Assays for detecting T cell immune subsets and methods of use thereof
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. A method for treating or delaying progression of cancer in a subject, comprising:
(a) measuring the number of CD4+OX40+Foxp3+ lymphocytes in a sample comprising cancer cells and lymphocytes obtained from the subject;
(b) determining the number of CD4+OX40+Foxp3+ lymphocytes in the sample, as compared with a reference; and
(c) if the number of CD4+OX40+Foxp3+ lymphocytes in the sample is higher than the reference, administering to the subject an effective amount of an agonist anti-human OX40 antibody, wherein the antibody comprises (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO:2, 8 or 9; (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO:3, 10, 11, 12, 13, or 14; (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO:4, 15 or 19; (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 the amino acid sequence of SEQ ID NO:7, 22, 23, 24, 25, 26, 27 or 28.
2. The method of claim 1 , wherein the number of CD4+OX40+Foxp3+ lymphocytes is a median, mean or average number of CD4+OX40+Foxp3+ lymphocytes in different regions of interest of the sample from the subject.
3. The method of claim 2 , wherein the number of CD4+OX40+Foxp3+ lymphocytes is normalized to total cells in the region of interest of the sample.
4. The method of any claim 1 , wherein the reference is based on the number of CD4+OX40+Foxp3+ lymphocytes in a sample comprising cancer cells and lymphocytes obtained from a cancer having the same type and/or stage as the cancer of the subject.
5. The method of claim 4 , wherein the reference is a median, mean, or average number of CD4+OX40+Foxp3+ lymphocytes in samples obtained from cancers having the same type and/or stage as the cancer of the subject.
6. The method of claim 1 , wherein the cancer is selected from the group consisting of non-small cell lung cancer, renal cell carcinoma, bladder cancer, ovarian cancer, glioblastoma, neuroblastoma, melanoma, triple-negative breast carcinoma, gastric cancer, colorectal cancer, and hepatocellular carcinoma.
7. A method for treating or delaying progression of cancer in a subject, comprising:
(a) measuring the number of OX40+, CD4+OX40+Foxp3+, or CD4+OX40+Foxp3− lymphocytes in a sample comprising metastatic cancer cells and lymphocytes obtained from the subject;
(b) determining the number of OX40+, CD4+OX40+Foxp3+, or CD4+OX40+Foxp3− lymphocytes in the sample, as compared with a reference; and
(c) if the number of OX40+, CD4+OX40+Foxp3+, or CD4+OX40+Foxp3-lymphocytes in the sample is higher than the reference, administering to the subject an effective amount of an agonist anti-human OX40 antibody, wherein the antibody comprises (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO:2, 8 or 9; (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO:3, 10, 11, 12, 13, or 14; (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO:4, 15 or 19; (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 the amino acid sequence of SEQ ID NO:7, 22, 23, 24, 25, 26, 27 or 28.
8. The method of claim 7 , wherein the number of OX40+, CD4+OX40+Foxp3+, or CD4+OX40+Foxp3− lymphocytes is a median, mean or average number of OX40+, CD4+OX40+Foxp3+, or CD4+OX40+Foxp3− lymphocytes in different regions of interest in the sample from the subject.
9. The method of claim 8 , wherein the number of OX40+, CD4+OX40+Foxp3+, or CD4+OX40+Foxp3− lymphocytes is normalized to total cells in the region in the sample.
10. The method of claim 7 , wherein the reference is based on the number of OX40+, CD4+OX40+Foxp3+, or CD4+OX40+Foxp3− lymphocytes in a sample comprising metastatic cancer cells and lymphocytes obtained from a cancer having the same type and/or stage as the cancer of the subject.
11. The method of claim 10 , wherein the reference is a median, mean, or average number of CD4+OX40+Foxp3+ lymphocytes in samples obtained from cancers having the same type and/or stage as the cancer of the subject.
12. The method of claim 7 , wherein the cancer is selected from the group consisting of non-small cell lung cancer, renal cell carcinoma, bladder cancer, ovarian cancer, glioblastoma, neuroblastoma, melanoma, triple-negative breast carcinoma, gastric cancer, colorectal cancer, and hepatocellular carcinoma.