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I n t e r n a t i o n a l C o n f e r e n c e o n
Immunology
Journal of Clinical Immunology and Allergy
ISSN 2471-304X
J u l y 0 5 - 0 7 , 2 0 1 8
V i e n n a , A u s t r i a
Immunology 2018
Page 50
W
e describe a unique strategy designed to identify dominant tumour antigens.
The antigen-discovery strategy is based on the finding that genes encoding
dominant tumour antigens (TAA) are expressed in a highly immunogenic form by
an allogeneic fibroblast cell line transfected with DNA from cancer cells. As the
transfected cells, express the products of multiple genes specifying an array of
tumour antigens, cells expressing genes specifying dominant tumor antigens that
induced therapeutic immune responses were identified for antigen discovery. As
only a small proportion of the transfected cell population was expected to have
incorporated gene-segments that specified TAA, a unique strategy was developed
that resulted in the identification of Cyp2e1, a derivative of cytochrome p450,
as an immune dominant tumor antigen in lung cancer cells and growth factor
receptor bound protein 10 (GRB10) as an immune dominant tumor antigen in
breast cancer cells. The strategy consisted of dividing aliquots of the suspension
of transfected cells into 10-15 small pools (initial inoculums 10E3) allowing the
cells from each pool to increase in number (to approximately 10E7) and then
dividing the transfected cell-populations from each pool into two portions. The
cells in the pool that induced immunity to lung cancer to the greatest (and as a
control) to the least extent were maintained frozen/viable for later recovery. The
remaining portion was co incubated with (mitomycin C-treated) lung cancer cells.
ELISPOT interferon gamma-release and 51Cr release cytotoxicity were used to
identify pools that stimulated immunity to the lung cancer cells to the greatest
and least extent. Frozen cells from these pools were reestablished in culture; the
cell-numbers were expanded and subdivided for additional rounds of positive
or negative selection. After further rounds, microarray was used to identify the
products of genes over-represented in the cell pool that stimulated the antitumor
immune response to the greatest extent. Cyp2e1, a derivative of cytochrome
p450, was identified as a dominant lung cancer antigen. As final confirmation of
the immunotherapeutic properties of the identified gene-product, a vaccine was
prepared by transfer of an expression vector specifying Cyp2e1 into an allogeneic
cell line followed by immunization of mice with squamous cell lung cancer. An
analogous strategy was used to identify dominant antigens expressed by breast
cancer cells. Growth factor receptor bound protein 10 (GRB10) was identified as a
dominant tumour antigen expressed by breast cancer cells.
Biography
Edward P. Cohen, M.D. is a graduate of Washington University
School of Medicine. He did his postgraduate education at the
National Institutes of Health, The University of Chicago and
the University of Colorado. He is the author of more than 150
peer review publications involving the formation of cellular can-
cer vaccines. His land mark paper with Jerrold Schwaber was
the first to describe the technique used to form monoclonal
antibodies. Below is an example of my paper. Schwaber J and
Cohen EP 1974 33. Pattern of Immunoglobulin Synthesis and
Assembly in a Human-Mouse Somatic Cell Hybrid Clone So-
matic Cell Hybrid Clone Proceedings of the National Academy
of Sciences of the United States of America. Vol. 71, No. 6 (Jun.,
1974), pp. 2203-2207.
epcohen@uic.eduNew strategy for the identification of tumor-
associated antigens that induce therapeutic
immune responses in tumor-bearing mice
Edward P Cohen, Tae Sung Kim and Amla Chopra
University of Illinois, Illinois
Edward P Cohen et al., Insights Allergy Asthma Bronchitis 2018, Volume: 4
DOI: 10.21767/2471-304X-C1-001