Journal: Nutrients

Article Title: Exclusive Enteral Nutrition: Clinical Effects and Changes in Mucosal Cytokine Profile in Pediatric New Inflammatory Bowel Disease

doi: 10.3390/nu11020414

Figure Lengend Snippet: MCP in biopsies from six children at IBD onset and after treatment harvested from colon (in some patients, it was not possible to analyze all MCPs).

Article Snippet: Probes were obtained from Applied Biosystems, TaqMan ® MGB probes, FAM™ dye-labeled: IL-1β (Hs00174097_m1), IL-4 (Hs00174122_m1), IL-6 (Hs00985639_m1), IL-10 (Hs00961622_m1), IL-12β (Hs001011518_m1), IL-22 (Hs01574154_m1), IL-23 (Hs00900828_g1), IL-36γ (Hs00219742_m1), TGF-β1 (Hs00998133_m1), TNF-α (Hs01113624_g1), IFN-γ (Hs00989291_m1), GM-CSF-2 (Hs00929873_m1) and ABL1 (Hs01104728_m1) according to the manufacturer protocol.

Techniques:

Journal: Nutrients

Article Title: Exclusive Enteral Nutrition: Clinical Effects and Changes in Mucosal Cytokine Profile in Pediatric New Inflammatory Bowel Disease

doi: 10.3390/nu11020414

Figure Lengend Snippet: Six cytokine profiles in mucosal biopsies according to mRNA levels measured by q-PCR at disease onset and after EEN treatment. ( A ). IFN-γ at onset and after EEN treatment. ( B ). CSF-2 at onset and after EEN treatment. ( C ). TNF-α at onset and after EEN treatment. ( D ). IL-10 at onset and after EEN treatment. ( E ). IL-12β at onset and after EEN treatment. ( F ). IL-23α at onset and after EEN treatment.

Article Snippet: Probes were obtained from Applied Biosystems, TaqMan ® MGB probes, FAM™ dye-labeled: IL-1β (Hs00174097_m1), IL-4 (Hs00174122_m1), IL-6 (Hs00985639_m1), IL-10 (Hs00961622_m1), IL-12β (Hs001011518_m1), IL-22 (Hs01574154_m1), IL-23 (Hs00900828_g1), IL-36γ (Hs00219742_m1), TGF-β1 (Hs00998133_m1), TNF-α (Hs01113624_g1), IFN-γ (Hs00989291_m1), GM-CSF-2 (Hs00929873_m1) and ABL1 (Hs01104728_m1) according to the manufacturer protocol.

Techniques:

Journal: Nutrients

Article Title: Exclusive Enteral Nutrition: Clinical Effects and Changes in Mucosal Cytokine Profile in Pediatric New Inflammatory Bowel Disease

doi: 10.3390/nu11020414

Figure Lengend Snippet: Real-time PCR values per patient pre- and post-EEN and mean values and SD for group per cytokine.

Article Snippet: Probes were obtained from Applied Biosystems, TaqMan ® MGB probes, FAM™ dye-labeled: IL-1β (Hs00174097_m1), IL-4 (Hs00174122_m1), IL-6 (Hs00985639_m1), IL-10 (Hs00961622_m1), IL-12β (Hs001011518_m1), IL-22 (Hs01574154_m1), IL-23 (Hs00900828_g1), IL-36γ (Hs00219742_m1), TGF-β1 (Hs00998133_m1), TNF-α (Hs01113624_g1), IFN-γ (Hs00989291_m1), GM-CSF-2 (Hs00929873_m1) and ABL1 (Hs01104728_m1) according to the manufacturer protocol.

Techniques: Real-time Polymerase Chain Reaction

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: Virulizin, a novel immunotherapy agent, activates NK cells through induction of IL-12 expression in macrophages

doi: 10.1007/s00262-005-0698-x

Figure Lengend Snippet: IL-12 mRNA expression in melanoma xenograft isolated from mice treated with Virulizin. CD-1 nude mice bearing human melanoma C8161 tumors were treated with 0.2 ml of PBS or Virulizin for 4 weeks, and then tumors were collected 1 h after the last injection. RNA was extracted from tumor tissue by TRIZol reagent. The levels of IL-12α, IL-12β and β-actin mRNA were determined by quantitative real-time RT-PCR. IL-12 mRNA expression was normalized to β-actin level. Each bar represents average fold change calculated from five to seven mice. * p<0.05 compared to PBS control

Article Snippet: Anti-mouse biotinylated IL-12β antibody was purchased from Becton Dickinson (Franklin Lakes, NJ, USA).

Techniques: Expressing, Isolation, Injection, Quantitative RT-PCR, Control

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: Virulizin, a novel immunotherapy agent, activates NK cells through induction of IL-12 expression in macrophages

doi: 10.1007/s00262-005-0698-x

Figure Lengend Snippet: Analysis of Intracellular IL-12 level in peritoneal macrophages by flow cytometry

Article Snippet: Anti-mouse biotinylated IL-12β antibody was purchased from Becton Dickinson (Franklin Lakes, NJ, USA).

Techniques: Flow Cytometry, Fluorescence

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: Virulizin, a novel immunotherapy agent, activates NK cells through induction of IL-12 expression in macrophages

doi: 10.1007/s00262-005-0698-x

Figure Lengend Snippet: Increased level of IL-12β in sera of mice treated with Virulizin. CD-1 nude mice were treated with 0.2 ml of PBS or Virulizin for 3 days, and serum was collected 1 h after the last injection. IL-12 β in serum was determined by quantitative ELISA as described in Materials and Methods section. Each bar shows average levels calculated from ten mice. * p<0.05 compared to PBS control

Article Snippet: Anti-mouse biotinylated IL-12β antibody was purchased from Becton Dickinson (Franklin Lakes, NJ, USA).

Techniques: Injection, Enzyme-linked Immunosorbent Assay, Control

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: Virulizin, a novel immunotherapy agent, activates NK cells through induction of IL-12 expression in macrophages

doi: 10.1007/s00262-005-0698-x

Figure Lengend Snippet: Increased IL-12 mRNA expression in peritoneal macrophages from mice treated with Virulizin. CD-1 nude mice were treated intramuscularly with 0.2 ml of PBS or Virulizin daily for 5 days. Thioglycollate medium (1.5 ml, 3% w/v) was injected i.p. 2 days prior to macrophages collection. RNA was extracted from the cells by TRIZol reagent and IL-12α, IL-12β as well as β-actin genes were quantified by real-time RT-PCR. IL-12 mRNA expression was normalized to β-actin level. Each bar represents average fold change in IL-12 levels calculated from five to seven mice. * p<0.05 compared to PBS control

Article Snippet: Anti-mouse biotinylated IL-12β antibody was purchased from Becton Dickinson (Franklin Lakes, NJ, USA).

Techniques: Expressing, Injection, Quantitative RT-PCR, Control

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: Virulizin, a novel immunotherapy agent, activates NK cells through induction of IL-12 expression in macrophages

doi: 10.1007/s00262-005-0698-x

Figure Lengend Snippet: Analysis of IL-12 intracellular level in peritoneal macrophages by flow cytometry. CD-1 nude mice were treated intramuscularly with 0.2 ml of PBS or Virulizin daily for 5 days. Thioglycollate medium (1.5 ml, 3% w/v) was injected i.p. 2 days prior to macrophages collection. The cells were stained with F4/80 antibody conjugated to FITC, biotinylated anti-IL-12 antibody and phycoerythrin (PE)-Cy5.5-conjugated streptavidin. Samples were examined by flow cytometry. a Histogram of flow cytometric analysis of the gated F4/80+ cells; b Percentage of IL-12α and β positive macrophages. The data are presented as the average of double positive cells (F4/80+ /IL-12+) (mean ± SE) calculated from three independent experiments in Table 1

Article Snippet: Anti-mouse biotinylated IL-12β antibody was purchased from Becton Dickinson (Franklin Lakes, NJ, USA).

Techniques: Flow Cytometry, Injection, Staining

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: Virulizin, a novel immunotherapy agent, activates NK cells through induction of IL-12 expression in macrophages

doi: 10.1007/s00262-005-0698-x

Figure Lengend Snippet: Neutralization of IL-12 in Virulizin-treated mice compromised NK cell cytotoxicity. CD-1 nude mice were treated with Virulizin or PBS daily for 3 days with or without one-time co-administration of 0.25 mg anti-IL-12 or control IgG per mouse. NK cells were isolated from the spleens one hour after the last injection, and co-cultured with YAC-1 target tumor cells to assess NK cell cytotoxicity using LDH release assay as shown in Fig. 2. The effector to target cell ratio used was 40:1. Each bar represents percent cytotoxicity (mean ± SE) calculated from three independent experiments

Article Snippet: Anti-mouse biotinylated IL-12β antibody was purchased from Becton Dickinson (Franklin Lakes, NJ, USA).

Techniques: Neutralization, Control, Isolation, Injection, Cell Culture, Lactate Dehydrogenase Assay