Activation of marrow infiltrating lymphocytes in hypoxic alternating with normoxic conditions

Abstract

In some aspects, the invention relates to compositions comprising marrow infiltrating lymphocytes (“MILs”). The MILs may be activated MILs. In some aspects, the invention relates to methods for activating MILs, comprising incubating MILs in an environment comprising less than 21% oxygen. In some aspects, the invention relates to methods for treating cancer in a subject, comprising administering to the subject a composition comprising activated MILs.

1 - 11 . (canceled) 
     
     
         12 . A method for making a composition comprising a population of marrow infiltrating lymphocytes (“MILs”), comprising incubating the MILs in an environment comprising less than 21% oxygen, wherein the composition comprises a population of MILs that expresses CD3; and at least about 40% of the cells in the composition are MILs from the population of MILs that expresses CD3. 
     
     
         13 . A method for activating marrow infiltrating lymphocytes (“MILs”), comprising incubating MILs in an environment comprising less than 21% oxygen. 
     
     
         14 - 34 . (canceled) 
     
     
         35 . A marrow infiltrating lymphocyte (MIL) formulation, comprising a composition of a population of MILs prepared by the method of  claim 12 . 
     
     
         36 . A therapeutically effective marrow infiltrating lymphocyte (MIL) formulation grown under hypoxic conditions in vitro. 
     
     
         37 - 38 . (canceled) 
     
     
         39 . The method of  claim 12 , wherein the composition comprises a population of MILs that expresses interferon gamma (“IFNγ”); and at least about 2% of the cells in the composition are MILs from the population of MILs that expresses IFNγ. 
     
     
         40 . The method of  claim 12 , wherein the population of MILs that expresses CD3 comprises a plurality of MILs that expresses interferon gamma (“IFNγ”); and at least about 2% of the cells in the composition are MILs from the plurality of MILs that expresses IFNγ. 
     
     
         41 . The method of  claim 12 , wherein the composition comprises a population of MILs that expresses CXCR4; and at least about 98% of the cells in the composition are MILs from the population of MILs that expresses CXCR4. 
     
     
         42 . The method of  claim 12 , wherein the composition comprises a population of MILs that expresses CD4. 
     
     
         43 . The method of  claim 42 , wherein the population of MILs that expresses CD4 comprises a plurality of MILs that expresses CXCR4; and at least about 98% of the cells in the composition are MILs from the plurality of MILs that expresses CXCR4. 
     
     
         44 . The method of  claim 42 , wherein the population of MILs that expresses CD4 comprises a plurality of MILs that expresses 4-IBB; and
 at least about 21% of the cells in the composition are MILs from the plurality of MILs that expresses 4-IBB.   
     
     
         45 . The method of  claim 12 , wherein the composition comprises a population of MILs that expresses CDS. 
     
     
         46 . The method of  claim 45 , wherein the population that expresses CDS comprises a plurality of MILs that expresses CXCR4; and at least about 98% of the cells in the composition are MILs from the plurality of MILs that expresses CXCR4. 
     
     
         47 . The method of  claim 45 , wherein the population of MILs that expresses CDS comprises a plurality of MILs that expresses 4-IBB; and at least about 21% of the cells in the composition are MILs from the plurality of MILs that expresses 4-IBB. 
     
     
         48 . The method of  claim 12 , wherein the composition comprises a population of MILs that expresses 4-IBB; and at least about 21% of the cells in the composition are MILs from the population of MILs that expresses 4-IBB.