human dendritic cell culture and bacterial infection

... pP{UAST} vector [18] Fly strains and bacterial infections Oregon-R and ywDD1; cnbw wild-type flies were used throughout this study [19] Flies were maintained on a standard cornmeal medium at 25 °C ... Procise cLC) Cell culture experiments S2 cells were purchased from Invitrogen, and maintained in Schneider’s medium supplemented with 10% (v/v) fetal bovine serum; 60 mgÆL)1 penicillin and 100 mgÆL)1 ... tube containing two layers of caesium chloride (5% and 40%), and ultracentrifuged for 16 h at 36 000 g at 15 °C The virus band was collected and DCV was recovered by centrifugation (25 000 g,...

Ngày tải lên: 08/03/2014, 08:20

... Rho GTPase activity in dendritic cells J Immunother 2008, 31(5):491-499 Shurin GV, Amina N, Shurin MR: Cancer therapy and dendritic cell immunomodulation In Dendritic Cells in Cancer Edited by: ... followed by syngeneic dendritic cell injection in the mouse: findings and implications for human treatment Urology 2007, 70(6 Suppl):36-41 Shurin MR: Preparation of human dendritic cells for tumor ... were added to cells and incubated for 30 minute at 4°C Murine FITC-IgG and PE-IgG were used as isotype controls Data analysis was performed using the CellQuest and WinMDI software and the results...

Ngày tải lên: 18/06/2014, 15:20

... with10% FBS and antibiotics Appropriate controls included uninfected cells, and cells treated with co-receptor antagonists TAK 779 (9.14 μmol/ http://www.aidsrestherapy.com/content/6/1/27 ml) and AMD ... AIDS Research and Reference Reagent Program for providing us cells and chemokine blockers The findings and conclusions in this article have not been formally disseminated by the Food and Drug Administration ... in Cameroon in terms of co-receptor usage and comparing cell culture based methods with V3 sequence based prediction algorithms for virus phenotyping and co-receptor usage of complex, emerging...

Ngày tải lên: 10/08/2014, 05:21

... BM-cells from femurs and tibias were washed and cultured overnight in 6-well plates (TPP, Trasadingen, Schwitzerland) at × 106 cells/ml in ml culture medium/well Culture medium (CM) was RPMI-1640 with ... England) and antibiotics The next day, non-adherent cells were harvested and resuspended in CM containing 10 ng/ml GM-CSF plus 20 ng/ml IL-4 (both from Peprotech, Rocky Hill, NJ, USA) and cultured ... Banchereau J, Steinman RM: Dendritic cells and the control of immunity Nature 1998, 392:245-52 Gad M, Claesson MH, Pedersen AE: Dendritic cells in peripheral tolerance and immunity APMIS 2003,...

Ngày tải lên: 11/08/2014, 10:23

... the cell suspension transferred into preheated (37°C) culture medium and centrifuged to remove the DMSO It was resuspended in culture medium and then transferred into cell culture flasks and cultured ... the cells was performed at 85 to 95% confluency as fibroblasts exhibit contact inhibition (unpublished observations) and were passaged 1:2 to provide cell- cell contacts between the cells Cell culture ... the cells were grown to 85 to 95% confluence, then trypsinized, counted and 50,000 cells were seeded into five fresh cell culture six-wells (passage 1) Each day, one well was trypsinized and...

Ngày tải lên: 12/08/2014, 12:20

... penicillin, and 100 μg/ml streptomycin, and then incubated in plastic cell cultures plates for h at 37°C in a 5% CO2 atmosphere Culture plates were then washed thrice with RPMI 1640 medium and nonadherent ... detaching, CD4+ T cells were stained with CD25 MicroBeads (Miltenyi Biotec) and CD4+CD25+T cells were positively and negatively selected Separation was controlled by FCM and Spleen CD4+ T cells were ... sensitized and challenged with OVA, OVA- pcDNA3.1-vaccinated mice sensitized and challenged with OVA, and OVA-Fc-pcDNA3.1-vaccinated mice sensitized and challenged with OVA * and ** stand for the...

Ngày tải lên: 14/08/2014, 19:22

... that binds to CD40 and enhances adenoviral infection of mouse and human DC and B lineage cells Materials and methods Cell lines Epstein–Barr virus- (EBV-) transformed human B cells were provided ... AdV-GFP infection of human DC, as well as human and murine B cells DC, LCL, L10A, Blasts: these labels refer, respectively, to day human DC, EBV-immortalized human B cells, murine B lymphoma L10A cells ... homology to CD40 ligand (data not shown) and that enhances adenoviral infection of CD40-positive cells of both human and murine origin, including DC and B cells resistant to infection with an unmodified...

Ngày tải lên: 20/02/2014, 11:20

... clonal expansion of T-cells bearing human Vb3, Vb12, Vb17 and Vb19 [20] Others showed proliferation of human T-cells bearing Vb1, Vb2, Vb3, Vb5.2, Vb12, Vb15 and Vb17 domains and found differences ... used to study T -cell proliferation capacity with human peripheral blood mononuclear cells (PBMCs) The binding of SSA and mutant C26S to soluble TCR b-chain molecules Vb1, Vb2.1 and Vb5.2 was measured ... wtSSA–Vb5.2 and SSA(C26S)– Vb5.2 Different concentrations of human Vb2.1 and Vb1 TCRs were also used to measure the binding to the immobilized SSAm and SSA(C26S) Vb1 showed a pattern of association and...

Ngày tải lên: 07/03/2014, 16:20

... PVP-coated AgNPs block HIV-1 cell- free and cell- associated infection and act as a virucidal agent As previously described, PVP-coated AgNPs are an interesting virucidal candidate Therefore, we investigated ... transmission of cellassociated infection in the cervical culture model We evaluated the time required for 0.1 and 0.15 mg/mL doses of PVP-coated AgNPs to block HIV-1IIIB infection of cell- associated ... using the SigmaPlot 10.0 software infection by HIV-IIIB and HIV-AZT-RV cell- free viruses as well as cell- associated infection at doses that were not toxic to the human cervical tissue In addition,...

Ngày tải lên: 11/08/2014, 00:22

... μg/ml) and unmethylated DNA (5 μg/ml) Cells were then pulsed with NL4-3/Balenv and co-cultured with autologous CD4+ T cells Finally cell- free supernatants were harvested at 2, and days post-coculture ... O: Dendritic cells and HIV-specific CD4+ T cells: HIV antigen presentation, T -cell activation, and viral transfer Blood 2006, 108:1643-1651 Pauza CD, Price TM: Human immunodeficiency virus infection ... treated with TLR2 and ligands for hours and washed twice Then, cells were pulsed with NL4-3/Balenv for 15, 30 and 60 at 37°C Next, the virus -cell mixture was washed extensively with PBS and trypsinized...

Ngày tải lên: 12/08/2014, 23:20

... infected cell and a target cell induces the accumulation of the viral proteins Gag and Env, viral RNA and microtubules, and the viral complex subsequently transfers into the target cell [49] ... HTLV-I-infected cells and cancer Lymphoid malignancy with a T -cell origin is rare compared with B -cell malignancy ATL shares hematological, pathological and immunological features with cutaneous T -cell ... such as chemokine receptors and adhesion molecules Skin-homing memory T-cells uniformly express CCR4, and its ligands are thymus and activation-regulated chemokine (TARC) and macrophagederived chemokine...

Ngày tải lên: 13/08/2014, 09:21

... mouse and human pDCs cluster together and apart from cDCs (Figure 3) Importantly, when analyzing the DC datasets alone, mouse CD8α and human BDCA3 cDCs on the one hand, and mouse CD11b and human ... primarily in T cells and often to a lower extent in NK cells; the mouse Klrb1c (nk1.1) gene or the human KIR genes in NK cells; Cd19 in B cells; the mouse Siglech and Bst2 genes or the human LILRA4 ... Myeloid cells Figure Clustering of mouse IKDCs and human CD16 cells Clustering of mouse IKDCs and human CD16 cells Hierarchical clustering with complete linkage was performed on the indicated cell...

Ngày tải lên: 14/08/2014, 08:20

... mDCs and moDCs [12] and moDCs and KG-1 cells [13], but no studies have compared the proteomes of all three cell types In this study we compare the proteomes of mDCs, moDCs and KG-1 cells, and ... to moDCs and KG-1 cells refereed research Figure Cell culture work-flow Cell culture work-flow Overview of the methods used for isolation/generation of mDCs, moDCs and DC-like KG-1 cells deposited ... inflammatory (IL-1β, IL-6 and IL-8) and regulatory (IL-10) cytokines, compared to moDCs and KG-1 cells Levels of IL-1β, IL-6, IL-8 and IL-10 were dose dependent, and following 24 h culture with either...

Ngày tải lên: 14/08/2014, 18:20

... after week of culture ( 20% of cells positive for iron) and reached a plateau after weeks of culture ( 60% of cells positive, Fig 2A) In all subsequent experiments, HepG2 cells were cultured in ... the cell surface of HepG2 cells and culture in iron-rich media did not influence their expression (Fig 4B) CD1d protein and mRNA are upregulated in iron-loaded HepG2 cells Immunocytochemistry and ... protein and mRNA transcripts HepG2 cells were cultured as indicated in legend of Fig and cells processed for immunocytochemistry, immunoprecipitation and mRNA studies (A) Cytospins of HepG2 cells...

Ngày tải lên: 19/02/2014, 16:20

... after 2–3 weeks and re-cloned by serial dilution The stable cells transfected with sense and antisense PKB were named S-PKB/H7721 and AS-PKB/H7721 Cell culture and treatment Cells were cultured for ... via PKB and SLex (Eur J Biochem 270) 3797 cells was performed by electroporation using Gene Pulser at 250 V/0.4 cm and 1000 lF [26] Cells were selected by G418 and the neomycin-resistant cells ... filter) [29–31] Cells (1 · 105) in 0.1 mL were added, and the incubation time was extended to 36 h Then the cells were fixed, stained and countered as described above, and the number of cells that...

Ngày tải lên: 21/02/2014, 00:20

... presented here indicate for the first time that human dendritic cells contain anandamide, 2-AG and PalEtn The amounts of anandamide and 2-AG in immature dendritic cells were similar to the ones detected ... The most abundant band was the one at  59 kDa, which was present in both human immature dendritic cells and in rat Ó FEBS 2002 The endocannabinoid system in human dendritic cells (Eur J Biochem ... that 2-AG originating from human dendritic cells might contribute to the important immune function played by these cells after activation and during bacterial infections and allergic responses This...

Ngày tải lên: 22/02/2014, 07:20

... demonstrated on child and adult heads [22] and, thus, the epidermal 3D model is a relevant skin cellular model, complementary to isolated cells In this study, we used human cutaneous cells and hRE to test ... chosen and used to induce apoptosis in the various cell types and a 200 mJÆcm)2 single dose to induce HSP expression in the various cell types and in the hRE Briefly, the cultures (primary cells ... (GSM-900, SHAM or UVB) and time in culture The number of hRE per condition (GSM or SHAM) was seven after weeks in ALI culture, four after weeks in ALI culture and six after weeks in ALI culture The Mann–Whitney...

Ngày tải lên: 07/03/2014, 11:20

... regulation of cancer cell proliferation and growth For instance, let-7 can inhibit proliferation of lung and liver cancer cells by targeting multiple cell cycle oncogenes [28] and miR-34b and miR-34c ... ESR1 and GAPDH are included in Table S2 Cell lines and cultures Five breast carcinoma cell lines were obtained from the Cell Resource Center of Peking Union Medical College (Beijing, China) and ... (ESR1–3¢-UTR–WT) and pGL3m–ESR1–3¢-UTR–MUT (ESR1–3¢-UTR–MUT) in HEK293T and MCF-7 cells cotransfected with pcDNA-3.0–miR-22 or pcDNA-3.0 and synthetic miR-22 duplex or control RNA duplex ,and in MDA-MB-231 cells...

Ngày tải lên: 22/03/2014, 21:20

... FEBS M K Andersson et al P2¢ specificity of the human mast cell chymase Results Production and purification of recombinant HC mutants Two single mutants of HC, Arg143 fi Gln and Lys192 fi Met, and a ... serum and cellular proteins, imidazole, and EK, they were purified over a heparin–Sepharose column The heparin–Sepharose-purified fractions were separated on SDS ⁄ PAGE gels, and no contaminating bands ... of the human mast cell chymase M K Andersson et al the substrate The marked difference in the importance of Lys40, Arg143 and Lys192 in determining substrate specificity between peptides and long...

Ngày tải lên: 22/03/2014, 21:21

... trypsin, breaks the cell- cell and cell- substrate links and creates a single -cell suspension Subculture of a primary culture into a secondary culture produces a cell line.” Once a cell- line is created, ... dissociated cells in culture Tissue culture Maintenance of tissue explants in culture Cell line A culture that is subcultured beyond the initial primary culture phase Finite cell line A cell line ... A and Flandermeyer, R R (1977) Inter- and intraspecies contamination of human breast tumor cell lines HBC and BrCa5 and other cell cultures Science 195, 1343–1344 Gey, G O., Coffman, W D., and...

Ngày tải lên: 10/04/2014, 11:11