polyclonal neutralizing antibodies against hmgb1  (Tokyo Chemical Industry)

Journal: Journal of Molecular Medicine (Berlin, Germany)

Article Title: Molecular targets in bone cancer pain: a systematic review of inflammatory cytokines

doi: 10.1007/s00109-024-02464-2

Figure Lengend Snippet: Comprehensive overview of the key characteristics of the included studies

Article Snippet: [ ] , Wistar rats (adult, female, 180–200 g) , Walker 256 TIBIA , HMGB1 , Intrathecal administration • polyclonal neutralizing antibodies against HMGB1 , ▪ 50% PWT , Spinal cord lumbar SDH HMGB1, IL-1β (WB) , After TCI ↑ HMGB1 ↑ IL-1β ↑ mechanical allodynia After blocking HMGB1 ↓IL-1β ↓ mechanical allodynia.

Techniques: Expressing, Activation Assay, Blocking Assay, Enzyme-linked Immunosorbent Assay, Binding Assay, Recombinant, Injection, Plasmid Preparation, Small Interfering RNA, Over Expression, Translocation Assay, Clinical Proteomics, Staining

Journal: Frontiers in Immunology

Article Title: Anti-High Mobility Group Box 1 Neutralizing-Antibody Ameliorates Dextran Sodium Sulfate Colitis in Mice

doi: 10.3389/fimmu.2020.585094

Figure Lengend Snippet: HnAb treatment attenuates the severity of dextran sulfate sodium (DSS)-induced colitis mice. (A) The disease activity index (DAI) gradually increased from day 4 onward in the HnAb-treated and vehicle treated DSS-treated mice. (B) Histological examination displays colitis severity. Scale bar: 80 μm (upper panels), 40 μm (lower panels). (C) Histological grading of colitis on day 8. There was a significant difference of scores when DSS/IgY-treated mice vs. the control, DSS/IgY-treated mice vs. DSS/HnAb-treated mice, DSS/HnAb-treated mice vs. the control. (D) The expression of MPO was displayed by immunohistochemistry (IHC). Scale bar: 80 μm (upper panels), 40 μm (lower panels). (E) The expression levels of MPO in the colon were evaluated by IHC using a semi-quantitative scoring system (see Materials and Methods section). A significant difference between the scores of DSS/IgY-treated mice vs. the control, DSS/IgY-treated mice vs. DSS/HnAb-treated mice. No significant difference was found between DSS/HnAb-treated and control mice. (F) The messenger RNA (mRNA) expression levels of IL-1β, IFN-γ, and TNF-α in the colonic tissues of the three groups were detected by RT-PCR. The expression of these three cytokines was increased in the colonic tissues of colitis mice, and HnAb administration resulted in reduced expression level. (G) The expression of HMGB1 was assessed by IHC. Scale bar: 80 μm (upper panels), 40 μm (lower panels). (H) Semiquantiative score for HMGB1 expression by IHC. A significant difference was observed between HMGB1 expression in DSS/IgY-treated vs. control colonic tissue, DSS/IgY-treated vs. HnAb treated colonic tissue, and DSS/HnAb-treated vs. control colonic tissue. (I) HMBG1 mRNA expression levels were detected by RT-PCR. The expression of HMGB1 was increased in the colonic tissues of colitis mice, and HnAb administration resulted in reduced expression level. (J, K) HMGB1 levels in nuclear and cytoplasmic lysate detected by Western blotting and semiquantiative scoring as described in the Materials and Methods . HMGB1 level was reduced in nucleus and cytoplasm of DSS/HnAb-treated compared to DSS/IgY treated colon. Data in (B, D, G, J) were representative of 5 independent experiments. Data in (A, C, E, F, H, I, K) were presented as mean ± SEM of 5 independent experiments. * P < 0.05, ** P < 0.01, *** P < 0.001, ns, not significant, by two-way ANOVA with Tukey’s post-test (A) , by one-way ANOVA with Tukey’s post-test (C, E, F, H, I, K) .

Article Snippet: Simultaneously, half of them were treated with HnAb, a polyclonal chicken IgY against murine HMGB1 neutralizing-antibody, (200 μg/mouse, Shino-Test Corporation, Tokyo, Japan) and others were treated with anti-IgY, a control chicken IgY antibody (200 μg/mouse, Shino-Test Corporation, Tokyo, Japan), via intraperitoneal injection on the 0th, 3rd, and 6th days of DSS feeding.

Techniques: Activity Assay, Expressing, Immunohistochemistry, Reverse Transcription Polymerase Chain Reaction, Western Blot

Journal: Frontiers in Immunology

Article Title: Anti-High Mobility Group Box 1 Neutralizing-Antibody Ameliorates Dextran Sodium Sulfate Colitis in Mice

doi: 10.3389/fimmu.2020.585094

Figure Lengend Snippet: HnAb treatment enhances adherent mucus layer and tight junction integrity during dextran sulfate sodium (DSS) colitis induction. (A) The mucus layer was delineated by immunofluorescence staining for mucin 2. The mucus layer in DSS/HnAb-treated colon was relatively intact, while no complete mucus layer structure could be found in DSS/IgY-treated colon. Scale bar: 80 μm (upper panels), 40 μm (lower panels). (B) Tight junction structure was observed by transmission electron microscope. Tight junction structure was relatively intact in DSS/HnAb-treated comparing to DSS/IgY-treated colon. (C) A significant increase for both ZO-1 and claudin-5 mRNA levels, but not for occludin mRNA levels was seen in DSS/HnAb-treated compared to DSS/IgY-treated colon ( P <0.05). (D, E) Likewise, a significant increase for ZO-1, claudin-5, and occludin was seen in DSS/HnAb-treated vs. DSS/IgY-treated colon by Western analysis. Data in (A, B, D) were representative of five independent experiments. Data in (C, E) were presented as mean ± SEM of five independent experiments. ** P < 0.01, *** P < 0.001, ns, not significant, by one-way ANOVA with Tukey’s post-test (C, E) .

Article Snippet: Simultaneously, half of them were treated with HnAb, a polyclonal chicken IgY against murine HMGB1 neutralizing-antibody, (200 μg/mouse, Shino-Test Corporation, Tokyo, Japan) and others were treated with anti-IgY, a control chicken IgY antibody (200 μg/mouse, Shino-Test Corporation, Tokyo, Japan), via intraperitoneal injection on the 0th, 3rd, and 6th days of DSS feeding.

Techniques: Immunofluorescence, Staining, Transmission Assay, Microscopy, Western Blot

Journal: Frontiers in Immunology

Article Title: Anti-High Mobility Group Box 1 Neutralizing-Antibody Ameliorates Dextran Sodium Sulfate Colitis in Mice

doi: 10.3389/fimmu.2020.585094

Figure Lengend Snippet: Macrophages and CD86+ macrophages are decreased in DSS/HnAb-treated colon. (A) Macrophages were observed in transmission electron microscopic images. Few macrophages could be found in lamina propria in the control, while an increased number was present in dextran sulfate sodium (DSS)/IgY con. DSS/HnAb treatment reduced the number of lamina propria macrophages. (B) Macrophages were identified by the high expression of F4/80 and CD11b. (C) Activation markers CD86, CD11c, CD206 were used to count macrophages by FACS analysis. (D) Number of macrophages in % of total counted cells was significantly higher in DSS/IgY treated vs. control colon ( P <0.001) as well as vs. DSS/HnAb-treated colon ( P <0.001), but not in DSS/HbAg-treated vs. control colon. (E) Significant increase of CD86+ macrophages, but not CD11c+ and CD206 macrophages in DSS/HbAg-treated vs. control colon. Data in (A–C) were representative of five independent experiments. Data in (D, E) were presented as mean ± SEM of five independent experiments. * P < 0.05, *** P < 0.001, ns, not significant, by one-way ANOVA with Tukey’s post-test (D, E) .

Article Snippet: Simultaneously, half of them were treated with HnAb, a polyclonal chicken IgY against murine HMGB1 neutralizing-antibody, (200 μg/mouse, Shino-Test Corporation, Tokyo, Japan) and others were treated with anti-IgY, a control chicken IgY antibody (200 μg/mouse, Shino-Test Corporation, Tokyo, Japan), via intraperitoneal injection on the 0th, 3rd, and 6th days of DSS feeding.

Techniques: Transmission Assay, Expressing, Activation Assay

Journal: Frontiers in Immunology

Article Title: Anti-High Mobility Group Box 1 Neutralizing-Antibody Ameliorates Dextran Sodium Sulfate Colitis in Mice

doi: 10.3389/fimmu.2020.585094

Figure Lengend Snippet: Concomitant dextran sulfate sodium (DSS)/HnAb treatment attenuates TLR4 and MyD88 upregulation during DSS induction of colitis. (A) Significant decrease for both MyD88 and TLR4 messenger RNA (mRNA), but not for TLR2 and TLR9 when HnAb treatment comparing to IgY treatment for DSS-induced colitis mice. (B, C) TLR4 and MyD88 protein was also dramatically reduced in DSS/HnAb-treated vs. DSS/IgY-treated colonic tissue ( P < 0.001). Data in (A, C) were presented as mean ± SEM of five independent experiments. * P < 0.05, ** P < 0.01, *** P < 0.001, ns, not significant, by one-way ANOVA with Tukey’s post-test (A, C) .

Article Snippet: Simultaneously, half of them were treated with HnAb, a polyclonal chicken IgY against murine HMGB1 neutralizing-antibody, (200 μg/mouse, Shino-Test Corporation, Tokyo, Japan) and others were treated with anti-IgY, a control chicken IgY antibody (200 μg/mouse, Shino-Test Corporation, Tokyo, Japan), via intraperitoneal injection on the 0th, 3rd, and 6th days of DSS feeding.

Techniques:

Journal: Frontiers in Immunology

Article Title: Anti-High Mobility Group Box 1 Neutralizing-Antibody Ameliorates Dextran Sodium Sulfate Colitis in Mice

doi: 10.3389/fimmu.2020.585094

Figure Lengend Snippet: TLR4 deficiency and HnAb treatment haven an additive effect in the attenuation of dextran sulfate sodium (DSS)-induced colitis. (A) Time course of DAI increase during DSS colitis induction in wild type (WT) and TLR4-/- mice with and w/o concomitant HnAB treatment. (B) Body weight loss was decreased with TLR4 deficiency. No significant decrease of body weight loss was found after HnAb-treated DSS-induced colitis comparing with the control in TLR4-/- mice. (C) Mucosal damage shown by HE staining in the different genotype/treatment groups. Scale bar: 80 μm (left panels), 40 μm (right panels). (D) Immunohistochemistry (IHC) HMGB1 expression in the different genotype/treatment groups. Scale bar: 80 μm (left panels), 40 μm (right panels). (E) Lowest histological score in DSS/HnAb-treated TLR4-/- mice, followed by DSS/HnAb-treated WT colon. Lower histological score in DSS/IgY-treated TLR4-/- compared to DSS/IgY-treated WT colon. (F) IHC staining showed that HMGB1 was released to cytoplasm and extracellular site in DSS-induced mice colon. Administration of HnAb could partially block HMGB1 redistribution into extracellular region. Data in (C, D) were representative of five independent experiments. Data in (A, B, E, F) were presented as mean ± SEM of five independent experiments. ** P < 0.01, *** P < 0.001, ns, not significant, by two-way ANOVA with Tukey’s post-test (A) , one-way ANOVA with Tukey’s post-test (B, E, F) .

Article Snippet: Simultaneously, half of them were treated with HnAb, a polyclonal chicken IgY against murine HMGB1 neutralizing-antibody, (200 μg/mouse, Shino-Test Corporation, Tokyo, Japan) and others were treated with anti-IgY, a control chicken IgY antibody (200 μg/mouse, Shino-Test Corporation, Tokyo, Japan), via intraperitoneal injection on the 0th, 3rd, and 6th days of DSS feeding.

Techniques: Staining, Immunohistochemistry, Expressing, Blocking Assay