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Artificial APC

Tatsunori Goto, Tetsuya Nishida, Erina Takagi, Kotaro Miyao, Daisuke Koyama, Reona Sakemura, Ryo Hanajiri, Keisuke Watanabe, Nobuhiko Imahashi, Seitaro Terakura, Makoto Murata, Hitoshi Kiyoi
Programmed death-ligand 1 (PD-L1) binds to programmed death-1 (PD-1) on activated T cells and contributes to T-cell exhaustion. PD-L1 expressed on antigen-presenting cells (APCs) could be thought to inhibit the induction of Ag-specific cytotoxic T lymphocytes (CTLs) by transducing negative signal into T cells; however, the roles of PD-L1 on APCs have not yet been well examined. Therefore, we evaluated the roles of PD-L1 on APCs in the induction of Ag-specific CTLs. CD3 T cells isolated from cytomegalovirus (CMV)-seropositive healthy donors were stimulated with mature dendritic cells pulsed with CMV pp65-derived HLA-restricted peptides in the presence of anti-PD-L1 blocking antibody...
October 2016: Journal of Immunotherapy
Alyssa L Siefert, Michael J Caplan, Tarek M Fahmy
Antigen-presenting cells (APCs) sense microorganisms via pathogen-associated molecular patterns (PAMPs) by both extra- and intracellular Toll-like Receptors (TLRs), initiating immune responses against invading pathogens. Bacterial PAMPs include extracellular lipopolysaccharides and intracellular unmethylated CpG-rich oligodeoxynucleotides (CpG). We hypothesized that a biomimetic approach involving antigen-loaded nanoparticles (NP) displaying Monophosphoryl Lipid A (MPLA) and encapsulating CpG may function as an effective "artificial bacterial" biomimetic vaccine platform...
August 2016: Biomaterials
Sarine Markossian, Alexei Arnaoutov, Nakhle S Saba, Vladimir Larionov, Mary Dasso
Most solid tumors are aneuploid, carrying an abnormal number of chromosomes, and they frequently missegregate whole chromosomes in a phenomenon termed chromosome instability (CIN). While CIN can be provoked through disruption of numerous mitotic pathways, it is not clear which of these mechanisms are most critical, or whether alternative mechanisms could also contribute significantly in vivo. One difficulty in determining the relative importance of candidate CIN regulators has been the lack of a straightforward, quantitative assay for CIN in live human cells: While gross mitotic abnormalities can be detected visually, moderate levels of CIN may not be obvious, and are thus problematic to measure...
July 2, 2016: Cell Cycle
Longhao Sun, Hao Guo, Ruoyu Jiang, Li Lu, Tong Liu, Zhixiang Zhang, Xianghui He
Professional antigen-presenting cells (APCs) are potent generators of tumor antigen-specific cytotoxic T lymphocytes (CTLs) for adoptive immunotherapy; however, generation of APCs is cumbersome, expensive, and subject to the tumor microenvironment. Artificial APCs (aAPCs) have been developed as a cost-effective alternative to APCs. We developed a cellular aAPC that efficiently generated alpha-fetoprotein (AFP)-specific CTLs. We genetically modified the human B cell lymphoma cell line BJAB with a lentiviral vector to establish an aAPC called BA15...
April 5, 2016: Oncotarget
Jitka Danadova, Natalie Matijescukova, Anna Mac Gillavry Danylevska, Martin Anger
Optimal culture conditions are essential for successful IVM of mammalian oocytes and for their further development into an embryo. In the present study we used live cell imaging microscopy to assess the effects of suboptimal culture temperature on various aspects of IVM, including duration of meiosis I, dynamics of polar body extrusion, chromosome congression, anaphase-promoting complex/cyclosome (APC/C) activation and aneuploidy. The data showed that even a small deviation from the optimal incubation temperature causes marked changes in the duration and synchronicity of meiosis, APC/C activity and the frequency of chromosome congression and segregation errors...
March 9, 2016: Reproduction, Fertility, and Development
Anthony Garnier, Mohamad Hamieh, Aurélie Drouet, Jérôme Leprince, Denis Vivien, Thierry Frébourg, Brigitte Le Mauff, Jean-Baptiste Latouche, Olivier Toutirais
Owing to their multiple immune functions, CD4(+) T cells are of major interest for immunotherapy in chronic viral infections and cancer, as well as for severe autoimmune diseases and transplantation. Therefore, standardized methods allowing rapid generation of a large number of CD4(+) T cells for adoptive immunotherapy are still awaited. We constructed stable artificial antigen-presenting cells (AAPCs) derived from mouse fibroblasts. They were genetically modified to express human leukocyte antigen (HLA)-DR molecules and the human accessory molecules B7...
August 2016: Immunology and Cell Biology
Anita Wnętrzak, Kazimierz Łątka, Katarzyna Makyła-Juzak, Joanna Zemla, Patrycja Dynarowicz-Łątka
Outer layer of cellular membrane contains ordered domains enriched in cholesterol and sphingolipids, called 'lipid rafts', which play various biological roles, i.e., are involved in the induction of cell death by apoptosis. Recent studies have shown that these domains may constitute binding sites for selected drugs. For example alkylphosphocholines (APCs), which are new-generation antitumor agents characterized by high selectivity and broad spectrum of activity, are known to have their molecular targets located at cellular membrane and their selective accumulation in tumor cells has been hypothesized to be linked with the alternation of biophysical properties of lipid rafts...
August 2015: Molecular Membrane Biology
Hyun-Woo Cho, Su-Yeon Kim, Dae-Hee Sohn, Min-Ji Lee, Mi-Young Park, Hyun-Jung Sohn, Hyun-Il Cho, Tai-Gyu Kim
Human γδ T cells play important roles in the regulation of infection and cancer. To understand the roles of costimulatory signals in activation and expansion ex vivo, Vγ9Vδ2 T cells were grown with artificial APCs that express CD83, 4-1BB ligand, and/or CD32, which allowed a loading of αCD3 and αCD28 antibodies. The costimulatory signals through CD80, 4-1BB, and CD83 ligand in low levels of IL-2 triggered an explosive ex vivo proliferation of Vγ9Vδ2 T cells capable of secreting high levels of IL-2, IFN-γ, and TNF-α...
April 2016: Journal of Leukocyte Biology
Markus Axmann, Gerhard J Schütz, Johannes B Huppa
T-cells are remarkably specific and effective when recognizing antigens in the form of peptides embedded in MHC molecules (pMHC) on the surface of Antigen Presenting Cells (APCs). This is despite T-cell antigen receptors (TCRs) exerting usually a moderate affinity (µM range) to antigen when binding is measured in vitro(1). In view of the molecular and cellular parameters contributing to T-cell antigen sensitivity, a microscopy-based methodology has been developed as a means to monitor TCR-pMHC binding in situ, as it occurs within the synapse of a live T-cell and an artificial and functionalized glass-supported planar lipid bilayer (SLB), which mimics the cell membrane of an Antigen presenting Cell (APC) (2)...
October 30, 2015: Journal of Visualized Experiments: JoVE
Alexander A Celik, Thomas Kraemer, Trevor Huyton, Rainer Blasczyk, Christina Bade-Döding
Human leukocyte antigen (HLA)-E molecules are potent inhibitors of NK cell-mediated killing. Low in polymorphisms, two alleles are widely expressed among diverse populations: HLA-E*01:01 and HLA-E*01:03. Both alleles are distinguished by one SNP resulting in the substitution Arg107Gly. Both alleles present a limited set of peptides derived from class I leader sequences physiologically; however, HLA-E*01:01 presents non-canonical peptides in the absence of HLA class I molecules. To further assess the functional differences between both alleles, we analyzed the peptide repertoire of HLA-E*01:03 by applying soluble HLA technology followed by mass-spectrometric peptide sequencing...
January 2016: Immunogenetics
S Davoodi, R F Cooke, A C C Fernandes, B I Cappellozza, J L M Vasconcelos, R L A Cerri
The aim of this study was to test the effect of expression of estrus at artificial insemination (AI) on endometrium, conceptus, and CL gene expression of beef cows. Thirty-six multiparous nonlactating Nelore cows were enrolled on an estradiol- and progesterone (P4)-based timed AI protocol (AI = Day 0) and then slaughtered for the endometrium, CL, and conceptus collection on Day 19. The animals were retrospectively grouped on the basis of cows that (1) showed signs of estrus near AI (n = 19; estrus) and (2) did not show any signs of estrus (n = 17; nonestrus)...
March 1, 2016: Theriogenology
Quan Wang, W E Moerner
Oligomerization plays a critical role in shaping the light-harvesting properties of many photosynthetic pigment-protein complexes, but a detailed understanding of this process at the level of individual pigments is still lacking. To study the effects of oligomerization, we designed a single-molecule approach to probe the photophysical properties of individual pigment sites as a function of protein assembly state. Our method, based on the principles of anti-Brownian electrokinetic trapping of single fluorescent proteins, step-wise photobleaching, and multiparameter spectroscopy, allows pigment-specific spectroscopic information on single multipigment antennae to be recorded in a nonperturbative aqueous environment with unprecedented detail...
November 10, 2015: Proceedings of the National Academy of Sciences of the United States of America
Yuchen Fan, James J Moon
Recent studies have demonstrated great therapeutic potential of educating and unleashing our own immune system for cancer treatment. However, there are still major challenges in cancer immunotherapy, including poor immunogenicity of cancer vaccines, off-target side effects of immunotherapeutics, as well as suboptimal outcomes of adoptive T cell transfer-based therapies. Nanomaterials with defined physico-biochemical properties are versatile drug delivery platforms that may address these key technical challenges facing cancer vaccines and immunotherapy...
2015: Vaccines
Leticia Cunha Amaral Gonzaga de Almeida, Diana Gabriela Soares, Fernanda Almeida Azevedo, Marjorie de Oliveira Gallinari, Carlos Alberto de Souza Costa, Paulo Henrique dos Santos, André Luiz Fraga Briso
This study evaluated the color change, cytotoxicity and hydrogen peroxide (HP) diffusion caused by different home bleaching protocols: 10% carbamide peroxide (CP) for 3 or 1.5 h, 6% hydrogen peroxide for 1.5 h or 45 min. To quantify the peroxide penetration, disks of bovine teeth were placed in artificial pulp chambers (APCs) containing acetate buffer, which was collected for evaluation in a spectrophotometer. For analysis of cytotoxicity, specimens were adapted in APCs containing culture medium, which subsequently was applied on MDPC-23 odontoblast-like cells for 1 h...
July 2015: Brazilian Dental Journal
Eric N Reither, Kenneth C Land, Sun Y Jeon, Daniel A Powers, Ryan K Masters, Hui Zheng, Melissa A Hardy, Katherine M Keyes, Qiang Fu, Heidi A Hanson, Ken R Smith, Rebecca L Utz, Y Claire Yang
Previously, Reither et al. (2015) demonstrated that hierarchical age-period-cohort (HAPC) models perform well when basic assumptions are satisfied. To contest this finding, Bell and Jones (2015) invent a data generating process (DGP) that borrows age, period and cohort effects from different equations in Reither et al. (2015). When HAPC models applied to data simulated from this DGP fail to recover the patterning of APC effects, B&J reiterate their view that these models provide "misleading evidence dressed up as science...
November 2015: Social Science & Medicine
Karlo Perica, Joan Glick Bieler, Christian Schütz, Juan Carlos Varela, Jacqueline Douglass, Andrew Skora, Yen Ling Chiu, Mathias Oelke, Kenneth Kinzler, Shibin Zhou, Bert Vogelstein, Jonathan P Schneck
Adoptive immunotherapy (AIT) can mediate durable regression of cancer, but widespread adoption of AIT is limited by the cost and complexity of generating tumor-specific T cells. Here we develop an Enrichment + Expansion strategy using paramagnetic, nanoscale artificial antigen presenting cells (aAPC) to rapidly expand tumor-specific T cells from rare naïve precursors and predicted neo-epitope responses. Nano-aAPC are capable of enriching rare tumor-specific T cells in a magnetic column and subsequently activating them to induce proliferation...
July 28, 2015: ACS Nano
Mohammad Arif, George Opit, Abigail Mendoza-Yerbafría, Shefali Dobhal, Zhihong Li, Zuzana Kučerová, Francisco M Ochoa-Corona
Several species of the genus Liposcelis are common insect pests that cause serious qualitative and quantitative losses to various stored grains and processed grain products. They also can contaminate foods, transmit pathogenic microorganisms and cause allergies in humans. The common occurrence of multi-species infestations and the fact that it is difficult to identify and discriminate Liposcelis spp. make accurate, rapid detection and discriminatory tools absolutely necessary for confirmation of their identity...
2015: PloS One
D G Soares, C A Brito, R H B Tavares da Silva, A P D Ribeiro, J Hebling, C A de Souza Costa
AIM: To evaluate the transdentinal cytotoxicity of resin-based luting cements (RBLCs), with no HEMA in their composition, to odontoblast-like cells. METHODOLOGY: Human dentine discs 0.3 mm thick were adapted to artificial pulp chambers (APCs) and placed in wells of 24-well plates containing 1 mL of culture medium (DMEM). Two categories of HEMA-free RBLCs were evaluated: group 1, self-adhesive Rely X Unicem (RU; 3M ESPE), applied directly to the dentine substrate; and group 2, Rely X ARC (RARC; 3M ESPE), applied to dentine previously acid-etched and treated with a bonding agent...
June 2016: International Endodontic Journal
Devrim Oz-Arslan, Eliza Tsitoura, Dorothea Kazazi, Vlasis Kouvatsis, Alberto L Epstein, Penelope Mavromara
Dendritic cells (DCs) turn into the most potent antigen-presenting cells following a complex transforming process, which leads to their maturation. Herpes simplex virus-1 (HSV-1) amplicon vectors represent highly versatile viral vector platforms with the ability to transduce immature DCs at exceedingly high efficiencies, while the efficiency of infection of mature DCs is significantly low. However, the bacterial artificial chromosome (BAC)-dependent (BD) amplicon vectors tested so far do not result in the maturation of mouse bone marrow-derived DCs (BMDCs) in vitro...
June 2015: Human Gene Therapy Methods
Gijsbert P van Nierop, Josef Mautner, Johanna G Mitterreiter, Rogier Q Hintzen, Georges M G M Verjans
BACKGROUND: The association between Epstein-Barr virus (EBV) and multiple sclerosis (MS) may involve intrathecal EBV-specific T-cell responses targeting the virus or indirectly, autoantigens. OBJECTIVE: Compare the prevalence and fine-specificity of EBV-specific T-cells in the cerebrospinal fluid (CSF) of patients with MS (n = 12), clinically-isolated syndrome (CIS) (n = 17) and other neurological diseases (OND) (n = 13). METHODS: Intrathecal EBV-specific T-cell reactivity was assayed using CSF-derived T-cell lines (CSF-TCL) and autologous EBV-transformed B-cells (autoBLCL) as antigen-presenting cells (APC)...
March 2016: Multiple Sclerosis: Clinical and Laboratory Research
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