Journal:

Article Title: Expression of Integrin-? E by Mucosal Mast Cells in the Intestinal Epithelium and Its Absence in Nematode-Infected Mice Lacking the Transforming Growth Factor-? 1 -Activating Integrin ? v ? 6

doi:

Figure Lengend Snippet: Immunocytochemical detection of MMCs in epithelial whole mounts and frozen sections of jejunum from T. spiralis-infected BALB/c mice. A to D: Whole mounts of jejunal epithelium were probed with anti-mMCP-1 (pseudo-colored red) or isotype-matched control antibodies together with anti-IgE (pseudo-colored green) or isotype-matched control antibodies and counterstained with DRAQ5 (pseudo-colored blue). IgE surface labeling was almost exclusively restricted to mast cells exhibiting strong intracellular mMCP-1 staining in samples probed with mMCP-1- and IgE-specific antibodies (A and D). Single-negative controls for IgE (B) and mMCP-1 (C) were used to confirm labeling specificity. Frozen sections of jejunum from uninfected (E) and T. spiralis-infected (F) BALB/c mice were probed with rabbit anti-tryptase (pseudo-colored green), anti-IgE (pseudo-colored red), and anti-integrin-α6 antibodies (pseudo-colored blue). No IgE or tryptase-positive cells were detected in uninfected animals (E). Abundant IgE+ve cells were detected in the samples from infected mice, the majority of which were located intraepithelially (F). Although significant numbers of intraepithelial IgE+ve cells were tryptase-negative (F, arrowheads), most IgE+ve cells detected in the lamina propria expressed tryptase (F, arrow). Staining specificity was confirmed using single-positive controls for tryptase (G), integrin-α6 (H), and IgE (I), in which the two other antigen-specific antibodies were replaced with control IgG. Identical excitation and exposure settings were used to acquire images of multilabeled and control samples. Scale bar length (μm) is provided in the bottom left corner of each image.

Article Snippet: Purified monoclonal rat IgG 2a anti-human integrin-α 6 (clone GoH3), control rat IgG 2a (clone R35-95), control rat IgG 1 (clone R3-34), and FITC-conjugated control rat IgG 1 (clone R3-34) were purchased from BD Biosciences, Cowley, UK.

Techniques: Infection, Labeling, Staining

Journal:

Article Title: Expression of Integrin-? E by Mucosal Mast Cells in the Intestinal Epithelium and Its Absence in Nematode-Infected Mice Lacking the Transforming Growth Factor-? 1 -Activating Integrin ? v ? 6

doi:

Figure Lengend Snippet: IgE+ve cells in the jejunal mucosa of T. spiralis-infected mice express integrin-αEβ7. Frozen sections of jejunum from infected BALB/c mice were triple-labeled with anti-integrin-αE or anti-integrin-β7 together with anti-IgE and anti-integrin-α6 (see Figure 4). Using integrin-α6 staining to differentiate the epithelium (filled bars) and lamina propria (open bars), IgE+ve cells mm−2 (A) and the relative frequencies of IgE+ve cells expressing integrin-αE or integrin-β7 (B) were determined for each location using five randomly selected 0.31-mm2 fields of view per sample. The mean percentage of intraepithelial IgE+ve cells ± 1 SEM is provided in parentheses (A). Data are expressed as mean values ± 1 SEM (n = 4). Differences observed between the epithelial and lamina propria compartments were significant at P < 0.01 (**).

Article Snippet: Purified monoclonal rat IgG 2a anti-human integrin-α 6 (clone GoH3), control rat IgG 2a (clone R35-95), control rat IgG 1 (clone R3-34), and FITC-conjugated control rat IgG 1 (clone R3-34) were purchased from BD Biosciences, Cowley, UK.

Techniques: Infection, Labeling, Staining, Expressing

Journal:

Article Title: Expression of Integrin-? E by Mucosal Mast Cells in the Intestinal Epithelium and Its Absence in Nematode-Infected Mice Lacking the Transforming Growth Factor-? 1 -Activating Integrin ? v ? 6

doi:

Figure Lengend Snippet: Immunofluorescent detection of integrin-αE and -β7 expression by IgE+ve cells and PCNA-specific staining of toluidine blue-positive mast cells in the jejunal mucosa of S129 β6−/− and β6+/+ mice. A to J: Frozen sections of jejunum were probed with monoclonal rat antibodies specific for integrin-α6 (pseudo-colored blue), IgE (pseudo-colored green), and integrin-β7 or -αE (pseudo-colored red). Representative images are shown from uninfected β6+/+ (A) and β6−/− (B) mice stained for IgE, integrin-α6, and integrin-β7; uninfected β6+/+ (C) and β6−/− (D) mice stained for IgE, integrin-α6, and integrin-αE; T. spiralis-infected β6+/+ (E) and β6−/− (F) mice stained for IgE, integrin-α6, and integrin-β7; and T. spiralis-infected β6+/+ (G) and β6−/− (H) mice stained for IgE, integrin-α6, and integrin-αE. Arrowheads in D and H highlight occasional cells that exhibit low-level integrin-αE immunoreactivity in sections from β6−/− mice (inset in D has been contrast enhanced). Higher magnification images from T. spiralis-infected β6+/+ (I) and β6−/− (J) mice stained for IgE, integrin-α6, and integrin-αE are annotated with arrows (intraepithelial IgE+ve cells) and arrowheads (lamina propria IgE+ve cells). Note that although large numbers of IgE+ve cells are present in the mucosa of T. spiralis-infected β6−/− mice, they are predominantly located in the lamina propria. Single-positive controls from β6+/+ mice for integrin-β7 (K), integrin-αE (L), IgE (M), and integrin-α6 (N), in which two of the three antigen-specific antibodies were substituted with isotype-matched controls, were used to confirm staining specificity. Identical excitation and exposure settings were used to acquire images A to D. Constant excitation and exposure settings were also maintained during acquisition of images E to N. Bright-field images of toluidine blue-labeled mast cells probed with anti-PCNA antibodies from T. spiralis-infected β6+/+ (O) and β6−/− (P) mice, arrow indicates a PCNA-positive mast cell in O. Scale bars, 25 μm.

Article Snippet: Purified monoclonal rat IgG 2a anti-human integrin-α 6 (clone GoH3), control rat IgG 2a (clone R35-95), control rat IgG 1 (clone R3-34), and FITC-conjugated control rat IgG 1 (clone R3-34) were purchased from BD Biosciences, Cowley, UK.

Techniques: Expressing, Staining, Infection, Labeling

Journal:

Article Title: Expression of Integrin-? E by Mucosal Mast Cells in the Intestinal Epithelium and Its Absence in Nematode-Infected Mice Lacking the Transforming Growth Factor-? 1 -Activating Integrin ? v ? 6

doi:

Figure Lengend Snippet: IgE+ve cell recruitment and integrin-αEβ7 expression in the jejunal mucosa of T. spiralis-infected β6−/− and β6+/+ S129 mice. Frozen sections of jejunum from infected β6−/− (open bars) and β6+/+ (filled bars) S129 mice were triple-labeled with anti-integrin-αE or anti-integrin-β7 together with anti-IgE and anti-integrin-α6 (see Figure 4). Data are presented as mean values ± 1 SEM (n = 3) for IgE+ve cells mm−2 (A); the percentage of IgE+ve cells located intraepithelially (B); and the relative frequencies of IgE+ve cells expressing integrin-αE (C) or integrin-β7 (D). Differences observed between β6−/− and β6+/+ S129 mice were significant at P < 0.05 (*) and P < 0.01 (**).

Article Snippet: Purified monoclonal rat IgG 2a anti-human integrin-α 6 (clone GoH3), control rat IgG 2a (clone R35-95), control rat IgG 1 (clone R3-34), and FITC-conjugated control rat IgG 1 (clone R3-34) were purchased from BD Biosciences, Cowley, UK.

Techniques: Expressing, Infection, Labeling

Journal:

Article Title: Expression of Integrin-? E by Mucosal Mast Cells in the Intestinal Epithelium and Its Absence in Nematode-Infected Mice Lacking the Transforming Growth Factor-? 1 -Activating Integrin ? v ? 6

doi:

Figure Lengend Snippet: RT-PCR analysis of integrin-αE and -β7 transcript abundance and toluidine blue analysis of MMC distribution in the jejunal mucosa of T. spiralis-infected β6+/+ and β6−/− S129 mice. A: RT-PCR products for integrin-αE, integrin-β7, and the housekeeping gene GAPDH from reverse-transcribed total jejunal RNA from infected β6−/− (lanes 1 to 4) and β6+/+ (lanes 5 to 8) mice. B: RT-PCR product signal intensity expressed as a percentage of GAPDH signal intensity. Differences in PCR product abundance between β6+/+ and β6−/− S129 mice were significant at P < 0.05 (*). Mast cells were identified using toluidine blue staining (pH 0.5) in Carnoy’s-fixed/paraffin-embedded sections from β6+/+ (C) and β6−/− (D) S129 mice. Note that mast cells are present in greater numbers in β6−/− mice but are predominantly restricted to the lamina propria. Scale bars, 50 μm.

Article Snippet: Purified monoclonal rat IgG 2a anti-human integrin-α 6 (clone GoH3), control rat IgG 2a (clone R35-95), control rat IgG 1 (clone R3-34), and FITC-conjugated control rat IgG 1 (clone R3-34) were purchased from BD Biosciences, Cowley, UK.

Techniques: Reverse Transcription Polymerase Chain Reaction, Infection, Staining

Journal: Oncoimmunology

Article Title: Oncolytic measles virus induces tumor necrosis factor-related apoptosis-inducing ligand (TRAIL)-mediated cytotoxicity by human myeloid and plasmacytoid dendritic cells

doi: 10.1080/2162402X.2016.1261240

Figure Lengend Snippet: MV induces TRAIL expression and IFN-α secretion by pDCs and CD1c + DCs. pDCs were cultured with IL3, IL3+MV or the TLR7 agonist R837 (A). CD1c + DCs were cultured alone (−), with MV or R837 (B). The expression of surface markers by the indicated cells was determined by flow cytometry. IFN-α secretion was measured by ELISA. #, values are below the limit of detection of the kit (7 pg/mL). Results are expressed as the mean ± SEM of three independent experiments. * p < 0.05, one-way ANOVA (Kruskal–Wallis).

Article Snippet: DCs were also cultivated with the TLR7 agonist R837 (1 μg/mL, Invivogen), exogenous type I IFNs (rhIFN-α-2a and rhIFN-β-1a, 100 ng/mL, ImmunoTools) or the RIG-I agonist 5′-ppp-dsRNA LyoVec (10 μg/mL, Invivogen).

Techniques: Expressing, Cell Culture, Flow Cytometry, Enzyme-linked Immunosorbent Assay

Journal: Oncoimmunology

Article Title: Oncolytic measles virus induces tumor necrosis factor-related apoptosis-inducing ligand (TRAIL)-mediated cytotoxicity by human myeloid and plasmacytoid dendritic cells

doi: 10.1080/2162402X.2016.1261240

Figure Lengend Snippet: TRAIL expression depends on IFN-α secreted by DCs exposed to MV. pDCs (A) and CD1c + DCs (B) were pretreated or not with Ruxolitinib (Rux) before exposure to IL3+MV or MV respectively, or before exposure to R837. The secretion of IFN-α was measured by ELISA. #, values are below the limit of detection of the kit (7 pg/mL). The expression of TRAIL by the indicated cells was determined by flow cytometry. (C) pDCs and CD1c + DCs were pretreated or not with Rux before exposure to type I IFNs (rhIFN-α-2a and rhIFN-β-1a). TRAIL positive cells were quantified by flow cytometry. Results are expressed as the mean ± SEM of three independent experiments. * p < 0.05, Mann–Whitney test.

Article Snippet: DCs were also cultivated with the TLR7 agonist R837 (1 μg/mL, Invivogen), exogenous type I IFNs (rhIFN-α-2a and rhIFN-β-1a, 100 ng/mL, ImmunoTools) or the RIG-I agonist 5′-ppp-dsRNA LyoVec (10 μg/mL, Invivogen).

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Flow Cytometry, MANN-WHITNEY

Journal: Oncoimmunology

Article Title: Oncolytic measles virus induces tumor necrosis factor-related apoptosis-inducing ligand (TRAIL)-mediated cytotoxicity by human myeloid and plasmacytoid dendritic cells

doi: 10.1080/2162402X.2016.1261240

Figure Lengend Snippet: MV induces IFN-α and TRAIL expression following RLR activation in CD1c + DCs and RLR/TLR7 activation in pDCs. (A) pDCs were pretreated or not with IRS661 (TLR7 inhibitor) or with MRT67307 (TBK1 and IKK-ε inhibitor) and then cultured with IL3, IL3+MV or R837. (B) CD1c + DCs were pretreated or not with IRS661 or MRT67307 and then cultured alone (−) or with MV. (C) pDCs were cultured with IL3+MV or with IL3+UV-inactivated MV (MV UV*) and CD1c + DCs were cultured alone (−), with MV or MV UV*. (D) pDCs and CD1c + DCs were exposed to 5′-ppp-dsRNA LyoVec, a RIG-I agonist. (A–D) IFN-α secretion was measured by ELISA. #, values are below the limit of detection of the kit (7 pg/mL). The expression of TRAIL by the indicated cells was determined by flow cytometry. Results are expressed as the mean ± SEM of three independent experiments. (A, B) * p < 0.05, one-way ANOVA (Kruskal–Wallis). (C) * p < 0.05, Mann–Whitney test.

Article Snippet: DCs were also cultivated with the TLR7 agonist R837 (1 μg/mL, Invivogen), exogenous type I IFNs (rhIFN-α-2a and rhIFN-β-1a, 100 ng/mL, ImmunoTools) or the RIG-I agonist 5′-ppp-dsRNA LyoVec (10 μg/mL, Invivogen).

Techniques: Expressing, Activation Assay, Cell Culture, Enzyme-linked Immunosorbent Assay, Flow Cytometry, MANN-WHITNEY

Journal: Oncoimmunology

Article Title: Oncolytic measles virus induces tumor necrosis factor-related apoptosis-inducing ligand (TRAIL)-mediated cytotoxicity by human myeloid and plasmacytoid dendritic cells

doi: 10.1080/2162402X.2016.1261240

Figure Lengend Snippet: MV induces IFN-α and TRAIL expression after RIG-I activation in Gen2.2 cells. Gen2.2 cells established by transduction with lentiviruses expressing scrambled shRNA, shRNA against TLR7 (shTLR7) or RIG-I (shRIG-I) were cultured with IL3, IL3+MV or IL3+UV-inactivated MV (MV UV*). IFN-α secretion was measured by ELISA. #, values are below the limit of detection of the kit (7 pg/mL).The percentage of TRAIL positive Gen2.2 cells was determined by flow cytometry. Results are expressed as the mean ± SEM of three independent experiments. * p < 0.05, one-way ANOVA (Kruskal–Wallis).

Article Snippet: DCs were also cultivated with the TLR7 agonist R837 (1 μg/mL, Invivogen), exogenous type I IFNs (rhIFN-α-2a and rhIFN-β-1a, 100 ng/mL, ImmunoTools) or the RIG-I agonist 5′-ppp-dsRNA LyoVec (10 μg/mL, Invivogen).

Techniques: Expressing, Activation Assay, Transduction, shRNA, Cell Culture, Enzyme-linked Immunosorbent Assay, Flow Cytometry

Journal: Oncoimmunology

Article Title: Oncolytic measles virus induces tumor necrosis factor-related apoptosis-inducing ligand (TRAIL)-mediated cytotoxicity by human myeloid and plasmacytoid dendritic cells

doi: 10.1080/2162402X.2016.1261240

Figure Lengend Snippet: DCs activated with MV acquire a functional cytotoxic activity. (A) pDCs were pretreated or not with IRS661 or Rux and then cultured with IL3+MV, R837, type I IFNs (rhIFN-α-2a and rhIFN-β-1a) or 5′-ppp-dsRNA LyoVec. (B) CD1c + DCs were pretreated or not with IRS661 or Rux and then cultured with MV, UV-inactivated MV, R837, type I IFNs (rhIFN-α-2a and rhIFN-β-1a) or 5′-ppp-dsRNA LyoVec. (A, B) DCs were then added to the target Jurkat cells at an effector:target ratio of 20:1 and the percentage of specific lysis of Jurkat cells was determined by the measure of 51 Cr release in the supernatants. Results are expressed as mean ± SEM of at least three independent experiments and each condition was assessed in triplicates for each experiment. For the left panel: * p < 0.05, one-way ANOVA (Kruskal–Wallis); for the right panel: * p < 0.05, Mann–Whitney test.

Article Snippet: DCs were also cultivated with the TLR7 agonist R837 (1 μg/mL, Invivogen), exogenous type I IFNs (rhIFN-α-2a and rhIFN-β-1a, 100 ng/mL, ImmunoTools) or the RIG-I agonist 5′-ppp-dsRNA LyoVec (10 μg/mL, Invivogen).

Techniques: Functional Assay, Activity Assay, Cell Culture, Lysis, MANN-WHITNEY

Journal: Physiological Reports

Article Title: Exercise training reduces ventricular arrhythmias through restoring calcium handling and sympathetic tone in myocardial infarction mice

doi: 10.14814/phy2.13972

Figure Lengend Snippet: (A) The expression of microRNA‐1 (miR‐1) in the LV was elevated in MI‐Sed, and reduced by exercise and β‐blocker treatment. (B) There was no difference in the expression of miR‐133a between the groups ( n = 5/group). (C) There was no difference in expression of protein phosphatase 1 (PP1), while PP2A‐B56‐α (F‐10) expression was significantly decreased in MI‐Sed compared to that in Sham and reversed in MI‐Ex and MI‐βb (D) ( n = 6/group) * P < 0.05, versus Sham; # P < 0.05, versus MI‐Sed.

Article Snippet: The blots were then blocked with the primary antibodies as follows: phospho‐RyR2 (Ser 2814) (A010‐31; Badrilla, Leeds, UK), phospho‐RyR2 (Ser 2808) (A010‐30, Badrilla), RyR2 (PA5‐36121; Thermo Fisher Scientific), SERCA2a (2A7‐A1; Thermo Fisher Scientific), phospho‐Phospholamban (p‐PLN) (S16 + T17; ab62170; Abcam, Cambridge, UK), total Phospholamban (PLN) (ab2865; Abcam), phospho‐PKA (T197; ab75991; Abcam), PKA (ab187515; Abcam), phospho‐CaMKII (T286) (ab32678; Abcam), Oxidized‐CaMKII (Met281/282) (EMD Millipore, Darmstadt, Germany), total CaMKII (ab103840; Abcam), protein phosphatase 2A (PP2A)‐B56‐α (F‐10; sc‐271151; Santa Cruz, Dallas, TX), phospho‐Troponin‐I (p‐TNI) (Ser23/24) (#4004; Cell Signaling Technology, Danvers, MA), total Troponin‐I (TNI) (#4002; Cell Signaling Technology), PP1 (E‐9; sc‐7482; Santa Cruz), and β‐actin (4967s; Cell Signaling Technology).

Techniques: Expressing

Journal: Retrovirology

Article Title: Increased BST2 expression during simian immunodeficiency virus infection is not a determinant of disease progression in rhesus monkeys

doi: 10.1186/s12977-015-0219-8

Figure Lengend Snippet: BST2 mRNA induction by type I interferon. a Fold induction of relative BST2 mRNA in PBMC from three uninfected rhesus macaques after stimulation with human Interferon Alpha A (Alpha 2a) for 16 h. Data are expressed as fold increase over baseline after normalization to pre-treatment values. Error bars represent standard deviation, b relative mRNA copies of BST2 in PBMC (shown in copy numbers per 100 copies of GAPDH) are illustrated in relation to plasma IFN-alpha levels from blood samples of 18 uninfected rhesus macaques 24 h after inoculation of replication incompetent adenovirus or fowl pox vectors. The black dashed line indicates the detection limit of the ELISA and c whole blood MX1 mRNA levels correlate with BST2 mRNA determined in 38 SIVmac251 infected rhesus macaques at 24 wpi. Relative mRNA levels are depicted as log-transformed copy numbers per 100 copies of GAPDH. Each data point represents one animal. Regression line is shown; r , Spearman’s correlation coefficient; p , p value

Article Snippet: PBMC from three healthy animals were stimulated with 10, 50 and 100 ng Human Interferon Alpha A (Alpha 2a) (PBL Biomedical Laboratories) for 16 h. Fold induction of BST2 mRNA expression was calculated using GAPDH as internal control.

Techniques: Standard Deviation, Clinical Proteomics, Enzyme-linked Immunosorbent Assay, Infection, Transformation Assay