Identification of murine basal cell carcinoma antigens using SEREX
Weber, Eric J.
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Gorlin-Goltz Syndrome, also known as nevoidbasal cell carcinoma syndrome, is anautosomal dominant genetic disease resulting frommutations inthe PTCH gene. It is characterized by a varying number of basal cell carcinomas on the skin,as well as jaw cysts and rib and spinal bone abnormalities. Basal cell carcinoma is the most common cancer affecting Americans and is more prevalent in Caucasians due to fairer pigmentation (KimonisVE, Goldstein AM,PastakiaB, YangML,KaseR, DiGiovanna JJ, Bale AE, Bale SJ. (1997) Clinical manifestations in 105 persons with nevoid basal cell carcinoma syndrome. Am J Med Genet, 69 299-308). Basal cell carcinoma is an immunogenic cancer and therefore lends itself to treatment by a cancer vaccine. Such a vaccine would allow antibodies to recognize and respond to the growth of cancer cells. SEREX was used to identify murine antigens against which a vaccine could be created. PTCH knock-out and c57bl6 mice were twice vaccinated with basal cells. Tumors formed at the injection sites and both times the mice successfully repressed the tumors. Serum from these mice was collected and screened against a mouse testes cDNA library expressed in bacterial phage grown on a bacterial lawn. Antibodies in the serum attached to expressed proteins indicating an antigen/antibodypair, which were visualizedby stainingwith a secondary antibody. Positive plaques were selected, and the gene inserts were excised into a bacterial plasmid. The plasmids were then sequenced to identify the gene which encoded the antigenic protein.