Marrow infiltrating lymphocytes (mils) as a source of t-cells for chimeric antigen receptor (car) thereapy

Abstract

In some embodiments, marrow-infiltrating lymphocytes (“MILs”) comprising a chimeric antigen receptor (“CAR”) are provided. In some aspects, the embodiments relate to a method for making a recombinant MIL, comprising obtaining bone marrow comprising MILs; and transfecting, transforming, or transducing the MILs with a nucleic acid encoding a chimeric antigen receptor. In some aspects, the embodiments relate to a method for treating a condition in a subject, comprising administering to the subject a MIL comprising a CAR.

1 - 35 . (canceled) 
     
     
         36 . A method for making a recombinant marrow infiltrating lymphocyte (MIL), comprising:
 obtaining bone marrow comprising MILs; and   transfecting, transforming, or transducing the MILs with a nucleic acid encoding a chimeric antigen receptor (CAR) comprising a single-chain variable fragment (scFV) antigen-binding domain that can bind to an extracellular domain of CD19 or CD38; and   a transmembrane domain of CD19 or CD38, and   an intracellular domain selected from the group consisting of 4-1BB, CD3ζ, and combinations thereof.   
     
     
         37 . The method of  claim 36 , wherein the bone marrow is obtained from a subject. 
     
     
         38 . The method of  claim 37 , wherein the subject has a neoplasm, an autoimmune disease, or an infection caused by a pathogen. 
     
     
         39 . The method of  claim 37 , wherein the subject has an autoimmune disease. 
     
     
         40 . The method of  claim 37 , wherein the subject has an infection caused by a pathogen. 
     
     
         41 . The method of  claim 36 , further comprising activating the MILs and expanding the MILs. 
     
     
         42 . The method of  claim 36 , wherein the method comprises making a plurality of recombinant MILs. 
     
     
         43 . The method of  claim 36 , further comprising incubating the MILs under hypoxic conditions prior to transfecting, transforming, or transducing the MILs with the nucleic acid encoding the CAR. 
     
     
         44 . The method of  claim 43 , wherein the hypoxic conditions comprise about 0.5% to about 5% oxygen gas. 
     
     
         45 . The method of  claim 43 , wherein the hypoxic conditions comprise about 1% to about 2% oxygen gas. 
     
     
         46 . The method of  claim 43 , further comprising incubating the MILs under normoxic conditions after transfecting, transforming, or transducing the MILs with the nucleic acid encoding the CAR. 
     
     
         47 . The method of  claim 43 , further comprising contacting the MILs with anti-CD3/anti-CD28 beads while incubating the MILs under hypoxic conditions. 
     
     
         48 . The method of  claim 36 , wherein the MIL is CD3+, CD4+, CD8+, or a combination thereof. 
     
     
         49 . The method of  claim 36 , wherein the MIL is CD45RO+, C62L+, or CXCR4+. 
     
     
         50 . The method of  claim 36 , wherein the MIL is 4-1BB+. 
     
     
         51 . The method of  claim 36 , wherein the MIL is interferonγ+ and/or is CD138+. 
     
     
         52 . The method of  claim 36 , wherein the MIL is CD33+. 
     
     
         53 . The method of  claim 36 , wherein the MIL is CD34−.