Difference between revisions of "Part:BBa K321004"
 
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===Usage and Biology===
 
===Usage and Biology===
  
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In wild-type NK cells, Killer cell immunoglobulin-like receptors (KIRs) with  long cytoplasmic domains inhibit cell mediated cytotoxicity upon ligand binding. They possess a immune tyrosine-based inhibitory motif (ITIM) that when phosphorylated recruits phosphotases like SHP-1 that decrease the activation molecules involved in cell signaling.  
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In immune Natural Killer cells, Killer cell immunoglobulin-like receptors (KIRs) inhibit cell mediated cytotoxicity upon ligand binding. In their intracellular domains, these receptors possess immune tyrosine-based inhibitory (ITIM) motifs that when phosphorylated recruit phosphotases that decrease the activation molecules involved in immune cell signaling.
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ANDN gate
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In order to understand the ANDN gate, let us set up a hypothetical situation in which antigen A and antigen B are expressed on the membrane surface of healthy cells. Since cancer cells typically discard many surface proteins as a result of genetic mutation, we represent this discarded protein as antigen B in our scenario. Our ANDN gate is designed to address this issue by triggering cytotoxicity in the presence of antigen A and absence of antigen B. Therefore, cancerous cells that express antigen A and “hide” antigen B will be targeted. Healthy cells expressing both antigen A and B will not set off the activation of the ANDN gate. This concept is valuable for ensuring a level of specificity that prevents the overly indiscriminate activation of killer cells.
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===Function===
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This part was shown to inhibit the activation of killer cells by the UCSF 2010 team.  We used this part in our ANDN gate to increase the precision of killing toward cancer cells.  The figure below shows that this part performs its inhibitory function as expected.  On the X-axis are different types of target cells, which represent normal and cancer cells expressing different combinations of surface antigents in our experiment. The Y-axis shows the activation levels of killer cells engineered to express synthetic receptors, one of which bears this part as its intracellular domain.  The data show that in the presence of antigen B, which is recognized by the synthetic receptor bearing this part, killer cell activation is inhibited
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(red and striped bars) regardless of the presence of activating antigen A.  For more information please see our <html><a href="http://2010.igem.org/Team:UCSF/Project/Precision#andn">wiki</a></html>.
  
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We tested two different ANDN gate designs to determine their effects on target recognition. To achieve the level of specificity as described by our hypothetical situation, we have set two different antigen binding domains to recognize antigen A and antigen B, respectively. Attached to the domain that recognized antigen A was an ITAM-bearing intracellular chain, from either the CD3 zeta or Fc receptor gamma, that signaled for killer cell activation. The domain that recognized antigen B, the antigen found in healthy cells, was fused to the intracellular portion of the ITIM-based receptor KIR3DL1, which inhibits killer cell activation. As a result of this combination, target cells expressing only antigen A would trigger killer cell activation, and target cells that do not express antigen A would not. Target cells that express both antigens A and B would be unharmed due to ITIM inhibitory signals, meaning that the presence of antigen B overwrites the input of antigen A. In conclusion, only a specific combination of surface antigens can set off the chain of activation, resulting in increased precision of detection and cancer killing.
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<img src="https://static.igem.org/mediawiki/2010/8/80/UCSF_ANDNgate_results.png"/>
 
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<partinfo>BBa_K321004 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K321004 SequenceAndFeatures</partinfo>
  
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==References==
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Brewerton DA, Hart FD, Nicholls A, Caffrey M, James DC, Sturrock RD. Ankylosing spondylitis and HL‐A 27. Lancet 1973; 1: 904– 7.
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Crossref CAS PubMed Web of Science®Google Scholar
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Brown MA. Breakthroughs in genetic studies of ankylosing spondylitis. Rheumatology (Oxford) 2008; 47: 132– 7.
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Crossref CAS PubMed Web of Science®Google Scholar
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Gonzalez‐Roces S, Alvarez MV, Gonzalez S, Dieye A, Makni H, Woodfield DG, et al. HLA‐B27 polymorphism and worldwide susceptibility to ankylosing spondylitis. Tissue Antigens 1997; 49: 116– 23.
  
 
<!-- Uncomment this to enable Functional Parameter display  
 
<!-- Uncomment this to enable Functional Parameter display  

Latest revision as of 18:33, 20 October 2020

intracellular chain of KIR3DL1

Encodes the intracellular signaling chain of the KIR3DL1 immune receptor. Inhibits immune cell activation via the ITIM (immunoreceptor tyrosine-based inhibition motif) motif.


Usage and Biology

In immune Natural Killer cells, Killer cell immunoglobulin-like receptors (KIRs) inhibit cell mediated cytotoxicity upon ligand binding. In their intracellular domains, these receptors possess immune tyrosine-based inhibitory (ITIM) motifs that when phosphorylated recruit phosphotases that decrease the activation molecules involved in immune cell signaling.

Function

This part was shown to inhibit the activation of killer cells by the UCSF 2010 team. We used this part in our ANDN gate to increase the precision of killing toward cancer cells. The figure below shows that this part performs its inhibitory function as expected. On the X-axis are different types of target cells, which represent normal and cancer cells expressing different combinations of surface antigents in our experiment. The Y-axis shows the activation levels of killer cells engineered to express synthetic receptors, one of which bears this part as its intracellular domain. The data show that in the presence of antigen B, which is recognized by the synthetic receptor bearing this part, killer cell activation is inhibited (red and striped bars) regardless of the presence of activating antigen A. For more information please see our wiki.




Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]

References

Brewerton DA, Hart FD, Nicholls A, Caffrey M, James DC, Sturrock RD. Ankylosing spondylitis and HL‐A 27. Lancet 1973; 1: 904– 7. Crossref CAS PubMed Web of Science®Google Scholar

Brown MA. Breakthroughs in genetic studies of ankylosing spondylitis. Rheumatology (Oxford) 2008; 47: 132– 7. Crossref CAS PubMed Web of Science®Google Scholar

Gonzalez‐Roces S, Alvarez MV, Gonzalez S, Dieye A, Makni H, Woodfield DG, et al. HLA‐B27 polymorphism and worldwide susceptibility to ankylosing spondylitis. Tissue Antigens 1997; 49: 116– 23.