Generation of antibodies to tumor antigens and generation of tumor specific complement dependent cytotoxicity by administration of oncolytic vaccinia virus
The present invention relates to methods and compositions for use in inducing tumor-specific antibody mediated complement-dependent cytotoxic response in an animal having a tumor comprising administering to said animal a composition comprising a replication competent oncolytic virus wherein administration of the composition induces in the animal production of antibodies that mediate a CDC response specific to said tumor.
1. A method of identifying a tumor-specific antigen comprising
a) contacting (i) a plurality of expression vectors into which a cDNA library prepared from a cancer cell has been cloned with (ii) serum isolated from one or more human subjects having a tumor, said one or more subjects having been administered a replication competent oncolytic Wyeth or Western Reserve strain vaccinia virus in an amount effective to mediate a complement dependent cytotoxic (CDC) response specific to said tumor in the subject prior to isolating said serum; and
b) isolating antigens from said cDNA library that are recognized by the serum, wherein the method further comprises confirming the presence of CDC response-producing antibodies in said serum after the replication competent oncolytic virus has been administered and before contacting said serum with said plurality of expression vectors.
2. The method of claim 1 , wherein said antigens are recognized by polyclonal antibodies induced by treatment with said replication competent oncolytic virus.
3. The method of claim 2 , wherein said antigens are not recognized by serum from a human subject with the same tumor type that has not been administered said replication competent oncolytic virus.
4. The method of claim 1 , further comprising the step of cloning the cDNA library prepared from the cancer cell into expression vector(s).
5. The method of claim 1 , further comprising the step of isolating serum from blood collected from the subject to which the oncolytic virus has been administered.
6. The method of claim 1 , wherein the tumor and cancer cell are the same cancer type.
7. The method of claim 1 , wherein said serum comprises pooled serum from a plurality of human subjects with the same tumor type that have been administered said replication competent oncolytic virus.
8. The method of claim 1 , wherein the tumor is selected from astrocytoma, oligodendroglioma, meningioma, neurofibroma, glioblastoma, ependymoma, Schwannoma, neurofibrosarcoma, neuroblastoma, pituitary adenoma, medulloblastoma, head and neck cancer, melanoma, prostate carcinoma, renal cell carcinoma, pancreatic cancer, breast cancer, lung cancer, colon cancer, bladder cancer, liver cancer, bone cancer, rectal cancer, ovarian cancer, sarcoma, esophageal cancer, stomach cancer, cervical cancer, fibrosarcoma, squamous cell carcinoma, neurectodermal, thyroid tumor, Hodgkin's lymphoma, non-Hodgkin's lymphoma, malignant hepatoma, mesothelioma, epidermoid carcinoma, and tumorigenic diseases of the blood.
9. The method of claim 8 , wherein the tumor is selected from colon cancer, rectal cancer, hepatocellular carcinoma, renal cell carcinoma, bladder cancer, lung cancer, stomach cancer, pancreatic cancer, melanoma, ovarian cancer, head and neck cancer, esophageal cancer, sarcoma and mesothelioma.
10. The method of claim 1 , wherein the replication competent oncolytic Wyeth or Western Reserve strain vaccinia virus comprises a transgene.
11. The method of claim 1 , wherein the transgene comprises a heterologous nucleic acid sequence encoding GM-CSF, cytosine deaminase, or carboxyl esterase.
12. The method of claim 10 , wherein the heterologous nucleic acid sequence encodes GM-CSF.
13. The method of claim 11 , wherein the oncolytic vaccinia virus is JX-594.