Antigen screening system
Methods and compositions for identifying antigens of human lymphocytes are provided herein.
1. A method of evaluating an immune response of a human subject, the method comprising:
(a) providing a library comprising bacterial cells, wherein each bacterial cell of the library comprises a heterologous polypeptide;
(b) contacting the bacterial cells with a first plurality of human cells which includes non-professional human antigen presenting cells that internalize the bacterial cells;
(c) contacting the first plurality of human cells with a second plurality of human cells from the human subject which comprises human lymphocytes that are not selected for specifically responding to the antigen, under conditions in which lymphocytes are stimulated by polypeptides presented by the first plurality of human cells; and
(d) determining whether a lymphocyte of the second plurality of human cells is stimulated by a heterologous polypeptide presented by a cell of the first plurality of human cells, thereby evaluating an immune response of the human subject.
2. The method of claim 1 , wherein the first plurality of human cells is isolated from the human subject.
3. The method of claim 1 , wherein the heterologous polypeptides comprise polypeptides encoded by an infectious agent, and wherein stimulation of a lymphocyte of the second plurality of human cells by a polypeptide encoded by the infectious agent presented by a cell of the first plurality of human cells indicates that the human subject has an immune response to the infectious agent.
4. The method of claim 1 , wherein the heterologous polypeptides comprise polypeptides differentially expressed in cancer or a tumor cell.
5. The method of claim 4 , wherein stimulation of a lymphocyte of the second plurality of human cells by a polypeptide differentially expressed in cancer or a tumor cell presented by a cell of the first plurality of human cells indicates that the human subject has an immune response to the polypeptide differentially expressed in cancer or a tumor cell.
6. The method of claim 1 , wherein the heterologous polypeptides comprise polypeptides from a target cell of the human subject.
7. The method of claim 6 , wherein stimulation of a lymphocyte of the second plurality of human cells by a polypeptide from a target cell of the human subject presented by a cell of the first plurality of human cells indicates that the human subject has an immune response to the target cell of the human subject.
8. The method of claim 3 , wherein stimulation of a lymphocyte of the second plurality of human cells by a polypeptide encoded by the infectious agent presented by a cell of the first plurality of human cells indicates that the human subject has been exposed to the infectious agent.
9. The method of claim 3 , wherein the bacterial cells further comprise a cytolysin polypeptide.
10. The method of claim 9 , wherein the cytolysin polypeptide is listeriolysin O (LLO).
11. The method of claim 3 , wherein the first plurality of human cells is provided in an array, and wherein cells in each location of the array are contacted with a set of bacterial cells, each set comprising a different heterologous polypeptide.
12. The method of claim 3 , wherein the infectious agent is an intracellular infectious agent.
13. The method of claim 3 , wherein the infectious agent is an extracellular infectious agent.
14. The method of claim 3 , wherein the infectious agent is a virus.
15. The method of claim 3 , wherein the infectious agent is a bacterium.
16. The method of claim 3 , wherein the infectious agent is a fungus, protozoan, or helminth.
17. The method of claim 1 , wherein the first and second plurality of human cells comprise cells from peripheral blood.
18. The method of claim 1 , wherein the first plurality of human cells comprises dendritic cells.
19. The method of claim 17 , wherein the first plurality of human cells comprises peripheral blood cells from which T cells and dendritic cells have been depleted.
20. The method of claim 17 , wherein the first plurality of human cells comprises dendritic cells and peripheral blood cells from which T cells and dendritic cells have been depleted.
21. The method of claim 20 , wherein the first plurality of human cells comprises dendritic cells and peripheral blood cells from which T cells and dendritic cells have been depleted at a ratio of about 1:2, 1:3, 1:4, or 1:5.
22. The method of claim 3 , further comprising identifying a polypeptide that stimulates a lymphocyte of the second plurality of human cells as an antigen of the infectious agent.
23. The method of claim 22 , further comprising formulating the antigen or a nucleic acid encoding the antigen as an immunogenic composition.