Difference between revisions of "Team:Evry/Project/SurfaceDisplay"

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<center>
 
<center>
 
<img border="0" class='img-responsive' width="500" src="https://static.igem.org/mediawiki/2015/7/7e/Manquante2.png" alt="" /></center>
 
<img border="0" class='img-responsive' width="500" src="https://static.igem.org/mediawiki/2015/7/7e/Manquante2.png" alt="" /></center>
<center><p class="text-justify"><strong> Figure 2: Plasmids with the constructions :</strong> A) AGA1P  (B) AGA2P-OVA1-DEC205 (C) AGA2P-OVA1 (D) AGA2P-DEC205</p></center>
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<p class="text-justify"><strong> Figure 2: Plasmids with the constructions :</strong> A) AGA1P  (B) AGA2P-OVA1-DEC205 (C) AGA2P-OVA1 (D) AGA2P-DEC205</p>
  
 
<h3>Surface display results</h3>
 
<h3>Surface display results</h3>
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<p class="text-justify">Yeast detrimental effect could be observed even at MOI1 with fixed yeasts.  A viability assay on DC confirmed that many DCs were dead after 24h of co-incubation with yeasts (red dots on figure below).</p>
 
<p class="text-justify">Yeast detrimental effect could be observed even at MOI1 with fixed yeasts.  A viability assay on DC confirmed that many DCs were dead after 24h of co-incubation with yeasts (red dots on figure below).</p>
  
<img border="0" class='img-responsive' src="https://static.igem.org/mediawiki/2015/2/2a/DC_mortality.jpeg" alt="" />
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<p class="text-justify"><strong> Figure 9: Flow cytometry data of DCs coincubated at MOI1 with fixed yeasts during 24h. </strong> Red dots correspond to dead DC and purple dots to living DCs.</p>
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<div class="row">
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<div class="col-md-7"><img border="0" class='img-responsive' src="https://static.igem.org/mediawiki/2015/2/2a/DC_mortality.jpeg" alt="" /></div>
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<div class="col-md-5">
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<p class="text-justify"><strong>Figure 9: Flow cytometry data of DCs coincubated at MOI1 with fixed yeasts during 24h.</strong></p>
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<p class="text-justify">  Red dots correspond to dead DC and purple dots to living DCs.</p>
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</div>
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</div>
  
 
<p class="text-justify">The ability of recombinant yeasts to elicit anti-tumor immune response in vivo was examined. In vivo assay on melanoma mice confirmed T-cell induction against the tumor antigen OVA1 (figure 10). Mice were transfected with the melanoma cell line B16-OVA at day 0, and yeasts were injected at day 10 inside a large tumor reaching 7 mm. In this experiment, a tetramer assay for blood CD8+ OVA1 was performed after mice sacrifice at day+18. Vaccination with yeast OVA1-DEC205/OVA2 resulted in a significant CD8+ OVA1 induction compared with PBS control and wild type yeast. This is coherent with in vitro DC immunophenotyping. Because of the large and heterogeneous tumor size, tumor regression cannot be measured accurately at this stage of late injection.</p>
 
<p class="text-justify">The ability of recombinant yeasts to elicit anti-tumor immune response in vivo was examined. In vivo assay on melanoma mice confirmed T-cell induction against the tumor antigen OVA1 (figure 10). Mice were transfected with the melanoma cell line B16-OVA at day 0, and yeasts were injected at day 10 inside a large tumor reaching 7 mm. In this experiment, a tetramer assay for blood CD8+ OVA1 was performed after mice sacrifice at day+18. Vaccination with yeast OVA1-DEC205/OVA2 resulted in a significant CD8+ OVA1 induction compared with PBS control and wild type yeast. This is coherent with in vitro DC immunophenotyping. Because of the large and heterogeneous tumor size, tumor regression cannot be measured accurately at this stage of late injection.</p>

Revision as of 01:38, 19 September 2015

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