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

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<p class="text-justify">To address both safety and efficiency, we selected S. cerevisiae for the following advantages:</p>
 
<p class="text-justify">To address both safety and efficiency, we selected S. cerevisiae for the following advantages:</p>
  
<il><p class="text-justify">A. S. cerevisiae is non pathogenic : phase I clinical trial with subcutaneous injection of heat-killed yeasts S. cerevisiae against hepatitis C showed no dose-related toxicity (14), demonstrating the safety of this vector for future human applications in cancer immunotherapy.</p></il>
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<p class="text-justify">A. S. cerevisiae is non pathogenic : phase I clinical trial with subcutaneous injection of heat-killed yeasts S. cerevisiae against hepatitis C showed no dose-related toxicity (14), demonstrating the safety of this vector for future human applications in cancer immunotherapy.</p>
 
<p class="text-justify">B. S. cerevisiae has a strong adjuvant effect, explaining why zymosan, an extract of S. cerevisiae cell wall, has been used to stimulate inflammation for 50 years (15). In particular, the mannose stimulate pro-inflammatory cytokines production in monocytes and dendritic cells, making the vector appropriate for APC targeting (16,17). </p>
 
<p class="text-justify">B. S. cerevisiae has a strong adjuvant effect, explaining why zymosan, an extract of S. cerevisiae cell wall, has been used to stimulate inflammation for 50 years (15). In particular, the mannose stimulate pro-inflammatory cytokines production in monocytes and dendritic cells, making the vector appropriate for APC targeting (16,17). </p>
 
<p class="text-justify">C. S. cerevisiae already proved its anti-tumor capacity : the first demonstration of recombinant yeast to induce adaptative immunity was shown in 2001 by Stubbs et al against ovalbumin tumor cells (18). Yeast expressing the mutated RAS protein inside their cytosol induced reduction of lung tumors in mice and a quarter of the tumors were eradicated (19). Tumor antigen MART-1 was also expressed in yeast cytosol and induced both CD4+ and CD8+ in mice. (20)</p>
 
<p class="text-justify">C. S. cerevisiae already proved its anti-tumor capacity : the first demonstration of recombinant yeast to induce adaptative immunity was shown in 2001 by Stubbs et al against ovalbumin tumor cells (18). Yeast expressing the mutated RAS protein inside their cytosol induced reduction of lung tumors in mice and a quarter of the tumors were eradicated (19). Tumor antigen MART-1 was also expressed in yeast cytosol and induced both CD4+ and CD8+ in mice. (20)</p>
 
<p class="text-justify">D. S. cerevisiae prior immunization with the wild type do not create a response neutralizing the antigen expressing yeast, allowing repeated injections of the vector. (21)</p>
 
<p class="text-justify">D. S. cerevisiae prior immunization with the wild type do not create a response neutralizing the antigen expressing yeast, allowing repeated injections of the vector. (21)</p>
  
<p class="text-justify">Surface display of tumor antigen for CD8+ cross-priming.
 
 
• We chose to express our antigen on the membranes of S. cerevisiae because surface displayed antigen is cross-presented much more efficiently than yeast cytosol antigen (22). This is due to a particular kinetics inside the early phagosome, allowing the external antigen to escape from the phagosome. Cross-presentation can be further enhanced by inserting linkers susceptible to Cathepsin S cleavage between the antigen and Aga2p, supporting the evidence that early antigen release is important for cross-presentation (22).
 
 
Enhancing cross-priming with the antibody anti-DEC205
 
 
• Our surface display antigen for ovalbumin was fused to DEC205 scFv. DEC205 is a lectin receptor expressed by some DCs subsets, including mouse spleen DC (23). It was shown that antibody targeting DEC-205, fused to tumor antigen, can induce T cell stimulation if administered with an additional stimulus triggering DC maturation, like anti-CD40 agonistic antibody (24). In the same way, immunization with DNA vectors encoding antigens fused to a DEC-205 scFv elicits a strong specific CD8+ responses in vivo (25). 
 
 
• The scFv of DEC205 was fused to our ovalbumin tumor antigen and surface displayed in order to get the yeast internalized in a DC endosome through DEC205 receptor, favoring CD8+ cross-presentation. We used the scFv instead of the whole antibody for the possibility to perform repeated immunisations without inducing deleterious host responses against the Fc part of the immunoglobulin chains.
 
 
Advantages of Yeast expressing DEC205 over DEC205 protein vaccines
 
 
• Pure protein vaccines with DEC205 are far less immunogenic than vaccine with micro-organisms mimicking pathogens and request an additional adjuvant. Moreover, the protein needs a prior step of antigen purification (26), leading us to develop this yeast surface display of DEC205 scFv fused to the antigen. The advantages of our system include better concentration of the yeast due to less diffusion than the protein DEC205 alone, the codelivery of both antigen and adjuvant, the possibility to target multiple DCs compartments at the same time (MHCI and MHC II) and the absence of purification step.</p>
 
  
  

Revision as of 21:37, 18 September 2015

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