Methods for the treatment of infections and tumors
PD-1 antagonists are disclosed that can be used to reduce the expression or activity of PD-1 in a subject. An immune response specific to an infectious agent or to tumor cells can be enhanced using these PD-1 antagonists in conjunction with an antigen from the infectious agent or tumor. Thus, subjects with infections, such as persistent infections can be treated using PD-1 antagonists. In addition, subjects with tumors can be treated using the PD-1 antagonists. In several examples, subjects can be treated by transplanting a therapeutically effective amount of activated T cells that recognize an antigen of interest and by administering a therapeutically effective amount of a PD-1 antagonist. Methods are also disclosed for determining the efficacy of a PD-1 antagonist in a subject administered the PD-1 antagonist. In some embodiments, these methods include measuring proliferation of memory B cells in a sample from a subject administered the PD-1 antagonist.
1. A method for diagnosing and treating a human subject having a persistent viral infection or cancer, said method comprising:
a) administering to the human subject a first dose of a PD-1 antagonist;
b) obtaining a first sample comprising peripheral blood mononuclear cells from the human subject;
c) quantifying both CD20 + CD27 + CD21 − memory B cell proliferation and CD20 + CD21 + CD27 − naïve B cell proliferation in the first sample
d) diagnosing the human subject as having effective treatment when an increased proliferation of CD20 + CD27 + CD21 − memory B cells and no significant increase in proliferation of CD20 + CD21 + CD27 − naïve B cells is detected from the first sample; and
e) administering second dose of the PD-1 antagonist to treat the persistent viral infection or the cancer in the human subject subsequently to step (d).
2. The method of claim 1 , wherein the subject has the persistent viral infection, and wherein the persistent viral infection is an HIV infection.
3. The method of claim 1 , wherein quantifying CD20 + CD27 + CD21 − memory B cell proliferation and/or CD20 + CD21 + CD27 − naïve B cell proliferation comprises a) measuring the expression of Ki67 using an antibody that specifically binds Ki67, b) measuring incorporation of bromodeoxyuridine into the CD20 + CD27 + CD21 − memory B cells, and/or c) using fluorescence activated cell sorting (FACS).
4. The method of claim 1 , wherein the subject has the cancer.
5. A method of inducing an immune response in a mammalian subject with a persistent infection with a virus or cancer, comprising:
a) administering to the subject a first dose of a PD-1 antagonist;
b) obtaining a first sample comprising peripheral blood mononuclear cells from the human subject;
c) quantifying CD20 + CD27 + CD21 − memory B cell proliferation and CD20 + CD21 + CD27 − naïve B cell proliferation in the first sample;
d) diagnosing the human subject as not having effective treatment when no significant increase in proliferation CD20 + CD27 + CD21 − memory B cells and no significant increase in proliferation of CD20 + CD21 + CD27 − naïve B cells is detected from the first sample; and
e) administering an increased second dose of the PD-1 antagonist to treat the persistent viral infection or the cancer in the human subject subsequently to step (d).
6. The method of claim 1 , further comprising
quantifying CD20 + CD27 + CD21 − memory B cell proliferation and CD20 + CD21 + CD27 − naïve B cell proliferation in a second sample from the subject following the administration of the second dose.
7. The method of claim 2 , wherein quantifying CD20 + CD27 + CD21 − memory B cell proliferation comprises measuring the expression of Ki67 using an antibody that specifically binds Ki67.
8. The method of claim 2 , wherein quantifying CD20 + CD27 + CD21 − memory B cell proliferation comprises measuring the incorporation of bromodeoxyuridine.
9. The method of claim 2 , wherein quantifying CD20 + CD27 + CD21 − memory B cell proliferation comprises the use of fluorescence activated cell sorting (FACS).
10. The method of claim 1 , wherein the subject has the persistent viral infection.
11. The method of claim 10 , wherein the subject is administered a viral antigen.
12. The method of claim 10 , wherein the viral infection is an infection with a hepatitis virus, a human immunodeficiency virus (HIV), a human T-lymphotrophic virus (HTLV), a herpes virus, an Epstein-Barr virus, or a human papilloma virus.
13. The method of claim 1 , wherein the PD-1 antagonist is an antibody that specifically binds PD-1, an antibody that specifically binds PD-L1, an antibody that specifically binds PD-L2, a small inhibitory anti-PD-1 RNAi, a small inhibitory anti-PD-L1 RNA, a small inhibitory anti-PD-L2 RNAi, an anti-PD-1 antisense RNA, an anti-PD-L1 antisense RNA, an anti-PD-L2 antisense RNA, a dominant negative PD-1 protein, a dominant negative PD-L1 protein, a dominant negative PD-L2 protein, a small molecule inhibitor of PD-1, or combinations thereof.
14. The method of claim 13 , wherein the PD-1 antagonist is an antibody that specifically binds PD-1, and wherein the antibody that specifically binds PD-1 is (1) a monoclonal antibody or a functional fragment thereof, (2) a humanized antibody or a functional fragment thereof, or (3) an immunoglobulin fusion protein.
15. The method of claim 13 , wherein the PD-1 antagonist is an antibody that specifically binds PD-L1, and wherein the antibody that specifically binds PD-L1 is (1) a monoclonal antibody or a functional fragment thereof, (2) a humanized antibody or a functional fragment thereof, or (3) an immunoglobulin fusion protein.
16. The method of claim 13 , wherein the PD1 antagonist is an antibody that specifically binds PD-L2, and wherein the antibody that specifically binds PD-L2 is (1) a monoclonal antibody or a functional fragment thereof, (2) a humanized antibody or a functional fragment thereof, or (3) an immunoglobulin fusion protein.
17. The method of claim 1 , wherein the subject is immunosuppressed.
18. The method of claim 1 , wherein the subject is assymptomatic.
19. The method of claim 1 , wherein the subject is human.
20. The method of claim 19 , wherein the subject has a human immunodeficiency virus (HIV) infection and wherein the PD-1 antagonist is an antibody that specifically binds PD-1.
21. The method of claim 1 , wherein the subject has a human immunodeficiency virus (HIV) infection, and wherein the PD-1 antagonist is an antibody that specifically binds PD-1.
22. The method of claim 1 , wherein the method further comprises measuring differentiation of the CD20 + CD27 + CD21 − memory B cells into antibody secreting cells.
23. The method of claim 1 , wherein the subject has a human immunodeficiency virus infection and wherein the method further comprises administering to the subject a therapeutically effective amount of an anti-retroviral agent.
24. The method of claim 1 , wherein the subject has a human immunodeficiency virus infection and wherein the method further comprises administering to the subject a therapeutically effective amount of a vaccine comprising a human immunodeficiency virus gp41, gp120 or polymerase.
25. The method of claim 5 , wherein the subject has the cancer.
26. The method of claim 5 , wherein the-virus is a hepatitis virus, a human immunodeficiency virus (HIV), a human T-lymphotrophic virus (HTLV), a herpes virus, an Epstein-Barr virus, or a human papilloma virus.
27. The method of claim 5 , wherein the infection with the virus is a hepatitis viral infection.
28. The method of claim 5 , wherein the infection with the virus is a human immunodeficiency viral (HIV) infection.
29. The method of claim 5 , wherein the PD-1 antagonist is an antibody that specifically binds PD-1, an antibody that specifically binds PD-L1, an antibody that specifically binds PD-L2, a small inhibitory anti-PD-1 RNAi, a small inhibitory anti-PD-L1 RNA, a small inhibitory anti-PD-L2 RNAi, an anti-PD-1 antisense RNA, an anti-PD-L1 antisense RNA, an anti-PD-L2 antisense RNA, a dominant negative PD-1 protein, a dominant negative PD-L1 protein, a dominant negative PD-L2 protein, a small molecule inhibitor of PD-1, or combinations thereof.
30. The method of claim 29 , wherein the PD1 antagonist is an antibody that specifically binds PD-1, and wherein the antibody that specifically binds PD-1 is (1) a monoclonal antibody or a functional fragment thereof, (2) a humanized antibody or a functional fragment thereof, or (3) an immunoglobulin fusion protein.
31. The method of claim 29 , wherein the PD1 antagonist is an antibody that specifically binds PD-L1, and wherein the antibody that binds PD-L1 is (1) a monoclonal antibody or a functional fragment thereof, (2) a humanized antibody or a functional fragment thereof, or (3) an immunoglobulin fusion protein.
32. The method of claim 29 , wherein the PD1 antagonist is an antibody that specifically binds PD-L2, and wherein the antibody that binds PD-L2 is (1) a monoclonal antibody or a functional fragment thereof, (2) a humanized antibody or a functional fragment thereof, or (3) an immunoglobulin fusion protein.
33. The method of claim 5 , wherein quantifying proliferation of the memory B cells comprises measuring expression of Ki67 using an antibody that specifically binds Ki67.
34. The method claim 5 , wherein quantifying proliferation of the memory B cells comprises measuring the incorporation of bromodeoxyuridine.
35. The method of claim 5 , wherein quantifying proliferation of the memory B cells comprises the use of fluorescence activated cell sorting (FACS).
36. The method of claim 1 , further comprising administering to the subject a therapeutically effective amount of an additional compound.
37. The method of claim 36 , wherein the additional compound is an antiviral compound, an antibacterial compound, an antifungal compound, an antiparasitic compound, an anti-inflammatory compound, or an analgesic.
38. The method of claim 5 , wherein the subject has a human immunodeficiency virus (HIV) infection, and wherein the PD-1 antagonist is an antibody that specifically binds PD-1.
39. The method of claim 5 , wherein isolating the CD20 + CD27 + CD21 − memory B cells comprises the use of fluorescence actived cell sorting.
40. The method of claim 5 , wherein the method further comprises measuring differentiation of the CD20 + CD27 + CD21 − memory B cells into antibody secreting cells.