Journal: Frontiers in Immunology

Article Title: Circulating and Salivary Antibodies to Fusobacterium nucleatum Are Associated With Cystic Pancreatic Neoplasm Malignancy

doi: 10.3389/fimmu.2020.02003

Figure Lengend Snippet: Circulating plasma antibody reactivity to oral commensals identified in pancreatic tumor lesions. Plasma diluent (1/300) from patients with histology verified pancreas tissues: IPMN with low grade dysplasia (LGD-IPMN), IPMN with high grade dysplasia or with invasive cancer (HGD-IPMN + Cancer), or benign tumors/non-IPMN (Control) was tested in duplicates in ELISA plates coated with indicated antigen in (A–F) (bacteria: 5 × 10E7 CFU/ml, peptide 10 μg/mL). The total plasma IgG level is shown in (G) . Internal control Mean OD absorbance values of duplicates are shown. Statistical analysis was performed using the Kolmogorov–Smirnov test, ** p < 0.01, *** p < 0.001, and **** p < 0.0001.

Article Snippet: Total levels of salivary IgA and plasma IgG antibodies were measured using Human IgA/IgG ELISA Kit (Novus Biologicals, Colorado, United States) according to the manufacturer’s instructions.

Techniques: Clinical Proteomics, Control, Enzyme-linked Immunosorbent Assay, Bacteria

Journal: Frontiers in Immunology

Article Title: Circulating and Salivary Antibodies to Fusobacterium nucleatum Are Associated With Cystic Pancreatic Neoplasm Malignancy

doi: 10.3389/fimmu.2020.02003

Figure Lengend Snippet: Salivary IgA antibody reactivity to oral commensals quantified by ELISA using saliva diluent (1/16) from indicated patient group. Salivary IgA reactivity to F. nucleatum , the Fap2 mimotope, and S. gordonii (A,B,F) in HG-IPMN/Cancer patients is significantly increased compared to other groups. The differences in antibody levels against bacterial antigens of G. adiacens, S. anginosus , and E. coli (C,D,E) were insignificant between groups. Total plasma IgA level did not differ between the groups (G) . Statistical analysis was performed using the Kolmogorov–Smirnov test, * p < 0.05 and ** p < 0.01.

Article Snippet: Total levels of salivary IgA and plasma IgG antibodies were measured using Human IgA/IgG ELISA Kit (Novus Biologicals, Colorado, United States) according to the manufacturer’s instructions.

Techniques: Enzyme-linked Immunosorbent Assay, Clinical Proteomics

Journal: Frontiers in Immunology

Article Title: Circulating and Salivary Antibodies to Fusobacterium nucleatum Are Associated With Cystic Pancreatic Neoplasm Malignancy

doi: 10.3389/fimmu.2020.02003

Figure Lengend Snippet: Correlation between circulating plasma (A) or salivary (B) antibody reactivity to F. nucleatum and Fap2 mimotope, or between circulating plasma antibody and salivary antibodies to either of antigen (C,D) . Competitive ELISA (E) with saliva samples with or without antigen pre-absorption as indicated for 2 h, then subjected to Fap2 mimotope ELISA test. Statistical analysis was performed using two-tailed Pearson correlation test and Wilcoxon test for two related samples. ** p < 0.01, ns = not significant.

Article Snippet: Total levels of salivary IgA and plasma IgG antibodies were measured using Human IgA/IgG ELISA Kit (Novus Biologicals, Colorado, United States) according to the manufacturer’s instructions.

Techniques: Clinical Proteomics, Competitive ELISA, Enzyme-linked Immunosorbent Assay, Two Tailed Test

Journal: Journal of Virology

Article Title: Infectivity of Hepatitis C Virus Is Influenced by Association with Apolipoprotein E Isoforms

doi: 10.1128/jvi.01063-10

Figure Lengend Snippet: FIG. 4. Suppression of LDLR and SR-BI results in reduced HCV infection. (A) Knockdown of the LDLR and SR-BI by siRNA. HuH7.5 cells were seeded onto 24-well plates and transfected with siRNA (si-control, si-LDLR, and/or si-SR-BI). Twenty-four hours after transfection, cell lysates were analyzed for the expression of the LDLR, SR-BI, and actin by Western blotting. (B) Analysis of HCV infectivity for HuH7.5 cells in which either the LDLR, SR-BI, or both were silenced. Cells were transfected with siRNA (si-control, si-LDLR, and/or si-SR-BI). Twenty-four hours after transfection, HCVcc produced from cells bearing ApoE2 (open bars) or ApoE3 (filled bars) was serially diluted and inoculated. Forty-eight hours after infection, infected cells were counted by fluorescence microscopy after staining with an anti-NS5A antibody. HCV/ApoE3, HCV bearing ApoE3; HCV/ApoE2, HCV bearing ApoE2. P values were determined by comparison (by Student’s t test) with si-control-treated cells. *, P 0.0005; **, P 0.005. (C) HuH7.5 cells were preincubated with control goat IgG (Ctrl-IgG), control rat preimmune serum (Ctrl-serum), or a goat anti-LDLR, rat anti-SR-BI, rat anti-claudin 1 (anti-CLDN1), or mouse anti-CD81 antibody for 1 h at 37°C before infection with serially diluted HCVcc from HuH7.5 cells expressing ApoE2 (open bars) or ApoE3 (filled bars). Forty-eight hours after infection, infected cells were stained with anti-NS5A antibodies and counted by fluorescence microscopy. The average values for three independent experiments are shown; error bars, standard deviations of the means. P values were determined by comparison (by Student’s t test) against cells treated with preimmune serum. *, P 0.0005; **, P 0.005.

Article Snippet: Rat anti-HA (3F10; Roche Applied Science), mouse anti-Flag (M2; Sigma), mouse anti-actin (AC-40; Sigma), goat anti- 1-antitrypsin (K15600G; Biodesign International), mouse anti-ApoE (13F45; Autogen Bioclear), goat anti-ApoE (AB947; Chemicon International), sheep anti-ApoB (K90086C; Biodesign International), mouse anti-core (CP11; Institute of Immunology), goat anti-LDLR (AF2148; R&D Systems), rabbit anti-SR-BI (EP1556Y; Abcam), mouse anti-CD81 (JS-81; BD Biosciences), and normal goat IgG (sc-2028; Santa Cruz Biotechnology) antibodies were purchased commercially.

Techniques: Infection, Knockdown, Transfection, Control, Expressing, Western Blot, Produced, Microscopy, Staining, Comparison

Journal: The Journal of Neuroscience

Article Title: Brain-Derived Neurotrophic Factor Increases in the Uninjured Dorsal Root Ganglion Neurons in Selective Spinal Nerve Ligation Model

doi: 10.1523/jneurosci.21-13-04891.2001

Figure Lengend Snippet: Figure 4. Effects of intrathecal injection (arrow) of anti-BDNF antibody (20 mg, closed circles) or normal sheep IgG (20 mg, open circles) on paw withdrawal response to noxious heat of the rats that received L5 SPNL. Difference scores were calculated by subtracting contralateral withdrawal latencies from ipsilateral withdrawal latencies and expressed as mean 6 SEM (n 5 8 for each treatment). Thermal hyperalgesia was abolished for at least 2.5 hr by anti-BDNF antibody injection, whereas normal sheep IgG injection had no significant effect on the behavior. *p , 0.05 versus before L5 SPNL ( pre ope); #p , 0.05 versus before injection ( pre inj.) by one-way ANOVA followed by Fisher’s PLSD.

Article Snippet: After an additional 3 d, the development of neuropathic pain was confirmed, and sheep antiBDNF antibody (20 mg in 20 ml of PBS; Chemicon, Temecula, CA) or normal sheep IgG (20 mg in 20 ml of PBS; Cappel, Aurora, OH) was injected through the intrathecal cannula.

Techniques: Injection

Journal: The Journal of Neuroscience

Article Title: Brain-Derived Neurotrophic Factor Increases in the Uninjured Dorsal Root Ganglion Neurons in Selective Spinal Nerve Ligation Model

doi: 10.1523/jneurosci.21-13-04891.2001

Figure Lengend Snippet: Figure 8. The effect of local application of anti-NGF antibody on thermal hyperalgesia in the L5 spinal nerve ligation (L5 SPNL) model. A small piece of gelatin sponge saturated with sheep anti-NGF antibody (100 mg, filled circles, n 5 5–9) or normal sheep IgG (100 mg, open circles, n 5 5) was placed on the surface of the L4 spinal nerve when the L5 SPNL was performed. The anti-NGF antibody prevented the development of ther- mal hyperalgesia for up to 6 d after surgery. Data are expressed as mean 6 SEM. *p , 0.05 versus before L5 SPNL ( pre) by one-way ANOVA followed by Fisher’s PLSD.

Article Snippet: After an additional 3 d, the development of neuropathic pain was confirmed, and sheep antiBDNF antibody (20 mg in 20 ml of PBS; Chemicon, Temecula, CA) or normal sheep IgG (20 mg in 20 ml of PBS; Cappel, Aurora, OH) was injected through the intrathecal cannula.

Techniques: Ligation

Journal: Immunobiology

Article Title: c-Rel is dispensable for the differentiation and functional maturation of M cells in the follicle-associated epithelium

doi: 10.1016/j.imbio.2016.09.008

Figure Lengend Snippet: Primers used for RT-qPCR analysis.

Article Snippet: For the detection of SpiB in paraformaldehyde-fixed sections, antigen retrieval was performed with citrate buffer (pH 7.0, 121 °C, 5 min) prior to immunostaining with sheep anti-mouse SpiB polyclonal antibody (R&D Systems).

Techniques:

Journal: Immunobiology

Article Title: c-Rel is dispensable for the differentiation and functional maturation of M cells in the follicle-associated epithelium

doi: 10.1016/j.imbio.2016.09.008

Figure Lengend Snippet: SpiB expression in the FAE is unaffected in the absence of c-Rel. (A) IHC analysis of SpiB expression (green) in the FAE of c-Rel −/− and wild-type (WT) control mice. Boxed areas in upper panels are shown at higher magnification in the lower panels. Broken lines indicate the FAE boundaries. Arrows, Spi-B + cell nuclei in the FAE. (B) Morphometric analysis showed that the number of SpiB + cells in the FAE of c-Rel −/− and WT were similar. (C) RT-qPCR analysis suggested there was no significant difference in the expression of Spib mRNA levels in Peyer’s patches from c-Rel −/− or WT mice. Gene expression data are normalised so that the mean level in samples from WT mice was 1.0. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: For the detection of SpiB in paraformaldehyde-fixed sections, antigen retrieval was performed with citrate buffer (pH 7.0, 121 °C, 5 min) prior to immunostaining with sheep anti-mouse SpiB polyclonal antibody (R&D Systems).

Techniques: Expressing, Control, Quantitative RT-PCR, Gene Expression