Difference between revisions of "Part:BBa K1993001"

Revision as of 16:41, 15 October 2016

CCR7

Chemokine receptors are found on the surface of certain cells that interact with chemokines. They have a 7-transmembrane (7TM) structure and couples to G-protein for signal transduction within a cell[1] (Figure 1). Following interaction with their specific chemokine ligands, chemokine receptors trigger a flux inintracellular calcium (Ca2+) ions which initiate the onset of chemotaxis that traffics the cell to a desired location (Figure 2).

Figure 1. typical structure of a chemokine receptor.

Figure 2. the mechanism of interaction between chemokine and chemokine receptor.

Under the circumstance of inflammation, various kinds of cytokines, including chemokines, are released by the lesions. Guided by the chemokines, cells expressing chemokine receptors move towards the lesions where they can function better. [2] What’s more, different diseases would release different pool of chemokines, which would recruit different effector cells.See our disease talbe-chemokine

Based on the chemotaxis theory, in order to enhance the homing ability of our marvelous mesenchymal stem cells(MSCs) since their lack of enough chemokine receptors on their cell surface, we, SYSU-MEDICINE, had constructed a series of chemokine receptors that cover different inflammatory diseases as far as possible. Among which, CCR7 is a significant chemokine receptor in Arthritis.See our disease talbe-chemokine

We got the gene from peripheral mononuclear blood cells (PMBCs) and purified it (Figure 3). Then we constructed it under the control of EF1-alpha by using Gateway technology.(Figure 4)

Figure 3. Purified of CCR7.

Figure 4. EF1-a CCR7.

We introduced that plasmid into MSCs and tested the expression of CCR7 in MSCs on the protein level (Figure 5)

Figure 5. Western Blot Of CCR7.

Then, we tested the chemotaxis of engineered MSCs by conducting Transwell assay against CCL17. To our excitement, our engineered MSCs had improved their homing ability with chemokine receptor CCR7(Figure 6).

Figure 6. transwell assay of CCR7.

References

[1]Allen, Samantha J.; Crown, Susan E.; Handel, Tracy M. (2007-01-01). "Chemokine: receptor structure, interactions, and antagonism". Annual Review of Immunology. 25: 787–820.

[2] Griffith J W, Sokol C L, Luster A D. Chemokines and chemokine receptors: positioning cells for host defense and immunity.[J]. Annual Review of Immunology, 2014, 32(1):659-702.

Sequence and Features