Cell-based microarrays and methods of use

摘要

The invention provides methods and compositions for rapid, sensitive, and highly specific detection of antigen-specific interactions between cytolytic T lymphocytes (CTLs) and antigen presenting cells (APCs). The invention also features compositions, including kits, for use in the methods of the invention.

主权项

1. A method for identifying a cytolytic T lymphocyte (CTL) antigen, the method comprising:
contacting a sample comprising a cytolytic T lymphocyte (CTL) with an array comprising a plurality of adherent recombinant antigen-presenting cells (APCs) adhered to a surface of the array at different, discrete locations on the array, wherein the adherent recombinant APCs express different recombinant polynucleotides encoding different polypeptides, and wherein the different, discrete locations on the array correlate with the different recombinant polynucleotides expressed by the adherent recombinant APCs; washing the array; and detecting the presence or absence of caspase activity in the plurality of adherent recombinant APCs by detecting the presence of a fluorescent signal generated from a fluorogenic caspase substrate present in said adherent recombinant APCs; wherein the presence of a fluorescent signal in an adherent recombinant APC is indicative of an antigen-specific interaction between the CTL and the adherent recombinant APC and indicates the recombinant polynucleotide of the adherent recombinant APC encodes a polypeptide that comprises a CTL antigen.