anti mhc a  (Thermo Fisher)

Journal: Molecular and Cellular Biology

Article Title: Pten Null Prostate Epithelium Promotes Localized Myeloid-Derived Suppressor Cell Expansion and Immune Suppression during Tumor Initiation and Progression

doi: 10.1128/MCB.00090-14

Figure Lengend Snippet: Prostate-specific Gr-1 + CD11b + cell expansion is associated with suppression of dendritic cell and macrophage maturation. (A, left) Mature CD11c + MHCII + dendritic cells are present in both wild-type and mutant prostates, but their levels decrease precipitously

Article Snippet: Single-cell suspensions were stained with directly conjugated antibodies against CD45, Gr-1, CD11b, CD4, CD8, CD69, B220 (BD Biosciences), F4/80, CD19, CD11c, major histocompatibility complex class II (MHCII) (eBioscience), and Ly6C (BioLegend), according to the manufacturers' instructions.

Techniques: Mutagenesis

Journal: Mucosal immunology

Article Title: Regulatory T-cells are essential to promote proper CD4 T-cell priming upon mucosal infection

doi: 10.1038/mi.2016.19

Figure Lengend Snippet: Dendritic cell trafficking from the vaginal mucosa is impaired in the absence of Tregs Foxp3 DTR or control mice were injected with DT and infected intravaginally with HSV-2 186ΔKpn as above. a) Expression of CCL21 in the dLN of Treg-sufficient or Treg-depleted mice 2d after infection determined by qPCR analysis (relative to the housekeeping gene HPRT). b) FITC painting experimental layout. c) Representative flow plots showing the fraction of CD11b+/CD8− DCs in the dLN that stained positive for FITC 2d after infection. Cells were first gated on MHC II/CD11c+/CD11b+ and CD8− cells. d) Representative data showing the number of FITC+ DCs that had entered the dLN from the infected vaginal tract 2d after the mice were infected. e) Ex vivo antigen presentation experimental layout. f) The amount of IFNγ produced by anti-HSV-2 CD4 T-cells after 60h of co-culture with DCs sorted from Treg-sufficient or Treg-depleted mice 2d after infection. On the left, cells were cultured without addition of exogenous antigen, whereas on the right, cells were provided HSV-2 ex vivo during culture. All data come from or are representative of at least 2 similar, independent experiments with 3–5 mice in each group that showed similar results. Each data set was first screened for outliers using the ROUT test with a Q value of 0.2%. Statistical significance was then determined using an ordinary one-way ANOVA followed by a Tukey test to compare the mean of each group with the mean of all other groups.*: p ≤ 0.05, **: p ≤ 0.01, ***: p ≤ 0.001 ****: p ≤ 0.0001. Error bars show standard deviation.

Article Snippet: CD4 T-cells that were already enriched using the CD4+ T-cell negative selection kit (Stem Cell Technologies, Vancouver, Canada) were also stained using anti-CD4 and anti-MHC II antibodies (eBioscience, San Diego, CA) and sorted into a population of CD4+ and MHC II – cells to ensure that the CD4 T-cells were not contaminated with antigen presenting cells.

Techniques: Mouse Assay, Injection, Infection, Expressing, Real-time Polymerase Chain Reaction, Flow Cytometry, Staining, Ex Vivo, Produced, Co-Culture Assay, Cell Culture, Standard Deviation

Journal: BMC Ophthalmology

Article Title: Heightened expression of MICA enhances the cytotoxicity of NK cells or CD8+T cells to human corneal epithelium in vitro

doi: 10.1186/1471-2415-12-6

Figure Lengend Snippet: Contribution of MICA to NK cells and CD8 + T cells mediated killing of human corneal epithelium . ( A ) Human MICA-transfected corneal epithelium or empty vector transfected corneal epithelium were preincubated for 4 h with either an isotype control (white bars), anti-MICA antibody (black bars) or anti-MHC class I antibody (gray bars) before being added to IL-2-activated NK cells (effector) at a 3:1 (E:T) ratio. ( B ) Human MICA-transfected corneal epithelium or empty vector transfected corneal epithelium were preincubated for 4 h with either an isotype control (white bars), anti-MICA antibody (black bars) or anti-MHC class I antibody (gray bars) before being added to allogeneic human CD8 + T cells (effector) at a 10:1 (E:T) ratio. Nonparametric test comparing isotype and MICA antibody showed a statistical difference between groups for MICA-transfected cells (NK cells, P

Article Snippet: Anti-human MICA monoclonal antibodies (R & D Systems, Minneapolis, MN), anti-MHC class I (eBiosciences, San Diego, CA), isotype control mouse IgG (BD Biosciences, Mountain View, CA) and mouse anti-human keratin 3/12 (Abcam, Cambridge, UK) Abs were purchased.

Techniques: Transfection, Plasmid Preparation

Journal: BMC Immunology

Article Title: DC-STAMP knock-down deregulates cytokine production and T-cell stimulatory capacity of LPS-matured dendritic cells

doi: 10.1186/1471-2172-12-57

Figure Lengend Snippet: DC-STAMP silencing does not induce phenotypic changes in mBMDCs . Murine BMDCs were infected with shScr and shST1 lentivirus at day 6 and day 7 of culture. (A) Light field microscopy image of mBMDCs in culture 4 days post-infection. (B) Lentiviraly transduced mBMDCs stained with antibody against calreticulin (green) to visualize ER and DAPI staining of nucleus (blue) analyzed by CLSM. (C) Flow cytometric analysis of CD80, CD86 and MHC class II surface expression in CD11c + immature (-LPS) and LPS-matured (+LPS) non-infected (no virus), shScr and shST1 mBMDCs. Isotype controls are indicated by shaded area. Data shown are representative of two or three independent experiments.

Article Snippet: Flow cytometry For cell surface labeling, the following anti-mouse antibodies were used (from BD Biosciences, unless stated differently): PE-conjugated anti-CD86 (GL1), anti-CD80 (16-10A1), anti-CD40 (3/23), anti-CD8α (53-6.7), anti-MHC class II (M5/114.15.2, eBioscience); PerCP-conjugated anti-CD62L (Mel-14, BioLegend); APC-conjugated anti-CD11c (N418, BioLegend); APCCy7-conjugated anti-CD4 (L3T4).

Techniques: Infection, Microscopy, Staining, Confocal Laser Scanning Microscopy, Flow Cytometry, Expressing