Assays for detecting t cell immune subsets and methods of use thereof

Abstract

The present disclosure provides methods for measuring the number of CD4+ OX40+ Foxp3+ lymphocytes in a sample containing cancer cells and lymphocytes obtained from a subject by labeling lymphocytes that show CD4 expression in the sample, then labeling lymphocytes that show OX40 expression in the sample, then labeling lymphocytes that show Foxp3 expression in the sample, then measuring the number of CD4+ OX40+ Foxp3+ lymphocytes in the sample. Further provided are methods for determining the prognosis of a subject, predicting responsiveness of a subject having cancer to an OX40 agonist treatment, and methods for treating or delaying progression of cancer based on the number of CD4+ OX40+ Foxp3+ lymphocytes in a sample.

1 - 24 . (canceled) 
     
     
         25 . A method for measuring the number of CD4+ OX40+ Foxp3+ lymphocytes in a sample comprising cancer cells and lymphocytes obtained from a subject, comprising the steps of:
 (a) labeling lymphocytes that show CD4 expression in the sample;   (b) labeling lymphocytes that show OX40 expression in the sample after step (a);   (c) labeling lymphocytes that show Foxp3 expression in the sample after step (b); and   (d) measuring the number of CD4+ OX40+ Foxp3+ lymphocytes in the sample after step (c).   
     
     
         26 - 62 . (canceled) 
     
     
         63 . The method of  claim 25 , further comprising treating the sample with a solution comprising EDTA or citrate buffer between steps (a) and (b) and/or between steps (b) and (c). 
     
     
         64 . The method of  claim 25 , wherein lymphocytes showing CD4 expression, OX40 expression, and Foxp3 expression are labeled by immunofluorescence staining. 
     
     
         65 . The method of  claim 64 , wherein lymphocytes showing OX40 expression are labeled using an antibody comprising (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO:2;
 (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO:3; (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO:4; (d) HVR-L1 comprising the amino acid sequence of SEQ ID NO:5; (e) HVR-L2 comprising the amino acid sequence of SEQ ID NO:6; and (f) HVR-L3 comprising an amino acid sequence selected from SEQ ID NO:7, 26 or 27.   
     
     
         66 . The method of  claim 64 , wherein quantifying the number of CD4+ OX40+ Foxp3+ lymphocytes in the sample comprises imaging said immunofluorescence staining. 
     
     
         67 . The method of  claim 25 , wherein, in the sample comprising cancer cells and lymphocytes, said cancer cells are from a colorectal cancer. 
     
     
         68 . The method of  claim 25 , wherein, in the sample comprising cancer cells and lymphocytes, said cancer cells are from a cancer selected from the group consisting of non-small cell lung cancer, renal cell carcinoma, bladder cancer, ovarian cancer, glioblastoma, neuroblastoma, melanoma, breast carcinoma, gastric cancer, and hepatocellular carcinoma. 
     
     
         69 . The method of  claim 68 , wherein the breast carcinoma is triple-negative breast carcinoma. 
     
     
         70 . The method of  claim 25 , wherein, in the sample comprising cancer cells and lymphocytes, said cancer cells are from a primary tumor. 
     
     
         71 . The method of  claim 25 , wherein, in the sample comprising cancer cells and lymphocytes, said cancer cells are from a metastasis. 
     
     
         72 . The method of  claim 25 , wherein the CD4+ OX40+ Foxp3+ lymphocytes are tumor-infiltrating lymphocytes.