bvdv infection alters toll like and tnf receptor signaling in bovine cells
Download
Skip this Video
Download Presentation
BVDV Infection Alters Toll-like and TNF-  Receptor Signaling in Bovine Cells

Loading in 2 Seconds...

play fullscreen
1 / 14

BVDV Infection Alters Toll-like and TNF-  Receptor Signaling in Bovine Cells - PowerPoint PPT Presentation


  • 96 Views
  • Uploaded on

BVDV Infection Alters Toll-like and TNF-  Receptor Signaling in Bovine Cells. J.D. Neill, R. Zuerner and J.F. Ridpath. Innate Immunity. First line of defense against invading pathogens Pathogens detected by sensors that are present in most cells

loader
I am the owner, or an agent authorized to act on behalf of the owner, of the copyrighted work described.
capcha
Download Presentation

PowerPoint Slideshow about ' BVDV Infection Alters Toll-like and TNF-  Receptor Signaling in Bovine Cells' - quincy-ayers


An Image/Link below is provided (as is) to download presentation

Download Policy: Content on the Website is provided to you AS IS for your information and personal use and may not be sold / licensed / shared on other websites without getting consent from its author.While downloading, if for some reason you are not able to download a presentation, the publisher may have deleted the file from their server.


- - - - - - - - - - - - - - - - - - - - - - - - - - E N D - - - - - - - - - - - - - - - - - - - - - - - - - -
Presentation Transcript
bvdv infection alters toll like and tnf receptor signaling in bovine cells

BVDV Infection Alters Toll-like and TNF- Receptor Signaling in Bovine Cells

J.D. Neill, R. Zuerner and J.F. Ridpath

innate immunity
Innate Immunity
  • First line of defense against invading pathogens
  • Pathogens detected by sensors that are present in most cells
  • Results in a rapid response aimed at eliminating the threat
  • Stimulates expression of genes encoding molecules that potentiate the response
innate immunity1
Innate Immunity
  • Toll-like receptors and intracellular soluble receptors
  • These receptors recognized pathogen-associated molecular patterns (PAMPs)
  • Stimulate expression of important response regulators:
    • Type I interferons, inflammatory cytokines (TNF-), chemokines, etc.
  • Induction of inflammatory response
    • Characterized by activation of NF-B
toll like and tnf receptors
Toll-Like and TNF- Receptors
  • Toll-like receptors (TLR):
    • Transmembrane proteins - 12 known
    • Detect specific PAMPS
    • TLR3 - dsRNA, Type I interferon, NF-B
    • TLR4 - lipopolysaccharide (LPS), NF-B
  • TNF- receptor
    • Ligated by TNF-, NF-B
activation of nf b and inflammation
Activation of NF-B and Inflammation
  • Activation of NF-B results in expression of large number of proteins:
    • P- and E-selectins in endothelial cells
    • A20
    • IL6, IL8
inhibition of innate immunity by bvdv
Inhibition of Innate Immunity by BVDV
  • Examined effect of BVDV infection on responses to LPS, dsRNA and TNF-
  • Bovine aortic endothelial cells (BAEC) and primary fetal testicular cells (Bte)
  • ncpBVDV 2 strain A13 isolated from commercial endothelial cells
  • Non-infected, 24-48 hr acute, chronic
indirect assay of nf b activation
Indirect Assay of NF-B Activation
  • Used real-time PCR to examine genes known to be upregulated by NF-B
    • P-selectin, E-selectin, A20, IL6 and IL8
  • Stimulated cells with LPS, dsRNA or TNF- for 4 hours
  • RNA isolated with TRIzol reagent
  • Real-time PCR using Superscript III SYBR green qt-PCR kit
  • Normalized to 2-microglobulin
  • Expression levels calculated by Liu and Saint

(Anal. Biochem. 302:52)

direct assay of nf b activation
Direct Assay of NF-B Activation
  • Plasmid containing minimal CMV promoter fused to NF-Bresponse elements upstream from luciferase gene
  • Plasmid introduced into Bte cells using lipofectin-mediated transfection - n=6
  • Cells stimulated with LPS, dsRNA or TNF- at 24 hours post-transfection for 4 hours
  • Cells were lysed and assayed for luciferase activity - relative light units
conclusions
Conclusions
  • Stimulation of BAE and Bte cells with TLR and TNFR ligands increased specific gene expression
  • BVDV2 infection negatively affected NF-B activation
  • Acute BVDV infection more effective at inhibition of NF-B activation
  • TLR3 (dsRNA) appears to be inhibited to greater degree
ad