yin, Rongjia Zhu, Shihua Wang, Robert Chunhua Zhao
BACKGROUND: Considering promising results in animal models and patients, therapeutic use of MSCs for immune disease is likely to undergo continued evaluation. Low-lever laser (LLL) has been widely applied to retard the inflammatory reaction. LLL treatment can potentially be applied in anti-inflammatory therapy followed by stem cell therapy. AIM OF THE STUDY: The purpose of this study was to investigate the effect of LLL (660 nm) on the inflammatory reaction induced by LPS in human adipose derived mesenchymal stem cells (hADSCs) and pertinent mechanism...
2017: PloS One
https://www.readbyqxmd.com/read/28584241/e-coli-promotes-human-v%C3%AE-9v%C3%AE-2%C3%A2-t-cell-transition-from-cytokine-producing-bactericidal-effectors-to-professional-phagocytic-killers-in-a-tcr-dependent-manner
#10
M Barisa, A M Kramer, Y Majani, D Moulding, L Saraiva, M Bajaj-Elliott, J Anderson, K Gustafsson
γδT cells provide immune-surveillance and host defense against infection and cancer. Surprisingly, functional details of γδT cell antimicrobial immunity to infection remain largely unexplored. Limited data suggests that γδT cells can phagocytose particles and act as professional antigen-presenting cells (pAPC). These potential functions, however, remain controversial. To better understand γδT cell-bacterial interactions, an ex vivo co-culture model of human peripheral blood mononuclear cell (PBMC) responses to Escherichia coli was employed...
June 5, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28544640/an-in%C3%A2-vitro-model-demonstrates-the-potential-of-neoplastic-human-germ-cells-to-influence-the-tumour-microenvironment
#11
B Klein, H-C Schuppe, M Bergmann, M P Hedger, B E Loveland, K L Loveland
Testicular germ cell tumours (TGCT) typically contain high numbers of infiltrating immune cells, yet the functional nature and consequences of interactions between GCNIS (germ cell neoplasia in situ) or seminoma cells and immune cells remain unknown. A co-culture model using the seminoma-derived TCam-2 cell line and peripheral blood mononuclear cells (PBMC, n = 7 healthy donors) was established to investigate how tumour and immune cells each contribute to the cytokine microenvironment associated with TGCT...
May 23, 2017: Andrology
https://www.readbyqxmd.com/read/28504020/propoxur-induced-oxidative-dna-damage-in-human-peripheral-blood-mononuclear-cells-protective-effects-of-curcumin-and-%C3%AE-tocopherol
#12
Tanzeel Ahmed, Vasu Goel, B D Banerjee
The present study enumerates the attenuating effects of curcumin and α-tocopherol against propoxur induced oxidative DNA damage in human peripheral blood mononuclear cells (PBMC). Cultured cells were isolated from peripheral blood of healthy volunteers, and were exposed to varying concentrations of propoxur (0-21 μg/ml) for 6, 12, and 24 h, and in combination with curcumin (9.2 μg/ml) or α-tocopherol (4.3 μg/ml) or both. Cytotoxic effect of propoxur was examined by MTT assay. The role of oxidative stress beneath the cytotoxicity of propoxur was evaluated by the measurement of reduced glutathione (GSH), malondialdehyde (MDA) and 8-hydroxy-2'-deoxyguanosine (8-OH-dG) levels in cell lysate...
May 15, 2017: Drug and Chemical Toxicology
https://www.readbyqxmd.com/read/28500235/beta-catenin-inhibitor-bc2059-is-efficacious-as-monotherapy-or-in-combination-with-proteasome-inhibitor-bortezomib-in-multiple-myeloma
#13
Ioanna Savvidou, Tiffany Khong, Andrew Cuddihy, Catriona McLean, Stephen Horrigan, Andrew Spencer
Currently available treatment options are unlikely to be curative for the majority of multiple myeloma (MM) patients, emphasizing a continuing role for the introduction of investigational agents that can overcome drug resistance. The canonical Wnt/beta-catenin signalling pathway, essential for self-renewal, growth and survival, has been found to be dysregulated in MM, particularly in advanced stages of disease. This provides the rationale for evaluating the novel beta-catenin inhibitor BC2059 as monotherapy and in combination with proteasome inhibitors in vitro and in vivo...
May 12, 2017: Molecular Cancer Therapeutics
https://www.readbyqxmd.com/read/28474861/group-2-innate-lymphoid-cells-are-elevated-and-activated-in-chronic-rhinosinusitis-with-nasal-polyps
#14
Julie A Poposki, Aiko I Klingler, Bruce K Tan, Pejman Soroosh, Homayon Banie, Gavin Lewis, Kathryn E Hulse, Whitney W Stevens, Anju T Peters, Leslie C Grammer, Robert P Schleimer, Kevin C Welch, Stephanie S Smith, David B Conley, Joseph R Raviv, James G Karras, Omid Akbari, Robert C Kern, Atsushi Kato
BACKGROUND: Chronic rhinosinusitis (CRS) with nasal polyps (CRSwNP) is characterized by type 2 inflammation with high levels of Th2 cytokines. Although T helper cytokines are released from T cells, innate lymphoid cells (ILC) are also known to produce high levels of the same cytokines. However, the presence of various types of ILC in CRS is poorly understood. OBJECTIVE: The objective of this study was to fully characterize the presence of all ILC subsets in CRS and to identify phenotypical differences of group 2 ILC (ILC2) in CRSwNP compared to ILC2 from non-type 2 inflamed areas...
September 2017: Immunity, Inflammation and Disease
https://www.readbyqxmd.com/read/28470343/pbmc-activation-via-the-erk-and-stat-signaling-pathways-enhances-the-anti-tumor-activity-of-staphylococcal-enterotoxin-a
#15
Xueting Liu, Liping Zeng, Zhongqiu Zhao, Jianxing He, Yang Xie, Lanyan Xiao, Shan Wang, Junyan Zhang, Zehong Zou, Ying He, Ailin Tao, Jianguo Zhang
Staphylococcal enterotoxin A (SEA) is well known as a superantigen and is highly potent in activating T lymphocytes. And it has been used clinically as an immunomodifier in the treatment of a number of tumors for years. However, the mechanism of its action remains largely unclear. In this study, SEA was found to significantly inhibit the proliferation and induce the death of human lung carcinoma A549 cells when co-cultured with human peripheral blood mononuclear cells (PBMCs). SEA could also induce the proliferation of human PBMCs and stimulate human PBMCs to release a wide range of cytokines that have broad anti-tumor activities such as IFN-γ, TNF-α, IL-2...
May 3, 2017: Molecular and Cellular Biochemistry
https://www.readbyqxmd.com/read/28411882/in-vitro-generation-of-interleukin-10-secreting-b-regulatory-cells-from-donor-adipose-tissue-derived-mesenchymal-stem-cells-and-recipient-peripheral-blood-mononuclear-cells-for-potential-cell-therapy
#16
Kunal S Gupte, Aruna V Vanikar, Hargovind L Trivedi, Chetan N Patel, Jignesh V Patel
BACKGROUND: Interleukin-10 secreting B-cells are a major subset of B-regulatory cells (B-regs), commonly recognized as CD19(+)/38(hi)/24(hi)/IL10(+). They carry out immunomodulation by release of specific cytokines and/or cell-to-cell contact. We have generated B-regs in-vitro from donor adipose tissue derived mesenchymal stem cells (AD-MSC) and renal allograft recipient (RAR) peripheral blood mononuclear cells (PBMC) for potential cell therapy. MATERIAL AND METHODS: Mononuclear cells separated by density gradient centrifugation from 50 ml anti-coagulated blood of 15-RAR and respective donors were analysed for baseline B-regs using appropriate antibodies...
February 2017: Biomedical Journal
https://www.readbyqxmd.com/read/28376277/dendritic-cells-from-rheumatoid-arthritis-patient-peripheral-blood-induce-th17-cell-differentiation-via-mir-363-integrin-%C3%AE-v-tgf-%C3%AE-axis
#17
F Pan, H Xiang, J Yan, L Hong, L Zhang, Y Liu, X Feng, C Cai
Dendritic cells (DCs) are critical regulators of immune responses. This study was to observe the effect of DCs from peripheral blood on the differentiation of Th17 in patients with rheumatoid arthritis (RA). Peripheral blood samples were collected from 30 patients with RA and 20 healthy controls, respectively. Flow cytometry results showed that in contrast to Treg cells, the proportion of Th17 cells in T cells and the Th17/Treg ratio were both increased in patients with RA. The RT-PCR results showed that Foxp3、ROR γt and miR-363 expression in PBMC of patients with RA were reduced, but the ITGAV expression was increased, which was negatively related to miR-363 expression...
June 2017: Scandinavian Journal of Immunology
https://www.readbyqxmd.com/read/28342803/antigen-specific-ctls-to-produce-autologous-cells-product-for-adoptive-cellular-therapy
#18
Sai Liu, Yi Shao, Jie Xu, Na Jiang, Yanchao Dai, Yu Wang, Huanqing Sun, Jianping Sun, Yonghong Zhang
As antiretroviral therapy provides long term viral suppression but no cure, alternative therapies such as adoptive cellular therapy have thus been investigated in the anti-AIDS field. OBJECTIVE: This study sought to establish a HLA-A02 specific CTL cell culture method with comparison of the effects of different cytokines used in CTL cultivation to decide the best cultivation environment. In order to produce CTLs with targeted HLA-A02 restricted antigen specificity for adoptive cellular therapy, we evaluated autologous PBMC cultivation in different cytokine environment to select a better expansion condition to produce qualified CTL production...
March 22, 2017: International Journal of Infectious Diseases: IJID
https://www.readbyqxmd.com/read/28331616/ex-vivo-akt-inhibition-promotes-the-generation-of-potent-cd19car-t-cells-for-adoptive-immunotherapy
#19
Ryan Urak, Miriam Walter, Laura Lim, ChingLam W Wong, Lihua E Budde, Sandra Thomas, Stephen J Forman, Xiuli Wang
BACKGROUND: Insufficient persistence and effector function of chimeric antigen receptor (CAR)-redirected T cells have been challenging issues for adoptive T cell therapy. Generating potent CAR T cells is of increasing importance in the field. Studies have demonstrated the importance of the Akt pathway in the regulation of T cell differentiation and memory formation. We now investigate whether inhibition of Akt signaling during ex vivo expansion of CAR T cells can promote the generation of CAR T cells with enhanced antitumor activity following adoptive therapy in a murine leukemia xenograft model...
2017: Journal for Immunotherapy of Cancer
https://www.readbyqxmd.com/read/28261215/tnf-blockade-maintains-an-il-10-phenotype-in-human-effector-cd4-and-cd8-t-cells
#20
Ceri A Roberts, Lucy E Durham, Veerle Fleskens, Hayley G Evans, Leonie S Taams
CD4(+) and CD8(+) effector T cell subpopulations can display regulatory potential characterized by expression of the prototypically anti-inflammatory cytokine IL-10. However, the underlying cellular mechanisms that regulate expression of IL-10 in different T cell subpopulations are not yet fully elucidated. We recently showed that TNF inhibitors (TNFi) promote IL-10 expression in human CD4(+) T cells, including IL-17(+) CD4(+) T cells. Here, we further characterized the regulation of IL-10 expression via blockade of TNF signaling or other cytokine/co-stimulatory pathways, in human T cell subpopulations...
2017: Frontiers in Immunology
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