Reagents and methods for identifying, enriching, and/or expanding antigen-specific t cells

Abstract

Antigen-specific T cells, including nave T cells, and including rare precursor cells are enriched and expanded in culture. Enrichment and expansion provides a platform for more effective immunotherapy by adoptive transfer, as well as platforms for personalizing immunotherapy by determining T cell reactivity with a library of candidate peptide antigens.

1 .  1 .- 83 . (canceled) 
     
     
         84 . A method for identifying a T cell antigen or component of a T cell repertoire, comprising:
 providing a sample comprising T cells from a subject,   screening the T cells against a library of paramagnetic particles each comprising on their surface an MHC-peptide antigen-presenting complex and a lymphocyte costimulatory ligand, wherein the particles are from about 10 to about 500 nm in diameter, wherein each particle presents a candidate peptide antigen,   magnetically activating T cells in the sample that are specific for the candidate peptide antigens by incubating the particles with the sample in the presence of a magnetic field,   separating cells associated with the paramagnetic particles from cells not associated with the paramagnetic particles,   culturing the cells associated with the paramagnetic particles, and identifying one or more candidate peptide antigens that cause activation of the subject's T cells, wherein activation of the subject's T cells is identified by measuring cytokine expression or intracellular signaling indicative of T cell activation.   
     
     
         85 . The method of  claim 84 , wherein the subject is a cancer patient. 
     
     
         86 . The method of  claim 85 , wherein the candidate peptides comprise peptide antigens predicted from genetic analysis of the patient's cancer. 
     
     
         87 . The method of  claim 84 , wherein activation of the subject's T cells is identified by cytokine expression, measured by immunochemistry or RT-PCR. 
     
     
         88 . The method of  claim 84 , wherein selected peptide antigens are loaded onto artificial antigen presenting cells and administered to the subject. 
     
     
         89 . The method of  claim 84 , wherein T cells that recognize the identified antigens are enriched and expanded ex vivo, and administered to the subject. 
     
     
         90 . The method of  claim 84 , wherein the subject is suspected of having an immune disease, and the paramagnetic particles present a library of candidate peptides indicative of autoimmune and/or immune disease. 
     
     
         91 . The method of  claim 84 , wherein the subject is suspected of having an infectious disease, and the paramagnetic particles, present a library of candidate peptides indicative of infectious diseases. 
     
     
         92 . The method of  claim 84 , wherein the incubation of the particles with the sample on the magnetic column is from 15 minutes to 5 hours. 
     
     
         93 . The method of  claim 84 , wherein the T cells associated with the paramagnetic particles are cultured with the paramagnetic particles, and cytokine expression is quantified within about 15 to 48 hours in culture. 
     
     
         94 . The method of  claim 84 , wherein the T cells associated with the paramagnetic particles are cultured with the paramagnetic particles, and intracellular signaling quantified after 15 minutes to 5 hours in culture.