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Sample & Assay Technologies
InflammaTools: Complete research tools to study Inflammation regulation
Raed Samara, Ph.D.R&D Scientist
Sample & Assay Technologies- 2 -
• SABiosciences is now a Company
Biology-focused solutions for pathway analysis
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Introduction: Inflammation
Definition: a protective tissue response to tissue damage or microbes, which serves to destroy, dilute, or wall off both the injurious agent and the injured tissues.
Microbes/InfectionTissue Damage
Acute Inflammation
Chronic InflammationInfection ClearanceTissue Homeostasis
Pre-cancer & CancerChronic Inflammatory Diseases
Cytokines & ChemokinesSignaling PathwaysImmune system composition
EpigeneticChanges
mRNAChanges
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Inflammation experimental design & QIAGEN
Gene Expression� RT-PCR
Epigenetics� miRNA� DNA methylation� Histone modifications
Functional Studies� Reporter assays� siRNA/shRNA
Protein Expression� Cytokine levels
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Basic principles of qRT-PCR: Overview
Real-Time PCR
Amplify and simultaneously quantify target DNA
Reverse Transcription Real-Time PCR
Amplify and simultaneously quantify mRNA
For more information and webinars on real-time PCR, visit:
www.sabiosciences.com/seminarlist.php
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Experimental overview: Gene expression analysis
Stimulate CellsRT-PCR Arrays or Assays
Data Analysis
Isolate RNA
Isolate DNA
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Gene expression in Inflammation
Definition: a protective tissue response to tissue damage or microbes, which serves to destroy, dilute, or wall off both the injurious agent and the injured tissues.
Microbes/InfectionTissue Damage
Infection ClearanceTissue Homeostasis
Pre-cancer & CancerChronic Inflammatory Diseases
Cytokines & ChemokinesSignaling PathwaysImmune system composition
EpigeneticChanges
mRNAChanges
Acute Inflammation
Chronic Inflammation
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Inflammation studies: Gene expression
The transcription factor cMyb is upregulated in naïve CD4 T cells.
Sample & Assay Technologies- 9 -
Inflammation studies: Gene expression
The transcription factor cMyb is upregulated in naïve CD4 T cells.
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Inflammation studies: Gene expression
The transcription factor cMyb is upregulated in naïve CD4 T cells. What is the effect of cMyb on naïve CD4 T cell maturation (to Th1, Th2, or Th3)?
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Inflammation studies: Gene expression
The transcription factor cMyb is upregulated in naïve CD4 T cells. What is the effect of cMyb on naïve CD4 T cell maturation (to Th1, Th2, or Th3)?
Experiment: Knockdown cMyb in human peripheral CD4+ T cells. Examine gene changes using the Human Th1-Th2-Th3 RT2 Profiler PCR Array (PAHS-034).
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Inflammation studies: Gene expression
The transcription factor cMyb is upregulated in naïve CD4 T cells. What is the effect of cMyb on naïve CD4 T cell maturation (to Th1, Th2, or Th3)?
Experiment: Knockdown cMyb in human peripheral CD4+ T cells. Examine gene changes using the Human Th1-Th2-Th3 RT2 Profiler PCR Array (PAHS-034).
Results:
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Inflammation studies: Gene expression
The transcription factor cMyb is upregulated in naïve CD4 T cells. What is the effect of cMyb on naïve CD4 T cell maturation (to Th1, Th2, or Th3)?
Experiment: Knockdown cMyb in human peripheral CD4+ T cells. Examine gene changes using the Human Th1-Th2-Th3 RT2 Profiler PCR Array (PAHS-034).
Results:
Conclusions: Knockdown of cMyb in human peripheral CD4+ T cells decreased the expression of Th2 cytokine genes, and negatively affected Th2 cell maturation in primary human peripheral blood T cells.
Sample & Assay Technologies- 14 -
Gene expression in Inflammation
Definition: a protective tissue response to tissue damage or microbes, which serves to destroy, dilute, or wall off both the injurious agent and the injured tissues.
Microbes/InfectionTissue Damage
Infection ClearanceTissue Homeostasis
Pre-cancer & CancerChronic Inflammatory Diseases
Cytokines & ChemokinesSignaling PathwaysImmune system composition
EpigeneticChanges
mRNAChanges
Acute Inflammation
Chronic Inflammation
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Inflammation studies: Gene expressionLimited material
Mucin depleted foci (MDF) are precancerous lesions of the colon that show signs of inflammation.
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Inflammation studies: Gene expressionLimited material
Mucin depleted foci (MDF) are precancerous lesions of the colon that show signs of inflammation.
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Inflammation studies: Gene expressionLimited material
Mucin depleted foci (MDF) are precancerous lesions of the colon that show signs of inflammation. What roles do TLRs play in MDF?
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Inflammation studies: Gene expressionLimited material
Mucin depleted foci (MDF) are precancerous lesions of the colon that show signs of inflammation. What roles do TLRs play in MDF?
Experiment: Treat mice with DMH for 15 weeks to induce MDF. Collect RNA from MDF and normal colon. Convert mRNA to cDNA. Pre-amplify cDNA with the Mouse TLR PreAMP primer mixes (PBM-0018). Analyze differential gene expression with the Mouse TLR RT2
Profiler PCR Arrays (PAMM-018)
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Inflammation studies: Gene expressionLimited material
Mucin depleted foci (MDF) are precancerous lesions of the colon that show signs of inflammation. What roles do TLRs play in MDF?
Experiment: Treat mice with DMH for 15 weeks to induce MDF. Collect RNA from MDF and normal colon. Pre-amplify cDNA with the Mouse TLR PreAMP primer mixes (PBM-0018). Analyze differential gene expression with the Mouse TLR RT2 Profiler PCR Arrays (PAMM-018)
Results:
Sample & Assay Technologies- 20 -
Inflammation studies: Gene expressionLimited material
Mucin depleted foci (MDF) are precancerous lesions of the colon that show signs of inflammation. What roles do TLRs play in MDF?
Experiment: Treat mice with DMH for 15 weeks to induce MDF. Collect RNA from MDF and normal colon. Pre-amplify cDNA with the Mouse TLR PreAMP primer mixes (PBM-0018). Analyze differential gene expression with the Mouse TLR RT2 Profiler PCR Arrays (PAMM-018)
Results:
Conclusion: MDF induces TLR2 gene expression in MDF compared to normal colon, suggesting a link between TLR2 and MDF.
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Inflammation experimental design & QIAGEN
Gene Expression� RT-PCR
Epigenetics� miRNA� DNA methylation� Histone modifications
Functional Studies� Reporter assays� siRNA/shRNA
Protein Expression� Cytokine levels
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Epigenetics: Overview
Structural Gene NFκB BS
Activated Transcription Factors
p53 BS
p53
NFκB
Protein “A”
mRNA ”A”
MeMeMe
Histones
Histone-DNA Interactions
DNA Methylation
Me AcMe Me MeMe
DNA Methylation
miRNAshRNAsiRNA
Transcription Initiation Complex
+
–
For more information and webinars on Epigenetics, v isit:www.sabiosciences.com/seminarlist.php
Sample & Assay Technologies- 23 -
miRNA reagents from QIAGEN
3’ UTR Reporters
Target Identification Expression
Isolation
Function
miRNA mimics & inhibitors
miScript miRNA PCR Arrays
miRNeasy
miRNAStudies
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miRNA in Inflammation
Definition: a protective tissue response to tissue damage or microbes, which serves to destroy, dilute, or wall off both the injurious agent and the injured tissues.
Microbes/InfectionTissue Damage
Infection ClearanceTissue Homeostasis
Pre-cancer & CancerChronic Inflammatory Diseases
Cytokines & ChemokinesSignaling PathwaysImmune system composition
EpigeneticChanges(miRNA)
mRNAChanges
Acute Inflammation
Chronic Inflammation
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Inflammation studies: miRNA function
Resolution of inflammation is a tightly controlled mechanism. The D-series resolvins (RvD1) are a class of players that mediate this resolution.
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Inflammation studies: miRNA function
Resolution of inflammation is a tightly controlled mechanism. The D-series resolvins (RvD1) are a class of players that mediate this resolution.
FASEB J. 25, 1–17 (2011)
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Inflammation studies: miRNA function
Resolution of inflammation is a tightly controlled mechanism. The D-series resolvins (RvD1) are a class of players that mediate this resolution. Can RvD1-mediated resolution happen through miRNAs?
FASEB J. 25, 1–17 (2011)
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Inflammation studies: miRNA function
Resolution of inflammation is a tightly controlled mechanism. The D-series resolvins (RvD1) are a class of players that mediate this resolution. Can RvD1-mediated resolution happen through miRNAs?
Experiment: Treat mice with zymosan A to induce acute inflammation (murine peritonitis), in the presence or absence of RvD1, for 4, 12, or 24 hours (representing onset, maximum, and resolution phases). Collect RNA from leukocytes. Analyze miRNA expression with the Mouse miRNome miRNA PCR Array (MIMM-216Z)
FASEB J. 25, 1–17 (2011)
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Inflammation studies: miRNA function
Resolution of inflammation is a tightly controlled mechanism. The D-series resolvins (RvD1) are a class of players that mediate this resolution. Can RvD1-mediated resolution happen through miRNAs?
Experiment: Treat mice with zymosan A to induce acute inflammation (murine peritonitis), in the presence or absence of RvD1, for 4, 12, or 24 hours (representing onset, maximum, and resolution phases). Collect RNA from leukocytes. Analyze miRNA expression with the Mouse miRNome miRNA PCR Array (MIMM-216Z)
Results:
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Inflammation studies: miRNA function
Resolution of inflammation is a tightly controlled mechanism. The D-series resolvins (RvD1) are a class of players that mediate this resolution. Can RvD1-mediated resolution happen through miRNAs?
Experiment: Treat mice with zymosan A to induce acute inflammation (murine peritonitis), in the presence or absence of RvD1, for 4, 12, or 24 hours (representing onset, maximum, and resolution phases). Collect RNA from leukocytes. Analyze miRNA expression with the Mouse miRNome miRNA PCR Array (MIMM-216Z)
Results:
Conclusion: RvD1 regulated resolution by controlling the expression of miR-219 and miR-146b. These miRNAs are the first identified miRNAs in resolvin resolution circuits.
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Epigenetics: Overview
Structural Gene NFκB BS
Activated Transcription Factors
p53 BS
p53
NFκB
Protein “A”
mRNA ”A”
MeMeMe
Histones
Histone-DNA Interactions
DNA Methylation
Me AcMe Me MeMe
DNA Methylation
miRNAshRNAsiRNA
Transcription Initiation Complex
+
–
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DNA methylation in Inflammation
Definition: a protective tissue response to tissue damage or microbes, which serves to destroy, dilute, or wall off both the injurious agent and the injured tissues.
Microbes/InfectionTissue Damage
Infection ClearanceTissue Homeostasis
Pre-cancer & CancerChronic Inflammatory Diseases
Cytokines & ChemokinesSignaling PathwaysImmune system composition
EpigeneticChanges
(DNA Methylation)
mRNAChanges
Acute Inflammation
Chronic Inflammation
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Inflammation studies: DNA methylation
ANCA Vasculitis is characterized by microvascular inflammation caused by activated neutrophils. These activated neutrophils aberrantly express PR3 and MPO, which are silenced in normal individuals.
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Inflammation studies: DNA methylation
ANCA Vasculitis is characterized by microvascular inflammation caused by activated neutrophils. These activated neutrophils aberrantly express PR3 and MPO, which are silenced in normal individuals.
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Inflammation studies: DNA methylation
ANCA Vasculitis is characterized by microvascular inflammation caused by activated neutrophils. These activated neutrophils aberrantly express PR3 and MPO, which are silenced in normal individuals.Does aberrant PR3 and MPO expression result from disrupted epigenetic silencing?
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Inflammation studies: DNA methylation
ANCA Vasculitis is characterized by microvascular inflammation caused by activated neutrophils. These activated neutrophils aberrantly express PR3 and MPO, which are silenced in normal individuals.Does aberrant PR3 and MPO expression result from disrupted epigenetic silencing?
Experiment: Isolate gDNA from neutrophils and treat with methylation-sensitive and methylation-dependent restriction enzymes (EpiTect Methyl DNA Restriction Kit 335451). Analyze DNA methylation by real-time PCR using primers specific for CpG islands in PR3 (MePH23428-1A) and MPO (MePH22382-1A).
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Inflammation studies: DNA methylation
ANCA Vasculitis is characterized by microvascular inflammation caused by activated neutrophils. These activated neutrophils aberrantly express PR3 and MPO, which are silenced in normal individuals.Does aberrant PR3 and MPO expression result from disrupted epigenetic silencing?
Experiment: Isolate gDNA from neutrophils and treat with methylation-sensitive and methylation-dependent restriction enzymes (EpiTect Methyl DNA Restriction Kit 335451). Analyze DNA methylation by real-time PCR using primers specific for CpG islands in PR3 (MePH23428-1A) and MPO (MePH22382-1A).
Results:
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Inflammation studies: DNA methylation
ANCA Vasculitis is characterized by microvascular inflammation caused by activated neutrophils. These activated neutrophils aberrantly express PR3 and MPO, which are silenced in normal individuals.Does aberrant PR3 and MPO expression result from disrupted epigenetic silencing?
Experiment: Isolate gDNA from neutrophils and treat with methylation-sensitive and methylation-dependent restriction enzymes (EpiTect Methyl DNA Restriction Kit 335451). Analyze DNA methylation by real-time PCR using primers specific for CpG islands in PR3 (MePH23428-1A) and MPO (MePH22382-1A).
Results:
Conclusion: Methylation of CpG islands is a mechanism that controls the expression of MPO, but not the expression of PR3.
The EpiTect Methyl System uses the MethylScreen™ Technology provided under license from Orion Genomics, LLC.
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Epigenetics: Overview
Structural Gene NFκB BS
Activated Transcription Factors
p53 BS
p53
NFκB
Protein “A”
mRNA ”A”
MeMeMe
Histones
Histone-DNA Interactions
DNA Methylation
Me AcMe Me MeMe
DNA Methylation
miRNAshRNAsiRNA
Transcription Initiation Complex
+
–
Sample & Assay Technologies- 40 -
Histone modification in Inflammation
Definition: a protective tissue response to tissue damage or microbes, which serves to destroy, dilute, or wall off both the injurious agent and the injured tissues.
Microbes/InfectionTissue Damage
Infection ClearanceTissue Homeostasis
Pre-cancer & CancerChronic Inflammatory Diseases
Cytokines & ChemokinesSignaling PathwaysImmune system composition
EpigeneticChanges
(Histone Modifications)
mRNAChanges
Acute Inflammation
Chronic Inflammation
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Inflammation studies: Histone modification
Th17 cells are a subset of CD4 T cells that differentiate from naïve T cells, secrete IL-17, and induce massive tissue inflammation. T cell factor 1 (TCF-1) is a transcription factor that plays an important role in T cell differentiation in the periphery.
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Inflammation studies: Histone modification
Th17 cells are a subset of CD4 T cells that differentiate from naïve T cells, secrete IL-17, and induce massive tissue inflammation. T cell factor 1 (TCF-1) is a transcription factor that plays an important role in T cell differentiation in the periphery.
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Inflammation studies: Histone modification
Th17 cells are a subset of CD4 T cells that differentiate from naïve T cells, secrete IL-17, and induce massive tissue inflammation. T cell factor 1 (TCF-1) is a transcription factor that plays an important role in T cell differentiation in the periphery. Does TCF-1 control the differentiation of Th17 cells, and how?
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Inflammation studies: Histone modification
Th17 cells are a subset of CD4 T cells that differentiate from naïve T cells, secrete IL-17, and induce massive tissue inflammation. T cell factor 1 (TCF-1) is a transcription factor that plays an important role in T cell differentiation in the periphery. Does TCF-1 control the differentiation of Th17 cells, and how?
Experiment: Isolate naïve T cells from spleens of mice. Induce Th17 cell differentiation. Collect chromatin and process with the EpiTect ChIP one-day kit (334471) for chromatin immunoprecipitation with anti-histone antibodies. Detect precipitated DNA with qRT-PCR.
Sample & Assay Technologies- 45 -
Inflammation studies: Histone modification
Th17 cells are a subset of CD4 T cells that differentiate from naïve T cells, secrete IL-17, and induce massive tissue inflammation. T cell factor 1 (TCF-1) is a transcription factor that plays an important role in T cell differentiation in the periphery. Does TCF-1 control the differentiation of Th17 cells, and how?
Experiment: Isolate naïve T cells from spleens of mice. Induce aTh17 cell differentiation. Collect chromatin and process with the EpiTect ChIP one-day kit for chromatin immunoprecipitation with anti-histone antibodies. Detect precipitated DNA with qRT-PCR.
Results:
Open chromatin = increased expression
Sample & Assay Technologies- 46 -
Inflammation studies: Histone modification
Th17 cells are a subset of CD4 T cells that differentiate from naïve T cells, secrete IL-17, and induce massive tissue inflammation. T cell factor 1 (TCF-1) is a transcription factor that plays an important role in T cell differentiation in the periphery. Does TCF-1 control the differentiation of Th17 cells, and how?
Experiment: Isolate naïve T cells from spleens of mice. Induce Th17 cell differentiation. Collect chromatin and process with the EpiTect ChIP one-day kit (334471) for chromatin immunoprecipitation with anti-histone antibodies. Detect precipitated DNA with qRT-PCR.
Results:
Conclusion: TCF-1 mediates the repression of IL-17 locus during T cell development by chromatin modifications.
Sample & Assay Technologies- 47 -
Inflammation experimental design & QIAGEN
Gene Expression� RT-PCR
Epigenetics� miRNA� DNA methylation� Histone modifications
Functional Studies� Reporter assays� siRNA/shRNA
Protein Expression� Cytokine levels
Sample & Assay Technologies- 48 -
Reporter Construct GFP/firefly luciferaseTATAbox
Tandem repeats of TRE
Transcriptional Regulatory Elements (TRE), which es tablish thespecificity of each reporter
Cignal Reporter Assays: Complete SolutionReporter assays: Overview
TF EGFP FL
Upstream SignalingEvents
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Reporter assays for Inflammation
Definition: a protective tissue response to tissue damage or microbes, which serves to destroy, dilute, or wall off both the injurious agent and the injured tissues.
Microbes/InfectionTissue Damage
Infection ClearanceTissue Homeostasis
Pre-cancer & CancerChronic Inflammatory Diseases
Cytokines & ChemokinesSignaling PathwaysImmune system composition
EpigeneticChanges
mRNAChanges
Acute Inflammation
Chronic Inflammation
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Inflammation studies: Reporter assay
Neisseria Meningitidis infections can be rapidly fatal due to acute inflammatory responses that are mediated by capsular polysaccharides (CPS), but the innate immunostimulatory activity of CPS is largely unknown.
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Inflammation studies: Reporter assay
Neisseria Meningitidis infections can be rapidly fatal due to acute inflammatory responses that are mediated by capsular polysaccharides (CPS), but the innate immunostimulatory activity of CPS is largely unknown.
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Inflammation studies: Reporter assay
Neisseria Meningitidis infections can be rapidly fatal due to acute inflammatory responses that are mediated by capsular polysaccharides (CPS), but the innate immunostimulatory activity of CPS is largely unknown. What signaling pathways are induced upon CPS recognition?
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Inflammation studies: Reporter assay
Neisseria Meningitidis infections can be rapidly fatal due to acute inflammatory responses that are mediated by capsular polysaccharides (CPS), but the innate immunostimulatory activity of CPS is largely unknown. What signaling pathways are induced upon CPS recognition?
Experiment: Reverse-transfect cells with 10 dual-luciferase reporter assays, individually, on a reporter array (Cignal Finder 10-pathway Reporter Array; CCA-108L). Induce transfected cells with CPS, and measure luciferase levels to determine which pathway(s) is(are) activated by CPS
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Inflammation studies: Reporter assay
Neisseria Meningitidis infections can be rapidly fatal due to acute inflammatory responses that are mediated by capsular polysaccharides (CPS), but the innate immunostimulatory activity of CPS is largely unknown. What signaling pathways are induced upon CPS recognition?
Experiment: Reverse-transfect cells with 10 dual-luciferase reporter assays, individually, on a reporter array (Cignal Finder 10-pathway Reporter Array; CCA-108L). Induce transfected cells with CPS, and measure luciferase levels to determine which pathway(s) is(are) activated by CPS
Results:
Sample & Assay Technologies- 55 -
Inflammation studies: Reporter assay
Neisseria Meningitidis infections can be rapidly fatal due to acute inflammatory responses that are mediated by capsular polysaccharides (CPS), but the innate immunostimulatory activity of CPS is largely unknown. What signaling pathways are induced upon CPS recognition?
Experiment: Reverse-transfect cells with 10 dual-luciferase reporter assays, individually, on a reporter array (Cignal Finder 10-pathway Reporter Array; CCA-108L). Induce transfected cells with CPS, and measure luciferase levels to determine which pathway(s) is(are) activated by CPS
Results:
Conclusions: NFkB is the major signaling pathway activated in response to CPS.
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shRNA/siRNA function
Structural Gene NFκB BS
Activated Transcription Factors
p53 BS
p53
NFκB
Protein “A”
mRNA ”A”
MeMeMe
Histones
Histone-DNA Interactions
DNA Methylation
Me AcMe Me MeMe
DNA Methylation
miRNAshRNAsiRNA
Transcription Initiation Complex
+
–
Sample & Assay Technologies- 57 -
Gene knockdown for Inflammation
Definition: a protective tissue response to tissue damage or microbes, which serves to destroy, dilute, or wall off both the injurious agent and the injured tissues.
Microbes/InfectionTissue Damage
Infection ClearanceTissue Homeostasis
Pre-cancer & CancerChronic Inflammatory Diseases
Cytokines & ChemokinesSignaling PathwaysImmune system composition
EpigeneticChanges
mRNAChanges
Acute Inflammation
Chronic Inflammation
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Inflammation studies: siRNA knockdown
Resident alveolar macrophages (AM) constitute the first line of defense against invading lung Streptococcus pneumoniae. Studies have shown that morphine-treated mice experience increased mortality and bacterial outgrowth and dissemination.
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Inflammation studies: siRNA knockdown
Resident alveolar macrophages (AM) constitute the first line of defense against invading lung Streptococcus pneumoniae. Studies have shown that morphine-treated mice experience increased mortality and bacterial outgrowth and dissemination.
The Journal of Immunology, 2008, 180: 3594–3600.
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Inflammation studies: siRNA knockdown
Resident alveolar macrophages (AM) constitute the first line of defense against invading lung Streptococcus pneumoniae. Studies have shown that morphine-treated mice experience increased mortality and bacterial outgrowth and dissemination. What is the mechanism by which morphine does this?
The Journal of Immunology, 2008, 180: 3594–3600.
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Inflammation studies: siRNA knockdown
Resident alveolar macrophages (AM) constitute the first line of defense against invading lung Streptococcus pneumoniae. Studies have shown that morphine-treated mice experience increased mortality and bacterial outgrowth and dissemination. What is the mechanism by which morphine does this?
Experiment: MH-S cells (murine AM cells) were transfected with either a negative control SureSilencing shRNA or a SureSilencing shRNA plasmid for mouse TLR9. Levels of MIP-2 were measured after treating transfected cells with morphine and Streptococcus pneumoniae
The Journal of Immunology, 2008, 180: 3594–3600.
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Inflammation studies: siRNA knockdown
Resident alveolar macrophages (AM) constitute the first line of defense against invading lung Streptococcus pneumoniae. Studies have shown that morphine-treated mice experience increased mortality and bacterial outgrowth and dissemination. What is the mechanism by which morphine does this?
Experiment: MH-S cells (murine AM cells) were transfected with either a negative control SureSilencing shRNA or a SureSilencing shRNA plasmid for mouse TLR9. Levels of MIP-2 were measured after treating transfected cells with morphine and Streptococcus pneumoniae.
Results:
Sample & Assay Technologies- 63 -
Inflammation studies: siRNA knockdown
Resident alveolar macrophages (AM) constitute the first line of defense against invading lung Streptococcus pneumoniae. Studies have shown that morphine-treated mice experience increased mortality and bacterial outgrowth and dissemination. What is the mechanism by which morphine does this?
Experiment: MH-S cells (murine AM cells) were transfected with either a negative control SureSilencing shRNA or a SureSilencing shRNA plasmid for mouse TLR9. Levels of MIP-2 were measured after treating transfected cells with morphine and Streptococcus pneumoniae.
Results:
Conclusion: Morphine reduces levels of Streptococcus pneumoniae -induced MIP-2 from AM cells in a TLR9-dependent manner.
Sample & Assay Technologies- 64 -
Inflammation experimental design & QIAGEN
Gene Expression� RT-PCR
Epigenetics� miRNA� DNA methylation� Histone modifications
Functional Studies� Reporter assays� siRNA/shRNA
Protein Expression� Cytokine levels
Sample & Assay Technologies- 65 -
Chemokine expression in Inflammation
Definition: a protective tissue response to tissue damage or microbes, which serves to destroy, dilute, or wall off both the injurious agent and the injured tissues.
Microbes/InfectionTissue Damage
Infection ClearanceTissue Homeostasis
Pre-cancer & CancerChronic Inflammatory Diseases
Cytokines & ChemokinesSignaling PathwaysImmune system composition
EpigeneticChanges
mRNAChanges
Acute Inflammation
Chronic Inflammation
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Inflammation studies: Chemokine levels
Diabetes mellitus (DM) is a major risk factor for tuberculosis (TB) but the defect in protective immunity responsible for this has not been defined.
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Inflammation studies: Chemokine levels
Diabetes mellitus (DM) is a major risk factor for tuberculosis (TB) but the defect in protective immunity responsible for this has not been defined.
The Journal of Immunology, 2010, 184: 6275–6282
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Inflammation studies: Chemokine levels
Diabetes mellitus (DM) is a major risk factor for tuberculosis (TB) but the defect in protective immunity responsible for this has not been defined. Could this be due to defective chemokine secretion leading to delayed priming of adaptive immunity?
The Journal of Immunology, 2010, 184: 6275–6282
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Inflammation studies: Chemokine levels
Diabetes mellitus (DM) is a major risk factor for tuberculosis (TB) but the defect in protective immunity responsible for this has not been defined. Could this be due to defective chemokine secretion leading to delayed priming of adaptive immunity?
Experiment: Lung homogenates from TB-infected Diabetic mice were tested for chemokine levels using the Multi-Analyte ELISArray Kit.
The Journal of Immunology, 2010, 184: 6275–6282
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Inflammation studies: Chemokine levels
Diabetes mellitus (DM) is a major risk factor for tuberculosis (TB) but the defect in protective immunity responsible for this has not been defined. Could this be due to defective chemokine secretion leading to delayed priming of adaptive immunity?
Experiment: Lung homogenates from TB-infected Diabetic mice were tested for chemokine levels using the ELISArray.
Results:
Sample & Assay Technologies- 71 -
Inflammation studies: Chemokine levels
Diabetes mellitus (DM) is a major risk factor for tuberculosis (TB) but the defect in protective immunity responsible for this has not been defined. Could this be due to defective chemokine secretion leading to delayed priming of adaptive immunity?
Experiment: Lung homogenates from TB-infected Diabetic mice were tested for chemokine levels using the Multi-Analyte ELISArray Kit.
Results:
Conclusion: Reduced levels of leukocyte chemoattractant factors including CCL2 and CCL5 at early timepoints post-infection could explain why diabetic mice are more prone to TB.
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Conclusions
QIAGEN offers many methods to study molecular and cellular mechanisms involved in Inflammation:
� Gene Expression� RT2 Profiler PCR Arrays & Assays� RT2 PreAMP Primer Mixes
� Epigenetics� miScript miRNA PCR System� EpiTect Methyl qPCR Arrays� EpiTect ChIP qPCR Arrays
� Functional Studies� Cignal Reporter Assays� SureSilencing shRNA Plasmid
� Protein expression� ELISArray
www.sabiosciences.com
Sample & Assay Technologies- 73 -
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Latest information on pathway and disease research, resources and demos.
Sample & Assay Technologies- 74 -
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