Serum from mice immunized in the context of Treg inhibition identifies DEK as a neuroblastoma tumor antigen.
BACKGROUND
We have developed a cell-based vaccine that features the expression of both CD80 and CD86 on the surface of a murine neuroblastoma cell line. The cellular immunity induced by this vaccine is enhanced by treatment with antibody that interferes with T-regulatory cell (Treg) function and we report here that immunization combined with interfering with Treg function also produces a profound serological effect. Serum from mice immunized with our cell-based vaccine in the context of Treg blockade was used to screen a cDNA expression library constructed from the parental neuroblastoma tumor cell line, AGN2a.


RESULTS
Serum from mice vaccinated in the context of Treg blockade identified a number of potentially oncogenic transcripts that may serve as important immune targets in a tumor-derived cDNA library screen. This novel approach identified far more candidates than could be seen with serum derived from vaccine-treated only, Treg-depleted only, or tumor-bearing mice. The most commonly identified tumor-associated antigen, using serum from immunized and Treg-depleted mice, was the DEK oncogene. Altered expression of the DEK oncogene has been implicated in a number of human cancers. Importantly, we were able to demonstrate that the DEK oncogene also induces a T cell response.


CONCLUSIONS
The use of post-vaccine immune serum in this report differs from previous approaches where serum collected at the time of cancer onset or diagnosis and was used for tumor antigen identification. We hypothesize that the use of diagnostic serum samples may be inadequate for the clinical translation of this approach, and that identification of protective immunogenic tumor antigens may require the use of serum from post-treatment or vaccinated subjects. The identification of DEK as a tumor-associated antigen capable of eliciting a T cell response validates our experimental approach and argues for the antigens we have identified here to be evaluated as targets of effector immunity and as vaccine candidates.
Zheng
Jin
J
Department of Pediatrics, Medical College of Wisconsin and the Children's Research Institute, Children's Hospital of Wisconsin, Milwaukee, WI 53226, USA. jzheng@mcw.edu <jzheng@mcw.edu>
Kohler
M Eric
ME
Chen
Qingrong
Q
Weber
James
J
Khan
Javed
J
Johnson
Bryon D
BD
Orentas
Rimas J
RJ
D000818:Animals; D000911:Antibodies, Monoclonal / Q000378:metabolism; D000951:Antigens, Neoplasm / Q000097:blood* / Q000276:immunology*; D018122:B7-1 Antigen / Q000276:immunology; D051940:B7-2 Antigen / Q000276:immunology; D015153:Blotting, Western; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology; D019496:Cancer Vaccines / Q000235:genetics / Q000276:immunology / Q000494:pharmacology*; D045744:Cell Line, Tumor; D018076:DNA, Complementary; D004268:DNA-Binding Proteins / Q000097:blood* / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay; D005455:Fluorescent Antibody Technique; D015723:Gene Library; D007111:Immunity, Cellular; D007114:Immunization; D051379:Mice; D009447:Neuroblastoma / Q000235:genetics / Q000276:immunology / Q000473:pathology / Q000628:therapy*; D020411:Oligonucleotide Array Sequence Analysis; D015513:Oncogene Proteins / Q000097:blood* / Q000276:immunology*; D000075223:Poly-ADP-Ribose Binding Proteins; D050378:T-Lymphocytes, Regulatory / Q000276:immunology* / Q000378:metabolism
D000911:Antibodies, Monoclonal; D000951:Antigens, Neoplasm; D018122:B7-1 Antigen; D051940:B7-2 Antigen; D019496:Cancer Vaccines; C519564:DEK protein, mouse; D018076:DNA, Complementary; D004268:DNA-Binding Proteins; D015513:Oncogene Proteins; D000075223:Poly-ADP-Ribose Binding Proteins