MHC-antigen arrays for detection and characterization of immune responses
View Patent ↗T cells are profiled with respect to their expression of antigen receptor. The cells are arrayed on a planar or three-dimensional substrate through binding to immobilized or partially diffused MHC-antigen complexes. The cells may further be characterized with respect to their ability to respond to external stimulus in the microenvironment. External stimuli include cell-cell interactions, response to factors, and the like.
1. A method of profiling T cells, the method comprising:
contacting a cell population comprising T cells with an array, wherein said array comprises multiple discrete regions of MHC-antigen complexes stably associated with the surface of a solid planar support, and wherein an MHC-antigen complex comprises monomers or multimers of an α MHC subunit, a β MHC subunit, and a peptide antigen bound in the cleft formed by the α and β subunits;
binding T cells to the MHC antigen complexes and
analyzing T cells bound to the array by evaluating phenotypic attributes.
2. The method according to claim 1 , wherein said array comprises a plurality of different MHC-antigen complexes.
3. A method of profiling T cells, the method comprising:
contacting a cell population comprising T cells with an array, wherein said array comprises multiple discrete regions of MHC-antigen complexes stably associated with the surface of a solid planar support coated with polyacrylamide, and wherein an MHC-antigen complex comprises monomers or multimers of an α MHC subunit, a β MHC subunit, and a peptide antigen bound in the cleft formed by the α and β subunits;
binding T cells to the MHC antigen complexes and
analyzing T cells bound to the array by evaluating phenotypic attributes.
4. The method according to claim 1 , wherein each of said regions comprises at least about 0.01 ng of said MHC-antigen complex.
5. The method according to claim 1 , wherein each of said regions comprises a plurality of different antigenic complexes.
6. The method according to claim 1 , wherein one or more of said MHC-antigen complexes comprises an antigen selected from the group consisting of tumor antigens; viral antigens, bacterial antigens; parasitic antigens; environmental antigens; allergens; and autoimmune antigens.
7. The method according to claim 1 , wherein said cell population comprises multiple T cell antigenic specificities.
8. The method according to claim 1 , wherein said cell population comprises multiple cell types.
9. The method according to claim 1 , wherein said cell population is selected from the group consisting of blood, lymph, cerebrospinal fluid and synovial fluid.
10. The method according to claim 9 , wherein said T cells are labeled with a detectable marker prior to said contacting step, and wherein said analyzing detects the presence of said marker.
11. The method according to claim 1 , wherein said MHC-antigen complexes comprise a library of antigenic peptides complexed with MHC.
12. A method of profiling T cells, the method comprising:
contacting a cell population comprising T cells with an array, wherein said array comprises multiple discrete regions of MHC-antigen complexes stably associated with the surface of a solid planar support, and wherein an MHC-antigen complex comprises monomers or multimers of an α MHC subunit, a β MHC subunit, and a peptide antigen bound in the cleft formed by the α and β subunits;
binding T cells to the MHC antigen complexes and
analyzing T cells bound to the array by determining a change in phenotype of said T cells after contacting with said array.
13. The method according to claim 12 , wherein said change in phenotype comprises secretion of a protein in response to antigenic stimulation.
14. A method of profiling T cells the method comprising:
contacting a cell population comprising T cells with an array, wherein said array comprises multiple discrete regions of MHC-antigen complexes stably associated with the surface of a solid planar support, wherein one or more of said discrete regions present on said array comprise cells and MHC-antigen complexes, and wherein an MHC-antigen complex comprises monomers or multimers of an α MHC subunit, a β MHC subunit, and a peptide antigen bound in the cleft formed by the α and β subunits;
binding T cells to the MHC antigen complexes and
analyzing T cells bound to the array by evaluating phenotypic attributes.
15. The method according to claim 14 , wherein said cells are antigen presenting cells.
16. The method according to claim 1 , further comprising the step of contacting said array with a second cell population, and determining the binding of cells present in said second cell population to said T cells bound to the array.
17. The method according to claim 1 , further comprising the step of contacting said array with a second cell population, and determining a change in the phenotype of said T cells bound to the array.
18. The method of claim 1 , further comprising the step of determining the expression of cell surface antigens by said T cells following said contacting step.
19. The method according to claim 1 , further comprising the step of releasing said cells from said array.
20. The method according to claim 1 , further comprising the step of expanding said T cells following said contacting step.
21. The method according to claim 20 , wherein said expanding step comprises contacting said T cells with one or more of growth factors, cytokines, cell adhesion molecules, and extracellular matrix material.
22. The method according to claim 1 , wherein said array comprises a plurality of spots having differing concentrations of MHC-antigen complexes.
23. The method according to claim 22 , further comprising the step of constructing a scaling curve for binding of T cells to said MHC-antigen complexes.
24. The method according to claim 1 , further comprising the step of determining binding of said cells to said MHC antigen complexes in a site specific analysis.