Methods, kits, and systems for scoring the immune response to cancer

Abstract

This disclosure describes methods, kits, and systems for scoring the immune response to cancer through examination of tissue infiltrating lymphocytes (TILs). Methods of scoring the immune response in cancer using tissue infiltrating lymphocytes include detecting CD3, CD8, CD20, and FoxP3 within the sample and scoring the detection manually or scoring the digital images of the staining with the aid of image analysis and algorithms.

1 . A method of scoring the immune response in cancer using tissue infiltrating lymphocytes comprising:
 contacting a sample with CD3, CD8, CD20, and FoxP3 specific binding moieties,   removing excess specific binding moieties from the sample,   detecting the specific binding moieties,   assessing a presence and a distribution for the specific binding moieties within the sample,   scoring cancer using tissue infiltrating lymphocytes by analyzing the presence and the distribution of the specific binding moieties.   
     
     
         2 . The method of  claim 1 , further comprising contacting the sample with a specific binding moiety for a tumor marker. 
     
     
         3 . The method of  claim 2 , wherein the specific binding moiety for the tumor marker is selected from antibodies to cytokeratin for epithelial tumors (carcinomas) and antibodies to S100 for melanomas. 
     
     
         4 . The method of  claim 1 , wherein the specific binding moiety for CD3 is anti-CD3 rabbit or mouse monoclonal antibodies. 
     
     
         5 . The method of  claim 1 , wherein the specific binding moiety for CD8 is anti-CD8 rabbit or mouse monoclonal antibodies. 
     
     
         6 . The method of  claim 1 , wherein the specific binding moiety for CD20 is anti-CD20 rabbit or mouse monoclonal antibodies. 
     
     
         7 . The method of  claim 1 , wherein the specific binding moiety for FoxP3 is anti-FoxP3 rabbit or mouse monoclonal antibodies. 
     
     
         8 . The method of  claim 1 , wherein contacting comprises contacting the sample with CD3, CD8, CD20, and FoxP3 specific binding moieties which are selected from rabbit or mouse monoclonal antibodies. 
     
     
         9 . The method of  claim 8 , wherein detecting comprises contacting the sample with an anti-species antibody specific to the rabbit or mouse monoclonal antibodies. 
     
     
         10 . The method of  claim 9 , wherein the anti-species antibody is haptenated, and wherein detecting the haptenated antibody further comprises contacting the sample with an anti-hapten antibody conjugated to an enzyme. 
     
     
         11 . The method of  claim 10 , wherein detecting further comprises contacting the sample with a detection compound. 
     
     
         12 . The method of  claim 11 , wherein the detection compound is selected from a chromogen or a fluorophore. 
     
     
         13 . The method of  claim 1 , wherein the specific binding moieties are haptenated, and wherein detecting the specific binding moieties further comprises contacting the sample with an anti-hapten antibody conjugated to an enzyme. 
     
     
         14 . The method of  claim 13 , wherein detecting further comprises contacting the sample with a detection compound. 
     
     
         15 . The method of  claim 14 , wherein the detection compound is selected from a chromogen or a fluorophore. 
     
     
         16 . A kit for scoring cancer comprising:
 a CD3 specific binding moiety and a CD3 detection reagent,   a CD8 specific binding moiety and a CD8 detection reagent,   a CD20 specific binding moiety and a CD20 detection reagent,   a FoxP3 specific binding moiety and a FoxP3 detection reagent,   contained in a plurality of vessels, and   instructions for scoring which describe detecting CD3, CD8, CD20, and   FoxP3 within a tissue sample and scoring based on a presence and a distribution of the detection reagents on the tissue.   
     
     
         17 . A system for scoring cancer by interrogating a tissue sample disposed upon a glass slide, the system comprising:
 reagents comprising a CD3 specific binding moiety, a CD3 detection reagent, a CD8 specific binding moiety, a CD8 detection reagent, a CD20 specific binding moiety, a CD20 detection reagent, a FoxP3 specific binding moiety, and a FoxP3 detection reagent,   an automated staining apparatus, and   an imaging apparatus;   
       wherein the automated staining apparatus is configured to deliver the reagents onto the slide in a manner such that a presence and distribution of CD3, CD8, CD20, and FoxP3, is specifically determined and the imaging apparatus is configured so that the presence and distribution of the CD3, CD8, CD20, and FoxP3 can be used to generate a score.